Korean Biosafety Law

Risk Assessment and Risk Management Plan
Application for licence for dealings involving an intentional release of GMOs into the environment
DIR 044/2003
Title: Agronomic assessment and seed increase of GM cottons ex ressing insecticidal genes !cry1Fa and cry1Ac" from Bacillus thuringiensis"
Applicant: Dow AgroSciences
May 2004
DIR 048/2003 Risk Assessment and Risk Management Plan
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AN !A
A"stralia New ealand !ood A"thority #now !SAN $
A%&MA A"stralian %esticides and &eterinary Medicines A"thority Bacillus thuringiensis variety aizawai
'ta 't( Bacillus thuringiensis variety kurstaki microgram per gram
ABBREVIATION
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ABBREVIATION
DIR 044/2003 Risk assessment and Risk management Plan
Office of the Gene Technology Regulator
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*he Gene Technology Act 2000 #the Act$ and the Gene Technology Regulations 2001 #the +eg"lations$ set o"t re,"irements which the Gene *echnology +eg"lator #the +eg"lator$ m"st follow when considering an application for a licence to intentionally release a genetically modified organism #GMO$ into the environment!or a licence to .e iss"ed/ the +eg"lator m"st .e satisfied that the release will not pose any ris(s to h"man health and safety or the environment that can not .e managed- As part of the eval"ation process/ section 01 of the Act re,"ires the +eg"lator to prepare a ris( assessment and ris( management plan #+A+M%$ for each licence application/ in cons"ltation with a wide range of e2pert gro"ps and sta(eholders3nder section 02 of the Act/ the +eg"lator is re,"ired to see( comment on the +A+M% from those cons"lted in its preparation and to invite s".missions from the p".lic- Matters raised relating to the protection of h"man health and safety or the environment are ta(en into acco"nt in finalising the +A+M%/ which then forms the .asis of the +eg"lator4s decision on whether/ or not/ to iss"e a licence*he Act is designed to operate in a cooperative legislative framewor( with other reg"latory a"thorities that have complementary responsi.ilities and specialist e2pertise- As well as enhancing coordinated decision ma(ing/ this arrangement avoids d"plication- *he OG*+ liaises closely with other reg"lators to ens"re the identification/ eval"ation and management of ris(s that may .e associated with development and "se of gene technology*he +eg"lator has made a decision to iss"e a licence in respect of application D5+ 044)2006 from Dow AgroSciences A"stralia %ty 7td #Dow AgroSciences$-
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Dow AgroSciences has applied for a licence #application D5+ 044)2006$ for the intentional release/ "nder limited and controlled conditions/ of three genetically modified #GM$ insecticidal)her.icide tolerant cotton lines!- Dow AgroSciences initially proposed to cond"ct trials over two s"mmer growing seasons and two winter growing seasons on "p to 20 sites covering a total c"m"lative area of "p to 10 hectares over 6 years #May 2004 to May 2008$ in New So"th 9ales/ :"eensland/ Northern *erritory and 9estern A"stralia- ;owever/ the company s".se,"ently amended their application to slightly increase the n"m.er and si<e of sites #"p to 60/ covering a total c"m"lative area of "p to 12-2 hectares over the same 6 years$ and withdrew the re,"est for trialing in Northern *erritory5n accordance with the provisions of section 1=0 of the Act/ Dow AgroSciences so"ght and received approval for some specific doc"ments/ which contain details of the gene constr"cts/
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*he term >line4 has .een "sed/ thro"gho"t this ris( assessment/ to denote cotton containing a specific genetic modification derived from specified individ"al transformation event)s #note that 9idestri(e? cotton contains 2 events$-
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gene se,"ence information and molec"lar characterisation of the inserted genetic materials/ as @onfidential @ommercial 5nformation #@@5$/ in connection with a previo"s licence application D5+ 040)2006- 9hile the +eg"lator is satisfied that the p".lic interest in the release as proposed did not o"tweigh the preA"dice that disclos"re wo"ld ca"se the applicant/ the @@5 was made availa.le to the vario"s prescri.ed e2pert gro"ps that were cons"lted on the preparation of the +A+M% for this application*wo of the three GM cotton lines proposed for release contain a single insecticidal gene/ either cry1Fa #line 2=1B24B268 or @ry1!a cotton$ or cry1Ac #line 6008B210B26 or @ry1Ac cotton$- *he third line #line 2=1B24B268)6008B210B26 or 9ideStri(e? cotton$ contains .oth cry1Fa and cry1Ac genes/ and was prod"ced .y conventional .reeding of the two singleBgene insecticidal cotton lines- *he insecticidal genes were derived from the common soil .acteri"m Bacillus thuringiensis #'t$ and e2press insecticidal proteins #'t to2ins$ that are to2ic to specific lepidopteran caterpillar insects/ incl"ding the maAor caterpillar pests of cotton*he cry1Fa and cry1Ac genes in these GM cotton lines are chimeric genes/ each containing parts of two other cry1 genes of 't- *he chimeric genes were developed to improve the level of e2pression of the encoded @ry1!a and @ry1Ac proteins in plants/ and enhance their sol".ility in the insect g"t- ;owever/ the encoded 't to2ins are very similar to the native @ry1!a and @ry1Ac proteins- 9ithin the f"nctional #core$ to2in/ the amino acid se,"ences of the native and chimeric proteins are CC-6D and CC-8D identical/ respectively- *herefore/ the species specificity of the introd"ced @ry1!a and @ry1Ac proteins to lepidopteran larvae is the same as native .acterial @ry1!a and @ry1Ac proteinsAll three lines also contain a her.icide tolerance selecta.le mar(er gene/ pat/ that confers tolerance to the her.icide gl"fosinate ammoni"m #the active ingredient of 7i.erty and 'astaEE her.icides$- *he mar(er gene was "sed in the la.oratory d"ring the development of the GMOs for identification and selection of plant tiss"es in which the insecticidal genes were also present- Dow AgroSciences does not intend to apply gl"fosinate ammoni"m her.icide to the GM cotton lines in the proposed releaseDow AgroSciences proposes to cond"ct a small scale/ m"ltiBsite trial in a range of A"stralian cotton growing regions- *he aims of the proposed release are: to test the efficacy #effectiveness$ of the twoBgene #cry1Fa)cry1Ac$ insecticidal cotton line/ 9ideStri(e? cotton/ against lepidopteran caterpillars pests of cotton as compared to its two parental lines #@ry1!a cotton and @ry1Ac cotton$F to eval"ate their respective agronomic performanceF and to collect data for developing an insect resistance management plan- Additionally/ the applicant proposes to meas"re the e2pression levels of the insecticidal proteins in cotton tiss"es and resid"es of these proteins in soil/ and to test the effect of GM cotton lines on nonB target organisms- Seed wo"ld also .e retained for s".se,"ent seasons of the trial or for potential f"t"re releases #s".Aect to f"rther approvals$Note that Dow AgroSciences4 decision whether to progress the commercialisation of these GM cotton lines in A"stralia will depend on the o"tcomes of these trials- 5f these GM cottons do not prove to .e s"ita.le for the A"stralian cotton prod"ction system d"ring the first two seasons of the trial/ Dow AgroSciences intends to stop the trial at the end of second season &
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#2004)00 s"mmer season$- *he efficacy data generated d"ring the initial stages of this trial will inform f"t"re research and potential commercialisation plans- *he GM cotton lines proposed for release are derived from the original American c"ltivar that was "sed in the initial .reeding program- 5f Dow AgroSciences wishes to cond"ct additional trials/ incl"ding .reeding the GM insecticidal traits into c"ltivars more s"ita.le for A"stralian conditions/ additional licence applications and approvals wo"ld .e re,"iredNone of the cotton plants from the release/ or their .yBprod"cts/ will .e "sed for animal feed or h"man food/ and seed not re,"ired for f"t"re plantings will .e destroyed- !ollowing harvest/ the applicant proposes that plant material remaining at the site will .e slashed and incorporated into the soil .y c"ltivation- Seed from pollen trap plants will also .e destroyedAny regrowth will .e controlled .y her.icide and)or c"ltivation- @ottonseed will .e sec"rely transported to prevent dissemination9ideStri(e? cotton is c"rrently .eing trialed .y Dow AgroSciences "nder limited and controlled conditions on two sites covering a total area of 0-04 ha in New So"th 9ales #D5+ 040)2006$- *here have .een no previo"s releases of either of the two singleBgene insecticidal GM cotton lines #cry1Fa or cry1Ac$ in A"straliaAll three GM cotton lines proposed for release in the c"rrent application have .een approved for field trials in the 3nited States since 2001- *hese cotton lines are also approved for field trial in Argentina in 2004%revio"sly/ other GM insecticidal cottons containing cry insecticidal gene#s$ derived from the same .acteri"m #B. thuringiensis$ have .een trialed e2tensively in A"stralia #eg- D5+s 000)2001/ 008)2001/ 00=)2001 and 00C)2001$/ as well as commercially released "nder licences D5+ 022)2002 #5NGA+DE cotton$ and D5+ 012)2001 #'ollgardE 55 cotton$;owever/ the chimeric cry1Ac gene in two of the GM cotton lines proposed for release in the c"rrent application differs slightly from the chimeric cry1Ac gene present in .oth 5NGA+DE and 'ollgardE 55 cottonsOther GM plants incorporating the pat gene and e2pressing %A* protein have also .een trialed in A"stralia #eg- D5+ 010)2001/ D5+ 010)2002 and D5+ 06=)2006$*here have .een no reports of adverse effects on h"man health or the environment res"lting from these releases-
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A +A+M% has .een prepared in relation to licence application D5+ 044)2006 from Dow AgroSciences in accordance with the Act/ the +eg"lations/ and the +is( Analysis !ramewor(*his framewor( was developed as part of the esta.lishment of the reg"latory arrangements in cons"ltation with the p".lic/ State/ *erritory and A"stralian Government agencies/ (ey sta(eholders and the Gene *echnology *echnical Advisory @ommittee/ and is availa.le at www.ogtr.gov.au)pdf)p".lic)raffinal-pdf. Details of the process that the +eg"lator m"st follow/ incl"ding the prescri.ed cons"ltation process on the application/ and the matters that she m"st consider in preparing a +A+M%/ are
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set o"t in Appendi2 = of the +A+M%- *he complete +A+M% can .e o.tained from the OG*+ .y contacting the Office on 1=00 1=1 060 or from its we. site: www.ogtr.gov.au*he ris( assessment considered information contained in the application #incl"ding information re,"ired .y the Act and the +eg"lations on the GMO/ the parent organism/ the proposed dealings and on potential impacts on h"man health and safety and the environment$/ c"rrent scientific (nowledge and s".missions received d"ring cons"ltation with e2pert gro"ps and a"thorities*hro"gh this process/ potential ha<ards to h"man health and safety or the environment that may .e posed .y the proposed release of the three GM insecticidal)her.icide tolerant cotton lines were identified- *hese have .een eval"ated to determine whether ris(s might arise/ .ased on the li(elihood of each ha<ard occ"rring and the li(ely impact of each ha<ard/ were it to .e realised*he identified potential ha<ards relate to: 0( toxicit0 and allergenicit0 to 12mans: co"ld these GM cottons .e more to2ic or allergenic than nonBGM cotton/ as a res"lt of the novel gene prod"cts or .eca"se of "nintended effectsG toxicit0 to non3target organisms: co"ld these GM cottons .e harmf"l to nonB target organisms as a res"lt of the novel gene prod"cts or .eca"se of "nintended effectsG 4eediness: co"ld the genetic modifications .e harmf"l to the environment .y increasing the potential for these GM cottons to esta.lish as pro.lem weedsG transfer of introd2ced genes to ot1er organisms: co"ld there .e adverse conse,"ences from potential transfer of the introd"ced genes to nonBGM cotton crops/ feral or native cottons/ or to other organismsG and insecticide resistance: co"ld target insects develop resistance to the insecticidal proteins prod"ced .y the introd"ced insecticidal genes in these GM cottonG
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*he A"stralian %esticides and &eterinary Medicines A"thority #A%&MA$ has a complementary reg"latory role in respect of this application d"e to its responsi.ility for agric"lt"ral chemical "se in A"stralia- 5nsecticidal GM plants fall "nder the Agricultural and eterinary !he"icals !ode Act 1##$ definition of an agric"lt"ral chemical prod"ct- !"rther information a.o"t the A%&MA4s assessment and approval process is contained in @hapter 1 and Appendi2 8 of the +A+M%!or commercial prod"cts/ the normal form of approval is thro"gh registration/ ."t the A%&MA may also iss"e permits for e2perimental wor( that allow restricted "se of an agric"lt"ral chemical/ for e2ample/ for a limited period of time or for a limited area- *he A%&MA can impose conditions of "se on .oth registrations and permits/ and m"st .e satisfied that the proposed "se wo"ld not present an "nd"e ris( to h"man health or the environment/ and the insecticide is efficacio"s- *he ha<ard of development of insecticide resistance in pests is also part of the A%&MA assessment of insecticide "se-
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Dow AgroSciences has s".mitted an application to the A%&MA for a research permit for the "se of the insecticidal genes in the GM cotton lines for the proposed trial- *he A%&MA and the OG*+ have wor(ed closely to ens"re the thoro"gh/ coordinated assessment of these parallel proposals/ and that the decisions .y .oth agencies coincide-
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5t is concl"ded the proposed release of three GM cotton lines does not pose significant ris(s to h"man health and safety and the environment as a res"lt of the genetic modification- *he +eg"lator has imposed licence conditions that will minimise potential e2pos"re of h"mans and other organisms to the GM cottons/ and limit the spread and persistence of the GMOs or the introd"ced genes while more data is gathered on the .ehavio"r and interactions of the GMOs in the environment- *he assessment of each potential ha<ard identified a.ove is s"mmarised "nder a separate heading .elow-
Toxicit0 or allergenicit0 to 12mans

*he GM insecticidal)her.icide tolerant cotton lines are "nli(ely to prove more to2ic or allergenic to h"mans via occ"pational e2pos"re than conventional cotton- ;"mans are commonly e2posed to the native .acterial forms of the proteins prod"ced from the introd"ced genes/ as the organisms from which they are derived are nat"rally widespread in the environment- None of the proteins have any (nown h"man to2icity or allergenicity- *here have .een no reports of to2ic or allergenic effects from previo"s releases of GM cotton lines containing insecticidal genes derived from the same .acteri"m- *he proposed release is limited in scale and conditions are imposed to limit the spread of the GMOs and the introd"ced genes*he applicant does not intend to "se any cottonseed prod"ced in the proposed release in h"man food or animal feed/ or to sell lint or linters for processing/ th"s limiting potential e2pos"re- !ood Standards A"stralia New ealand #!SAN $ is responsi.le for h"man food safety assessment/ and !SAN approval wo"ld .e needed .efore prod"cts from these GM cottons co"ld .e "sed in h"man food5t sho"ld .e noted that !SAN is c"rrently eval"ating an application for approval for food "se of oil and linters derived from one of the GM cotton lines/ 9ideStri(e? cotton/ proposed for release-
Toxicit0 to non3target organisms

*he GM insecticidal)her.icide tolerant cottons are "nli(ely to prove more to2ic to nonBtarget organisms than conventional cotton- As disc"ssed a.ove/ altho"gh the introd"ced genes are chimeric/ the encoded to2in proteins are very similar to the native to2ins- *herefore the specificity of the e2pressed to2in proteins to lepidopteran caterpillars is the same as native .acterial @ry1!a and @ry1Ac proteins- *hese proteins are nat"rally widespread in the environment and have no (nown to2icity to mammals/ .irds or fish- *he proposed release is limited in scale and conditions are imposed to limit the movement of the GMOs and the introd"ced genes/ and the applicant does not intend to "se any material from the release in animal feed-
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+es"lts from la.oratory and field st"dies in the 3SA indicate that the GM insecticidal)her.icide tolerant cotton lines proposed for release are not to2ic to nonBtarget inverte.rates in that environment- ;owever/ the applicant will .e re,"ired to provide information on the effect of the chimeric @ry1!a and @ry1Ac proteins on nonBtarget organisms "nder A"stralian field conditions .efore any application for a larger scale release of these GM cottons is eval"ated-
6eediness
*he ris( of the GM insecticidal)her.icide tolerant cotton lines esta.lishing as pro.lematic weeds/ as a res"lt of the proposed release in the areas of so"thern A"stralia is low and not li(ely to .e greater than that of nonBGM cotton- *his is .eca"se the germination and persistence of .oth GM and nonBGM cottons in so"thern A"stralia are limited .y the availa.ility of ade,"ate soil moist"re/ n"trients/ her.ivory .y nonBlepidopteran species #verte.rate and inverte.rate$/ fire/ plant competition and)or frosts7imited e2perimental data s"ggests that insecticidal GM cottons may have the potential to .e weedier than nonBGM cotton in northern A"stralia in certain ha.itats where there is ade,"ate soil moist"re and n"trients/ limited plant competition and protection from fire and no frosts5t is highly "nli(ely that the genetic modifications will affect the response of the GM cottons to these varia.les and/ there.y/ alter the weediness of the GM cottons- +es"lts from field st"dies in the 3SA indicate that the agronomic characteristics of the GM cottons which may relate to weediness potential have not .een altered- ;owever/ licence conditions have .een imposed to minimise the spread and persistence of these GM cottons in the environment-
Transfer of introd2ced genes to ot1er organisms

Some gene transfer from the GM insecticidal)her.icide tolerant cotton lines to other c"ltivated cottons wo"ld .e li(ely "nder "ncontrolled conditions/ altho"gh the overall fre,"ency of o"tBcrossing wo"ld .e very low as cotton is primarily selfBpollinating- *ransfer of introd"ced genes to other c"ltivated cotton wo"ld pose the same ris(s as the low ris(s posed .y the GM cottons themselves;er.ari"m records s"ggest that nat"ralised)feral cotton pop"lations may occ"r/ or might have occ"rred in the past/ in central and so"th eastern :"eensland/ in the northern Northern *erritory and in northern 9estern A"stralia- *he remnants of these pop"lations/ a few of which may .e within pollinating distance of commercial cotton crops/ has not yet .een confirmed- As part of the licence conditions for D5+ 022)2006/ a s"rvey of nat"ralised cotton pop"lations in :"eensland/ in locations s"ggested .y her.ari"m records/ is .eing cond"cted*he ris( of transferring the introd"ced genes to native cotton is negligi.le/ .eca"se of geographic separation and genetic incompati.ility7icence conditions have .een imposed to limit crossBpollination to compati.le cotton plants o"tside the release sites*he li(elihood of transfer of the introd"ced genes to other organisms is negligi.le .eca"se of well esta.lished genetic incompati.ility- Hven if s"ch transfer occ"rred it wo"ld .e "nli(ely to pose any ha<ard to h"man health and safety or the environment-
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As part of the OG*+4s ongoing commitment to the review of data/ specific research conditions have .een imposed in the licence- *he research is intended to confirm data from research on gene flow "nderta(en prior to the implementation of the Act and prior to the commercial release of GM cottons/ and in so doing validate the containment meas"res for field trials of GM cottons- 5n order to facilitate this review and data collection/ the +eg"lator has imposed a licence condition to esta.lish 400 metre research <ones aro"nd field trial sites in e2cess of one hectare- *he associated research program wo"ld .e developed in cons"ltation with the OG*+-
Insecticide resistance
Development of insects resistant to the insecticidal proteins in these three GM cotton lines might occ"r if they were grown on a larger scale witho"t "sing any management strategies;owever/ given the limited scope of the release in scale and time/ the li(elihood of this ris( res"lting from the release is negligi.le5n addition/ as these GM cottons fall "nder the Agricultural and eterinary !he"icals !ode Act 1##$ definition of an agric"lt"ral chemical prod"ct d"e to their prod"ction of insecticidal s".stances/ they are s".Aect to reg"lation .y the A"stralian %esticides and &eterinary Medicines A"thority #A%&MA$- *he ha<ard of development of insecticide resistance in pests was also considered .y the A%&MA in considering Dow AgroSciences4 permit application for the "se of the insecticidal genes in the GM cottons- *he A%&MA has the reg"latory responsi.ility for management of this ha<ard-
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As part of the eval"ation process for this licence application/ a ris( management plan has .een developed to address the ris(s identified #refer to concl"sions of the ris( assessment/ a.ove$- *his plan has .een given effect .y the licence conditions imposed- *he (ey licence conditions are o"tlined .elow-
Toxicit0 or allergenicit0 to 12mans

7icence conditions have .een imposed which re,"ire the applicant to: &( '( )( 8( *( limit the scale of the releaseF prevent entry of the GMOs and prod"cts derived from the GMOs into the h"man food s"pplyF destroy all GM materials not re,"ired for f"t"re trialsF sec"rely transport and store the GMOsF and report adverse impacts-
Toxicit0 to non3target organisms

7icence conditions have .een imposed which re,"ire the applicant to: !0( limit the scale of the releaseF !!( prevent GM cottonseed .eing "sed as stoc(feedF
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!0
!2( destroy all GM materials not re,"ired for f"t"re trialsF and !3( sec"rely transport and store the GMOs-
6eediness
7icence conditions have .een imposed which re,"ire the applicant to: !4( limit the scale of the releaseF !&( prevent cottonseed .eing "sed as stoc(feedF !'( either s"rro"nd the GM cottons .y a 20 m pollen trap of nonBGM cotton or ens"re that there are no other cotton crops or nat"ralised cotton pop"lations within 400 m of the GM cotton cropsF !)( sec"rely transport and store the GM cottonsF !8( clean the release sites and e,"ipment "sed at release sitesF and !*( monitor release sites after harvest and destroy vol"nteers-
Transfer of introd2ced genes to ot1er organisms

7icence conditions have .een proposed which re,"ire the applicant to: 20( limit the scale of the releaseF 2!( either s"rro"nd the GM cottons .y a 20 m pollen trap of nonBGM cotton or ens"re that there are no other cotton crops or nat"ralised cotton pop"lations within 400 m of the GM cotton cropsF 22( provision for a 400 m wide research <one aro"nd trial sites in e2cess of one hectare for the p"rpose of cond"cting research on gene flow/ and for development of an agreed research program to validate previo"s research on gene transfer containment meas"res for ongoing review of dataF 23( sec"rely transport and store the GM cottonsF 24( clean the release sites and e,"ipment "sed at release sitesF and 2&( monitor release sites after harvest and destroy vol"nteers-
Insecticide resistance
No conditions have .een imposed in relation to insecticide resistance management- *he A%&MA also has the reg"latory responsi.ility for this iss"e- *he applicant4s o.ligation to comply with any conditions imposed .y the A%&MA is noted in the licence-
General conditions
Any licence iss"ed .y the +eg"lator also contains a n"m.er of general conditions/ which are also relevant to ris( management- *hese incl"de/ for e2ample: 2'( identification of the persons or classes of person covered .y the licenceF
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2)( re,"irement that the applicant allow access to the release sites .y the +eg"lator/ or persons a"thorised .y the +eg"lator/ for the p"rpose of monitoring or a"ditingF and 28( a re,"irement to inform the +eg"lator if the applicant .ecomes aware of any additional information a.o"t ris(s to h"man health and safety or to the environment@hapter 2 of the ris( assessment and ris( management plan provides a ta."lated s"mmary of assessment concl"sions and corresponding management conditions- !"ll details of the licence conditions are provided in Appendi2 I-
Researc1 re72irements
+esearch conditions have .een imposed in the licence to collect data on: 2*( levels of e2pression of the insecticidal and her.icide tolerance genes in the GM cotton tiss"es "nder A"stralian field conditionsF 30( effect of the GM cottons on nonBtarget organisms "nder A"stralian field conditionsF 3!( potential for the introd"ced proteins to acc"m"late in the soil "nder A"stralian field conditionsF 32( efficacy of gene flow containment meas"res for any release sites in e2cess of one hectare/ within the associated +esearch one/ and for any release site for which a pollen trap is not re,"ired/ d"e to having no other cotton crops or pop"lations within 400 m-
Additional data
*he proposed limited and controlled release is a small scale/ m"ltiBsite trial over fo"r cotton growing seasons- 5f the applicant ma(es any f"t"re applications for larger scale releases of GM insecticidal)her.icide tolerant cotton lines/ more data are re,"ired to .e provided on: 33( agronomic characteristics of the GM cottons in relation to potential weediness "nder A"stralian field conditionsF 34( effect of the GM cottons on soil .iotaF 3&( "nintended effects of the genetic modification-
Monitoring and enforcement of com liance 80 t1e &GTR

As well as the legislative capacity to enforce compliance with licence conditions/ the +eg"lator has additional options for ris( management- *he +eg"lator can direct a licence holder to ta(e any steps the +eg"lator deems necessary to protect the health and safety of people or the environment- *he OG*+ also independently monitors releases that the +eg"lator has a"thorised- At least 20D of all field trial sites will .e inspected each year/ in accordance with a monitoring and compliance strategy .ased on ris( profiling #which ta(es into acco"nt .iological/ seasonal/ geographical and ecological ris( factors$ to determine whether licence holders are complying with the licence conditions/ or whether there are any "nforseen pro.lems-
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!3
+/APT$R 9 #A+5GR&-'D
!+ *his chapter provides .ac(gro"nd information a.o"t the application and previo"s releases of relevant genetically modified organisms #GMOs$ into the environment-
($+TI&' 9 T/$ APP)I+ATI&'

2+ *he OG*+ has received an application #licence application n"m.er D5+ 044)2006$ from Dow AgroSciences A"stralia %ty 7td #Dow AgroSciences$ for the intentional release of three genetically modified #GM$ insecticidal)her.icide tolerant cottons into the environment/ on a limited scale and "nder controlled conditions- Jey information on the application is given .elow:
AGR&'&MI+ A(($((M$'T A'D ($$D I'+R$A($ &* GM +&TT&'( $,PR$((I' G I'($+TI+I DA) G$'$( !CRY1FA A'D CRY1AC" *R&M BACILLUS THURINGIE NSIS D&6 AGR&(+I$'+$( A-(TRA)IA PT. )TD )&+5$D #AG :02 *R$'+/( *&R$(T '(6 9;40
%,a-te. !
Ba/kg.#0nd
!4
MON NAME OF THE PARENT ORGANIS M:
+&TT&' GOSSY IU! HIRSUTU! )< I'($+TI+IDA) A'D /$R#I+ID$ T&)$RA'+ $
NTIFIC NAME OF THE PARENT ORGANIS M:
3'( chimeric cry1Fa gene

from the .acteri"m Bacillus thuringiensis #insecticidal$
FIED TRAIT(S):
TITY OF THE GENE(S) RESPONS IBLE FOR THE MODIFIED TRAIT(S):
3)( chimeric cry1Ac gene

from the .acteri"m Bacillus thuringiensis #insecticidal$
38( pat gene from

%trepto"yces viridochro"ogenes #her.icide tolerance)selecta.le mar(er$
%,a-te. !
Ba/kg.#0nd
!&
OSED LOCATION(S)
(/IR$( &* #A)&''$= #A'A'A= #A-/I'IA= $M$RA)D= MI))M$RR A'= M-RI))A= PITT(6&R T/= 6AGGAM# A= 6AM#& A'D 6ARR&& I' >-$$'()A 'D? (/IR$( &* #&-R5$= +ARRAT/& &)= G-''$DA/ = M&R$$ P)AI'(= 'ARRA#RI= 6A)G$TT A'D 6ARR$' I' '$6 (&-T/ 6A)$(? (/IR$( &* 6.'D/AM /$A(T 5IM#$R)$ . A'D D$R#./6$ (T 5IM#$R)$ . I' 6$(T$R' A-(TRA)I A< MA. 2004 @ MA. 200;
OSED TIME OF RELEASE '& &* (IT$( T&TA) AR$A !/A" (-MM$R 200:/;
%,a-te. ! Ba/kg.#0nd
!'
92<20
%,a-te. !
Ba/kg.#0nd
!)
3+ 5n the original application/ Dow AgroSciences proposed to cond"ct the trial over fo"r cotton growing seasons on "p to 20 sites covering a total c"m"lative area of 10 hectares- ;owever/ Dow AgroSciences s".se,"ently amended its application to slightly increase the n"m.er and si<e of sites #"p to 60/ covering a total c"m"lative area of "p to 12-2 hectares over the same 6 years$ and withdrew the re,"est for trialing in Northern *erritory4+ 5n accordance with the provisions of section 1=0 of the Gene Technology Act 2000 #the Act$/ Dow AgroSciences so"ght and received approval for details of the gene constr"cts/ gene se,"ence information and molec"lar characteristics of the inserted genetic materials as @onfidential @ommercial 5nformation #@@5$ in connection with a previo"s licence application #D5+ 040)2006$- 9hile the +eg"lator is satisfied that the p".lic interest in the release did not o"tweigh the preA"dice that disclos"re wo"ld ca"se the applicant/ the @@5 was made availa.le to the vario"s prescri.ed e2pert gro"ps that were cons"lted on the preparation of the ris( assessment and ris( management plan for this application-
(ection 9<9 T1e ro osed dealings

&+ Dow AgroSciences proposes to cond"ct a small scale/ m"ltiBsite trial in a range of cotton growing regions- *he aims of the proposed release are to test the efficacy #effectiveness$ of a GM cotton line/ 9ideStri(e? cotton/ containing two insecticidal genes #cry1Fa and cry1Ac$ against lepidopteran caterpillars pests of cotton/ as compared to its two parental lines #line 6008B210B26 or @ry1Ac cotton and line 2=1B 24B268 or @ry1!a cotton$F to eval"ate their respective agronomic performanceF and to collect data for development of an insecticide resistance management plan- 5n addition/ the applicant proposes to meas"re the e2pression levels of the insecticidal proteins in cotton tiss"es and resid"es of these proteins in soil/ and to test the effect of the GM cotton lines on nonBtarget organisms- Seed wo"ld also .e retained for s".se,"ent trial seasons for the proposed release or for potential f"t"re releases #s".Aect to f"rther approvals$- None of the cotton plants from the release/ or their .yB prod"cts/ wo"ld .e "sed for h"man food and animal feed/ nor wo"ld lint or linters .e sold for processing'+ Note that Dow AgroSciences has not yet decided whether to commercialise these GM cotton lines in A"stralia- 5f these GM cottons do not prove to .e s"ita.le for the A"stralian cotton prod"ction system d"ring the first two seasons of the trial/ Dow AgroSciences intends to stop the trial at the end of second season #2004)00 s"mmer season$- *he efficacy data generated d"ring the initial stages of this trial will inform f"t"re research and potential commercialisation plans- *he GM cotton lines proposed for release are derived from the original American c"ltivar that was "sed in the initial .reeding program- 5f Dow AgroSciences wishes to cond"ct additional trials/ incl"ding .reeding the GM insecticidal traits into c"ltivars more s"ita.le for A"stralian conditions/ additional licence applications and approvals wo"ld .e re,"ired-
%,a-te. !
Ba/kg.#0nd
!8
(ection 9<2 Parent organism

)+ *he parent organism is c"ltivated cotton #Gossypiu" hirsutu" 7-$/ which is e2otic to A"stralia and is grown as an agric"lt"ral crop in New So"th 9ales and :"eensland and on a trial .asis in 9estern A"stralia and the Northern *erritory- More detailed information on cotton can .e fo"nd in a review doc"ment >*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"stralia4 #OG*+ 2002$ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cotton- *his doc"ment is availa.le at www.ogtr.gov.au-
(ection 9<3 Genetic modification and its effect

8+ Dow AgroSciences proposes to release three GM cotton lines/ two of which contain a single insecticidal gene/ cry1Fa #line 2=1B24B268 or @ry1!a cotton$ or cry1Ac #line 6008B210B26 or @ry1Ac cotton$- *he third line/ 9ideStri(e? cotton/ was prod"ced .y conventional .reeding of these two singleBgene insecticidal lines- *herefore 9ideStri(e? cotton contains .oth cry1Fa and cry1Ac genes*+ *he insecticidal genes in these GM cotton lines are derived from the common soil .acteri"m Bacillus thuringiensis #'t$- ;owever/ the cry1Fa and cry1Ac genes are chimeric genes/ ie- they contain parts of two other cry1 genes of 't/ ."t the proteins they e2press retain the species specific to2icity of native @ry1!a and @ry1Ac proteins for lepidopteran caterpillar insects/ incl"ding the maAor caterpillar pests of cotton #see Appendi2 1 Section 6 for details$- *he chimeric genes were developed to achieve improved e2pression in plants and sol".ility of the 't to2ins in the insect g"t!0+ 5n addition to the chimeric cry1Fa and)or cry1Ac genes/ these cotton lines contain a selecta.le mar(er gene #pat$ from the common soil .acteri"m Strepto"yces viridochro"ogenes- *he pat gene encodes the %A* #phosphinothricin acetyl transferase$ protein/ which confers tolerance to the her.icide gl"fosinate ammoni"m #the active ingredient of 7i.ertyE and 'astaE her.icides$- *he mar(er gene was "sed in the la.oratory d"ring the development of the GMOs for identification and selection of plant tiss"es in which the insecticidal genes were also present- Dow AgroSciences does not intend to apply the her.icide gl"fosinate ammoni"m to the GM cotton lines d"ring the proposed release!!+ Short reg"latory se,"ences #promoters and terminators$ that control e2pression of the introd"ced genes are also present in the GM cottons- *hese are derived from a plant/ &ea "ays #corn$/ and from a common .acteri"m/ Agro'acteriu" tu"e(aciensAltho"gh A. tu"e(aciens is a plant pathogen/ the reg"latory se,"ences comprise only a small part of its total genome/ and are not in themselves capa.le of ca"sing disease!2+ Additional information on the chimeric cry1Fa/ cry1Ac genes/ the pat gene/ and molec"lar characterisation of the inserted genetic materials and the new proteins e2pressed .y the GM cottons is provided in Appendi2 1-
(ection 9<4 Met1od of gene transfer

!3+ *he two cry1 genes were each introd"ced separately into the American cotton variety G@010/ in com.ination with one copy of the pat gene/ .y Agro'acteriu"Bmediated
%,a-te. ! Ba/kg.#0nd
!*
DNA transformation # am.rys(i 1CC2$- *he genes were introd"ced into the genome of cotton from plasmid vectors carried .y A. tu"e(aciens- *hese vectors were >disarmed4/ lac(ing the genes that encode the t"mo"rBind"cing f"nctions of A. tu"e(aciens #see Appendi2 1/ Section 4 for details$!4+ *he GM cotton plants containing the single insecticidal gene #cry1Fa or cry1Ac$/ or their progeny from selfBpollination/ were then crossed and repeatedly .ac(crossed/ to another elite American commercial cotton variety/ %S@600 #the >rec"rrent parent4 in the .reeding program$ to generate the two >parental4 GM cotton lines #referred to as @ry1!a cotton and @ry1Ac cotton$- *he two insecticidal traits were then com.ined .y conventional cross .reeding .etween these two GM cotton lines to generate the third GM cotton line/ referred to as 9ideStri(e? cotton- *h"s the GM 9ideStri(e? cotton line contains two insecticidal genes and two copies of the her.icide tolerance pat gene-
($+TI&' 2 PR$%I&-( R$)$A($( A'D I'T$R'ATI&'A) APPR&%A)(
(ection 2<9 PreAio2s releases in A2stralia

!&+ One of the GM cotton lines/ 9ideStri(e? cotton/ proposed for release is c"rrently .eing trialed "nder limited and controlled conditions on two sites covering a total area of 0-04 hectares in New So"th 9ales #D5+ 040)2006$- *here have .een no previo"s releases of either of the two singleBgene insecticidal GM cotton lines #@ry1!a and @ry1Ac cottons$ in A"stralia!'+ %revio"sly/ other GM cottons containing cry insecticidal genes derived from the same .acteri"m #Bacillus thuringiensis$ have .een trialed e2tensively in A"stralia #eg- D5+ s 000)2201/ 008)2001/ 00=)2001 and 00C)2001$/ as well as commercially released "nder licences D5+ 022)2002 #5NGA+DE cotton$ and D5+ 012)2001 #'ollgardE 55 cotton$- *he chimeric cry1Ac gene in two of the GM cotton lines proposed for release in the c"rrent application differs slightly from the chimeric cry1Ac gene present in 5NGA+DE and 'ollgardE 55 cottons!)+ Other GM plants e2pressing %A* proteins have also .een trialed in A"stralia #eg- D5+ 010)2001/ D5+ 010)2002 and D5+ 06=)2006$-
(ection 2<2 A
roAals 80 &t1er A2stralian GoAernment Agencies
!8+ *he OG*+ is responsi.le for assessing the ris(s to h"man health and the environment associated with development and "se of gene technology- Other government reg"latory re,"irements may .e relevant in respect of the release of GMOs/ and the "se of prod"cts of the GMOs- 5n this instance the responsi.ilities of the A"stralian %esticides and &eterinary Medicines A"thority #A%&MA$ and !ood Standards A"stralia New ealand #!SAN $ are relevant-
2<2<9
*ood (tandards A2stralia 'e4 Bealand
!*+ !SAN is responsi.le for h"man food safety assessment and la.elling/ incl"ding GM food- !SAN 4s approval wo"ld need to .e o.tained .efore any materials from the GM cotton lines proposed for release "nder this application co"ld .e "sed for h"man food%,a-te. ! Ba/kg.#0nd
20
20+ @"rrently/ !SAN is eval"ating an application for approval for food "se of oil and linters derived from 9ideStri(e? cotton- ;owever/ it sho"ld .e noted that none of the prod"cts from this release will .e "sed for h"man food2!+ GM corn containing @ry1!a core to2in has .een approved for food "se in A"stralia since 200622+ !"rther information is availa.le from !SAN : *ood (tandards A2stralia 'e4 Bealand %O 'o2 I1=8 @an.erra Mail @entre A@* 2810 %hone: #02$ 82I1 2222 !a2: #02$ 82I1 22I= HBmail: infoKfoodstandards-gov-a" http)**www.(oodstandards.gov.au
2<2<2
A2stralian Pesticides and %eterinar0 Medicines A2t1orit0
23+ *he A%&MA has a complementary reg"latory role in respect of this application d"e to its responsi.ility for agric"lt"ral chemical "se in A"stralia- 5nsecticidal GM plants fall "nder the Agricultural and eterinary !he"icals !ode Act 1##$ definition of an agric"lt"ral chemical prod"ct d"e to their prod"ction of insecticidal s".stances encoded .y the introd"ced genes- Any "se of her.icide on the GM cotton is also s".Aect to A%&MA reg"lation- !or commercial prod"cts/ the normal form of approval is thro"gh registration/ ."t the A%&MA may also iss"e permits allowing restricted "se of an agric"lt"ral chemical/ for e2ample for a limited period of time or on a limited area24+ Dow AgroSciences has s".mitted an application to the A%&MA for a research permit for the "se of the insecticidal genes in the GM cottons for the proposed trialDow AgroSciences does not intend to apply gl"fosinate ammoni"m her.icide to the GM cottons in the proposed release- *he A%&MA and the OG*+ have wor(ed closely to ens"re the thoro"gh/ coordinated assessment of these parallel proposals/ and that the decisions .y .oth agencies coincide2&+ 5n considering applications for registrations or permits/ the A%&MA also considers and/ if necessary/ imposes conditions in relation to a n"m.er of iss"es that are o"tside the scope of the Gene *echnology +eg"lator4s assessment/ s"ch as the efficacy of the prod"ct and resistance development2'+ !"rther information is availa.le from the A%&MA:
A2stralian Pesticides and %eterinar0 Medicines A2t1orit0 2!
%,a-te. !
Ba/kg.#0nd
%O 'o2 H240 J5NGS*ON A@* 2804 %hone: #02$ 82I2 0=02 !a2: #02$ 82I2 4I06 Hmail: contactKapvma-gov-a" http)**www.apv"a.gov.au
(ection 2<4 International a
roAals
2)+ All three GM insecticidal)her.icide tolerant cotton lines proposed for release "nder the c"rrent application have .een released for field trials in the 3nited States of America since 2001- A c"rrent H2perimental 3se %ermit has .een iss"ed .y the 3nited States Hnvironmental %rotection Agency #3S H%A$- *he 3S H%A has also iss"ed tolerance e2emptions for resid"es of the @ry1!a and @ry1Ac insecticidal proteins and their genes in all raw agric"lt"ral commodities when applied)"sed as plantBincorporated protectants #3S H%A 1CCIF 3S H%A 2006$- Dow AgroSciences has also .een cond"cting a field trial of the GM cottons in Argentina since 2004 #http)**www.sagpya."econ.gov.ar*12*ingles*release.ht"$-
%,a-te. !
Ba/kg.#0nd
22
+/APT$R 2 (-MMAR. &* RI(5 A(($((M$'T A'D RI(5 MA'AG$M$'T P)A'

28+ *he Act and the +eg"lations re,"ire that ris(s associated with dealings with GMOs are identified and assessed as to whether they can .e managed to protect h"man health and safety and the environment #see Appendi2 =$-
($+TI&' 9 I((-$( RAI($D I' (-#MI((I&'( &' T/$ APP)I+ATI&' A'D T/$
RI(5 A(($((M$'T A'D RI(5 MA'AG$M$'T P)A'
2*+ @omments received in response to the cons"ltation with e2pert gro"ps and a"thorities on the preparation of the ris( assessment and ris( management plan #+A+M%$ "nder section 00 of the Act and with the same sta(eholders and the p".lic on the +A+M%/ "nder section 02 of the Act #see Appendi2 =$/ were very important in finalising the plan/ which formed the .asis of the +eg"lator4s final decision on the application30+ 9ritten s".missions in relation to D5+ 044)2006 received from the agencies and a"thorities s"ggested the following iss"es relating to ris(s to h"man health and safety or the environment/ which have .een addressed in the +A+M%: 3*( the potential to2icity and allergenicity of GM cottons #Appendices 2 and 6 refer$F 40( the possi.ility that the GM cottons may .e harmf"l to the environment .eca"se of inherent weediness or increased potential for weediness/ partic"larly in favo"ra.le ha.itats s"ch as adAacent to nat"ral or artificial waterways #Appendi2 4 refers$F 4!( the e2tent of crossBpollination from GM cottons to other cotton crops #Appendi2 0/ section 2 refers$F 42( the possi.ility that the new genes introd"ced into the cottons can transfer to other organisms with adverse conse,"ences #Appendi2 0 refers$F 43( the emergence of insects resistant to the insecticidal proteins in the GM cottons #Appendi2 8 refers$F 44( meas"res to limit the "nintended dispersal in the environment of GM cottonseed and GM pollen #@hapter 2 and Appendi2 I refer$F and 4&( additional data re,"irements for the f"t"re development of the GMOs #@hapter 2 refers$3!+ *he +eg"lator received two s".missions from the p".lic on this application- A s"mmary of these written s".missions is provided in Appendi2 C- *he (ey iss"es raised .y the p".lic that related to h"man health and safety or the environment were: 4'( effect of the GM cottons on h"man and animal health and safetyF and 4)( effect of the GM cottons on soil organisms and the environment32+ One of the p".lic s".missions also raised .enefits of insecticidal GM cottons/ partic"larly red"ced pesticide "se/ which are o"tside the scope of eval"ation cond"cted "nder the Act and therefore have not .een considered as part of the assessment process%,a-te. 2 0mma.1 #$ Risk Assessment and Risk Management Plan
23
33+ 5n accordance with section 08 of the Act/ the +eg"lator has ta(en into acco"nt all iss"es raised in written s".missions that related to h"man health and safety and to the environment in finalising the ris( assessment and ris( management plan- *hese iss"es were considered caref"lly and weighed against the .ody of c"rrent scientific information in reaching the concl"sions set o"t in this doc"ment-
($+TI&' 2 *I'A)I(ATI&' &* T/$ RI(5 A(($((M$'T A'D RI(5 MA'AG$M$'T

P)A'
34+ *he +eg"lator has cond"cted a ris( assessment in relation to the proposed dealings and prepared a ris( management plan in accordance with the Act and the +eg"lations*he ris( assessment process "sed a +is( Analysis !ramewor( developed in cons"ltation with the p".lic and (ey sta(eholders #availa.le from the OG*+ we.site www.ogtr.gov.au$- A n"m.er of ha<ards were identified that may .e posed .y the proposed dealings- *he ris(s posed .y these ha<ards were assessed as .eing either >negligi'le4/ >very low+/ >low4/ >"oderate+/ >high4 or >very high+ .y considering: 48( the li(elihood of the ha<ards occ"rringF and 4*( the li(ely conse,"ences #impact$ of the ha<ards/ were they to .e realised3&+ *he following ta.le #*a.le 1$ lists each of the potential ha<ards that were considered d"ring the ris( assessment process in the ,azard -denti(ication col"mn and s"mmarises the assessment of each ha<ard "nder the col"mn headed Risk- A comprehensive ris( assessment of each identified ha<ard is provided in Appendices 2 B 8/ as crossBreferenced in the col"mn headed %u""ary o( Risk Assess"ent3'+ 9here it is considered that ris( management may .e necessary to protect the health and safety of h"mans and)or the environment/ the Risk .anage"ent col"mn identifies the selected methods and the reasons they were chosen- A ris( management plan for the proposed dealing is given effect .y specific conditions imposed in the licence*hese conditions are s"mmarised in the final col"mn/ headed /icence !onditions/ and detailed in Appendi2 I-
($+TI&' 3 R$($AR+/ R$>-IR$M$'T(

3)+ *he licence conditions incl"de the re,"irement that the applicant collect and provide to the +eg"lator f"rther information regarding: 38+ levels of e2pression of the insecticidal and her.icide tolerance genes in the GM cottons tiss"es "nder A"stralian field conditionsF &0( effect of the GM cottons on nonBtarget organisms "nder A"stralian field conditionsF &!( potential for the introd"ced proteins to acc"m"late in the soil "nder A"stralian field conditionsF and &2( efficacy of gene flow containment meas"res for any release sites in e2cess of one hectare/ within the associated +esearch one/ and for any release site for which a pollen trap is not re,"ired/ d"e to having no other cotton crops or pop"lations within 400 m-
%,a-te. 2
0mma.1 #$ Risk Assessment and Risk Management Plan
24
($+TI&' 4 ID$'TI*I+ATI&' &* I((-$( T& #$ ADDR$(($D *&R *-T-R$

R$)$A($(
3*+ *he proposed limited and controlled release is a small scale/ m"ltiBsite trial over fo"r cotton growing seasons- 5f the applicant ma(es any applications for f"t"re larger scale releases of GM insecticidal)her.icide tolerant cotton lines/ more data wo"ld .e re,"ired to .e provided on: &3( agronomic characteristics of the GM cottons in relation to potential weediness "nder A"stralian field conditionsF &4( effect of the GM cottons on soil .iotaF and &&( "nintended effects of the genetic modification40+ 5t sho"ld .e noted that collection of the a.ove data d"ring the proposed release is not re,"ired to ens"re the management of ris(s to h"man health and safety and the environment- *he ris( management meas"res s"mmarised in *a.le 1/ and given effect .y the licence conditions/ will effectively manage any s"ch ris(s4!+ 5t sho"ld also .e noted that the "se of prod"cts of the GM cottons in food wo"ld re,"ire approval from !SAN -
($+TI&' : D$+I(I&' &' T/$ APP)I+ATI&'

42+ Details of the matters that the +eg"lator m"st consider in ma(ing a decision are provided in Appendi2 =- 5t is important to note that the legislation re,"ires the +eg"lator to .ase the licence decision on whether ris( posed .y the dealings are a.le to .e managed so as to protect h"man health and safety and the environment43+ 5t is concl"ded that the proposed release of the GM cotton lines does not pose significant ris(s to h"man health and safety and the environment as a res"lt of the genetic modification- *he +eg"lator has imposed licence conditions to minimise potential e2pos"re of h"mans and other organisms to the GM cottons/ and to limit the spread and persistence of the GMOs or the introd"ced genetic materials while additional data is collected on the .ehavio"r and interactions of the GMOs in the A"stralian environment- Detailed ris( analyses .ased on the availa.le scientific information are provided in Appendices 2B8 in s"pport of this concl"sion*herefore/ the +eg"lator has iss"ed licence D5+ 044)2006 in respect of this application-
%,a-te. 2
2&
Ta8le 9
GM cottons: 7epidoptera: 't to2ins:
(2mmar0 of risk assessment and risk management lan !incl2ding im osed licence conditions"
the genetically modified cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cotton$ proposed for releasethe caterpillar insect pests targeted .y the GM cottons .elong to this order of insectsthe @ry1!a and @ry1Ac proteins may also .e referred to as a 't to2ins/ .eca"se they are some of the many protein to2ins prod"ced .y the .acteria Bacillus thuringiensis #'t$ in nat"rethe protein encoded .y the introd"ced pat gene/ which provides tolerance to the her.icide gl"fosinate ammoni"mnot applica.le-
%A*: N)A: Hazard Identifi ati!n
Ri"# (combine s 'likelihoo d' & 'impact') L!,
D!e" Ri"# S$%%ar& !f Ri"# A""e""%ent (refer to appendices for details) Re'$ire Mana(e%ent ) -es
.*
Ri"# Mana(e%ent Method(s) and Reason(s) for selection I" Ri"# Mana(e d)
Li en e !nditi!n" (See Appendix 7 for detailed licence conditions)
See A--endi. /
0*
TO*ICI TY AND ALLER GENIC ITY FOR H+MA NS: Fo od
products of the G cotton lines !ill not be used for human food or animal feed" or processed for lint or oil production# the introduced core proteins are %er& similar to the nati%e bacterial proteins !hich are alread& !idespread in the en%ironment and present in human food# the introduced proteins are of %er& lo! oral toxicit&# the introduced proteins are not kno!n to be aller)enic" nor do the& ha%e properties characteristic of kno!n aller)enic proteins# and compositional anal&ses ha%e not indicated an& differences bet!een the G and non+G cottonseed" other than the presence of the introduced proteins,
/*
Pre0ent "eed fr!% enterin( 1$%an f!!d "$--2&: pre%ents exposure throu)h food# De"tr!& a22 "eed n!t re'$ired f!r -!""i32e f$t$re tria2": pre%ents unintended exposure# and En"$re "e $re tran"-!rt and "t!ra(e !f retained "eed: pre%ents unintended exposure,
-es
0*
Pr!1i3it entr& int! 1$%an f!!d "$--2&: no materials from the G 1s to be used in human food or stock feed# De"tr!& "eed: destro& all seed not re3uired for possible future trials# Se $re tran"-!rt and "t!ra(e: har%ested material to be securel& !rapped for transport# G cottonseed must be transported !ithin a primar&" sealed container that is packed in a secondar& unbreakable container# store in sealed container !ithin a locked facilit& that is si)ned to indicate G cottonseed is stored !ithin,
$*
2*
$0*
'* (*
7*
**
%,a-te. 2
2'
L!, TO*ICI TY FOR OTHE R ORGA NISMS : a m mal s an d !il dlif e" incl udi n) bir ds an d fish
See A--endi. 4
$$*
-es
$7*
the introduced core proteins are %er& similar to the nati%e bacterial proteins !hich are alread& !idespread in the en%ironment" includin) in soil" on plants and on fresh produce" in the microor)anisms from !hich the )enes !ere deri%ed# the release is small in si4e and limited in duration# the toxicit& of 5r&$Fa and 5r&$Ac proteins is specific to lepidopteran caterpillar lar%ae# the 6A7 protein is not kno!n to be toxic to an& or)anism# exposure of li%estock and !ildlife to the G cotton lines !ould be lo!" and no materials from the release are proposed to be used in stockfeed# and toxicit& studies !ith purified proteins and 8t microbial preparations indicate that the 5r&$Fa and 5r&$Ac proteins are not toxic to mammals" birds or fish,
Li%it " a2e !f re2ea"e: decreases likelihood of exposure# Pre0ent "eed fr!% 3ein( $"ed a" "t! #feed: pre%ents exposure of animals# De"tr!& a22 "eed n!t re'$ired f!r f$rt1er tria2": pre%ents unintended exposure# and En"$re "e $re tran"-!rt and "t!ra(e GM %ateria2: pre%ents unintended exposure,
-es
'$*
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# Pre0ent "eed fr!% 3ein( $"ed a" "t! #feed: no material from the G 1s to be used in stock feed, De"tr!& "eed: destro& all seed not re3uired for possible future trials# and Se $re tran"-!rt and "t!ra(e: har%ested material to be securel& !rapped for transport# G cottonseed must be transported !ithin a primar&" sealed container that is packed in a secondar& unbreakable container# store in sealed container !ithin a locked facilit& that is si)ned to indicate G cottonseed is stored !ithin,
$0*
''*
$2*
'(*
$'* $(*
'**
'0*
$** $.*
$/*
%,a-te. 2
2)
TO*ICI TY FOR OTHE R ORGA NISMS : 9o n+ tar )et in% ert ebr ate s" incl udi n) soil d! elli n) or) ani sm s
L!,
See A--endi. 4
'.*
-es
($*
the introduced core proteins are %er& similar to the nati%e bacterial proteins !hich are alread& !idespread in the en%ironment" includin) in soil" on plants and on fresh produce" in the microor)anisms from !hich the )enes !ere deri%ed# the release is small in si4e and limited in duration# the toxicit& of the 5r&$Fa and 5r&$Ac proteins is specific to lepidopteran caterpillar lar%ae# the 6A7 protein is not kno!n to be toxic to an& or)anism# laborator& and field studies carried out in the :SA indicate that populations of non+tar)et in%ertebrates are unlikel& to be affected b& 5r&$Fa" 5r&$Ac and 6A7 proteins# and althou)h the risk is lo!" further information is re3uired on toxicit& for Australian non+tar)et or)anisms,
Li%it " a2e !f re2ea"e: decrease the likelihood of exposure,
-es
((*
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# Re'$ire re"ear 1: in%esti)ate potential toxicit& to non+tar)et or)anisms under Australian field conditions,
(** ('*
'/* '7*
F$rt1er Re"ear 1: in%esti)ate potential toxicit& to non+tar)et or)anisms under Australian field conditions,
'0* '2*
(0*
%,a-te. 2
28
TO*ICI TY FOR OTHE R ORGA NISMS : ic ro+ or) ani sm s
L!,
See A--endi. 4
(.*
-es
(0*
the introduced core proteins are %er& similar to the nati%e bacterial proteins !hich are alread& !idespread in the en%ironment" includin) in soil" on plants and on fresh produce" in the microor)anisms from !hich the )enes !ere deri%ed# the release is small in si4e and limited in duration# and althou)h the introduced proteins are unlikel& to ha%e ad%erse effect on soil microor)anisms" more information is re3uired on toxicit& for soil microor)anisms under Australian field conditions,
Li%it " a2e !f re2ea"e: decrease the likelihood of exposure,
-es
*0*
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# Re'$ire re"ear 1: in%esti)ate the potential for the introduced proteins to accumulate in soil under Australian conditions,
*$* (2*
F$rt1er re"ear 1: in%esti)ate persistence of the insecticidal proteins in soil under Australian conditions,
(/* (7*
%,a-te. 2
2*
5EEDI NESS:
L!,
See A--endi. 6
*'*
-es
*0*
cotton does not possess characteristics commonl& associated !ith !eediness" and is not kno!n to be a problematic !eed in an& en%ironment# the )enetic modifications in the G cotton lines are unlikel& to affect these characteristics# other G insecticidal cottons (tar)eted to lepidopteran caterpillar pests) )ro!n commerciall& in Australia ha%e not become problematic !eeds# ma;or constraints on !eediness of both G and non+G cottons are !ater a%ailabilit&" nutrient a%ailabilit&" plant competition" herbi%or& b& non+lepidopteran species" frost and fire# the presence of the insecticidal )ene(s) in the G cottons ma& confer a selecti%e ad%anta)e if the cotton is limited b& lepidopteran insects" ho!e%er" the risk is lo!# and althou)h the t!o traits (insecticidal and herbicide tolerance) ma& ha%e an additi%e effect on competiti%eness" neither of these traits indi%iduall& has been found to be si)nificant for !eediness,
Li%it " a2e !f t1e re2ea"e: decreases likelihood of escape, Eit1er "$rr!$nd t1e GM !tt!n" ,it1 a -!22en tra- !r i"!2ate fr!% !t1er !tt!n r!-" !r nat$ra2i"ed !tt!n -!-$2ati!n": minimises spread of the introduced )enes be&ond the release sites %ia pollen flo!, En"$re "e $re tran"-!rt and "t!ra(e GM %ateria2: pre%ents spread of G plant material outside the release sites, C2ean e'$i-%ent $"ed ,it1 t1e GMO": pre%ents spread of G plant material outside the release sites, Pre0ent !tt!n"eed 3ein( $"ed a" "t! #feed: pre%ent dispersal of cotton seed, De"tr!& an& 0!2$nteer": pre%ents persistence,
-es
.**
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# P!22en tra- !r i"!2ati!n: each release site must be surrounded b& a '0 m pollen trap of non+G cotton# or be at least *.0 m a!a& from an& other cotton crops and naturalised cotton populations, Se $re tran"-!rt and "t!ra(e: har%ested material to be securel& !rapped for transport# G cottonseed must be transported !ithin a primar&" sealed container that is packed in a secondar& unbreakable container# store in sealed container !ithin a locked facilit& that is si)ned to indicate G cottonseed is stored !ithin, C2ean e'$i-%ent: e3uipment must be cleaned before it is used for an& other purpose, <f the G cottons are )rinned" the )in must be cleaned immediatel& follo!in) its use" before an& other cotton is )inned, Pre0ent "eed fr!% 3ein( $"ed a" "t! #feed: no cottonseed to be used as stockfeed, De"tr!& 0!2$nteer": the release sites must be inspected after har%est at least once e%er& t!o months for at least $' months and an& cotton %olunteers destro&ed before flo!erin),
*2*
..*
*(*
***
./*
.0*
*.*
.$*
*/*
.7*
.'*
*7*
.(*
.0*
.2*
%,a-te. 2
30
GENE TRAN SFER: 6lants= 1th er cott on cro ps
L!,
See A--endi. 7
/0* /$*
-es
/'*
cotton is mostl& self+pollinated# and )ene transfer to other cotton crops or %olunteer cotton !ould not pose an& risks additional to the lo! risks posed b& the G cottons themsel%es,
/(*
Li%it " a2e !f t1e re2ea"e: decreases potential transfer, Eit1er "$rr!$nd t1e GM !tt!n" ,it1 a -!22en tra- !r i"!2ate fr!% !t1er !tt!n r!-" !r nat$ra2i"ed !tt!n -!-$2ati!n": minimises spread of the introduced )enes be&ond the release sites %ia pollen flo!, En"$re "e $re tran"-!rt and "t!ra(e !f retained "eed: pre%ents spread of G plant materials outside the release sites, C2ean e'$i-%ent $"ed ,it1 t1e GMO": pre%ents spread of G plant materials into the en%ironment outside the release sites, De"tr!& an& 0!2$nteer": pre%ents persistence,
-es
/0*
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# P!22en tra- !r i"!2ati!n: each release site must be surrounded b& a '0 m pollen trap of non+G cotton# or be at least *.0 m a!a& from an& other cotton crops and naturalised cotton populations, Se $re tran"-!rt and "t!ra(e: har%ested material to be securel& !rapped for transport# G cottonseed must be transported unless contained !ithin a primar&" sealed container that is packed in a secondar& unbreakable container# store in sealed container !ithin a locked facilit& that is si)ned to indicate G cottonseed is stored !ithin, C2ean e'$i-%ent: e3uipment must be cleaned before it is used for an& other purpose, <f the G cottons are )rinned" the )in must be cleaned immediatel& follo!in) its use" before an& other cotton is )inned, De"tr!& 0!2$nteer": the release sites must be inspected after har%est at least once e%er& t!o months for at least $' months and an& cotton %olunteers destro&ed before flo!erin), Re'$ire re"ear 1: pro%ide for a *00 m research 4one surroundin) release sites in excess of $ ha" for the purpose of conductin) research on )ene flo! and conduct research on efficac& of )ene flo! containment measures for sites !ith no pollen traps but *.0 m a!a& from other cotton populations,
/2*
70*
/**
/.*
7$*
//*
7'* /7*
F$rt1er re"ear 1: in order to inform the on)oin) re%ie! of the data on )ene transfer and %alidate the efficac& of containment measures,
7(*
%,a-te. 2
3!
Hazard Identifi ati!n
Ri"# Mana(e%ent Ri"# (combines 'likelihood' & 'impact') S$%%ar& !f Ri"# A""e""%ent (refer to appendices for details) D!e" Ri"# Re'$ire Mana(e%ent ) See A--endi. 7
7** 7.*
Li en e !nditi!n" I" Ri"# Mana(e d) (See Appendix 7 for detailed licence conditions)
Met1!d(") and Reason(s) for selection
GENE TRANSF ER:
L!,
-es
7/*
cotton is mostl& self+pollinated# and )ene transfer to naturalised cotton !ould not pose an& risks additional to the risks posed b& the G cottons themsel%es,
77*
Li%it " a2e !f t1e re2ea"e: decreases potential transfer, Eit1er "$rr!$nd t1e GM !tt!n" ,it1 a -!22en tra- !r i"!2ate fr!% !t1er !tt!n r!-" !r nat$ra2i"ed !tt!n -!-$2ati!n": minimises spread of the introduced )enes be&ond the release sites %ia pollen flo!, En"$re "e $re tran"-!rt and "t!ra(e !f retained "eed: pre%ents spread plant material outside the release sites, C2ean e'$i-%ent $"ed ,it1 t1e GMO": pre%ents spread of G plant material into the en%ironment outside the release sites, De"tr!& an& 0!2$nteer": pre%ents persistence,
-es
0$*
Li%it " a2e: restrict area to $',' ha o%er four cotton )ro!in) seasons# P!22en tra- !r i"!2ati!n: each release site must be surrounded b& a '0 m pollen trap of non+G cotton# or be at least *.0 m a!a& from an& other cotton crops and naturalised cotton populations, Se $re tran"-!rt and "t!ra(e: har%ested material to be securel& !rapped for transport# G cottonseed must not be transported unless contained !ithin a primar&" sealed container that is packed into a secondar& unbreakable container# onl& transport to the extent necessar& to store# store in sealed container !ithin a locked facilit& that is si)ned to indicate G cottonseed is stored !ithin, C2eanin( e'$i-%ent: e3uipment must be cleaned before it is used for an& other purpose, <f G cotton is )inned" the )in must be cleaned immediatel& follo!in) its use" before an& other cotton is )inned, De"tr!& 0!2$nteer": the release site must be monitored after har%est at least once e%er& t!o months for at least $' months and an& cotton %olunteers destro&ed before flo!erin),
0'*
6lants= Feral (naturalis ed) cotton
0(*
70*
72*
00*
0**
0.*
%,a-te. 2
32
GENE TRANSF ER: 6lants
Ne(2i(i32e
See A--endi. 7
0/*
9o
9>A
9>A
9one re3uired
)enetic incompatibilit& and )eo)raphical isolation from nati%e populations pre%ent the production of fertile h&brids,
9ati% e cotto ns GENE TRANSF ER: 6lants

07*
Ne(2i(i32e
See A--endi. 7
00*
9o
9>A
9>A
9one re3uired
?ell+established )enetic incompatibilit& pre%ents successful cross+pollination !ith other plant species,
1th er )en era
%,a-te. 2
33
Hazard Identifi ati!n
Ri"# Mana(e%ent Ri"# (combines 'likelihood' & 'impact') S$%%ar& !f Ri"# A""e""%ent (refer to appendices for details) D!e" Ri"# Re'$ire Mana(e%ent ) See A--endi. 7
02*
Li en e !nditi!n" I" Ri"# Mana(e d) (See Appendix 7 for detailed licence conditions)
Met1!d(") and Reason(s) for selection
GENE TRANSF ER: icro + or)an isms
Ne(2i(i32e
9o
9>A
9>A
9one re3uired
the introduced )enes in the G cottons are deri%ed from" and are similar to" nati%e bacterial )enes that are alread& !idespread in the en%ironment" and are readil& a%ailable for transfer from these sources %ia demonstrated natural mechanisms# and )ene transfer from plants to microor)anisms has not been demonstrated under natural conditions" and the likelihood of such transfer is )reatl& exceeded b& the likelihood of transfer from other sources of these )enes, 9o 9>A 9>A 9one re3uired
20*
GENE TRANSF ER: Anim als" inclu din) huma ns
Ne(2i(i32e
See A--endi. 7
2$*
the introduced )enes in the G cottons are deri%ed from" and are similar to" nati%e bacterial )enes that are alread& !idespread in the en%ironment# transfer or the introduced )enes !ould be unlikel& to pose a ha4ard to human health and safet& or to the en%ironment# products from the G cottons are not intended for stockfeed or human food# most animals a%oid feedin) on G or non+G cotton plants# and e%en if the G cottons !ere appro%ed b& FSA9@ for use in food" the cotton b&+products used in food do not contain A9A#
2'*
2(*
2**
2.*
%,a-te. 2
34
RESISTA NCE: <nsec ticide
N8A
See A--endi. 9
2/*
A6B A is responsible for assessin) and mana)in) this risk
ana)ed b& the A6B A
A6B A !ould impose appropriate conditions
9one re3uired Cicence notes the re3uirement to adhere to an& A6B A conditions" includin) an& insecticide resistance mana)ement strate)&,
APP$'DI, 9 I'*&RMATI&' A#&-T T/$ GM&(

44+ 5n preparing the ris( assessment and ris( management plan/ the +eg"lator is re,"ired "nder section 4C #2$ of the Act to consider the properties of the parent organism and the effects of genetic modification4&+ *his Appendi2 addresses these matters and provides detailed information a.o"t the GMOs proposed for release/ the parent organism/ the genetic modification process/ the genes that have .een introd"ced and the new proteins that are e2pressed in the GM cotton lines4'+ 5t sho"ld .e noted that some specific Dow AgroSciences doc"ments/ which contain details of the plasmid vectors/ gene se,"ence information and molec"lar characterisation of the inserted genetic materials/ have previo"sly .een declared as @onfidential @ommercial 5nformation #@@5$ "nder section 1=0 of the Act/ in connection with a previo"s Dow AgroSciences4 licence application/ D5+ 040)2006;owever/ the @@5 was made availa.le to the prescri.ed e2pert gro"ps/ which were cons"lted in the preparation of the ris( assessment and ris( management plan-
($+TI&' 9 (-MMAR. I'*&RMATI&' A#&-T T/$ GM&(

4)+ Dow AgroSciences proposes to release three GM cotton lines/ two of which contain a single insecticidal gene/ chimeric cry1Fa #line 2=1B 24B268 or @ry1!a cotton$ or chimeric cry1Ac #line 6008B210B26 or @ry1Ac cotton$- *he third line/ 9ideStri(e? cotton/ was prod"ced .y conventional .reeding of these two singleBgene insecticidal lines- *herefore 9ideStri(e? cotton contains .oth cry1Fa and cry1Ac genes- *he insecticidal genes are derived from the common soil .acteri"m Bacillus thuringiensis #'t$ and e2press insecticidal proteins #'t to2ins$ that are to2ic to specific lepidopteran caterpillar insects/ incl"ding the maAor caterpillar pests of cotton48+ All three lines also contain a her.icide tolerance selecta.le mar(er gene/ pat/ that confers tolerance to the her.icide gl"fosinate ammoni"m #the active ingredient of 7i.ertyE and 'astaE her.icides$- *he mar(er gene was "sed in the la.oratory d"ring the development of the GMOs for identification and selection of plant tiss"es in which the insecticidal genes were also present4*+ Short reg"latory se,"ences #promoters and terminators$ that control e2pression of the introd"ced genes are also present in these GM cottons- *hese se,"ences are derived from a plant/ &ea "ays #corn$/ and a common .acteri"m/ Agro'acteriu" tu"e(aciens- Altho"gh A. tu"e(aciens is a plant pathogen/ the reg"latory se,"ences comprise only a small part of its total genome/ and are not in themselves capa.le of ca"sing disease%,a-te. 2 0mma.1 #$ Risk Assessment and Risk Management Plan
3&
&0+ *he characteristics of the introd"ced genes and their prod"cts/ and methods "sed to introd"ce the genes into cottons are disc"ssed in Sections 6 L 4 of this Appendi2-
($+TI&' 2 T/$ PAR$'T &RGA'I(M

&!+ A comprehensive review of the parent organism/ Gossypiu" hirsutu" 7- #c"ltivated cotton$/ is provided in the doc"ment >*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"stralia4 #OG*+ 2002$ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cottons- *his doc"ment can .e accessed at www.ogtr.gov.au.
($+TI&' 3 T/$ I'TR&D-+$D G$'$( A'D T/$IR PR&D-+T(
(ection 3<9 T1e cry1Fa and cry1Ac genes

&2+ *he cry1Fa and cry1Ac genes in the GM insecticidal)her.icide tolerant cotton lines are chimeric genes/ each com.ining parts of three different cry genes isolated from Bacillus thuringiensis #'t$- *he part of the chimeric cry1Fa gene which corresponds to the active core to2in is derived from the native cry1Fa gene of 't variety aizawai #'ta$- *he part of the chimeric cry1Ac gene which corresponds to the active core #f"nctional$ to2in is derived from the native cry1Ac gene of 't variety kurstaki #'t($ strain ;DI6- *he remainder of each of these genes/ encoding the car.o2yBterminal portion of the proteins which is cleaved off in the insect g"t/ is derived from parts of the cry1!a0 and cry1A'1 genes&3+ *he car.o2ylBterminal potion of the @ry1 proteins is not essential for to2icity- 5ts f"nction appears to .e in the maintenance of the "n"s"al sol".ility of the @ry1 proteins #7"thy L H.ersold 1C=1$- ;owever it is a highly conserved region/ a property which allows coB assem.ly of different @ry1 proteins into the same crystal #;ofte L 9hiteley 1C=C$- *he chimeric genes were developed to improve the level of e2pression in plants and the sol".ility of the encoded 't to2ins in the insect g"t- *he coding se,"ence of the chimeric genes has .een f"rther modified to achieve optimal e2pression in plants/ witho"t affecting the encoded protein se,"ences&4+ *he chimeric cry1Fa and cry1Ac genes each encode a protein to2in #@ry protein or 't to2in$/ @ry1!a and @ry1Ac/ respectively which are very similar to native @ry1!a and @ry1Ac proteins- 9ithin the core to2in/ the amino acid se,"ences of the native and chimeric proteins are CC-6D and CC-8D identical/ respectively/ and retain the species specificity of to2icity to larvae of lepidopteran insects #moths and ."tterflies$ characteristic of native @ry1!a and @ry1Ac proteins&&+ *he chimeric cry1Ac gene in the GM cotton lines differs from the cry1Ac gene present in .oth 5NGA+DE and 'ollgardE 55 cottons/ which is a chimeric gene derived from cry1Ac and cry1A' genes of 't- 5NGA+DE and 'ollgardE 55 are approved for commercial release in A"stralia "nder licences D5+ 022)2002 and D5+ 012)2002/ respectively #see www.ogtr.gov.au$-
%,a-te. 2
3'
&'+ H2pression of the chimeric cry1Fa gene is controlled .y the #4O@S$mas 24 promoter/ a synthetic promoter derived from the mannose synthase gene #"as$ promoter and octopine synthase gene #ocs$ enhancer of A. tu"e(aciens #Ni et al- 1CC0$- *he m+NA termination region is provided .y the .idirectional polyadenylation signal of A. tu"e(aciens open reading frame 20 #'ar(er et al- 1C=6$&)+ H2pression of the chimeric cry1Ac gene in these GM cottons is controlled .y the 3.i,"itin promoter of &ea "ays #corn$ #@hristensen et al- 1CC2$- *he m+NA termination region is also provided .y the .idirectional polyadenylation signal of Agro'acteriu" tu"e(aciens open reading frame 20 #'ar(er et al- 1C=6$-
(ection 3<2 &rigin and 2se of insecticidal Bacillus thuringiensis

&8+ 5nformation on the origin and "se of B. thuringiensis #'t$ is incl"ded here to provide .ac(gro"nd information on 't and 't prod"cts- 't is a gram positive/ sporeBforming soil .acteri"m that is ".i,"ito"s in the environment- 5t has .een fo"nd in dead insects/ insect .reeding environments/ stored grain/ soil and leaf s"rfaces #D"lmage 1C=2$- %rod"cts consisting of .acterial spores and @ry proteins have .een applied to crops for over 40 years in the 3SA #9al(er et al- 2006$- Discovered in the early 20th cent"ry in Mapan .y S- 5shiwata/ the .acteri"m was first isolated from diseased flo"r moth larvae in *h"ringia/ Germany .y H- 'erliner #see 9al(er et al- 2006$- 7ater/ HJ"rsta( of !rance and ;- D"lmage of the 3S Department of Agric"lt"re isolated and acc"m"lated the first significant collection of strains- *hese strains that were active against lepidopteran insects were gro"ped together and given the s".species name kurstaki #D"lmage 1C=2$&*+ 't first .ecame availa.le as a commercial insecticide in !rance in 1C6= #Neppl 2000$ and was "sed commercially in the 3SA in 1C0=*he first 3S Hnvironment %rotection Agency registration of 't occ"rred in 1C81 #Starnes L 7i" 1CC6$- *he discovery of other 't s".species/ s"ch as B. thuringiensis israelensis that is active against mos,"itoes and .lac(flies #Gold.erg L Margalit 1CII$ and B. thuringiensis tene'rionis with activity against some coleopteran insects #eg- .eetles$ #Jrieg et al- 1C=6$ showed the .road insecticidal potential of 't s".species- Hach s".species demonstrated a certain level of insecticidal specificity/ s"ggesting that 't was ideal for integrated pest management #Glare and O4@allaghan/ 2000$- *h"s 't preparations have .ecome the most widely "sed micro.ial insecticides #acco"nting for over C0D of all commercial sales$ #Glare and O4@allaghan/ 2000$'0+ 't was introd"ced to A"stralia in late 1CI0s #*ea(le 1CC1$- *he first registration of 't was in So"th A"stralia in 1C=0 for "se in that State #www.apv"a.gov.au$-
(ection 3<3 Insecticidal +r0 roteins !#t toxins" and t1eir mode of action
'!+ *he @ry proteins/ also referred to as Bendoto2ins/ insecticidal crystal proteins or proto2ins are one class of to2ins prod"ced .y 'tD"ring spor"lation/ 't prod"ces a parasporal crystal composed of one or more @ry proteins- *he formation of the parasporal crystal
%,a-te. 2 0mma.1 #$ Risk Assessment and Risk Management Plan
3)
disting"ishes B. thuringiensis from other Bacillus species- *he @ry proteins of each B. thuringiensis s".species are often to2ic to specific insect genera'2+ *he @ry proteins are classified according to their degree of amino acid homology #;ofte L 9hiteley 1C=CF @ric(more et al- 2002$/ which also determines their target specificity- !or e2ample the @ry1 family of to2ins/ incl"ding the @ry1!a and @ry1Ac to2ins #from the native cry1Fa and cry1Ac genes$/ are highly specific against lepidopteran insects #moths and ."tterflies$ #Macintosh et al- 1CC0F @ham.ers et al1CC1F !ederici 2006$- *he @ry1!a and @ry1Ac to2ins have a different ."t overlapping spectr"m of to2icity to lepidopteran insect species'3+ *he species specificity of the @ry protein res"lts from a series of steps which m"st occ"r .efore the to2icity is realised- !irstly/ the @ry protein crystal re,"ires al(aline conditions/ as in the larval insect g"t/ with p; val"es of 10 or a.ove/ to .e sol".le- *he @ry protein/ which is in the form of a proto2in/ m"st then .e partially digested .y a specific protease in the insect g"t to create its >active4 #to2ic$ form #core to2in$- *he active core to2in m"st then diff"se thro"gh the midg"t mem.rane and .ind with specific receptors fo"nd on the midg"t epitheli"m s"rface in order to e2ert its to2ic activity #;ofmann et al- 1C==F &an +ie et al- 1C=CF Jarim et al- 2000$'4+ 'inding of activated @ry to2in to the insect epithelial g"t cell receptors leads to formation of pores in the cell mem.rane/ allowing lea(age of intracell"lar contents #for e2ample potassi"m ions$ into the g"t l"men and water into the cell #Sacchi et al- 1C=8F Hnglish L Slatin 1CC2F Jnowles L Dow 1CC6$- *he larval g"t epithelial cells swell d"e to osmotic press"re and r"pt"re- *he g"t .ecomes paralysed .eca"se of changes in the electrolyte and p; .alance and the insects stop eating and die #Gold."rg et al- 1CC0$'&+ *he specific set of conditions re,"ired for @ry1 to2icity is only fo"nd in lepidopteran insect larvae- Specific proteases cleave off the car.o2ylBterminal domain as well as appro2imately 2= amino acids from the aminoBterminal end/ of the @ry1!a and @ry1Ac proteins/ leaving an active proteaseBresistant core of appro2imately 800 amino acids #'ietlot et al- 1C=CF @homa L Japlan 1CC0F Narva et al2001.F Narva et al- 2001a$''+ NonBtarget insects/ mammals #incl"ding h"mans$/ .irds and fish do not possess the specific receptors for the @ry proteins and therefore are not s"scepti.le to the to2ic effects #!ederici 2006$')+ *he .iological activity of the @ry1!a and @ry1Ac proteins e2pressed in the GM cotton lines has .een demonstrated in the la.oratory and in field trials in the 3nited States of America #3SA$/ on to.acco ."dworm #,eliothis virescens$ and pin( .ollworm #1ectinophora gossypiella$ #%ellow 2001$-
%,a-te. 2
38
(ection 3<4 T1e "at gene

'8+ *he pat gene in the GM cotton lines is derived from the .acteri"m %trepto"yces viridochro"ogenes #Stra"ch et al- 1C==F 9ohlle.en et al1C==$- *he coding se,"ence of the pat gene has .een modified to achieve optimal e2pression in plants/ witho"t affecting the encoded protein se,"ence'*+ %trepto"yces spp- are saprophytic/ soil .acteria/ not considered pathogens of plants/ h"mans or other animals #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$- 5n %. viridochro"ogenes-/ the pat gene provides resistance to the tripeptide anti.iotics nat"rally prod"ced .y a small n"m.er of actinomycete .acteria #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$: 'ialaphos #phosphinothricylB7BalanylB7Balanine$/ prod"ced .y %. viridochro"ogenes and %. hygroscopicus #the .acterial species from which the pat and related 'ar genes/ respectively/ are derived$F and %hosalacine #phosphinothricylB7BalanylB7Ble"cine$/ prod"ced .y 2itasatosporia phosalacinea)0+ *here are two versions of the pat gene in the GM cotton lines/ each "nder control of a different promoter- *hese promoters are identical to those controlling the two cry1 genes a.ove/ namely the 3.i,"itin promoter of &ea "ays #corn$ #@hristensen et al- 1CC2$ and the #4O@S$mas 24 synthetic promoter/ derived from the mannose synthase gene promoter and octopine synthase gene enhancer of A. tu"e(aciens #Ni et al- 1CC0$- *he m+NA termination region for .oth is provided .y the .idirectional polyadenylation signal of A. tu"e(aciens open reading frame 20 #'ar(er et al- 1C=6$- 5n the @ry1Ac cotton line #e2pressing of cry1Ac "nder control of the 3.i,"itin promoter$/ pat is "nder control of the #4O@S$mas 24 promoter/ whereas in the @ry1!a cotton line #e2pressing of cry1Fa "nder control of the #4O@S$mas 24 promoter$/ pat is "nder control of the 3.i,"itin promoter-
(ection 3<:
T1e PAT rotein
)!+ *he pat gene encodes the en<yme phosphinothricin acetyl transferase #%A*$/ which deto2ifies the her.icide gl"fosinate ammoni"m #also referred to as phosphinothricin$ #9ohlle.en et al- 1C==$- *he gl"fosinate ammoni"m tolerance trait was introd"ced into the GM cotton lines as a selecta.le mar(er to identify GM plant cells and GM plants d"ring their development in the la.oratory/ as well as potentially ena.ling the "se of gl"fosinate ammoni"m as a her.icide to control weeds in the crop- ;owever/ the applicant does not intend to apply gl"fosinate ammoni"m to the GM cottons in the proposed trial)2+ Gl"fosinate ammoni"m acts as a her.icide .y inhi.iting the plant en<yme gl"tamine synthetase/ res"lting in ammonia acc"m"lation/ inhi.ition of amino acid synthesis and inhi.ition of photosynthesis/ leading to severe damage to plant tiss"es/ "ltimately (illing the plant #%line 1CCC$- Gl"fosinate ammoni"m is the active ingredient of a n"m.er of proprietary her.icides incl"ding 'astaE/ !inaleE and 7i.ertyE3*
%,a-te. 2
($+TI&' 4 M$T/&D &* G$'$ TRA'(*$R

)3+ *he two chimeric cry1 genes were each introd"ced separately into an American commercial cotton variety/ G@010/ in com.ination with one copy of the pat gene/ .y Agro'acteriu"Bmediated DNA transformation # am.rys(i 1CC2$)4+ A. tu"e(aciens is a common gramBnegative soil .acteri"m that ca"ses crown gall disease in a wide variety of plants- %lants can .e genetically transformed #modified$ .y the transfer of DNA #*BDNA/ located .etween specific .order se,"ences on a resident plasmid$ from A. tu"e(aciens/ thro"gh the mediation of genes from the vir #vir"lence$ region of *i plasmids)&+ Disarmed Agro'acteriu" strains have .een constr"cted specifically for plant transformation- *he disarmed strains do not contain the genes #iaa./ iaa, and ipt$ responsi.le for the overprod"ction of a"2in and cyto(inin/ which are re,"ired for t"mo"r ind"ction and rapid call"s growth #Jlee L +ogers 1C=C$)'+ *he plasmids "sed contain well characterised DNA segments re,"ired for their replication and selection in .acteria/ and for transfer of DNA from Agro'acteriu" and its integration into the plant cell genome #'evan 1C=4F 9ang et al- 1C=4$- Agro'acteriu"Bmediated transformation has .een widely "sed in A"stralia and overseas for introd"cing new genes into plants witho"t ca"sing any .iosafety pro.lems))+ !ollowing coBc"ltivation with A. tu"e(aciens carrying the plasmid with the gene constr"ct/ cotton cells were c"lt"red in the presence of gl"fosinate ammoni"m to select for those cells containing inserted gene constr"ct #since the pat gene confers tolerance to gl"fosinate ammoni"m$- S".se,"ently cotton plants containing the individ"al insecticidal genes were regenerated from these GM cells)8+ *he chimeric cry1Ac gene #"nder control of the 3.i,"itin promoter$ with one pat gene #"nder control of the #4O@S$mas24 promoter$ were introd"ced into cotton from plasmid pMN@6008/ leading to a genetic modification or genetic >transformation4/ referred to as transformation event 6008B210B26 or @ry1Ac cotton)*+ *he chimeric cry1Fa gene #"nder control of the #4O@S$mas24 promoter$ with one pat gene #"nder control of the 3.i,"itin promoter$ were introd"ced into cotton cells from plasmid pAGM2=1/ leading to transformation event 2=1B24B268 or @ry 1!a cotton80+ *he two GM cotton plants containing the single insecticidal traits/ or their progeny from self pollination/ were then crossed/ and repeatedly .ac(crossed/ to another elite American commercial cotton variety/ %S@600 #the >rec"rrent parent4 in the .reeding program$*he two transformation events #2=1B24B268 and 6008B210B26$ were then com.ined .y conventional .reeding to generate the third GM cotton line/ referred to as 9ideStri(e? cotton- *h"s the GM 9ideStri(e? cotton line contains two insecticidal genes/ chimeric cry1Fa and chimeric cry1Ac and two copies of the her.icide tolerance pat gene-
%,a-te. 2
40
($+TI&' : +/ARA+T$RI(ATI&' A'D (TA#I)IT. &* T/$ I'($RT$D G$'$TI+ MAT$RIA)(

8!+ So"thern .lot analysis/ "sing pro.es from each gene and from reg"latory se,"ences #promoters and termination region$/ demonstrates that the transformation event 6008B210B26 contains one intact copy of the insecticidal gene/ chimeric cry1Ac/ and one intact copy of the her.icide tolerance gene/ pat- *ransformation event 2=1B24B268 contains one intact copy of the insecticidal gene/ chimeric cry1Fa/ and one intact copy pl"s an additional small fragment of the pat gene #Green 2002F Green et al- 2002.F Green et al- 2002a$82+ *he DNA se,"ences from each transformation event have also .een confirmed .y DNA se,"ence analysis #Song 2002aF Song 2002.$83+ *he gene constr"ct in each transformation event have .een shown to .e sta.le over several generations/ .oth .y phenotype #insecticidal and gl"fosinate ammoni"m tolerance$ and So"thern .lot analysis/ adhering to Mendelian inheritance ratios #Narva et al- 2001.F Narva et al- 2001a$84+ *he GM 9ideStri(e? cotton contains all of the introd"ced genetic material of transformation events 2=1B24B268 and 6008B210B26 #Green 2002$-
($+TI&' ; $,PR$((I&' &* T/$ I'TR&D-+$D PR&T$I'(

8&+ H2pression levels of @ry1!a/ @ry1Ac and %A* proteins in vario"s plant tiss"es/ as well as in processed cottonseed fractions/ from plants of these three cotton lines grown in the field in the 3SA/ have .een determined .y en<yme lin(ed imm"nosor.ent assay #H75SA$ #%hillips et al- 2002$8'+ Data for mean protein levels from samples collected from these GM cotton lines grown in 2001 at si2 sites in five States of the 3SA #only two sites for nectar$ is presented in *a.les 1 and 2- *he @ry1!a protein was detected in @ry1!a cotton and 9ideStri(e? cotton at a low level in all tiss"es and processed fractions e2cept nectar/ meal and oil- *he highest mean val"e of its e2pression was 20-6 g protein)g tiss"e dry weight #e,"ivalent to ppm$ fo"nd in whole plants #com.ined a.ove gro"nd tiss"es$ of 9ideStri(e? cotton at pollination stage- *he @ry1Ac protein in @ry1Ac cotton and 9ideStri(e? cotton was e2pressed at lower levels than @ry1!a protein in all tiss"es and fractions e2cept for pollen- *he %A* protein was detected in most of the samples of @ry1!a cotton and 9ideStri(e? cotton/ ."t was not detecta.le in most tiss"es and fractions of @ry1Ac cotton #*a.les 1 and 2$8)+ H2pression of the introd"ced proteins in these GM cotton lines thro"gho"t the season was also e2amined at two sites #*a.le 6$H,"ivalent tiss"e was collected at different stages of plant growth #eg- yo"ng leaves collected in areas of new plant growth at appro2imately 1/ 2/ 6 and 4 months/ or open flowers every 2 O 6 wee(s from first flowering$- *he @ry1Ac and %A* proteins were e2pressed thro"gho"t the season witho"t any significant changes in e2pression level- At only one site the @ry1!a protein was fo"nd at s".stantially higher levels in yo"ng leaves of @ry1!a cotton and 9ideStri(e? cotton later in the season #*a.le 6$%,a-te. 2 0mma.1 #$ Risk Assessment and Risk Management Plan
4!
($+TI&' C I'($+TI+IDA) $**I+A+. &* T/$ GM +&TT&'(

88+ *he insecticidal efficacy of the GM cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ has .een assessed in field trials cond"cted at different locations in the 3SA in 2001 #%ellow 2001$8*+ *hree trials "sed either artificial or nat"ral infestation with to.acco ."dworm #,eliothis virescens$- *he performance of these GM cottons was compared to nonBGM cotton with and witho"t chemical spray control for the insect pest- N"m.ers of insect larvae and damage to s,"ares and .olls were assessed thro"gho"t the growing season*0+ Hach of these cotton lines was fo"nd to perform .etter than nonBGM cotton that was not sprayed to control lepidopteran insect pests/ and at least as well as sprayed nonBGM cotton- 5n some cases the GM cottons o"tBperformed sprayed nonBGM cotton*!+ One trial was cond"cted "sing artificial infestation with pin( .ollworm #1ectinophora gossypiella$- 7arvae/ p"pae and e2it holes were co"nted 2 wee(s after inoc"lation with eggs- *he @ry1Ac cotton and the 9ideStri(e? cotton provided nearly complete control of larvae/ while @ry1!a cotton was similar to nonBGM cotton*2+ 7ater trials cond"cted with 9ideStri(e? cotton in comparison sprayed nonBGM cotton/ variety %S@600/ showed that this GM cotton can provide effective control of maAor insect pests of cotton at vario"s locations in the 3SA/ incl"ding to.acco ."dworm 3,eliothis virescens$/ cotton .ollworm #,elicoverpa zea$ and pin( .ollworm #1ectinophora gossypiella$- 9ideStri(e? cotton was also effective against .eet armyworm #%podoptera e4igua$/ so"thern armyworm #%. eridania$ fall armyworm #%. (rugiperda$/ soy.ean looper #1seudoplusia includens$ and ca..age looper #Trichoplusia ni$- *his s"ggests that the 9ideStri(e? cotton can provide effective control of a wide range of lepidopteran pests of cotton #%ellow 2002$-
%,a-te. 2
42
DIR 044/2003 Risk Assessment and Risk Management Plan Office of the Gene Technology Regulator
Ta8le 9
$x ression leAels of t1e introd2ced roteins in Aario2s tiss2es of t1e t1ree GM cotton lines
Protein $x ression
a Ti #Mean g protein)g dry weight or fresh weight. standard ss deviation$ 2 e
@ ry 1 A c
@ ry 1 ! a
%A*
9 @ 9 @ 9 @ @ id ry id ry id ry ry e 1 e 1 e 1 1 St A St ! St A ! ri c ri a ri c a (e co (e co (e co co ? tt ? tt ? tt tt o o o o n n n n
A--endi2 !
In$#.mati#n a"#0t t,e 3MOs
43
N o" ng le af #6 B8 w ee ($ *e r m in al le af #C B 16 w ee ($
1=2 08
1C2 0I
8=1 68
84= 66
046 016
N D
046 012
161 04
144 00
=1C 60
I8I 06
026 011
N D
021 012
A--endi2 !
44
000 00I
00I 00C
416 11
016 11
004 021
#0 -0 8$
d
04I 01I
Se ed
. a
g )g is e,"ivalent to parts per million #ppm$+es"lts .ased on fresh tiss"e weight for pollen/ seed and nectar/ dry weight for all other tiss"es/ representing the mean of samples collected from si2 sites9hole plant refers to com.ined a.oveBgro"nd plant tiss"e!ig"res in .rac(ets represent val"es .elow the validated limit of ,"antitation for the assay-
ND: not detected-
A--endi2 !
4&
Ta8le 2 )eAels of t1e introd2ced roteins in rocessed GM cottonseed fractions

+ ot to n se e d * r a ct io n
PR&T$I' $,PR$((I&' !mean g protein)ga fresh weight"
@ @ ry ry 1 1 A ! c a
%A*
A--endi2 !
4'
9 id e St ri ( e ?
00 1
@ ry 1 A c c ot to n
04 1
9 id e St ri ( e ?
6C
@ ry 1 ! a c ot to n
60
9 id e St ri ( e ?
0I =
@ @ ry ry 1 1 A ! c a c c ot ot to to n n
#0 -2 6$
.
J er n el
08 I
N D + ef in e d oi l
a
N D
N D
N D
N D
N D
N D
g )g is e,"ivalent to parts per million #ppm$-
!ig"res in .rac(ets represent val"es .elow the validated limit of ,"antitation for the assay-
A--endi2 !
4)
ND: not detected-
Ta8le 3 $x ression leAels of t1e introd2ced roteins in t1e t1ree GM cotton lines oAer time
Protein $x ression
Ti ss 2e
(t ag e
(i
#Mean g protein)ga dry weight.$
@r y1 Ac
@r y1 !a
%A*
9i @r 9i @r 9i @r @r de y1 de y1 de y1 y1 Str Ac Str !a Str Ac !a i( i( i( co co e e e co co ? tto ? tto ? tto tto n n n n

No "n g 6B 8 we e( s 1 2-8 2-C I-6 =-I 0-0 1 N D 0-0 1
lea f
1-=
2-4
C-4
11
0-4 1
0-1 2
0-6 8
A--endi2 !
48
P 1 mo nth
1-C
2-0
8-I
0-4
0-4 0
N D
0-6 =
1-4
1-=
8-I
I-1
0-4 1 0-6 8
#00= $c #00I $c #00I $c 0-1 2
0-4 4 0-6 0
P 2 mo nth
1-1
0-I I
C-=
I-0
2-0
1-8
60
1C
0-4 0 0-6 0
0-6 0 0-1 =
P 6 mo nth
0-8 I
1-2
I-I
=-8
2 !l ow er !ir st flo we r 1
2-6 2-1
2-I 2-1
41 4-=
2= 4-6
0-4 = 0-6 4
0-2 0 N D
0-6 8 0-2 I
A--endi2 !
4*
2 P 2B 6 we e( s 1
2-2 2-0
2-2 1-C
0-8 4-4
0-6 4-0
0-4 6 0-6 C
N D N D
0-4 1 0-6 1
2 P 4B 8 we e( s 1
1-= 0-C 1
1-I 1-6
8-0 2-8
0-6 2-I
0-0 C 0-2 =
N D N D
0-0 C 0-2 =
2 'o ll Ha rly .ol l 1
1-C 0-0 8
2-0 0-I 0
0-I 6-0
8-0 4-4
0-4 8 0-2 1
N D N D
0-4 8 0-2 6
0-4 I
0-0 =
1-4
1-0
#01C $
N D
#01I $c
A--endi2 !
&0
P 2B 6 we e( s
0-4 4
0-8 8
8-6
8-I
0-4 2
N D
0-6 1
2 P 4B 8 we e( s 1
0-6 6 0-6 I
0-4 1 0-4 8
I-8 0-0
0-1 I-6
0-2 8 0-0 2
N D N D
0-2 0 0-4 4
0-4 0
0-4 6
4-=
4-8
0-6 4
#008 $c
#028 $c
g )g is e,"ivalent to parts per million #ppm$Hach fig"re represents the mean val"e of C replicate samples!ig"res in .rac(ets represent val"es .elow the validated limit of ,"antitation for the assay-
ND: not detected-
A--endi2 !
&!
($+TI&' D +&MPARI(&' &* GM A'D '&'3GM +&TT&' +/ARA+T$RI(TI+(

*3+ *he demonstrated agronomic and compositional similarity .etween the GM 9ideStri(e? cotton/ two single insecticidal gene lines #@ry1!a or @ry1Ac cotton$ and nonBGM cotton indicates that no significant pleiotropic or epistatic effects #that is/ "nintended effects of a genetic change on other/ apparently "nrelated/ plant genes or plant characteristics$ have occ"rred-
(ection D<9 Agronomic erformance

*4+ Agronomic performance and varietal selection trials were cond"cted in the 3SA d"ring 2002- *he three GM cotton lines were grown at 62 locations alongside the nonBGM rec"rrent parental variety #%S@600$ "sed in its .reeding #%ellow 2006$*&+ *hirty agronomic characteristics were analysed from "p to 20 locations- Growth characteristic meas"res incl"ded germination/ field emergence/ vegetative vigo"r/ growth ha.it #plant height/ n"m.er of .ranches/ fr"iting positions etc-$/ flowering period/ reprod"ctive potential #.oll retention/ open .olls/ seed cotton Qseed em.edded in the lintR weight per .oll etc-$ and lint yield- !i.re ,"ality indicators incl"ded length/ strength/ micronaire #a fineness indicator$/ "niformity and colo"r*'+ !or most of these meas"rements there was no statistically significant difference .etween the GM cotton lines and the %S@600 cotton variety- !or some there was a small ."t statistically significant difference/ however the val"es for the GM cotton were well within the normal range for nonBGM cottons*)+ !i.re characteristics were meas"red from 14 s".Blines of the GM 9ideStri(e? cotton/ each grown at 11 locations- All s".Blines contained the same GM events #iethe com.ined events/ 6008B210B26 and 2=1B24B268$ and were crossed with the same rec"rrent parent #%S@600$ ."t (ept separate for three s".se,"ent generations of .reeding*8+ *here was considera.le variation .etween the GM cotton s".Blines/ pres"ma.ly reflecting differences in the .ac(gro"nd genetic ma(eB"p/ arising from the com.ination of the original genetically modified parental cotton variety #G@010$ and the rec"rrent parent in the .reeding program #%S@600$- *he report states that this level of variation is typical for s".Blines within a given pedigree in a cotton .reeding program- ;owever most fi.re characteristics meas"rements were not significantly different .etween the s".Blines of GM cotton lines and the nonBGM rec"rrent parent**+ 5t sho"ld .e noted that the genetic difference .etween the original parent and the rec"rrent parent is considera.le- G@010 is a high fi.re ,"ality variety adapted to completely irrigated @alifornian conditions/ while %S@600 is a moderate fi.re ,"ality variety adapted to nonBirrigated midBso"th region of the 3SA!00+ 5t sho"ld also .e noted that sho"ld Dow AgroSciences decide to attempt the commercialisation of these GM cottons/ the GM insecticidal traits wo"ld need to .e .red into c"ltivars more s"ita.le for the A"stralian conditions- *hese trials wo"ld re,"ire additional applications and approvalsA--endi2 ! In$#.mati#n a"#0t t,e 3MOs
&2
(ection D<2
+om ositional anal0sis
!0!+ *he res"lts of e2tensive compositional analyses of cottonseed/ processed cottonseed fractions and fresh cotton tiss"e #%hillips et al- 2002$ in a st"dy in the 3SA demonstrated that the levels of the important n"tritional and antiBn"tritional components in the three cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ are compara.le to those in the similar nonBGM variety and to esta.lished val"es for commercial cotton varieties-
D<2<9
+ottonseed
!02+ Ash/ total fat/ moist"re/ protein/ car.ohydrate/ calories and fi.re in cottonseed were analysed for all three lines- *he only statistically significant difference .etween 9ideStri(e? cotton seed and nonBGM seed was for cr"de fi.re #10-6D for 9ideStri(e? cotton and 1I-0D for nonBGM cotton$- 'oth of these val"es are within the range reported in the literat"re for nonBGM cotton seed!03+ *he levels of the minerals calci"m/ copper/ iron/ magnesi"m/ manganese/ moly.den"m/ phosphor"s/ potassi"m/ sodi"m/ s"lf"r and <inc were analysed- No statistically significant differences .etween the three GM cottonseed and nonBGM cottonseed were fo"nd/ and all minerals were within reported literat"re ranges!04+ No statistically significant differences in amino acid content .etween the three GM and nonBGM cottonseed were fo"nd/ and all val"es were within or close to reported literat"re ranges #'er.erich et al- 1CC8$!0&+ No statistically significant differences in fatty acid content .etween the three GM and nonBGM cottonseed were fo"nd- %".lished val"es are only availa.le for a few fatty acids/ and are slightly higher than those fo"nd in .oth the GM cotton lines and nonBGM cottonseed in this st"dy!0'+ @otton tiss"e/ partic"larly the seeds/ can .e to2ic if ingested in e2cessive ,"antities .eca"se of the presence of antiBn"tritional and to2ic factors incl"ding gossypol and cyclopropenoid fatty acids #incl"ding dihydrosterc"lic/ sterc"lic and malvalic acids$- No statistically significant differences in these components .etween the three GM and nonBGM cottonseed were fo"nd/ and all val"es were within reported literat"re ranges- No aflato2ins/ to2icants which may .e prod"ced .y infecting f"ngi/ were detected in any sample-
D<2<2
Processed fractions
!0)+ Ash/ total fat/ moist"re/ protein/ car.ohydrate/ calories and fi.re were analysed in cottonseed processed fraction samples/ incl"ding (ernels/ h"lls/ toasted meal and refined oil- 5ndivid"al analysis res"lts were compara.le for all three GM cotton lines and nonBGM samples and were within reported ranges!08+ ;"lls and meal were analysed for minerals #calci"m/ copper/ iron/ magnesi"m/ manganese/ moly.den"m/ phosphor"s/ potassi"m/ sodi"m/ s"lf"r and <inc$- No
A--endi2 !
&3
statistically significant differences .etween fractions from these three GM cotton lines and nonBGM fractions were fo"nd!0*+ No statistically significant differences in amino acid content of toasted meal from the three GM cotton lines and nonBGM cotton were fo"nd/ and all val"es were within or close to reported literat"re ranges!!0+ +efined cottonseed oil was analysed for fatty acid content- No statistically significant differences .etween these three GM cotton lines and nonBGM oil were fo"nd/ and all val"es were within reported literat"re ranges- +efined oil was also analysed for tocopherol isomers/ which are nat"rally occ"rring antiBo2idants- +es"lts for GM and nonBGM oil were very similar- *he level of alpha tocopherol #vitamin H$ in the cottonseed oil for the GM cotton lines @ry1!a/ @ry1Ac and 9ideStri(e? cottons #001/ 002 and 010 g)g respectively$/ and nonBGM cotton #04C g)g$ was higher than the availa.le literat"re val"e #620 g)g$!!!+ Jernel/ meal and refined oil were analysed for gossypol- +es"lts for these three GM cotton lines and nonBGM fractions were compara.le and similar to literat"re val"es- +efined oil was also analysed for antiBn"tritional cyclopropenoid fatty acids7evels in oil from these GM lines and nonBGM cottonseed were compara.le and similar to or .elow literat"re val"es-
D<2<3
+otton leaAes and s72ares
!!2+ AntiBn"trients gossypol and polyphenols were analysed in terminal leaves and s,"ares from the three GM cotton lines and nonBGM cotton grown at two sites- *otal gossypol was higher in the terminal leaves of @ry1Ac cotton and 9ideStri(e? cotton lines than that of nonBGM cotton at .oth sites ."t very similar in s,"ares- *otal polyphenols were higher in terminal leaves of @ry1!a cotton and 9ideStri(e? cotton lines than that of nonBGM cotton at only one site- %olyphenols were very similar for GM and nonBGM s,"ares-
($+TI&' E R$($AR+/ R$>-IR$M$'T(

!!3+ *he proposed release of three GM cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ is a small scale/ m"ltiBsite trial over fo"r cotton growing seasons- +esearch conditions have .een imposed in the licence iss"ed for this release/ incl"ding a re,"irement to collect data on the levels of e2pression of the insecticidal and her.icide tolerance genes in cotton tiss"es "nder A"stralian field conditions!!4+ 5f the applicant ma(es any applications for f"t"re larger scale releases of the GM cotton lines/ more data wo"ld .e re,"ired to demonstrate the a.sence of "nintended effects of the genetic modification "nder A"stralian field conditions-
A--endi2 !
&4
APP$'DI, 2 T&,I+IT. A'D A))$RG$'I+IT. T& /-MA'(

!!&+ 3nder section 01 of the Act/ the +eg"lator is re,"ired to consider ris(s to h"man health and safety and the environment in preparing the ris( assessment and ris( management plan- *his Appendi2 considers potential ha<ards that may .e posed to h"man health and safety as a res"lt of any to2icity or allergenicity of the GMOs or its novel proteins!!'+ 5t sho"ld .e noted that GM cottons containing cry insecticidal genes derived from the same .acteri"m have .een e2tensively trialed and commercially released in A"stralia since 1CC8 and 2002 with no reported adverse effects to h"mans #refer to D5+s 022)2002 and 012)2002 respectively/ www.ogtr.gov.au$- @"rrently/ one of the GM cotton lines proposed for release/ 9ideStri(e? cotton/ is .eing trialed "nder limited and controlled conditions in New So"th 9ales #D5+ 040)2006$!!)+ 5n addition/ GM cottons containing the her.icide tolerance pat gene have .een trialed "nder licence D5+ 010)2002 and are .eing trialed "nder limited and controlled conditions "nder licence D5+ 06=)2006-
($+TI&' 9 'AT-R$ &* T/$ P&T$'TIA) T&,I+IT. &R A))$RG$'I+IT.

/ABARD
!!8+ A to2ic response to a chemical is shown .y the cascade of reactions res"lting from e2pos"re to a dose of chemical s"fficient to ca"se direct cell"lar or tiss"e inA"ry/ or otherwise inhi.it normal physiological processes #!elsot 2000$- Allergic responses are imm"ne system reactions/ res"lting from stim"lation of a specific gro"p of anti.odies (nown as 5gH or sensitisation of specific tiss"e .o"nd lymphocytes #!AO L 9;O 2000F *aylor L 7ehrer 1CC8$- Allergic responses have a wellBdefined etiology #ie- .iochemical ca"se$ that is ,"ite different from to2icity!!*+ An allergic response can have severe conse,"ences for an individ"al/ for e2ample/ anaphyla2is is a shoc( syndrome ca"sed .y a massive release of histamine and other allergic mediators from even min"te e2pos"res to an allergen in a sensitised individ"al- !ood proteins are common ca"ses of anaphyla2is/ especially pean"t and shell fish #!ric( 1CC0F !ric( 1CC0$!20+ @"rrent scientific (nowledge s"ggests that common food allergens tend to .e resistant to degradation .y heat/ acid/ and proteases #Astwood et al- 1CC8$- *his is .eca"se it is necessary that a protein is s"fficiently sta.le to reach and cross the m"cosal mem.rane for it to stim"late an allergenic response following oral ingestion!2!+ *he GM insecticidal)her.icide tolerant cotton lines differ from conventional cotton in the e2pression of two or three additional proteins- *hese are either one or .oth of the insecticidal @ry1!a and @ry1Ac proteins/ and the %A* en<yme that confers tolerance to the her.icide gl"fosinate ammoni"m #see Appendi2 1 for details of protein e2pression in the GMOs$- *he potential for these cottons to .e to2ic or allergenic to h"mans d"e to either e2pression of the novel gene prod"cts or .eca"se of "nintended effects of the genetic modification is considered in this Appendi2-
A--endi2 2
T#2i/it1 and alle.geni/it1 t# ,0mans
&&
!22+ *he A%&MA has a complementary reg"latory role in respect of this application d"e to its responsi.ility for agric"lt"ral chemical "se in A"stralia/ incl"ding insecticides and her.icides/ "nder the Agricultural and eterinary !he"icals !ode Act 1##$ #refer to @hapter 1 for details$- *he GM insecticidal)her.icide tolerant cottons are s".Aect to reg"lation .y the A%&MA d"e to their prod"ction of insecticidal proteins- Any "se of her.icide on the GM cottons is also s".Aect to A%&MA reg"lation- As part of their assessment of chemical "se/ the A%&MA considers any potential h"man health effects/ for e2ample/ ris(s arising thro"gh occ"pational e2pos"re or resid"es in food- Dow AgroSciences does not intend to apply gl"fosinate ammoni"m her.icide #eg- 7i.erty and 'asta her.icides$ to the GM cottons in the proposed release- *h"s ris(s associated with the "se of the her.icides in connection with the GMOs are not considered here-
($+TI&' 2 )I5$)I/&&D &* T/$ T&,I+IT. &R A))$RG$'I+IT. /ABARD

&++-RRI'G
!23+ 5n assessing the li(elihood of adverse impacts d"e to to2icity or allergenicity of the GM cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ on h"man health and safety/ the following factors were considered: &'( the inherent to2icity and allergenicity of nonBGM cotton #OG*+/ 2002$F &)( the potential e2pos"re to the GM cottons/ to their prod"cts and to the new proteins which are e2pressed in the GM cottons/ the @ry1!a/ @ry1Ac and %A* proteinsF &8( the potential e2pos"re to the @ry1!a/ @ry1Ac and %A* proteins from other so"rces in the environmentF &*( the potential to2icity and allergenicity of the new proteins e2pressed in the GM cottonsF and '0( the potential to2icity and allergenicity of the GM insecticidal)her.icide tolerant cottons-
(ection 2<9 Toxicit0 and allergenicit0 of conAentionall0 8red non3GM cotton

!24+ @otton is a well esta.lished field crop with a long history of safe "se- A comprehensive review of conventional nonBGM cotton/ incl"ding information on its to2icity and allergenicity/ is provided in the doc"ment >*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"stralia4 #OG*+ 2002$ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cotton*his doc"ment can .e accessed at www.ogtr.gov.au- 5nformation on nonBGM cotton is incl"ded here to esta.lish a .ase line for comparison with the GM cottons .eing considered in this ris( assessment!2&+ @otton tiss"e/ partic"larly the seeds/ can .e to2ic if ingested in large ,"antities .eca"se of the presence of to2ic and antiBn"tritional factors/ incl"ding gossypol and cyclopropenoid fatty acids #eg- dihydrosterc"lic/ sterc"lic and malvalic acids$-
A--endi2 2
&'
!2'+ %rocessed cotton fi.re contains CC-=D cell"lose and is widely "sed in pharmace"tical and medical applications .eca"se of its very low allergenicity@ottonseed oil has .een in common "se since the middle of the nineteenth cent"ry and achieved G+AS #Generally +ecognised As Safe$ stat"s "nder the 3nited States !ederal !ood Dr"g and @osmetic Act .eca"se of its common "se prior to 1C0= #AN !A 2002$!2)+ @otton pollen is large/ stic(y and not transported easily .y wind #OG*+ 2002$/ therefore its potential to act as an air.orne allergen is e2tremely low- ;owever/ inhalation of cotton d"st .y mill wor(ers can ca"se .yssinosis/ an asthmaBli(e condition/ in sensitive individ"als- %reventative meas"res s"ch as the "se of facemas(s have .een s"ccessf"l in lowering the incidence of this condition-
(ection 2<2 $x os2re of eo le to t1e GM insecticidal/1er8icide tolerant cottons

!28+ *he applicant proposes to destroy the GM cottons prod"ced in the release/ apart from some seed which will .e retained for f"rther planting as part of this proposed release or for possi.le f"t"re releases #s".Aect to separate approvals$- Since it is not intended that any prod"ct of the release will .e "sed in h"man food or animal feed/ there will .e no opport"nity for h"man e2pos"re to these GM cottons thro"gh food*herefore potential ha<ards to h"mans thro"gh food do not warrant detailed disc"ssion here- 5f prod"cts from these GM cottons were proposed to .e "sed in food/ approval wo"ld need to .e o.tained from !SAN !2*+ 5t sho"ld .e noted that !SAN is c"rrently eval"ating an application for approval for food "se of oil and linters derived from one of the GM cotton lines/ 9ideStri(e? cotton/ for release!30+ *here will also .e no opport"nity for h"mans to .e e2posed to the GM cottons thro"gh cotton lint in clothing or other ho"sehold prod"cts/ or thro"gh cottonseed oil in cosmetics- *herefore potential ris(s to h"mans as a res"lt of s"ch e2pos"re to GM cotton prod"cts will not .e disc"ssion f"rther here!3!+ %otential e2pos"re of people to the GM cottons will .e thro"gh:
'!( wor(ing with GM cottons #on cotton farms/ in cotton processing facilities$F '2( living in or near the areas where GM cottons are grown #general environmental e2pos"re/ eg- people .reathing cotton pollen$F
2<2<9
$x os2re to t1e GM cottons t1ro2g1 4orking 4it1 cotton and liAing near cotton lantations
!32+ ;"mans wor(ing with cotton plants wo"ld .e e2posed primarily to the o"ter wa2y c"ticle layer at the plant s"rface/ to the seed coat or to the cotton fi.res/ all of which are essentially free of proteins- H2pos"re to proteins #incl"ding the new proteins e2pressed in the GM cottons$ or to other cell"lar components of the cotton plants will only occ"r if plant cells are r"pt"red- Hven if the cells r"pt"re/ e2pos"re to
A--endi2 2
&)
the new proteins e2pressed in the GM cottons will .e very low/ as these proteins are only present at low levels in the GM cotton tiss"es #see Appendi2 1 for details$!33+ @otton pollen is large/ stic(y and not transported easily .y wind #OG*+ 2002$/ therefore limiting possi.le e2pos"re to cotton pollen as a potential air.orne allergen*he introd"ced proteins are e2pressed at low levels in the GM cotton tiss"es/ incl"ding cotton pollen #see Appendi2 1 for details$- Dermal e2pos"re of wor(ers to cotton pollen is possi.le/ ."t the amo"nts to which wor(ers may .e e2posed is e2pected to .e very low!34+ *he primary processing of seed cotton #seed em.edded in the lint$ at cotton gins/ and the ."l( handling of cottonseed and cotton fi.re/ can create and stir "p fine d"st and lint particles- 3se of personal protective e,"ipment .y e2posed wor(ers is commonplace in s"ch facilities to prevent respiratory irritations- Since cotton lint contains no DNA or protein and fi.re characteristics of the GM cottons are the same as nonBGM cotton #see Appendi2 1$/ lint from the GM cottons is not more li(ely to ind"ce adverse responses in wor(ers than is lint from nonBGM cotton- %rocessing of the GM seed cotton will only occ"r on a small scale/ for preparation of seed for possi.le f"t"re trials!3&+ After harvest/ seed cotton may .e dispersed .eyond the limits of the farms where it is grown d"ring transportation to ginning facilities- Specific licence conditions have .een imposed to re,"ire cleaning of e,"ipment "sed in connection with the release/ and harvested material to .e sec"rely wrapped .efore transporting away from the release sites/ so as to prevent the GM cotton seed escaping into the wider environment #see Appendi2 I for details$- Seeds not re,"ired for f"t"re releases m"st .e destroyed-
(ection 2<3 &t1er so2rces of +r09*a= +r09Ac and PAT roteins in t1e enAironment
!3'+ *he @ry1!a/ @ry1Ac and %A* proteins are widespread in the environment/ thro"gh the presence of the .acteria to which they are native- Altho"gh the introd"ced cry1 genes in the GM cotton lines for release are chimeric/ the encoded @ry proteins are very similar to the native @ry proteins #CC-6D similarity in amino acid se,"ence of core to2in .etween native and chimeric @ry1!a and CC-8D .etween native and chimeric @ry1Ac$/ and retain the species specific insecticidal to2icity to lepidopteran caterpillar pest- As disc"ssed in Section 6-6 of Appendi2 1/ all the @ry1 to2ins are highly specific to lepidopteran insects #Macintosh et al- 1CC0F @ham.ers et al- 1CC1F !ederici 2006$!3)+ *he native @ry1!a and @ry1Ac proteins are nat"rally prod"ced .y the .acteri"m Bacillus thuringiensis #'t$/ varieties aizawai #'ta$ and kurstaki #'t($/ respectively+elated @ry proteins are also prod"ced .y other varieties of 't #see Appendi2 1$/ as well as in other GM cottons which are .eing grown commercially in A"stralia #'ollgardE 55 and 5NGA+DE cottons/ "nder licences D5+ 012)2001 and D5+ 026)2002/ see www.ogtr.gov.au$- 't spores and their crystal #@ry$ to2ins are fo"nd widely in soils/ on plant leaves and in grain stores #Meadows 1CC6$A--endi2 2 T#2i/it1 and alle.geni/it1 t# ,0mans
&8
!38+ *he presence of @ry1 proteins in food has increased over the past 60 years d"e to the commercial "se of 't micro.ial sprays to protect food crops/ incl"ding organic crops/ from insect attac( #AN !A 1CCC$- 5nsecticidal prod"cts containing 't( or 'ta as the active ingredient are registered in A"stralia/ incl"ding for "se on cotton/ vegeta.les/ vines and fr"it trees #A%&MA/ see www.apv"a.gov.au$- *h"s resid"es of 't proteins/ incl"ding @ry 1!a and @ry1Ac/ can .e present on a wide variety of fresh foods s"ch as ca..age/ lett"ce and tomato/ with no reported to2ic or allergic responses!3*+ %A* en<ymes are prod"ced nat"rally .y the common soil .acteria %trepto"yces viridochro"ogenes and %. hygroscopicus/ encoded .y pat and 'ar genes respectively #9ohlle.en et al- 1C==F Stra"ch et al- 1C==$- %trepto"yces spp. are saprophytic/ soilB .orne .acteria and are not considered pathogens of plants/ h"mans or other animals #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$- A search of the Gen'an( data.ase reveals that other genes encoding %A* or similar en<ymes are present in a wide variety of .acteria- *he class of en<ymes to which %A* .elongs/ acetyltransferases/ are common en<ymes in all microorganisms/ plants and animals%A* en<ymes have also .een e2pressed in GM crop plants trialed in A"stralia #egcotton/ "nder licences D5+ 010)2002 and D5+ 06=)2006F and canola/ "nder D5+ 010)2001$-
(ection 2<4 Toxicit0 and allergenicit0 of t1e introd2ced roteins 2<4<9 Toxicit0
!40+ St"dies "sing the p"rified forms of the introd"ced proteins have .een cond"cted/ as the very low e2pression of these proteins in GM cotton means it is generally not possi.le to feed test animals the ,"antity of the plant material necessary to prod"ce a specific effect- ;owever/ it is possi.le to test the mammalian to2icity of the p"rified proteins at m"ch higher concentrations than present in the GM plants!4!+ As disc"ssed in Section 6-1 of Appendi2 1 and in Section 2-6 a.ove/ the @ry1!a and @ry1Ac proteins encoded .y the introd"ced chimeric genes are similar to the native @ry1!a and @ry1Ac proteins and retain the specificity of the native proteins-
@+N1A@
!42+ %"rified native 't( @ry1Ac protein/ at single oral doses of "p to 4600 mg)(g .ody weight/ prod"ced no adverse effects in mice #Naylor 1CC6aF Naylor 1CC6.$Several st"dies on ac"te oral to2icity of 't micro.ial preparations/ containing @ry1Ac/ in rats and ra..its revealed no adverse effects at doses of "p to tho"sands of milligrams per (ilogram #'ar.era 1CC0F @arter L 7igget 1CC4F Mc@lintoc( et al1CC0F Spencer et al- 1CC8$- *hese st"dies reported no treatmentBrelated effects on s"rvival/ .ody weight/ food cons"mption/ clinical o.servations or gross pathology findings!43+ A twoByear chronic rat feeding st"dy was "nderta(en with 't( micro.ial prod"cts at doses of "p to =400 mg)(g of .ody weight)day- A decrease in weight gain was o.served in female rats at the highest dose ."t/ in the a.sence of any other adverse
A--endi2 2 T#2i/it1 and alle.geni/it1 t# ,0mans
&*
findings #eg- s"rvival/ clinical o.servations or pathology$/ this was not considered to indicate @ry protein to2icity #Mc@lintoc( et al- 1CC0$!44+ *wo separate st"dies on h"mans fo"nd no o.serva.le health effect of an oral dose of 1000 mg of 't( micro.ial spores per day for 6 or 0 days #Mc@lintoc( et al- 1CC0F 'et< et al- 2000$!4&+ 5n a recent review/ .ased on the information from the ac"te to2icity)pathogenicity st"dies on 't strains/ Jo"gh #Jo"gh 2006$ also concl"ded that the 't microorganisms are not pathogenic or to2ic to mammals!4'+ *he 3nited States Hnvironment %rotection Agency #3S H%A$ considers @ry1Ac protein to .e nonBto2ic to mammals and has esta.lished an e2emption from resid"e tolerance re,"irements #H%A 2000$- 5n A"stralia/ the A%&MA has also determined that a ma2im"m resid"e limit #M+7$ for h"man food and animal feed is not necessary for the @ry1Ac protein e2pressed in GM cottons or on fr"it and vegeta.les sprayed with 't( micro.ial prod"cts/ indicating that it is of no to2icological significance #see *he M+7 Standard/ ta.le 0 at: www.apv"a.gov.au*residues*"rl5standard.sht"l$-
@+N1!A
!4)+ Given the very close amino acid se,"ence similarity .etween @ry1!a and other @ry proteins/ s"ch as @ry1Ac/ @ry1!a is e2pected to have a similar to2icological profile- Some of the st"dies a.ove related to 'ta/ the strain of 't/ which nat"rally e2presses @ry1!a #Mc@lintoc( et al- 1CC0F 'et< et al- 2000$- !48+ Data s".mitted to the 3S H%A indicates no adverse effects in rats after oral administration of greater than 10= 'ta micro.ial spores #Mc@lintoc( et al- 1CC0$!4*+ *he 3S Department of Agric"lt"re #3SDABA%;5S 2001$/ ;ealth @anada #;ealth @anada 2002$ and !SAN #!SAN 2006$ have reported that the p"rified @ry1! protein is of low ac"te oral to2icity in mice #having an 7D00/ the dose fo"nd to (ill 00D of test animals/ of greater than 0I8 mg)(g .ody weight$- *he 3S H%A also reported chimeric @ry1! protein e2pressed in 9idestri(e? cotton to .e of low ac"te oral to2icity and considered it "nnecessary to esta.lish a ma2im"m permissi.le level for resid"es prod"ced in cotton #3S H%A 2006$- 5n A"stralia/ the A%&MA has also determined that an M+7 is not necessary for 'ta #e2pressing native @ry1!a$ micro.ial prod"cts on food prod"cing and nonBfood prod"cing crops #see *he M+7 Standard/ ta.le 0 at: www.apv"a.gov.au*residues*"rl5standard.sht"l$- !SAN has approved corn e2pressing the core to2in of @ry1!a for food "se since 2006 #see Food %tandards !ode 1-0-2 at www.(oodstandards.gov.au*(oodstandardscode*$
%A*
!&0+ Hvidence indicates that the %A* protein is not to2ic to either h"mans or other animals- *he potential for %A* to .e to2ic has .een addressed via ac"te to2icity st"dies "sing %A* protein having several e2tra amino acids at the NBterminal end of the protein as an aid in its p"rification #Merriman 1CC8$- Mice #0 male and 0 female$ were given a single oral gavage dose of %A* protein at 2000 mg)(g .odyweight-
A--endi2 2
'0
'ody weights of the test animals were determined prior to dosing #day 0$ and on days I and 14 after dosing/ and the animals were o.served daily for any clinical a.normalities or mortality- No mortality occ"rred d"ring the st"dy- !ollowing sched"led e"thanasia of test animals on day 14/ no gross internal findings were o.served- 'ased on this test/ the ac"te oral 7D00 was estimated to .e greater than 2000 mg of %A*)(g .ody weight!&!+ 5n addition/ a st"dy .y %fister et al #1CC8/ cited in 'remmer L 7eist 1CC8$ investigated the to2icity of p"rified %A* protein in a repeat dose oral to2icity st"dy in rats- Gro"ps of five male and five female rats were fed %A* protein for 14 days at levels of 0/ 0-0 or 0D of their diet #e,"ivalent to 0/ I0I and IIC2 mg)(g .ody weight)day$- *he highest concentration is appro2imately 100/000 times the %A* concentration in the GM cotton tiss"e- No clinical signs of to2icity or mortality were o.served d"ring the st"dy and no significant differences were o.served in white .lood cell co"nts or spleen and thym"s weights/ and there were no histological changes in the organs of the imm"ne system that were e2amined!&2+ %A* protein/ as e2pressed in a variety of GM plants/ has .een assessed .y a n"m.er of reg"latory .odies in A"stralia/ 3SA/ @anada/ and H"rope #AN !A 2001cF @anadian !ood 5nspection Agency 1CC8F H"ropean Scientific @ommittee on %lants 2000F !SAN 2006F ;ealth @anada 1CCIF ;ealth @anada 2000F !DA 1CC0F !DA 1CCIF !DA 2001$- *he %A* en<yme was fo"nd to .e of very low to2icity- *he 3nited States Hnvironmental %rotection Agency/ for e2ample/ has determined that %A* is e2empt from the re,"irement to esta.lish a ma2im"m permissi.le level for resid"es in plants #H%A 1CCI$- !SAN has approved a corn e2pressing the %A* protein for food "se #see Food %tandards !ode 1-0-2 at www.(oodstandards.gov.au*(oodstandardscode*$
2<4<2
Allergenicit0
!&3+ Altho"gh there are no predictive assays availa.le to assess the allergenic potential of proteins/ m"ch is (nown a.o"t the .iochemical events associated with allergic reactions/ as well as the (inds of proteins that ca"se pro.lems #*aylor L 7ehrer 1CC8F Metcalfe et al- 1CC8$!&4+ %redictions of allergenicity have .een .ased on se,"ence/ str"ct"ral and .iochemical comparisons with (nown allergens- %rotein allergens "s"ally share a n"m.er of characteristics #Davies 1C=8F !lavell et al- 1CC2F !"chs et al- 1CC6aF !"chs et al- 1CC6.F !"chs et al- 1CC6aF *aylor 1CC0F Metcalfe et al- 1CC8F !"chs L Astwood 1CC8F AN !A 2001.F Jim.er et al- 1CCC$/ incl"ding the following: '3( molec"lar weight ranges .etween 10BI0 (DF '4( typically glycosylatedF '&( sta.le in the mammalian digestive systemF ''( sta.le d"ring high temperat"res involved in coo(ing or processingF and ')( present as the maAor protein component in the specific foods-
A--endi2 2
'!
!&&+ None of the introd"ced proteins in the GM insecticidal)her.icide tolerant cottons are derived from (nown so"rces of allergens/ nor are they present as maAor components of the GM cotton plants #see Appendi2 1$!&'+ *aylor #1CC2$ has shown in do".leB.lind place.oBcontrolled food challenges that individ"als who are allergic to pean"ts or s"nflower seeds are a.le to cons"me oil derived from these seeds witho"t it eliciting an allergic response- *aylor #1CC2$ also s"ggests that when a protein is present in food at levels well .elow 1 mg per serving/ the ha<ard for allergenic cons"mers is minimal- *h"s cons"mers wo"ld .e highly "nli(ely to develop allergic responses as a res"lt of the "se of oil or linters derived from GM insecticidal)her.icide tolerant cottons in food-
@+N1A@
!&)+ *he @ry1Ac protein is heat la.ile and rapidly degraded/ in 60 seconds/ "nder sim"lated mammalian gastrointestinal conditions #!"chs et al- 1CC6a$- *he @ry1Ac protein does not display characteristics common to (nown food allergen proteinsSearches of allergen se,"ence data.ases have shown no significant matches of the @ry1 proteins to (nown allergens #Metcalfe et al- 1CC8$!&8+ 9hile there have .een reports in the 3S claiming allergic reactions to 't micro.ial prod"cts in topical insecticidal sprays/ these are not d"e the @ry1Ac proteinA s"rvey cond"cted among farm wor(ers who pic(ed vegeta.les treated with 't micro.ial prod"cts indicated that e2pos"re to 't prod"cts may lead to allergic s(in sensitisation/ however there was no clinical allergic disease in any of the wor(ersMost reactions in these wor(ers were shown to .e d"e to other constit"ents of the 't sprays/ and there was no evidence of anti.odies specific to the endoto2in proteins of the 't sprays #'ernstein et al- 1CCC$- *he 3S H%A has also determined that reports of reactions to 't micro.ial prod"cts have .een d"e to nonB@ry proteins prod"ced d"ring fermentation or to other ingredients added to the insecticidal form"lations #H%A 2001$-
@+N1!A
!&*+ *he @ry1!a protein is e2pected to have a similar to2icological profile/ incl"ding potential for allergenicity/ as @ry1Ac and other related @ry proteins!'0+ !SAN #!SAN 2006$/ the 3SH%A #3S H%A 2006$ and the 3SDABA%;5S #3SDABA%;5S 2001$ have reported that the @ry1! protein as e2pressed in GM cotton and corn was rapidly degraded in vitro "nder conditions sim"lating mammalian digestion and the amino acid se,"ence of the protein has no significant homology with (nown allergens or protein to2ins- *herefore/ the @ry1!a protein is "nli(ely to elicit an allergenic response-
%A*
!'!+ *he %A* protein is not a (nown allergen and is not derived from a (nown so"rce of allergens- Altho"gh its molec"lar weight of S22 (D is within the range of molec"lar weights "s"ally shown .y allergens/ it lac(s glycosylation sites #'remmer
A--endi2 2
'2
L 7eist 1CC8$ and many of the other characteristics which are common to plant food allergens #H%A 1CCIF @anadian !ood 5nspection Agency 1CC=aF AN !A 2001a$!'2+ A st"dy in which %A* protein was s".Aected to sim"lated gastric conditions #low p; pl"s the proteolytic en<yme pepsin$ reported that the protein was degraded within seconds #9ehrmann et al- 1CC8$- Other st"dies have shown that the %A* en<yme was inactivated within one min"te when s".Aected to typical mammalian stomach conditions and was inactivated d"ring processing of GM canola seed #e2pressing the %A* en<yme$ into feed ingredients #H"ropean Scientific @ommittee on %lants 1CC=$!'3+ 5n addition/ a st"dy .y 'remmer L 7eist #'remmer L 7eist 1CC8$ investigated the allergenicity of p"rified %A* protein in a repeated highBdose st"dy in rats- *he st"dy did not reveal any imm"noto2ic allergenic effects .ased on a n"m.er of screening parameters-
(ection 2<: Toxicit0 and allergenicit0 assessment of t1e GM insecticidal/1er8icide tolerant cottons
!'4+ *he insecticidal and the her.icide tolerance genes operate thro"gh independent/ "nrelated .iochemical mechanisms- *here is no evidence of any interaction .etween the two genes/ their proteins or their meta.olic pathways/ and no reason to e2pect that this is li(ely to occ"r!'&+ *he composition of cottonseed from these three GM insecticidal)her.icide tolerant cotton lines is within the normal range for cottonseed in terms of the concentration of protein/ oils/ car.ohydrate and ash/ and amino acid and fatty acid profiles- *he levels of (nown to2ic and antiBn"tritional factors in the GM cotton seed (ernel/ meal and oil/ incl"ding gossypol and cyclopropenoid fatty acids/ are also within the range of nonBGM cotton controls #see Appendi2 1$- *his s"ggests that no "nintended effects have occ"rred as a res"lt of the genetic modifications in the GM cottons!''+ *he availa.le information for the introd"ced proteins shows that they are of low ac"te oral to2icity and "nli(ely to .e allergens- !"rthermore/ e2pos"re to the GM cottons as a res"lt of the proposed release wo"ld .e low- *h"s/ the ris( of adverse effects as a res"lt of to2icity or allergenicity of the GM cottons are li(ely to .e low-
($+TI&' 3 +&'+)-(I&'( R$GARDI'G T&,I+IT. &R A))$RG$'I+IT.

!')+ 5t is considered that the ris( of GM insecticidal)her.icide tolerant cottons #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ .eing to2ic or allergenic for h"mans is low .eca"se: '8( prod"cts of the GM cotton lines will not .e "sed for h"man food or animal feed/ or processed for lint or oil prod"ctionF '*( e2pos"re to the introd"ced proteins thro"gh wor(ing with cotton plants is very lowF )0( cotton pollen is not wind dispersed and therefore not li(ely to .e an air.orne allergenF
A--endi2 2
'3
)!( processing of the GM seed cottons will only occ"r on a small scale #for preparation of seed for f"t"re plantings$ and lint from GM cottons is not more li(ely to ind"ce adverse responses in wor(ers than is lint from nonBGM cottonF )2( the introd"ced core proteins are very similar to native .acterial proteins which are already widespread in the environment and present in h"man foodF )3( the introd"ced proteins are of very low oral to2icityF )4( the introd"ced proteins are not (nown to .e allergenic/ nor do they have properties characteristic of (nown allergenic proteinsF and )&( compositional analyses have not indicated any differences .etween the GM and nonBGM cotton/ other than the presence of the introd"ced proteins and the intended plant traits!'8+ *he licence holder is re,"ired to report any adverse effects on h"man health and safety #for e2ample allergic reactions as a res"lt of occ"pational e2pos"re to the GM cottons$ or to the environment-
A--endi2 2
'4
APP$'DI, 3 T&,I+IT. T& '&'3TARG$T &RGA'I(M(

!'*+ 3nder section 01 of the Act/ the +eg"lator is re,"ired to consider ris(s to h"man health and safety and the environment in preparing the ris( assessment and ris( management plan- *his Appendi2 considers potential ha<ards that may .e posed thro"gh any potential to2icity of the GMOs or their novel proteins to nonBtarget organisms-
($+TI&' 9 'AT-R$ &* T/$ P&T$'TIA) T&,I+IT. /ABARD

!)0+ *he GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ differ from conventional cotton in the e2pression of two or three additional proteins- *hese are @ry1!a/ @ry1Ac insecticidal proteins and %A* en<yme that confers her.icide tolerance #see Appendi2 1 for details of protein e2pression in these GMOs$- *he potential for the GM cottons to .e to2ic to organisms/ other than the target 7epidopteran insect pests/ is considered- *his co"ld occ"r either d"e to e2pression of the novel gene prod"cts or .eca"se of "nintended effects of the genetic modifications!)!+ 5f the GM cottons are to2ic to nonBtarget organisms/ the potential ha<ards co"ld incl"de adverse impacts on: )'( livestoc( and wildlife/ incl"ding mammals/ fish and .irdsF ))( inverte.rates/ incl"ding .eneficial insects #pollinators/ parasitoids or predators of target insect pests$F and )8( microorganisms/ partic"larly soil microorganisms/ with direct impact on growth of crops on farms!)2+ *o2icity for the lepidopteran insects may also present indirect ha<ards/ with potential to harm the nat"ral environment #for e2ample/ adverse impacts on native .iodiversity$ thro"gh secondary effects on: )*( pop"lations of specialist parasitoids and predators that feed on lepidopteran insectsF and 80( pop"lations of organisms that are preyed on .y nonBtarget lepidopteran insects!)3+ As noted in @hapter 1 and disc"ssed in more details in @hapter 8/ the A%&MA has responsi.ility for agric"lt"ral chemical "se in A"stralia/ incl"ding insecticides and her.icides/ "nder the Agricultural and eterinary !he"icals !ode Act 1##$- GM insecticidal cottons are s".Aect to reg"lation .y the A%&MA d"e to their prod"ction of insecticidal s".stances- Any "se of her.icide on the GM cottons is also s".Aect to A%&MA reg"lation- As part of their assessment of chemical "se/ the A%&MA considers potential environmental effects/ for e2ample resid"es and to2icity to other organisms- Dow AgroSciences does not intend to apply gl"fosinate ammoni"m her.icide #eg- 7i.erty and 'asta her.icides$ to the GM cottons in the proposed release- *h"s ris(s associated with the "se of the her.icides in connection with the GMOs are not considered here-
A--endi2 3
T#2i/it1 t# n#n4ta.get #.ganisms
'&
($+TI&' 2 )I5$)I/&&D &* T/$ T&,I+IT. /ABARD &++-RRI'G

!)4+ 5n assessing the li(elihood of adverse impacts d"e to to2icity of these GM cotton lines/ a n"m.er of factors were considered incl"ding:
8!( the inherent to2icity of nonBGM cotton #OG*+ 2002$F 82( the potential e2pos"re to @ry1!a/ @ry1Ac and %A* proteins from other so"rces in the environmentF 83( information a.o"t the li(ely ro"tes of e2pos"re to these GM cottons and to the introd"ced proteins/ the @ry 1!a/ @ry1Ac and %A* proteinsF 84( the potential to2icity of the new proteins e2pressed in the GM cottons for partic"lar speciesF and 8&( the potential to2icity of the GM cottons for partic"lar species-
(ection 2<9 Toxicit0 of non3GM cotton

!)&+ @otton is a wellBesta.lished field crop with a long history of safe "se- A comprehensive review of conventional cotton/ incl"ding information on its to2icity/ is provided in the doc"ment T*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"straliaU #OG*+ 2002$ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cotton- *his doc"ment can .e accessed at www.ogtr.gov.au- 5nformation on nonBGM cotton is incl"ded here to esta.lish a .ase line for comparison with the GM cottons .eing considered in this ris( assessment!)'+ @otton tiss"e/ partic"larly the seeds/ can .e to2ic if ingested in large ,"antities .eca"se of the presence of to2ic and antiBn"tritional factors/ incl"ding gossypol and cyclopropenoid fatty acids #eg- dihydrosterc"lic/ sterc"lic and malvalic acids$!))+ Mammals avoid feeding on cotton plants d"e to .oth the gossypol content and the morphology of the plant- *he presence of gossypol and cyclopropenoid fatty acids in cottonseed limits the "se of whole cottonseed as a protein s"pplement in animal feed/ e2cept for cattle which are less affected .y these components- 5nactivation or removal of these components d"ring processing ena.les the "se of some cotton seed meal for catfish/ po"ltry and swine- *he meal and h"lls of cottonseed can also .e "sed for cattle feed- 5ts "se as stoc(feed is limited/ nonetheless/ to a relatively small proportion of the diet and it m"st .e introd"ced grad"ally/ to avoid potential to2ic effects!)8+ 'est Management %ractices for the A"stralian cotton ind"stry prohi.its the "se of cotton trash and st"..le as a feed for animals/ d"e to resid"es of other pesticides that co"ld .e fo"nd in the cotton trash and st"..le-
A--endi2 3
''
(ection 2<2 &t1er so2rces of t1e introd2ced roteins in t1e enAironment

!)*+ As disc"ssed in Section 6-1 of Appendi2 1 and Section 2-6 of Appendi2 2/ @ry1!a/ @ry1Ac and %A* proteins are widespread in the environment/ thro"gh the presence of the .acteria to which they are native- Altho"gh the introd"ced cry1 genes in GM cotton lines proposed for release are chimeric/ the encoded core to2ins are very similar to the native @ry proteins #CC-6D similarity in amino acid se,"ence of core to2in .etween native and chimeric @ry1!a and CC-8D .etween native and chimeric @ry1Ac$/ and retain the species specific insecticidal to2icity to lepidopteran caterpillar pest!80+ @ry1!a and @ry1Ac proteins are nat"rally prod"ced .y the common .acteri"m B. thuringiensis/ varieties aizawai #'ta$ and kurstaki #'t($/ respectively- +elated @ry to2ins are also prod"ced .y other varieties of B. thuringensis #see Appendi2 1$/ as well as .y other GM cottons which are .eing grown commercially in A"stralia #'ollgardE 55 and 5NGA+DE cottons/ "nder licences D5+ 012)2001 and D5+ 022)2002 respectively/ see www.ogtr.gov.au$- @ommercial 't( and 'ta micro.ial form"lations #prod"ced .y the fermentation of the same strains of .acteria from which these genes were derived$ also contain these proteins/ and have .een "sed over the past 40 years to protect food crops #see Appendi2 1/ Section 6-2$/ incl"ding organic crops from insect attac(- 't spores and their @ry to2ins are fo"nd widely in .oth agric"lt"ral and nat"ral environments/ incl"ding in soils/ on plant leaves/ in grain stores and in dead insects #Meadows 1CC6$ and on a variety of fresh foods #s"ch as lett"ce and tomato$ #AN !A 1CCC$!8!+ %A* proteins as en<ymes are prod"ced nat"rally .y the common soil .acteria %trepto"yces viridochro"ogenes and %. hygroscopicus/ encoded .y pat and 'ar genes respectively #9ohlle.en et al- 1C==F Stra"ch et al- 1C==$- %trepto"yces spp. are saprophytic/ soilB.orne .acteria and are not considered pathogens of plants/ h"man or other animals #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$- A search of the Gen'an( DNA se,"ence data.ase reveals that other genes encoding %A* or similar en<ymes are present in a variety of other .acteria- *he class of en<ymes to which %A* .elongs/ acetyltransferases/ are common en<ymes in all microorganisms/ plants and animals-
(ection 2<3 Potential toxicit0 1aFard for liAestock and 4ildlife= incl2ding mammals= 8irds and fis1 2<3<9 $x os2re to t1e GM cottons
!82+ None of the cotton plants from the proposed release or their .yBprod"cts will .e "sed as stoc(feed- As disc"ssed in section 2-1/ most mammals avoid feeding on cotton/ nonBGM or GM/ d"e to its prod"ction of to2ic/ antiBn"trient s".stances and plant morphology- *he applicant proposes to destroy all materials prod"ced in the release/ apart from some cottonseed for "se in s".se,"ent seasons "nder the proposed release or in possi.le f"t"re releases #s".Aect to f"rther application and assessment processes$-
A--endi2 3
')
!83+ 5n the field/ seed cotton is present as large lintBcovered seeds that are "nattractive to avian species #OG*+ 2002$/ so .irds are not li(ely to .e e2posed to the insecticidal protein e2pressed in the seeds of the GM cottons!84+ @ottonseed or pollen does not enter a,"atic ha.itats in any significant ,"antity #OG*+ 2002$/ and therefore the level of e2pos"re of a,"atic species to the GM cottons will .e low!8&+ 5n addition/ the proposed release is small in si<e and limited in d"ration/ f"rther limiting the potential for e2pos"re of stoc( and wildlife to the GM cottons!8'+ After harvest/ seed cotton may .e dispersed .eyond the limits of the farms where it is grown d"ring transportation to ginning facilities- Specific licence conditions have .een imposed to re,"ire cleaning of e,"ipment "sed in connection with the release/ and harvested material to .e sec"rely wrapped .efore transporting away from the release sites/ so as to prevent the GM cotton seed escaping into the wider environment #see Appendi2 I for details$- Seeds not re,"ired for f"t"re releases m"st .e destroyed-
2<3<2
Toxicit0 of t1e GM cottons for liAestock and 4ildlife
!8)+ As disc"ssed a.ove/ the native @ry1!a/ @ry1Ac and other related @ry proteins are widespread in the environment- *o e2ert a to2ic effect/ the @ry to2ins/ which are e2pressed in the form of proto2ins/ m"st first .e digested .y specific proteases in the insect g"t and then diff"se thro"gh the midg"t mem.rane and .ind to specific receptors on the midg"t epitheli"m s"rface #for more details of the specificity and mode of action refer to Appendi2 1/ Section 6$- Mammals/ .irds and fish do not possess these receptors and therefore are not s"scepti.le to the specific to2ic effects of the @ry to2ins!88+ St"dies with high doses of p"rified @ry1!a and @ry1Ac proteins/ or of 't micro.ial preparations containing these proteins/ in mice/ rats and ra..its have revealed no adverse to2ic effects/ as disc"ssed in detail in Appendi2 2/ Section 2-4!8*+ !eeding st"dies with other GM insecticidal cottons have also indicated no adverse effects of the e2pressed @ry proteins- Northern 'o.white :"ail fed raw cottonseed meal/ derived from either 5NGA+DE #containing cry1Ac gene$ or conventional cotton/ at "p to 10D #w)w$ of diet/ e,"ivalent to 100 seeds).ird)day/ for five days showed no significant differences in feed cons"mption or .ody weight #Gallagher et al- 2000/ Monsanto 3np".lished$- @ommercial catfish fed a 20D processed cottonseed meal/ either 'ollgardE 55 #containing cry1Ac and cry1A' genes$ or conventional/ showed no differences in s"rvival/ weight gain/ feed conversion ratio or fillet composition #7i L +o.inson 2000$!*0+ *he performance of cows fed controlled diets incl"ding cottonseed from either nonBGM cotton/ 5NGA+DE or 'ollgardE 55 cottons have .een compared #@astillo et al- 2001.F @astillo et al- 2001aF ;artnell et al- 2001/ Monsanto 3np".lished$- *here were no significant differences in .ody condition/ mil( yield or mil( composition .etween cows fed the alternative diets- 9estern .lot assays of the mil( tested negative for the new proteins e2pressed in 5NGA+DE cotton #;artnell et al- 2001/
A--endi2 3
'8
Monsanto 3np".lished$- A st"dy of dairy cows fed silage of GM corn containing the @ry1A. and %A* proteins fo"nd no meas"ra.le impact on shortBterm lactational performance and r"minal fi.re degradation relative to cows fed nonBGM corn silage #!olmer et al- 2000$!*!+ GM corn containing @ry1!a core to2in has .een approved for "se in food in A"stralia since 2006- No adverse effect of the GM crop on h"mans has .een reported- !SAN concl"ded that corn containing this cry1F gene is as safe and wholesome as from other commercial corn varieties #http)**www.(oodstandards.gov.au*5src(iles*A!F16.pd($!*2+ Hvidence shows that the %A* protein is not to2ic to any animal- As disc"ssed a.ove/ acetyltransferases/ the class of en<ymes to which %A* .elongs/ are present in all microorganisms/ plants and animals- *he potential for %A* to .e to2ic has .een addressed via ac"te and repeat dose to2icity st"dies in mice and rats/ witho"t any adverse findings #see Appendi2 2/ Section 2-4$!*3+ *he demonstrated agronomic and compositional similarity of the GM cotton lines to nonBGM cotton #see Appendi2 1/ Section =$ indicates that no significant "nintended effects have occ"rred-
(ection 2<4 Potential toxicit0 1aFard for inAerte8rates= incl2ding 8eneficial insects 2<4<9 $x os2re to t1e GM cottons
!*4+ NonBtarget inverte.rates may .e e2posed to the GM cottons and the introd"ced proteins/ either directly thro"gh feeding on the GM plants/ or indirectly thro"gh eating other organisms/ incl"ding the lepidopteran target organisms/ that feed on the plants+elative e2pos"re will .e greatest for other her.ivoro"s species feeding on the cotton plants- %ollinator species and vario"s ad"lt insects that feed on the pollen will .e e2posed to the proteins- Sap feeders/ s"ch as aphids/ will have minimal e2pos"re/ as the sap is composed primarily of s"gars and mineral salts dissolved in water!*&+ NonBtarget lepidopteran species may also .e e2posed to the GM cottons and may .e affected .y the @ry1!a and @ry1Ac proteins- As cotton is not the preferred food so"rce for nonBtarget 7epidopteran species/ their pop"lations wo"ld .e s"stained on other types of plants fo"nd aro"nd the release locations- !"rthermore/ the proposed release is small in si<e and limited in d"ration/ f"rther limiting the potential for e2pos"re of nonBtarget inverte.rates to the GM cottons-
2<4<2
Toxicit0 of t1e GM cottons for inAerte8rates
!*'+ *he @ry1!a and @ry1Ac proteins are to2ic specifically to a range of lepidopteran insect larvae/ incl"ding pest species of cotton: ,elicoverpa zea #cotton .ollworm$/ ,elicoverpa virescens #to.acco ."dworm$/ 1ectinophora gossypiella #pin( .ollworm$/ %podoptera e4igua #.eet armyworm$/ %. eridania #so"thern armyworm$/ %. (rugiperda #fall armyworm$/ 1seudoplusia includens #soy.ean looper$ and Trichoplusia ni #ca..age looper$-
A--endi2 3
'*
!*)+ Data provided .y the applicant show that there is no significant negative impact of the .acterially e2pressed @ry1!a and @ry1Ac proteins on vario"s nonBtarget arthropods #*a.le 1$- +eprod"ctive activity in Folso"ia candida #springtails or collem.ola$ was red"ced when fed @ry1Ac #22-8 g)ml diet$ in two e2periments/ .y a.o"t 00D and 20D- ;owever/ feeding @ry1Ac with @ry1!a #22-8 P I0C g)ml diet respectively$/ @ry1!a alone #I0C g)ml diet$/ or leaves of GM cotton e2pressing only @ry 1Ac #@ry1Ac cotton/ at 0D or 00D of diet$ did not affect reprod"ction- *he highest mean level of e2pression of these proteins in the GM cottons/ o.served in yo"ng leaves/ was 2-8 g)g @ry1Ac and 41 g)g @ry1!a #see Appendi2 1/ *a.le 6$Since there was no adverse effect from feeding @ry1Ac with @ry1!a/ or @ry1Ac e2pressed in GM cotton leaves/ it is "nli(ely that GM cottons will have a significant negative impact on collem.ola reprod"ction!*8+ !ield s"rveys of the GM 9idestri(e? cotton line cond"cted in the 3S indicate no significant adverse effects on nearly 200 nonBtarget arthropod species in sweep net and aerial trap samples #Mahill L Storer 2002$- ;owever/ c"rrently there is no information availa.le on the potential to2icity or other adverse effects of these GM cottons to nonBtarget organisms "nder A"stralian field conditions- As noted in @hapter 1/ Dow AgroSciences proposes to test the effects of the GM cotton lines on nonBtarget organisms- !"rther/ research conditions have .een imposed for collection of this information d"ring the proposed field trial-
A--endi2 3
)0
Ta8le 9 'on3target s ecies fo2nd to 8e insensitiAe to t1e +r09*a and +r09Ac roteins and GM material in la8orator0 anal0sis ( ecies +ommon 'ame Apis "elli(era ;oney .ee #larvae$ Proteins tested (o2rce %ollen e2pressing @ry1Ac)@ry1! a 'acterial derived protein 7isenia (etida Harthworm 'acterial derived protein Dose/2nit diet 200 mg pollen)ml
11-C4 g @ry1Ac P 1-C= g @ry1!a)ml 24I g @ry1Ac)g 10I g @ry1!a)g 24I g @ry1Ac P 10I g @ry1!a)g
,ippoda"ia convergens
7ady.ird .eetle
'acterial derived protein
22-0 g @ry1Ac)ml 600 g @ry1!a)ml 22-0 g @ry1Ac P 600 g @ry1!a)ml
!hrisoperia carnea
Green lacewing
48-= g @ry1Ac)g 0-2 g @ry1!a)g 48-= g @ry1Ac P 0-2 g @ry1!a)g
A--endi2 3
)!
8aphnia "agna
Daphnid
2-0 g @ry1Ac)ml 0-01 g @ry1!a)ml 2-0 g @ry1Ac P 0-01 g @ry1!a)ml
!**+ As disc"ssed a.ove #Section 2-2$/ acetyltransferases/ the class of en<ymes to which %A* .elongs/ are present in all microorganisms/ plants and animals- *his en<yme has .een e2tensively st"died/ and there is no indication that it is to2ic to any organism #e-g- earthworms or honey.ees$ #3SDABA%;5S 1CC=$- !"rther detail is given in the ris( assessment for D5+ 021)2002/ availa.le at www.ogtr.gov.au
(ection 2<: Potential toxicit0 1aFard for microorganisms= artic2larl0 soil microorganisms 2<:<9 $x os2re to t1e GM cottons
200+ Microorganisms/ partic"larly soil microorganisms/ will .e e2posed to the GM cotton plants and the introd"ced proteins d"ring growth and decomposition of plant material- 9hile cotton plants are living/ e2pos"re of soil microorganisms to the introd"ced proteins may occ"r as a res"lt of root e2"dations/ as has .een o.served in 't corn e2pressing @ry1A. #Sa2ena et al- 1CCC$F #Stot<(y 2000$ and 5NGA+D cotton e2pressing @ry1Ac #G"pta et al- 2002$- +oot .rea(age co"ld also lead to the release of the introd"ced proteins into soil- After the cotton is harvested/ the remaining plant resid"es will .e tilled into the soil- As noted in @hapter 1/ the applicant proposes to meas"re the e2pression levels of the insecticidal proteins in cotton leaves and s,"ares and resid"es of these proteins in soil 20!+ *he proposed release is small in si<e and limited in d"ration/ which will f"rther limit e2pos"re of microorganisms to the GM cottons- +esearch conditions have .een imposed to re,"ire collection of more information on the persistence of insecticidal proteins in the soil-
2<:<2
Toxicit0 of t1e GM cottons for microorganisms
202+ A n"m.er of st"dies on the effect of related @ry1 proteins/ e2pressed .y 't or in GM crops e2pressing @ry1 proteins #e-g- 5NGA+DE cotton$/ on soil microorganisms have shown no detrimental effects/ as disc"ssed in detail in Appendi2 6 of the ris( assessment and ris( management plan for D5+ 022)2002/ availa.le at www.ogtr.gov.au. 203+ As disc"ssed a.ove #Section 2-2$/ acetyltransferases/ the class of en<ymes to which %A* .elongs/ are present in all microorganisms/ plants and animals- *his en<yme has .een e2tensively st"died/ and there is no indication that it is to2ic to any organism #3SDABA%;5S 1CC=$A--endi2 3 T#2i/it1 t# n#n4ta.get #.ganisms
)2
($+TI&' 3 +&'+)-(I&'( R$GARDI'G T&,I+IT. T& '&'3TARG$T &RGA'I(M(

204+ 5t is considered that the ris( of the GM cottons .eing to2ic to nonBtarget organisms is low .eca"se: 8'( the introd"ced core proteins are very similar to native .acterial proteins which are already widespread in the environment/ incl"ding in soil/ on plants and on fresh prod"ce/ in the microorganisms from which the genes were derivedF 8)( the release is small in si<e and limited in d"rationF 88( the to2icity of @ry1!a and @ry1Ac proteins is specific to lepidopteran caterpillar larvaeF 8*( the %A* protein is not (nown to .e to2ic to any organismF *0( e2pos"re of livestoc( and wildlife to the GM cotton lines wo"ld .e low/ and no materials from the release are proposed to .e "sed in stoc(feedF *!( to2icity st"dies with p"rified proteins and 't micro.ial preparations indicate that the @ry1!a and @ry1Ac proteins are not to2ic to mammals/ .irds or fishF and *2( la.oratory and field st"dies done in the 3nited State of America indicate that pop"lations of nonBtarget inverte.rates are "nli(ely to .e affected .y @ry1!a/ @ry1Ac and %A* proteins-
($+TI&' 4 R$($AR+/ R$>-IR$M$'T(

20&+ *he proposed release of three GM cotton lines is a small scale/ m"ltiBsite trial over fo"r cotton growing seasons- *he licence holder is re,"ired to collect data on the potential to2icity or other adverse effects of these GM cotton lines on nonBtarget organisms and on persistence of the insecticidal proteins in soil "nder A"stralian field conditions-
A--endi2 3
)3
APP$'DI, 4 6$$DI'$((
20'+ 3nder section 01 of the Act/ the +eg"lator is re,"ired to consider ris(s to h"man health and safety and the environment in preparing the ris( assessment and the ris( management plan- 5n this Appendi2/ ris(s posed .y the proposed release to the environment are considered in relation to the potential for the GMOs to .ecome pro.lematic weeds-
($+TI&' 9 'AT-R$ &* T/$ 6$$DI'$(( /ABARD

20)+ *here are n"mero"s definitions of weeds incl"ding >a plant growing where it sho"ld not .e4- 9eeds .ecome a pro.lem to the comm"nity when their presence or a."ndance interferes with the intended "se of the land they occ"py- 9eeds may also represent a so"rce of food to vario"s organisms/ hence the introd"ction of weeds to an environment may also .ring a.o"t ecological change .y altering the str"ct"re of food we.s208+ 9eeds are tho"ght to share a n"m.er of life history characters that ena.le them to rapidly colonise and persist in ecosystems/ partic"larly those that are reg"larly dist"r.ed#+oy 1CC0F 9illiamson L !itter 1CC8$F - *hese characteristics incl"de: *3( a.ility to germinate/ s"rvive/ and reprod"ce "nder a wide range of environmental conditionsF *4( longBlived seed with e2tended dormancy periodsF *&( rapid seedling growthF *'( rapid growth to reprod"ctive stageF *)( long contin"o"s seed prod"ctionF *8( a.ility to self pollinate ."t not e2cl"sively a"togamo"sF **( "se of "nspecialised pollinators or wind when o"tcrossingF !00( high seed o"tp"t "nder favo"ra.le conditionsF !0!( special adaptations for long distance and short distance dispersalF and !02( .eing good competitors20*+ *he GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ differ from conventional cotton in the e2pression of two or three additional proteins/ the @ry1!a and)or @ry1Ac to2in and the %A* en<yme/ which confers tolerance to the her.icide gl"fosinate ammoni"m #see Appendi2 1 for details of protein e2pression in the GMOs$2!0+ *he possi.ility was considered that the GM cottons might have the potential to .e harmf"l to the environment/ .eca"se of inherent weediness or increased potential for weediness/ either d"e to e2pression of the novel gene prod"cts or as a res"lt of "nintended effects of the genetic modification-
A--endi2 4 5eediness
)4
2!!+ *his co"ld occ"r if the GM cottons displayed altered characteristics s"ch as increased fitness or increased fec"ndity- 5f the GM cottons were to spread in the environment as a weed/ this co"ld res"lt in impacts s"ch as loss of native .iodiversity or adverse effects on agric"lt"ral systems-
($+TI&' 2 )I5$)I/&&D &* T/$ 6$$DI'$(( /ABARD &++-RRI'G

2!2+ 5n assessing the li(elihood of adverse impacts d"e to weediness of GM cottons/ a n"m.er of factors were considered/ incl"ding: !03( the inherent weediness of conventionally .red nonBGM cottonF !04( the potential selective advantage conferred .y the introd"ced proteinsF and !0&( the potential weediness of GM insecticidal)her.icide tolerant cottons-
(ection 2<9 In1erent 4eediness of conAentional non3GM cotton

2!3+ Attri."tes of nonBGM cotton associated with potential weediness are disc"ssed in the doc"ment >*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"stralia4 #OG*+ 2002$ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cotton- *his doc"ment can .e accessed at www.ogtr.gov.au- 5n s"mmary/ the doc"ment concl"des that nonBGM cotton is not a pro.lematic weed in A"stralia/ .eca"se factors incl"ding soil moist"re/ n"trient limitation/ temperat"re and roadside management practices limit the esta.lishment and)or persistence of cotton seedlings- 5nformation on the weediness of nonBGM cotton is incl"ded here to esta.lish a .aseline for comparison with the GM cottons .eing considered2!4+ @otton is not considered to possess the characteristics commonly associated with s"ccessf"l weeds/ s"ch as seed dormancy/ long persistence in the soil/ germination "nder a .road range of environmental conditions/ rapid vegetative growth/ short lifecycle/ very high seed o"tp"t/ high seed dispersal and longBdistance seed dispersal #Jeeler 1C=0F Jeeler 1C=C$2!&+ Another important element in prediction of weediness is ta2onomic relationship/ considering weediness within a ta2on/ incl"ding its history of weediness in any part of the world #'ergelson et al- 1CC=F %anetta 1CC6F %helo"ng 1CC0$- @otton has .een grown for cent"ries thro"gho"t the world witho"t any reports that it is a serio"s weed pest- @otton is not considered to .e a pro.lematic weed in A"stralia #Groves et al2000F Groves et al- 2002$- *here are a.o"t 00 species of Gossypiu" #@raven et al1CC4F !ry2ell 1CC2$ of which only one #G. to"entosu"$ is listed as a weed in the 3SA #;olm et al- 1CCI$-
(ection 2<2 Potential selectiAe adAantage conferred 80 t1e introd2ced roteins 2<2<9 +r09*a and +r09Ac toxins
2!'+ *he @ry1!a and @ry1Ac proteins co"ld confer a selective advantage in areas where lepidopteran insect predation limits one or more of the (ey life stages of cotton)&
2!)+ Another GM insecticidal cotton/ 5NGA+DE cotton/ that is resistant to lepidopteran insect pests/ d"e to e2pression of a different version of the cry1Ac gene/ has .een commercially grown in A"stralia since 1CC8- *he potential weediness of 5NGA+D cotton has .een considered in the ris( assessment for application D5+ 022)2002- S"rveys of vol"nteer cotton in A"stralia and e2perimental research on the weedy potential of GM insecticidal)her.icide tolerant cottons in A"stralia consistently s"ggest that maAor factors limiting .oth the GM cottons and nonBGM cotton esta.lishment and s"rvival incl"de water and n"trient availa.ility/ her.ivory .y nonBlepidopteran species #verte.rate and inverte.rate$/ plant competition/ frost and fire #Hastic( 2002F !arrell L +o.erts 2002$-
2<2<2
PAT enF0me
2!8+ *he %A* protein co"ld only confer a selective advantage in the presence of gl"fosinate ammoni"m her.icide application- 5n an agric"lt"ral setting/ the GM cottons lines proposed for release will have increased fitness where gl"fosinate ammoni"m is applied for weed control- ;owever/ gl"fosinate ammoni"m is not generally "sed to control vol"nteer cotton plants in agric"lt"ral systems/ and has limited effectiveness on esta.lished cotton #ie- .eyond the seedling stage$@"ltivation or other her.icides are the main control strategies employed #A"stralian @otton @ooperative +esearch @entre 2002$-
2<2<3
+r09*a= +r09Ac and PAT +om8ination
2!*+ *he insecticidal and her.icide tolerance genes operate thro"gh independent/ "nrelated .iochemical mechanisms- *here is no evidence of any interaction .etween the genes/ their proteins or their meta.olic pathways/ and no reason to e2pect that this is li(ely to occ"r- Agronomic ,"alities of all three GM cotton lines are similar to those of nonBGM cotton/ apart from the intended her.icide tolerance and insecticidal characteristics/ s"ggesting that no "nintended effects have occ"rred as a res"lt of the genetic modifications #see Appendi2 1/ Section =$-
(ection 2<3 Potential 4eediness of t1e GM insecticidal/1er8icide tolerant cottons

220+ Many of the characteristics associated with weediness are also important agronomic characteristics- @onse,"ently these are assessed as part of the agronomic eval"ations d"ring the development of new cotton varieties/ incl"ding GM varietiesGrowth ha.it/ germination/ emergence in the field/ vigo"r/ flowering period and reprod"ctive potential of these three GM cotton lines are very similar to those of the nonBGM parental cotton variety "sed in its .reeding/ and are within the range typical for nonBGM cotton varieties #%ellow 2006$- *hese data s"ggest that the genetic modifications in these GM cotton lines have not led to any "nintended effects on characteristics typically associated with weediness-
)'
(ection 2<4 ( read of t1e GM cottons 8e0ond t1e release sites

22!+ D"ring and after planting and harvesting/ cottonseed may .e dispersed .eyond the limits of the trial sites on e,"ipment or d"ring transportation of harvested seed cottonSpecific licence conditions have .een imposed to re,"ire cleaning of e,"ipment "sed in connection with the release/ and harvested material to .e sec"rely wrapped .efore transporting away from the release sites/ so as to prevent the GM cotton seed escaping into the wider environment #see Appendi2 I for details$- Seeds not re,"ired for f"t"re releases m"st .e destroyed-
(ection 2<: Persistence of t1e GM cottons at t1e release sites

222+ !ollowing harvest of the seed cotton from the release sites/ the remaining plant material will .e slashed and incorporated into the soil- Some seed may fall to the gro"nd d"ring harvesting and also .e incorporated into the soil- @otton has little dormancy/ meaning seed will germinate with the arrival of favo"ra.le soil moist"re and temperat"re conditions- GM cotton vol"nteers are e2pected to germinate in the wet season #Novem.er to March$ following harvest223+ %ost harvest inspections of the release sites are re,"ired as a condition of the licence/ to ens"re vol"nteer cotton plants are destroyed .efore flowering and that the GM cottons are "na.le to persist in the environment-
($+TI&' 3 +&'+)-(I&'( R$GARDI'G 6$$DI'$((

224+ 5t is concl"ded that the ris( of the GM cottons esta.lishing as environmental weeds as a res"lt of the proposed release is low .eca"se: !0'( cotton does not possess characteristics commonly associated with weediness/ and is not (nown to .e a pro.lematic weed in any environmentF !0)( the genetic modifications in the GM cotton lines are not li(ely to affect these characteristicsF !08( other GM insecticidal cottons grown #targeted to lepidopteran caterpillar pests$ commercially in A"stralia have not .ecome pro.lematic weedsF !0*( maAor constraints on weediness of .oth GM and nonBGM cottons are water availa.ility/ n"trient availa.ility/ plant competition/ her.ivory .y nonBlepidopteran species/ frost and fireF !!0( the insecticidal gene#s$ in the GM cotton may confer a selective advantage if the cotton is limited .y lepidopteran insects/ however/ the ris( is lowF and !!!( altho"gh the two traits #insecticidal and her.icide tolerance$ may have an additive effect on competitiveness/ neither of these traits individ"ally has .een fo"nd to .e significant for weediness22&+ 5t is considered that the low ris( of the GM cotton lines esta.lishing as weeds can .e managed .y applying vario"s strategies to limit the spread and persistence of the GM cottons from the release sites- 7icence conditions have .een imposed to manage this ris(/ incl"ding a re,"irement to cond"ct post harvest inspections of the release ))
sites and destroy vol"nteer cotton plants .efore they flower #see @hapter 2 and Appendi2 I for details$-
($+TI&' 4 *-RT/$R R$($AR+/ R$>-IR$M$'T(

5f the applicant ma(es any applications for f"t"re larger scale releases of these GM cottons/ more data wo"ld .e re,"ired to .e collected on agronomic characteristics of the GM cottons in relation to potential weediness "nder A"stralian field conditions and the additive effects of the two traits #insecticidal and her.icide tolerance$ on weediness-
)8
APP$'DI, : TRA'(*$R &* I'TR&D-+$D G$'$( T& &T/$R &RGA'I(M(

22'+ 3nder section 01 of the Act/ the +eg"lator is re,"ired to consider ris(s to h"man health and safety and the environment in preparing the ris( assessment and the ris( management plan- *his Appendi2 considers potential ha<ards that may .e posed thro"gh the transfer of the introd"ced genes from the GM insecticidal)her.icide tolerant cottons to other organisms22)+ Gene transfer is the movement of genes .etween individ"als- 9ithin a species genes are ro"tinely e2changed .etween individ"als of s"ccessive generations thro"gh se2"al reprod"ction- ;y.rids can sometimes .e prod"ced .etween closely related species thro"gh se2"al reprod"ction altho"gh this may re,"ire significant assistance!or e2ample/ in plants cross pollination of wheat and rye in the la.oratory prod"ced triticale- 5n animal/ fertilisation of a mare .y a don(ey prod"ces a m"le- ;y.rid progeny may .e fertile or sterile/ meaning hy.ridisation may or may not lead to the introgression of new genes into a pop"lation228+ 9itho"t the application of gene technology/ gene transfer is not readily o.served .etween distantly related species/ e2cept for .acteria and vir"ses- ;owever gene transfer .etween se2"ally incompati.le organisms can occ"r- Detailed e2amination of DNA se,"ence similarities reveals that ancestral plants have occasionally e2changed small DNA fragments with distantly related organisms- 5n general there seems to have .een only very limited transfer of f"nctional genes from plants to other types of organisms22*+ *he li(elihood of a ha<ard arising as a res"lt of gene transfer from the GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideSri(e? cottons$ is dependent on a n"m.er of factors/ which m"st form a chain for a ha<ard to .e realised/ incl"ding: !!2( o ort2nit0 for gene transfer to occ"r s"ch that the recipient organism is e2posed to the genetic material of the donor in the form of pollen/ plant cells or DNAF !!3( occ2rrence of the genetic material of the donor .eing incorporated into the genetic material of the recipient organism at a site and in a config"ration that allows the gene to .e f"nctionalF !!4( ersistence of the transferred genetic material s"ch that the recipient organism is a.le to s"rvive/ reprod"ce and maintain the genetic modificationF and !!&( significance of the transferred genetic material s"ch that its presence and)or e2pression in the recipient organism will res"lt in an adverse impact on h"man health and safety/ or the environment230+ !or ease of reference/ the assessment of gene transfer to other organisms is presented in fo"r sections:
A--endi2 &
T.ans$e. #$ int.#d0/ed genes t# #t,e. #.ganisms
)*
!!'( (ection 9 details the nat"re and li(elihood of a ha<ard arising thro"gh transfer of the introd"ced genes from the GM cottons to other plants/ incl"ding other cotton cropsF !!)( (ection 2 details the nat"re and li(elihood of a ha<ard arising thro"gh transfer of the introd"ced genes from the GM cottons to microorganismsF !!8( (ection 3 details the nat"re and li(elihood of a ha<ard arising thro"gh transfer of the introd"ced genes from the GM cottons to animals/ incl"ding h"mansF and !!*( (ection 4 draws together the concl"sions from these sections23!+ A detailed assessment of the li(elihood of gene transfer from GM cottons to other organisms is also presented in the ris( assessments for licence applications D5+s 000)2001/ 008)2001/ 00=)2001/ 00C)2002/ 012)2002/ 010)2002/ 018)2002/ 01I)2002/ 022)2002/ 026)2002/ 020)2002/ 064)2006/ 060)2006/ 068)2006/ 06=)2006 and 040)2006-
($+TI&' 9 G$'$ TRA'(*$R *R&M T/$ GM +&TT&'( T& &T/$R P)A'T(
(ection 9<9 'at2re of t1e gene transfer 1aFard

232+ *ransfer of the introd"ced genes #cry1Fa9 cry1Ac and pat$ or reg"latory se,"ences #promoter and terminator regions$ from the GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideSri(e? cottons$ to other c"ltivated cotton plants/ vol"nteer cotton or nat"ralised #feral$ cotton wo"ld present the same ha<ards/ and have the same potential impacts/ as the presence of the genes in the GM cottons proposed for release- *hese ris(s are considered in Appendices 2 B 8;owever/ if s"ch a transfer occ"rred/ it wo"ld increase the possi.ility that these genes co"ld f"rther spread in the environment233+ 5f gene transfer to other plant species were to occ"r/ the ha<ards to the environment associated with any s"ch transfers co"ld .e highly varied/ .roadly depending "pon the nat"re of the genes and of the species to which transfer occ"rred*ransfer of the introd"ced genes or reg"latory se,"ences into other plant species/ in partic"lar to native flora/ may have adverse effects on .iodiversity if the recipient plants gained a selective advantage/ s"ch as enhanced s"rvival or reprod"ctive capacity-
9<9<9
Potential 1aFards from t1e introd2ced genes
T,7 cry1Fa AND cry1Ac #5NSH@*5@5DA7$ GHNHS

234+ %lants e2pressing these genes wo"ld .e to2ic to lepidopteran insects- *his co"ld confer a selective advantage on the plants or adversely affect s"rvival of lepidopteran insects and conse,"ently other organisms lin(ed to lepidopteran insects thro"gh food we.s-
A--endi2 &
80
*;H pat #;H+'5@5DH *O7H+AN@H$ GHNH

23&+ %lants e2pressing this gene co"ld .ecome tolerant to the her.icide gl"fosinate ammoni"m/ conferring a selective advantage on the plant in the presence of gl"fosinate ammoni"m "se #eg- 7i.ertyE and 'astaE her.icides$-
*;H 5N*+OD3@HD %+OMO*H+S AND O*;H+ +HG37A*O+N SH:3HN@HS

23'+ 5f these se,"ences were to .e transferred to other plants witho"t the associated genes of the GM cottons/ the e2pression of endogeno"s plant genes co"ld .e altered with "npredicta.le effects- *he impact co"ld .e highly varia.le and wo"ld .e dependent on any res"lting phenotypic change ind"ced23)+ Some of the introd"ced reg"latory se,"ences are derived from a plant pathogen #Agro'acteriu" tu"e(aciens$- ;owever these se,"ences are not pathogenic in themselves nor do they ca"se any disease symptoms in GM plants238+ All of the introd"ced reg"latory se,"ences operate in the same manner as do endogeno"s plant reg"latory elements- *he transfer of endogeno"s reg"latory elements to new genetic conte2ts occ"rs nat"rally in all plant genomes and co"ld also res"lt in "npredicta.le effects- *h"s the potential ha<ards from the introd"ced se,"ences are not different from those posed .y se,"ence transfer from nonBGM plants or se,"ence transfer occ"rring within the genome of a plant species-
(ection 9<2 )ikeli1ood of a 1aFard arising t1ro2g1 transfer of t1e introd2ced genes to ot1er lants 9<2<9 Transfer to ot1er c2ltiAated cotton and Aol2nteer cotton
23*+ @otton is primarily selfBpollinating/ however in a cropping sit"ation a low level of pollen transfer/ .y insect pollinators/ to near.y cotton plants is li(ely- @otton pollen dispersal st"dies consistently show that o"tcrossing is localised aro"nd the pollen so"rce and decreases significantly with distance- St"dies carried o"t in so"thern A"stralia .y 7lewellyn and !itt #1CC8$ and *homson #1CC8$ demonstrated that o"tcrossing was very rare #less than 0-01D$ or was not detected at a distance of 10 m from GM cotton plants/ and no o"tcrossing was detected at 20 m- !or a detailed consideration of the li(elihood of this occ"rring/ incl"ding an overview of the pollination .iology of cotton/ see the doc"ment T*he 'iology and Hcology of @otton #Gossypiu" hirsutu"$ in A"straliaU/ availa.le at www.ogtr.gov.au/ that was prod"ced in order to inform the ris( assessment processes for licence applications involving GM cotton240+ Gossypiu" 'ar'edense #pima cotton$ is also "sed for commercial cotton prod"ction/ ."t only to a very minor e2tent in A"stralia #7a(e *ando" and 'o"r(e/ NS9$- G. hirsutu" and G. 'ar'edense are closely related and hy.ridisation .etween the two species can occ"r/ yielding fertile progeny- ;y.rid progeny e2hi.it characteristics intermediate to the parents ."t typically with lower capacity to prod"ce fr"it- After several generations/ progeny of the hy.rids revert to the characteristics of one or other of the parents- G. 'ar'edense and hy.rids are not weedier or more
A--endi2 &
8!
diffic"lt to control than is G. hirsutu" #personal comm"nication/ 9arwic( Stiller L Greg @onsta.le/ @S5+O$24!+ *ransfer of the introd"ced genes or reg"latory se,"ences to other cotton wo"ld present the same low ha<ards as their presence in the GM cottons proposed for release #see Appendices 2 B 8$- ;owever/ if s"ch a transfer occ"rred/ it wo"ld increase the possi.ility that these genes co"ld persist and f"rther spread in the environment242+ *he li(elihood of a ha<ard arising thro"gh transfer of the introd"ced genes wo"ld .e f"rther minimised .eca"se of the small si<e and limited d"ration of the proposed release- 7icence conditions have .een imposed to manage this ris(- *hese incl"de a re,"irement to either s"rro"nd the GM cottons with a 20 m pollen trap of nonBGM cotton or to ens"re that there are no other cotton crops or nat"ralised cotton pop"lations within 400 m of the GM cotton crops- @onditions have also .een imposed to esta.lish a 400 m wide area of land #called a >+esearch one4$ aro"nd release sites in e2cess of 1 ha for the p"rpose of cond"cting research on gene flowDevelopment of an agreed research program to inform the ongoing review of data on gene flow and to validate the efficacy of containment meas"res is a condition of the licence #see @hapter 2 and Appendi2 I for details$-
9<2<2
Transfer to nat2ralised cotton
243+ *ransfer of the introd"ced genes to nat"ralised cotton co"ld also increase the li(elihood that the genes co"ld spread and)or persist in the environment #away from cotton farming systems$- ;er.ari"m records of G. hirsutu" and G. 'ar'adense s"ggest that nat"ralised pop"lations may occ"r/ or may have occ"rred in the past/ in central and so"th eastern :"eensland/ in northern Northern *erritory and northern 9estern A"stralia- *he remnants of these pop"lations/ a few of which may .e within pollinating distance of commercial cotton crops/ has not .een confirmed- As part of the licence conditions for D5+ 022)2002/ a s"rvey of nat"ralised cotton pop"lations in :"eensland/ in locations s"ggested .y her.ari"m records/ is .eing cond"cted244+ *ransfer of the introd"ced genes or reg"latory se,"ences to nat"ralised cotton plants wo"ld present the same low ha<ards as their presence in the GM cottons proposed for release #see Appendices 2 B 8$- ;owever/ if s"ch a transfer occ"rred/ it wo"ld increase the possi.ility that these genes co"ld persist and f"rther spread in the environment24&+ 7icence conditions are imposed to limit crossBpollination to plants o"tside the release sites #see Section 1-2-1 a.ove or @hapter 2 and Appendi2 I for details$-
9<2<3
Transfer to natiAe cottons and ot1er lant s ecies
24'+ A"stralian flora contains 1I native Gossypiu" species- All of the A"stralian Gossypiu" species are diploids #@/ G or J genomes$/ while the c"ltivated cottons are tetraploids #ADBgenomes$- *he native species with highest potential for hy.ridising with G. hirsutu" is G. sturtianu"- ;y.rids have .een prod"ced witho"t application of plant hormones/ when plants were planted in close pro2imity of each other-
A--endi2 &
82
;owever these hy.rids were sterile/ effectively eliminating any potential for introgression of G. hirsutu" genes into G. sturtianu" pop"lations #OG*+ 2002$24)+ *he centre of native Gossypiu" diversity in A"stralia is in northern 9estern A"stralia and the Northern *erritory- Most of the A"stralian Gossypiu" species have limited distri."tions and occ"r at considera.le geographic distance from c"ltivated cotton fields- *h"s gene transfer from these GM cotton lines to native cottons is prevented not only .y genetic incompati.ility ."t also .y geographic constraints to crossBpollination #OG*+ 2002$248+ *he fail"re of crossBpollination d"e to well esta.lished genetic incompati.ility also prevents gene transfer from these GM cotton lines to other plant species-
($+TI&' 2 G$'$ TRA'(*$R *R&M T/$ GM +&TT&' T& MI+R&&RGA'I(M(

24*+ *he transfer of genes from plants to microorganisms cannot occ"r thro"gh crossB pollination- ;ori<ontal gene transfer is defined as the transfer of genetic material from one organism #the donor$ to another organism #the recipient$ which is not se2"ally compati.le with the donor #@onner et al- 2006$- *here is growing evidence that hori<ontal gene transfer has .een a principal force in the evol"tion of .acteria #Nielsen 1CC=F Ochman et al- 2000F Smalla et al- 2000F Stanhope et al- 2001$2&0+ *he potential ha<ards associated with the introd"ced genes of the GM insecticidal)her.icide tolerant cottons transferring to microorganisms co"ld .e highly varied/ .roadly depending "pon the phenotype of the recipient and any changes to its s"rvival or reprod"ctive capacity- *he impact of any ha<ard arising thro"gh gene transfer wo"ld also depend on other so"rces of the introd"ced genes in the environment-
2<9<9
Potential 1aFards from t1e introd2ced genes
*;H cry1Fa AND cry1Ac #5NSH@*5@5DA7$ GHNHS

2&!+ Microorganisms e2pressing these genes wo"ld .e to2ic to lepidopteran insects*his co"ld impact on s"rvival of lepidopteran insects if the recipient microorganisms were ingested at high levels- Microorganism pop"lations co"ld also .e affected if to2icity to lepidopteran insects gave the recipient a s"rvival or reprod"ctive advantage-

2&2+ Microorganisms e2pressing this gene wo"ld gain resistance to gl"fosinate ammoni"m and to the tripeptide anti.iotics nat"rally prod"ced .y a small n"m.er of actinomycete .acteria #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$: 'ialaphos #phosphinothricylB7BalanylB7Balanine$/ prod"ced .y %. viridochro"ogenes and %. hygroscopicus #the .acterial species from which the pat and related 'ar genes/ respectively/ are derived$F and %hosalacine #phosphinothricylB 7BalanylB7Ble"cine$/ prod"ced .y 2itasatosporia phosalacinea-
A--endi2 &
83
2&3+ Neither .ialaphos or phosalacine are "sed in h"man or veterinary therapy%trepto"yces spp- are saprophytic soil .acteria/ and not considered pathogens of plants/ h"mans/ or other animals #Organisation for Hconomic @oBoperation and Development #OH@D$ 1CCC$- *h"s transfer of this gene is "nli(ely to present a ha<ard in relation to pathogenesis2&4+ Anti.iotic prod"ction .y nonBpathogenic .acteria has .een implicated in s"ppression of some plant diseases #'rimecom.e et al- 2001$- *h"s transfer of the pat gene to new .acterial species/ incl"ding plant pathogens/ co"ld potentially impact on micro.ial pop"lations or plant disease s"scepti.ility- ;owever/ e2pression of the pat or the related 'ar gene in a variety of crop plants #for e2ample canola and corn$/ over several years of agronomic performance testing and commercial c"ltivation/ has not .een lin(ed to increased s"scepti.ility to disease/ as assessed .y reg"latory a"thorities of other co"ntries/ incl"ding the Animal and %lant ;ealth 5nspection Service #A%;5S$ of the 3- S- Department of Agric"lt"re #eg- 3SDABA%;5S 2006$ and the @anadian !ood 5nspection Agency #eg- @anadian !ood 5nspection Agency 1CC=.$/ also see Gene files #2002$ for lin(s to f"rther safety assessments-
%+OMO*H+S AND O*;H+ +HG37A*O+N SH:3HN@HS

2&&+ 5f these se,"ences were to .e transferred to microorganisms witho"t the associated genes of GM cottons/ the e2pression of endogeno"s genes co"ld .e altered with "npredicta.le effects- *he impact co"ld .e highly varia.le and wo"ld .e dependent on any res"lting phenotypic change2&'+ Some of the introd"ced reg"latory se,"ences are derived from a plant pathogen #Agro'acteriu" tu"e(aciens$- ;owever these se,"ences are not pathogenic in themselves nor do they ca"se any disease symptoms in GM plants2&)+ All of the introd"ced reg"latory se,"ences operate in the same manner as do endogeno"s plant reg"latory elements- *he transfer of endogeno"s reg"latory elements to a new genetic conte2t co"ld also res"lt in "npredicta.le effects- *h"s the li(elihood of a ha<ard arising d"e to transfer of the introd"ced se,"ences is no different to that of se,"ence transfer from nonBGM plants-
2<9<2
&t1er so2rces of t1e introd2ced genes in t1e enAironment
2&8+ 5nformation on other so"rces of the introd"ced genes in the environment is disc"ssed here to provide .aseline information on the prevalence and transfer of these genes in the a.sence of the GM cotton2&*+ All of the introd"ced genes in the GM cotton are already widespread in the environment/ .eing derived from common soil .acteria- *he reg"latory se,"ences are derived from a plant and a common .acteri"m-
*;H cry1Fa A:8 cry1Ac #5NSH@*5@5DA7$ GHNHS

2'0+ Altho"gh the introd"ced cry1 genes in GM cotton lines proposed for release are chimeric/ the encoded @ry proteins are very similar to the native @ry proteins #CC-6D similarity in amino acid se,"ence of core to2in .etween native and chimeric @ry1!a 84
A--endi2 &
and CC-8D .etween native and chimeric @ry1Ac$/ and retain the species specific insecticidal to2icity to lepidopteran caterpillar pest- *he cry1Fa and cry1Ac genes occ"r nat"rally in the common soil .acteria B. thuringiensis #'t$- 't has .een isolated from a wide range of so"rces s"ch as forest/ soil/ grain d"st/ .at d"ng/ sea water and dead insects #Martin L *ravers 1C=C$2'!+ Many @ry to2in genes are not carried in chromosomal DNA/ ."t are encoded on e2traBchromosomal DNA/ (nown as plasmids- %lasmids are (nown to .e e2changed .etween .acterial species in nat"re .y processes (nown as conA"gation and transformation- *he native cry1Ac gene has .een identified on a plasmid of 't variety kurstaki/ strain ;DBI6 #7erecl"s et al- 1CC6$- 5t has .een demonstrated in the la.oratory that 't strains can interchange to2inBencoding plasmids with other 't strains and with other .acterial species #Glare L OV@allaghan 2000$- ;ori<ontal gene transfer may also occ"r .y transd"ction mediated .y .acteriophages #Glare L OV@allaghan 2000$-

2'2+ *he her.icide tolerance pat gene was originally isolated from the common soil .acteri"m %. viridochro"ogenes/ and another gene/ 'ar/ which also encodes a %A* en<yme is nat"rally present in %. hygroscopicus- *hese .acteria are not considered pathogenic to plants/ h"mans or other animals #Organisation for Hconomic @oB operation and Development #OH@D$ 1CCC$ and are widespread in nat"ral and agric"lt"ral environments2'3+ A search of the Gen'an( DNA se,"ence data.ase reveals that other genes encoding %A* or similar en<ymes are present in a variety of other .acteria-
(ection 2<2 )ikeli1ood of a 1aFard arising t1ro2g1 transfer of t1e introd2ced genes to microorganisms
2'4+ *he li(elihood of gene transfer creating a ha<ard for h"man health and safety or the environment depends on the characteristics of introd"ced gene se,"ences/ as well as on the li(elihood of the transfer itself2'&+ Most instances of hori<ontal gene transfer have .een identified thro"gh analyses of gene se,"ences #9oro.ey L ;olmes 1CCCF Ochman et al- 2000$- 5n general/ gene transfers are detected over evol"tionary time scales of millions of years #7awrence L Ochman 1CC=$- Most gene transfers have .een from vir"s to vir"s #7ai 1CC2$/ or .etween .acteria #Ochman et al- 2000$2''+ 5n contrast/ transfers of plant genes to other organisms s"ch as .acteria/ f"ngi or vir"ses are e2ceedingly rare #Greene L Allison 1CC4F Nielsen et al- 1CC=F Nielsen et al- 2000F %ittard 1CCIF Schoel< L 9intermantel 1CC6F 9oro.ey L ;olmes 1CCCF Ao(i L Syono 1CCCF ;arper et al- 1CCCF Mayo L Molly 1CC1$- *he transfer of plant genes to .acteria and vir"ses has .een o.served in la.oratory and glassho"se e2periments #Nielsen et al- 1CC=F %ittard 1CCIF 9oro.ey L ;olmes 1CCCF Greene L Allison 1CC4F Nielsen et al- 2000F Schoel< L 9intermantel 1CC6$- ;owever/ in all cases this was achieved only "nder controlled conditions with the presence of related gene se,"ences
A--endi2 & T.ans$e. #$ int.#d0/ed genes t# #t,e. #.ganisms
8&
#homologo"s recom.ination$/ and "sing powerf"l selection methods to detect e2tremely rare gene transfer events-
2<2<9
#acteria
2')+ *hree different mechanisms of hori<ontal gene transfer in .acteria have .een descri.ed: transd"ction/ conA"gation/ and transformation #Nielsen 1CC=$2'8+ *ransd"ction is a .acterial cellBvir"s interaction that can mediate gene transfer .etween .acteria in the environment #e-g- on plant leaf s"rfaces/ in soil or water$&ir"ses that f"nction in more than one species are (nown/ ."t vir"ses that f"nction in .oth plants and .acteria/ and there.y facilitate hori<ontal gene transfer from plants to .acteria have not .een identified #Nielsen et al- 1CC=$2'*+ @onA"gation is a mechanism of cellBtoBcell interaction that can mediate gene transfer .etween .acteria in the environment #e-g- in soil/ on plant s"rfaces/ in water etc$2)0+ @onA"gation is (nown to occ"r fre,"ently .etween compati.le .acteria with the transfera.le genes "s"ally residing on plasmids- *ransfer of chromosomal genes is m"ch less fre,"ent/ e2cept for some high fre,"ency recom.ination strains@onA"gative gene transfer has .een regarded as the most fre,"ently occ"rring mechanism of hori<ontal gene transfer .etween .acteria #Sprag"e 1CC1F Ama.ileB @"evas L @hic"rel 1CC6$- ;owever/ mechanisms that s"pport conA"gative gene transfer from higher plants to .acteria #e-g- transposons that f"nction in .oth plants and pro(aryotes$ are not (nown #Nielsen 1CC=$2)!+ Gene transfer .y transformation res"lts from the "pta(e of na(ed DNA .y .acteria/ and has .een shown to occ"r in environments s"ch as in soil and in water #7oren< L 9ac(ernagel 1CC4F Streips 1CC1$- Most st"dies descri.ing nat"ral transformation have .een cond"cted in vitro #7oren< L 9ac(ernagel 1CC4F Streips 1CC1$ ."t often are of little relevance to most nat"ral terrestrial environments2)2+ Nat"ral transformation is a mechanism .y which transfer of DNA from plants to microorganisms co"ld have occ"rred d"ring evol"tion #'ertolla L Simonet 1CCC$ and is the mechanism that is most li(ely to contri."te to a hori<ontal gene transfer from GM plants to .acteria #Smalla et al- 2000$- Nat"ral transformation ena.les competent .acteria to generate genetic varia.ility .y ta(ing "p and integrating free DNA that is present in their s"rro"ndings- *his "pta(e of DNA does not necessarily depend on DNA se,"ence/ th"s indicating the potential for gene transfer from divergent donor organisms #Nielsen 1CC=$2)3+ A n"m.er of steps and conditions wo"ld need to .e f"lfilled for f"nctional nat"ral transformation to occ"r #'ertolla L Simonet 1CCC$/ many of which are highly "nli(ely/ ma(ing the overall li(elihood of gene transfer/ and of res"lting ha<ard/ e2tremely low: !20( release of t1e D'A molec"les from plant cells into the environmentF !2!( ersistence of the free DNA in the environmentF
A--endi2 &
8'
!22( resence of .acterial genotypes capa.le of developing competence for nat"ral transformationF !23( a ro riate 8iotic and a8iotic conditions for the development of the competent stateF !24( 2 take of DNA fragmentsF !2&( c1romosomal integration via recom.ination or a"tonomo"s replication of the transforming DNAF !2'( ex ression of the genes .y the recipient .acteri"mF and !2)( selectiAe adAantage to fi2 #maintain$ the transferred DNA in the gene pool of the recipient species2)4+ *h"s hori<ontal gene transfer from plants to .acteria has not .een demonstrated "nder nat"ral conditions #Syvanen 1CCC$ and deli.erate attempts to ind"ce s"ch transfers have so far failed #eg- SchlWter et al- 1CC0F @oghlan 2000$- *ransfer of plant DNA to .acteria has .een demonstrated only "nder highly artificial la.oratory conditions/ .etween homologo"s se,"ences and "nder conditions of selective press"re #Mercer et al- 1CCCF Ge.hard L Smalla 1CC=F De &ries L 9ac(ernagel 1CC=F De &ries et al- 2001$/ and even then only at a very low fre,"ency-
2<2<2
%ir2ses
2)&+ *here is a theoretical possi.ility of recom.ination .etween se,"ences that have .een introd"ced into the genome of GM plants and the genome of vir"ses that infect the plants #;o et al- 2000F ;odgson 2000aF ;odgson 2000.$- +ecom.ination .etween viral genomes and plant DNA has only .een o.served at very low levels/ and only .etween homologo"s se,"ences "nder conditions of selective press"re/ egregeneration of infectio"s vir"s .y complementation of a defective vir"s .y viral se,"ences introd"ced into a GM plant genome #Greene L Allison 1CC4F *eycheney L *epfer 1CCC$- 9ith homologo"s se,"ences the conse,"ent ris( of adverse effects arising from gene transfer is red"ced .eca"se with highly similar se,"ences the li(elihood of any recom.inants e2pressing novel properties is low2)'+ *h"s the li(elihood of gene transfer leading to ha<ardo"s conse,"ences is e2tremely low/ and greatly e2ceeded .y the li(elihood of transfer from other so"rces of these genes and reg"latory se,"ences #see Section 2-1-2$-
2<2<3
*2ngi
2))+ !"ngi are (nown to .e transforma.le/ and hori<ontal gene transfer from plants to plantBassociated f"ngi has .een claimed- 3pta(e of DNA from the host plant .y 1las"odiophora 'rassicae #'ryngelsson et al- 1C==F '"hariwalla L Mithen 1CC0$ and "pta(e of the hygromycin gene from a GM plant .y Aspergillus niger #;offman et al- 1CC4$ have .een reported- ;owever/ sta.le integration and inheritance of the plant DNA in the genome of these f"ngi has not .een s".stantiated .y e2perimental evidence #Nielsen 1CC=$-
A--endi2 &
8)
2)8+ *h"s the li(elihood of gene transfer leading to ha<ardo"s conse,"ences is e2tremely low/ and greatly e2ceeded .y the li(elihood of transfer from other so"rces of these genes and reg"latory se,"ences #see Section 2-1-2$-
($+TI&' 3 G$'$ TRA'(*$R *R&M T/$ GM +&TT&' T& A'IMA)(

2)*+ *he potential ha<ards associated with the introd"ced genes in the GM insecticidal)her.icide tolerant cottons transferring to animals/ incl"ding h"mans/ co"ld .e highly varied/ .roadly depending "pon the phenotype of the recipient and any changes to the s"rvival or reprod"ctive capacity of it or its progeny-
3<9<9
Potential 1aFards from introd2ced genes
*;H cry1Fa AND cry1Ac #5NSH@*5@5DA7$ GHNHS

280+ Animals co"ld .ecome to2ic to lepidopteran insects- *his is not li(ely to pose any conse,"ences for lepidopteran insects/ nor wo"ld s"ch a transfer confer a selective advantage or disadvantage to the animal-

28!+ *he e2pression of the %A* en<yme in an animal wo"ld not .e e2pected to prod"ce any adverse meta.olic effects/ since %A* has e2tremely high s".strate specificity for 7B phosphinothricin #7B%%*$/ and cannot acetylate any other amino acids or proteins #9ehrmann et al- 1CC8F @anadian !ood 5nspection Agency 1CC0$-
%+OMO*H+S AND O*;H+ +HG37A*O+N SH:3HN@HS

282+ 5f these se,"ences were to .e transferred to animals witho"t the associated genes of the GM cottons/ the e2pression of endogeno"s genes co"ld .e altered with "npredicta.le effects- *he impact co"ld .e highly varia.le and wo"ld .e dependent on the res"lting phenotypic change ind"ced283+ Some of the introd"ced reg"latory se,"ences are derived from plant pathogens #Agro'acteriu" tu"e(aciens$- ;owever these se,"ences are not pathogenic in themselves nor do they ca"se any disease symptoms in GM plants284+ All of the introd"ced reg"latory se,"ences operate in the same manner as do endogeno"s plant reg"latory elements- *he transfer of endogeno"s reg"latory elements to a new genetic conte2t co"ld also res"lt in "npredicta.le effects- *h"s the li(elihood of a ha<ard arising d"e to transfer of the introd"ced se,"ences from the GM cottons is no different to that arising from nonBGM plants-
(ection 3<2 )ikeli1ood of a 1aFard arising t1ro2g1 transfer of t1e introd2ced genes to animals
28&+ *he most significant ro"te for entry of foreign DNA into animals/ incl"ding h"mans/ wo"ld .e thro"gh food/ as it passes thro"gh the gastrointestinal tract- *he epithelial lining of the gastrointestinal tract is e2posed to foreign DNA released from food- Microorganisms colonise the whole length of the gastrointestinal tract/ aiding
A--endi2 & T.ans$e. #$ int.#d0/ed genes t# #t,e. #.ganisms
88
the digestive process- ;owever/ the proportion of DNA which may derive from the introd"ced genes of GM plants in the animal diet is e2tremely low- !or e2ample/ 'eever and Jemp #2000$ estimated that in a diet comprising 40D GM mai<e/ the introd"ced genes wo"ld represent 0-00042D of total dietary DNA inta(e28'+ A recent st"dy showed that the cry1A' gene for GM mai<e was not deteca.le in the peripherical .lood monon"clear cells and tiss"es of calves 0 to 1= ho"rs after the calves were fed with the GM mai<e/ indicating the gene was not .eing transferred into those cells or tiss"es #@howdh"ry et al- 2004$28)+ *here are few p".lished st"dies eval"ating the s"rvival of transgene in the h"mans- A st"dy cond"cted .y Netherwood #2004$ with GM soya showed that the introd"ced gene #epsps$ did not s"rvive passage thro"gh the intact gastrointestinal tract of h"man s".Aects fed GM soya/ s"ggesting the gene was not .eing transferred to h"mans #Netherwood et al- 2004$288+ *he fate of DNA in the digestive tract of vario"s animals has .een st"died and is disc"ssed in detail in the ris( assessments for D5+ 021)2002 and D5+ 22)2002- *hese ris( assessments concl"ded that the li(elihood of transfer via food is e2tremely low/ and no greater than the li(elihood of transfer from other so"rces of the introd"ced genes in the environment #see Section 2-1-2 of this Appedi2$28*+ No prod"cts from the GM cottons in the proposed field trial will .e "sed for h"man food or animal feed- Most animals avoid feeding on cotton d"e to its nat"ral to2icity and morphology #OG*+ 2002$- *h"s the li(elihood of gene transfer to animals/ incl"ding h"mans/ is negligi.le- !"rthermore/ any "pta(e of plant DNA is li(ely to occ"r in nonBreprod"ctive #somatic$ cells s"ch as imm"ne system of g"t epitheli"m cells/ and the introd"ced gene wo"ld .e transmitted to the cells of any progeny- 5t is worth noting that cottonseed oil and linters are the only fraction of cotton plants "sed in h"man food- Since these prod"cts are free of DNA/ even if prod"cts of the GM cottons were approved .y !SAN for "se in food/ h"mans wo"ld not .e e2posed to DNA of these GM cottons via food/ e2cl"ding the possi.ility of gene transfer to h"man cells in the g"t-
($+TI&' 4 +&'+)-(I&'( R$GARDI'G G$'$ TRA'(*$R T& &T/$R &RGA'I(M(
(ection 4<9 +oncl2sions regarding gene transfer to ot1er lants

2*0+ 5t is considered that ris(s arising thro"gh gene transfer from the GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ to other plants are low .eca"se: !28( gene transfer to other cotton crops/ vol"nteer or nat"ralised cotton wo"ld not pose any ris(s additional to the low ris(s posed .y the GM cottons themselvesF !2*( genetic incompati.ility prevents gene transfer to native cotton species and other plant species2*!+ *he licence conditions have .een imposed to f"rther limit gene transfer to other c"ltivated/ vol"nteer and nat"ralised cotton- *hese incl"de a re,"irement to either
A--endi2 &
8*
s"rro"nd the GM cottons with a 20 m pollen trap of nonBGM cotton or to ens"re that there are no other cotton crops or nat"ralised cotton pop"lations within 400 m of the GM cotton crops- Additional conditions have .een imposed to esta.lish a 400 m wide area of land #called a >+esearch one4$ aro"nd release sites in e2cess of 1 ha for the p"rpose of cond"cting research to inform the ongoing review of data on gene flow and to validate the efficacy of containment meas"res #see @hapter 2 and Appendi2 I for details$-
(ection 4<2 +oncl2sions regarding gene transfer to microorganisms

2*2+ 5t is considered that ris(s arising thro"gh transfer of the introd"ced genes from GM cottons to microorganisms are negligi.le/ and do not re,"ire management/ .eca"se: !30( the introd"ced genes in the GM cottons are derived from/ and are similar to/ native .acterial genes that are already widespread in the environment and are readily availa.le for transfer from these so"rces via demonstrated nat"ral mechanismsF and !3!( gene transfer from plants to microorganisms has not .een demonstrated "nder nat"ral conditions/ and the li(elihood of s"ch transfer is greatly e2ceeded .y the li(elihood of transfer from other so"rces of these genes-
(ection 4<3 +oncl2sions regarding gene transfer to animals= incl2ding 12mans

2*3+ 5t is considered that ris(s thro"gh transfer of the introd"ced genes from GM cottons to animals/ incl"ding h"mans/ are negligi.le/ and do not re,"ire management/ .eca"se: !32( the introd"ced genes in the GM cottons are derived from/ and are similar to/ native .acterial genes that are already widespread in the environmentF !33( transfer of the introd"ced genes wo"ld .e "nli(ely to pose a ha<ard to h"man health and safety or to the environmentF !34( prod"cts from the GM cotton lines are not intended for stoc(feed or h"man foodF !3&( most animals avoid feeding on GM or nonBGM cotton plantsF and !3'( even if the GM cottons were approved .y !SAN for "se in food/ the cotton .yB prod"cts "sed in food do not contain DNA-
($+TI&' : R$($AR+/ R$>-IR$M$'T(

2*4+ *he proposed release of the GM cotton is a small scale/ m"ltiBsite trial over fo"r cotton growing seasons- *he licence holder is re,"ired to esta.lish a 400 m wide area of land #called a >+esearch one4$ aro"nd release sites in e2cess of 1 ha for the p"rpose of cond"cting research on gene flow- Development of an agreed research program to inform the ongoing review of data on gene flow and to validate the efficacy of containment meas"res is a condition of the licence #see @hapter 2 and Appendi2 I for details$-
A--endi2 &
*0
A--endi2 &
*!
APP$'DI, ; D$%$)&PM$'T &* I'($+TI+ID$ R$(I(TA'T P$(T(

2*&+ 3nder section 01 of the Act/ the +eg"lator is re,"ired to consider ris(s to h"man health and safety and the environment in preparing the ris( assessment and the ris( management plan- 5n this Appendi2/ ris(s posed .y the proposed release to the environment are considered in relation to the potential for development of insecticide resistance among target pests2*'+ 5t sho"ld .e noted that Dow AgroSciences does not intend to apply gl"fosinate ammoni"m her.icide #e-g- 7i.ertyE and 'astaE her.icides$ to the GM cotton lines proposed for the release/ therefore iss"es relating to ris(s of development of her.icide resistant weeds are not considered in this Appendi2-
($+TI&' 9 R$G-)ATI&' &* AGRI+-)T-RA) +/$MI+A)( I' A-(TRA)IA

2*)+ +eg"lation of agric"lt"ral chemicals/ incl"ding her.icides and insecticides/ is principally the responsi.ility of the A"stralian %esticides and &eterinary Medicines A"thority #A%&MA$ "nder the Agricultural and eterinary !he"icals !ode Act 1##$ #the Ag &et @ode Act$- *he GM cotton lines proposed for release fall "nder the Ag &et @ode Act definition of an agric"lt"ral chemical prod"ct/ d"e to its prod"ction of an insecticidal s".stance#s$/ and are th"s s".Aect to reg"lation .y the A%&MA2*8+ *he A%&MA operates the national system that eval"ates/ registers and reg"lates agric"lt"ral and veterinary chemical prod"cts- Any changes to the "se of a prod"ct that is already on the mar(et m"st also .e referred to the A%&MA- !or commercial prod"cts/ the normal form of approval is thro"gh registration/ ."t the A%&MA may also iss"e permits for e2perimental wor( that allow restricted "se of an agric"lt"ral chemical/ for e2ample/ for a limited period of time or for a limited area2**+ 5n considering applications for registration or permits/ as well as considering potential health and environmental impacts/ the A%&MA also considers a n"m.er of iss"es that are o"tside the scope of the Gene *echnology +eg"lator4s assessment/ s"ch as efficacy and the trade implications of resid"es- *he ha<ard of development of resistance to agric"lt"ral chemicals is part of the A%&MA4s assessment of agric"lt"ral chemical "se- *he A%&MA can impose conditions on the "se of chemical prod"cts in .oth registrations and permits- *hese conditions can incl"de restrictions on "se/ implementation of a resistance management plan/ and ongoing reporting on compliance300+ *he Gene Technology Act 2000 re,"ires the reg"lator to cons"lt the A%&MA in relation to the assessment of licence applications involving intentional release of GMOs to the Hnvironment- *he Gene Technology 3!onse;uential A"end"ents< Act 2000 places a reciprocal o.ligation "pon the A%&MA to cons"lt the Gene *echnology +eg"lator in relation to certain decisions regarding registrations and permits for an agric"lt"ral chemical that is or contains a genetically modified prod"ct30!+ *he A%&MA and the OG*+ have wor(ed closely to ens"re the thoro"gh/ coordinated assessment of these parallel proposals/ and that the decisions .y .oth
A--endi2 '
De6el#-ment #$ inse/ti/ide .esistant -ests
*2
agencies coincide- !"rther information a.o"t the A%&MA4s assessment and approval processes can .e o.tained from www.apv"a.gov.au-
($+TI&' 2 'AT-R$ &* T/$ I'($+TI+ID$ R$(I(TA'+$ /ABARD

302+ 5f the GM insecticidal)her.icide tolerant cotton lines #@ry1!a/ @ry1Ac and 9ideStri(e? cottons$ were c"ltivated e2tensively/ ,elicoverpa ar"igera/ ,. punctigera and other s"scepti.le lepidopteran insect pest species that feed on the GM cottons wo"ld .e placed "nder selection press"re for resistance to the @ry1!a and @ry1Ac insecticidal proteins- 5f resistance were to develop in target pests/ the insecticidal efficacy of these GM cottons wo"ld .e adversely affected/ potentially atten"ating any .enefits of these GM cottons and of other applications of these insecticidal proteins-
($+TI&' 3 )I5$)I/&&D &* T/$ I'($+TI+ID$ R$(I(TA'+$ /ABARD

&++-RRI'G
303+ D"e to the small scale and limited d"ration of the proposed release/ the li(elihood of insects developing resistance to the insecticidal proteins in these GM cottons is negligi.le304+ Dow AgroSciences has s".mitted an application to the A%&MA for a research permit for the "se of the insecticidal genes in GM cotton lines for the proposed release- *he ha<ard of development of insecticide resistance in pests is also .eing assessed .y the A%&MA in considering Dow AgroSciences4 permit application- *he A%&MA wo"ld impose conditions if it considered this necessary to manage any identified ris(-
($+TI&' 4 +&'+)-(I&' R$GARDI'G I'($+TI+ID$ R$(I(TA'+$

30&+ *he ris( of development of insecticide resistance in pests is negligi.le d"e to the small scale and limited d"ration of the proposed release- *his ris( is .eing assessed .y the A%&MA in considering Dow AgroSciences4 permit application for the "se of the insecticidal genes as insecticides in the GM cottons30'+ *herefore/ the +eg"lator has not imposed specific conditions in relation to management of insecticide resistance/ however the re,"irement to comply with any conditions imposed .y the A%&MA is noted in the licence-
A--endi2 '
De6el#-ment #$ inse/ti/ide .esistant -ests
*3
APP$'DI, C )I+$'+$ +&'DITI&'( Gene Techn#l#gy Regulati#n in Australia

The Gene Technology Act 32000< and corresponding State and territory legislation form a s".stantial part of a range of integrated reg"latory meas"res relevant to controlling genetically modified organisms #GMOs$ and their "se*he Gene *echnology +eg"lator is re,"ired to cons"lt with/ and ta(e into acco"nt advice from/ a range of reg"latory a"thorities on ris(s to h"man health and safety and the environment in assessing applications for dealings involving the intentional release of GMOs into the A"stralian environment-
N#te in relati#n t# a""r#$al #% genetically &#'i%ie' %##'s %#r hu&an c#nsu&"ti#n

!ood Standards A"stralia New ealand #!SAN $ is responsi.le for h"man food safety assessment- !SAN approval wo"ld need to .e o.tained .efore any parts of GM cottons/ s"ch as oil and linters derived from GM cottonseed/ co"ld .e "sed as h"man food- *his licence contains a condition that prohi.its this "se-
N#te in relati#n t# insectici'e resistance &anage&ent

*he Gene Technology 3!onse;uential A"end"ents< Act 2000 re,"ires the A"stralian %esticides and &eterinary Medicines A"thority #A%&MA$/ to cons"lt the Gene *echnology +eg"lator for the p"rposes of ma(ing certain decisions/ incl"ding the imposition of conditions of "se regarding registration or iss"ing a permit for a chemical prod"ct that is/ or contains a genetically modified prod"ct*he GMOs referred to in this licence fall "nder the Agricultural and eterinary !he"icals !ode 1##$ definition of agric"lt"ral chemical prod"cts/ d"e to their prod"ction of insecticidal s".stances/ and are therefore s".Aect to reg"lation .y the A%&MA- *he A%&MA assesses the ha<ard of development of insecticide resistance as part of its eval"ation process and wo"ld impose conditions to manage any identified ris(s- *herefore/ the conditions of this licence do not relate to management of insecticide resistance/ and do not replace any conditions set .y the A%&MA-
A--endi2 )
7i/en/e /#nditi#ns
*4
($+TI&' 9
I'T$RPR$TATI&'( A'D D$*I'ITI&'(
*his licence does not a"thorise dealings with GMOs that are otherwise prohi.ited as a res"lt of the operation of State legislation declaring areas to .e GM/ GM free/ or .oth/ for mar(eting p"rposes-
5n this licence:
#a$ 9ords and phrases "sed in this licence have the same meaning as they do in the Act and the +eg"lationsF #.$ 9ords importing a gender incl"de any other genderF #c$ 9ords in the sing"lar incl"de the pl"ral and words in the pl"ral incl"de the sing"larF #d$ 9ords importing persons incl"de a partnership and a .ody whether corporate or otherwiseF #e$ +eferences to any stat"te or other legislation #whether primary or s".ordinate$ are a reference to a stat"te or other legislation of the @ommonwealth of A"stralia as amended or replaced from time to time and e,"ivalent provisions/ if any/ in corresponding State law/ "nless the contrary intention appearsF #f$ 9here any word or phrase is given a defined meaning/ any other part of speech or other grammatical form in respect of that word has a corresponding meaningF and #g$ Specific conditions prevail over standard conditions to the e2tent of any inconsistency5n this licence:
GActH means the Gene Technology Act 2000 #@th$ and e,"ivalent provisions in corresponding State lawF
G+leanH #or G+leanedH$/ as the case re,"ires/ means: #a$ in relation to a 7ocation or other area/ the Destr"ction of the plants and %lant Material in that 7ocation or area/ to the reasona.le satisfaction of the +eg"latorF or #.$ in relation to H,"ipment/ the removal and Destr"ction of plants and %lant Material from the H,"ipment/ to the reasona.le satisfaction of the +eg"latorF
G+ottonH means plants of the species Gossypiu" hirsutu" 7F
A--endi2 )
7i/en/e /#nditi#ns
*&
GDestro0H/ #or GDestro0edH or GDestr2ctionH$ means/ as the case re,"ires/ (illed .y one or more of the following methods: #a$ stal( p"llingF or #.$ "prooting .y plo"ghingF #c$ root c"ttingF or #d$ ."rningF or #e$ treatment with her.icideF or #f$ hand weedingF
:ote) =As the case re;uires+ has the e((ect that9 depending on the circu"stances9 one or "ore o( these techni;ues "ay not 'e appropriate.
G$72i mentH incl"des machinery/ harvesters/ seeders/ storage e,"ipment/ transport e,"ipment #eg .ags/ containers/ tr"c(s$/ ginning facilities/ clothing and tools "sed in connection with this licence?
GGMH means genetically modifiedF
GGM&sH means the genetically modified organism or organisms a"thorised for release .y this licenceF
GIsolation BoneH means the area of land/ e2tending o"twards 00 metres in all directions from the o"ter edge of a 7ocation?
G)ocationH means an area of land where the GMOs are planted and grownF
GPlant MaterialH means via.le parts of GMOs and %ollen *rap plants/ incl"ding seed/ st"..le/ pollen/ whether from the plant itself or derived from or prod"ced .y the plantF
G'at2ral 6ater4a0sH means waterways other than irrigation channels/ holding dams or storage ponds "sed to collect water r"noff from irrigated areasF
A--endi2 )
7i/en/e /#nditi#ns
*'
G&GTRH means the Office of the Gene *echnology +eg"latorF
GPollen Tra H means the area of land e2tending o"twards 20 metres in all directions from the o"ter edge of a 7ocationF
GPollen Tra
lantH means @otton from a %ollen *rapF
GResearc1 BoneH means an area of land e2tending o"twards at least 400 metres in all directions from the o"ter edge of a 7ocationF
GReg2latorH means the Gene *echnology +eg"latorF
G(eedH means whole @otton seed from the GMOs or %ollen *rap plants/ incl"ding seed cotton/ f"<<y seed and .lac( seedF
G(ign3offH means a notice in writing from the +eg"lator/ in respect of a place/ that inspection conditions no longer apply in respect of that placeF
G%ol2nteer lantsH means progeny of the GMOs or %ollen *rap plants and regrowth of @otton plantsF
A--endi2 )
7i/en/e /#nditi#ns
*)
($+TI&' 2
G$'$RA) +&'DITI&'(
D2ration of )icence
!+ *his licence remains in force "ntil it is s"spended/ cancelled or s"rrendered- No dealings with GMOs are a"thorised d"ring any period of s"spension-
/older of )icence
2+ *he holder of this licence #>the licence holder4$ is Dow AgroSciences A"stralia %ty 7td.
ProIect (2 erAisor
3+ *he %roAect S"pervisor in respect of this licence is identified at Attachment A4+ *he licence holder m"st immediately notify the +eg"lator in writing if any of the contact details of the %roAect S"pervisor change-
'o dealings 4it1 GM&s exce t as a2t1orised 80 t1is licence

&+ %ersons covered .y this licence m"st not deal with the GMOs e2cept as e2pressly permitted .y this licence-
GM&s coAered 80 t1is licence

'+ *he GMOs covered .y this licence are descri.ed at Attachment '
Permitted dealings
)+ *he permitted dealings with the GMOs are to plant and grow the GMOs and to cond"ct e2periments on the GMOs that are grown- *he permitted dealings incl"des the possession/ storage/ s"pply/ "se/ transport and disposal of the GMOs for the p"rpose of any of the permitted dealings with the GMOs/ or in the co"rse of any of these dealings-
Persons coAered 80 t1is GM& licence

8+ *he persons covered .y this licence are the licence holder and employees/ agents or contractors of the licence holder and other persons who are/ or have .een/ engaged to "nderta(e any activity in connection with GMOs or places that are referred to in this licence-
Informing eo le of t1eir o8ligations

*+ *he licence holder m"st inform any person covered .y this licence/ to whom a partic"lar condition of this licence applies/ of the following: #a$ the partic"lar condition #incl"ding any variations of it$F #.$ the cancellation or s"spension of the licenceF #c$ the s"rrender of the licence-
A--endi2 )
7i/en/e /#nditi#ns
*8
!0+ *he licence holder m"st provide the +eg"lator/ on the +eg"lator4s written re,"est/ signed statements from persons covered .y this licence that the licence holder has informed those people of the conditions of this licence that apply to them-
)icence 1older to notif0 of circ2mstances t1at mig1t affect s2ita8ilit0

!!+ *he licence holder m"st immediately/ .y notice in writing/ inform the +eg"lator of: #a$ any relevant conviction of the licence holder occ"rring after the commencement of this licenceF #.$ any revocation or s"spension of a licence or permit held .y the licence holder "nder a law of the A"stralian Government/ a State or a foreign co"ntry/ .eing a law relating to the health and safety of people or the environmentF #c$ any event or circ"mstances occ"rring after the commencement of this licence that wo"ld affect the capacity of the holder of his licence to meet the conditions in it-
Additional information to 8e giAen to t1e Reg2lator

!2+ 5t is a condition of a licence that the licence holder inform the +eg"lator if the licence holder: #a$ .ecomes aware of additional information as to any ris(s to the health and safety of people/ or to the environment/ associated with the dealings a"thorised .y the licenceF or #.$ .ecomes aware of any contraventions of the licence .y a person covered .y the licenceF or #c$ .ecomes aware of any "nintended effects of the dealings a"thorised .y the licence-
Peo le dealing 4it1 GM&s m2st allo4 a2diting and monitoring of t1e dealing
!3+ 5f a person is a"thorised .y this licence to deal with GMOs and a partic"lar condition of this licence applies to the dealing .y that person/ the person m"st allow the +eg"lator/ or a person a"thorised .y the +eg"lator/ to enter premises where the dealing is .eing "nderta(en/ for the p"rposes of a"diting or monitoring the dealing-
Remaining an accredited organisation

!4+ *he licence holder m"st/ at all times/ remain an accredited organisation in accordance with the Act and comply with its instr"ment of accreditation-
A--endi2 )
7i/en/e /#nditi#ns
**
($+TI&' 3
(P$+I*I+ +&'DITI&'(
!+ *he permitted dealings with the GMOs/ other than disposal of the GMOs/ may only .e "nderta(en .etween May 2004 and M"ly 20082+ *he GMOs m"st only .e grown in accordance with the restrictions set o"t in Attachment @- Attachment @ sets o"t/ for each growing season #in which the GMOs may .e grown$: 8a9 those shires where the GMOs may .e grownF 8"9 the ma2im"m n"m.er of 7ocationsF and 8/9 the ma2im"m com.ined area of all 7ocations3+ *he licence holder m"st .e a.le to access and control a 7ocation where the GMOs are grown to the e2tent necessary to comply with this licence/ for the d"ration of the life of the licence-
'otification of lanting of t1e GM&s

4+ *he licence holder m"st provide a notice in writing to the +eg"lator each time a crop of the GMOs are planted at a 7ocation&+ *he notice m"st set o"t: #a$ *he date on which planting of the GMOs commencedF #.$ *he 7ocation4s G%S coordinates and either a street address or other directions to the 7ocationF #c$ *he period d"ring which the licence holder considers the GMOs are li(ely to flowerF and #d$ *he period d"ring which the licence holder considers the 7ocation will .e @leaned #after the GMOs have .een grown there$'+ *he notice m"st .e provided to the +eg"lator within 14 days of the date on which planting of the GMOs commenced-
'otice of +leaning of )ocation

)+ *he licence holder m"st provide a notice in writing to the +eg"lator when a 7ocation is @leaned p"rs"ant to this licence8+ *he notice m"st .e provided to the +eg"lator within 14 days of the date on which @leaning the 7ocation concl"ded-
)ocation m2st 8e s2rro2nded 80 a Pollen Tra or an Isolation Bone

*+ Hvery 7ocation where a crop of the GMOs is grown m"st .e s"rro"nded .y either: 8a9 a %ollen *rapF or 8"9 an 5solation one-
A--endi2 )
7i/en/e /#nditi#ns
!00
Isolation Bones ma0 onl0 8e 2sed in limited sit2ations

!0+ A 7ocation may only .e s"rro"nded .y an 5solation one if: 8a9 *here are no nat"ralised cotton pop"lations #Gossypiu" hirsutu" or G. 'ar'adense$ within 400 m of the 7ocationF and 8"9 *here are no cotton crops #Gossypiu" hirsutu" or G. 'ar'adense$ within 400 m of the 7ocation #other than crops of the GMOs grown p"rs"ant to this licence$-
+onditions a8o2t Pollen Tra s

!!+ A %ollen *rap m"st contain nonBgenetically modified @otton that is grown in s"ch a way as to reasona.ly promote a dense and vigoro"s growth and flowering of the nonB genetically modified @otton at the same time as the GMOs!2+ *he edge of the %ollen *rap that is farthest from the GMOs m"st not .e within 00 metres of a Nat"ral 9aterway!3+ Once planted/ %ollen *rap plants and %lant Material from those plants m"st .e handled and controlled as if they are GMOs and %lant Material from the GMOs- #ie-/ once planted/ %ollen *rap plants and %lant Material from those plants are GMOs for the p"rposes of this licence and s".Aect to other applica.le conditions elsewhere in this licence$!4+ A %ollen *rap m"st .e a.le to .e accessed and controlled .y the licence holder to an e2tent that is commens"rate with the licence holder4s rights to access and control the 7ocation within it-
+onditions a8o2t Isolation Bones

!&+ No cotton of any (ind may .e grown in an 5solation one!'+ No flowering cotton plants may .e present in an 5solation one while the GMOs are .eing grown at the 7ocation and any cotton plant that is fo"nd to .e present in an 5solation one m"st .e immediately Destroyed!)+ 5f any cotton crop not grown p"rs"ant to this licence occ"rs within 400 m of a 7ocation with an 5solation one while the GMOs are .eing grown at the 7ocation/ either the cotton crop or the GMOs in the 7ocation m"st .e Destroyed prior to flowering- 5f GMOs are Destroyed p"rs"ant to this condition/ they are ta(en to have .een harvested for the p"rposes of this licence
)ocations of more t1an 9 1ectare in siFe m2st 8e s2rro2nded 80 a Researc1 Bone

!8+ Hach 7ocation of 1 hectare or more in si<e m"st .e s"rro"nded .y a +esearch one "nless the licence holder has a notice in writing from the +eg"lator that a +esearch one in connection with the 7ocation is not re,"ired!*+ A +esearch one m"st .e a.le to .e accessed and controlled .y the licence holder to the e2tent necessary to ena.le the licence holder to meet its o.ligations "nder this licence to cond"ct research in the +esearch oneA--endi2 ) 7i/en/e /#nditi#ns
!0!
(eed and ot1er Plant Material ma0 8e collected

20+ 7eaf tiss"e from the GMOs may .e collected from a 7ocation for the p"rpose of cond"cting e2periments on it2!+ 7eaf tiss"e from the GMOs that is collected may only .e transported off the 7ocation to: 8a9 a loc(ed facility on the same property as the 7ocation that is signed so as to indicate GM %lant Material is stored within the facilityF or 8"9 a facility certified .y the +eg"lator to physical containment level 2 #%@2$22+ 7eaf tiss"e that is collected from the 7ocation may .e stored in a loc(ed facility on the same property as the 7ocation that is signed so as to indicate GM %lant Material is stored within the facility- 7eaf tiss"e stored in the facility m"st .e stored in a sealed container23+ After any e2periments with leaf tiss"e from the GMOs are completed/ the leaf tiss"e m"st .e incinerated-
+ro s of t1e GM&s m2st 8e eit1er 1arAested or Destro0ed

24+ 9ithin C months of .eing planted/ crops of the GMOs m"st .e either harvested or Destroyed2&+ 5f the GMOs are harvested/ they m"st .e harvested separately from any other @otton other than %ollen *rap plants-
+onditions in relation to t1e +leaning of )ocations and Pollen Tra s after eac1 cro of GM&s is gro4n
2'+ After the GMOs are harvested or Destroyed at a 7ocation/ the 7ocation and the %ollen *rap aro"nd it #if any$ m"st .e @leaned2)+ A 7ocation m"st .e @leaned within 14 days of harvest or Destr"ction of the GMOs in it/ whichever occ"rs first28+ A %ollen *rap m"st .e @leaned within 14 days of the @leaning of the 7ocation within it-
/arAested cro s of t1e GM&s ma0 8e ginned

2*+ Seed harvested from the GMOs and %ollen *rap plants may .e ginned- 5f it is ginned/ it m"st .e ginned separately from any other @otton- 5f it is not ginned it m"st .e Destroyed30+ !ollowing ginning/ Seed m"st .e: 8a9 stored in a sealed container/ within a loc(ed facility that is signed so as to indicate that GM Material is stored within the facilityF 8"9 e2portedF 8/9 Destroy it .y ."rningF or 8d9 transported to a facility certified .y the +eg"lator to physical containment level 2 #%@2$-
A--endi2 )
7i/en/e /#nditi#ns
!02
General conditions in relation to t1e +leaning of all ot1er laces and $72i ment 2sed in connection 4it1 t1is licence
3!+ 5f: 8a9 an area or place other than a 7ocation or %ollen *rap is "sed in connection with this licenceF or 8"9 H,"ipment is "sed in connection with the GMOs/ %ollen *rap plants or %lant MaterialF then that area/ place or H,"ipment m"st also .e @leaned32+ @leaning m"st occ"r immediately or as soon as practica.le after the "se and .efore it is "sed for any other p"rpose33+ 5f H,"ipment is @leaned/ the area in which the H,"ipment is @leaned m"st also .e @leaned- #5t is not necessary for H,"ipment to .e @leaned only at a 7ocation-$ 34+ On the re,"est of the +eg"lator/ the +eg"lator m"st .e provided with written doc"mentation of the proced"res in place to ens"re contin"ing compliance with these @leaning conditions-
General conditions t1at a l0 41ereAer ins ections m2st 8e 2ndertaken for t1e existence of %ol2nteer lants
3&+ After a 7ocation is @leaned/ the following places m"st .e inspected for the e2istence of &ol"nteer plants: 8a9 the 7ocationF 8"9 the %ollen *rap #if any$F 8/9 the 5solation one #if any$F 8d9 irrigation channels and drains thro"gh which water flows to and from the 7ocation and the %ollen *rapF 8e9 any areas "sed to @lean H,"ipment3'+ 5nspection m"st .e performed .y a person who is a.le to recognise &ol"nteer plants3)+ *he res"lts of inspection activities m"st .e recorded in a log.oo(- *he log.oo( m"st .e availa.le on re,"est for e2amination or photocopying .y the OG*+- *he findings of the inspections as recorded in the log.oo( m"st .e incl"ded in the licence holder4s ann"al report to the +eg"lator- *he log.oo( m"st contain at least the following: #a$ details of the areas inspectedF #.$ details of the date of inspectionF #c$ the names of the person or persons who "ndertoo( the inspection and details of the e2perience/ training or ,"alification that ena.led them to recognise &ol"nteer plantsF #d$ the n"m.er of &ol"nteer plants o.served/ if anyF #e$ details of the development stages reached .y the &ol"nteer plants/ if anyF and
A--endi2 )
7i/en/e /#nditi#ns
!03
#f$ details of methods "sed to Destroy &ol"nteer plants/ if any. 38+ Any &ol"nteer plant identified m"st .e Destroyed prior to the plant flowering3*+ 3nless this licence provides otherwise/ a place m"st .e inspected at least once every 80 days/ either "ntil the GMOs are once again planted at the 7ocation p"rs"ant to this licence/ or "ntil the +eg"lator has iss"ed a signBoff40+ 5f: 8a9 inspections have .een ro"tinely completed in a place for a period of a yearF and 8"9 inspection records for that place show that no &ol"nteers have .een o.served in the most recent 8 month inspection periodF the licence holder may ma(e written application to the +eg"lator that these inspection conditions no longer apply in respect of that place4!+ 5nspection conditions do not apply in respect of a place if the +eg"lator has iss"ed a signB off in respect of that place-
Restrictions in relation to areas and lants after t1e GM&s are gro4n
42+ !rom the time a 7ocation has .een harvested "ntil s"ch time as the licence holder has received a signBoff/ no @otton of any (ind may .e grown in the 7ocation or its %ollen *rap "nless it is grown p"rs"ant to this licence43+ !rom the time a 7ocation has .een harvested "ntil s"ch time as the licence holder has received a signBoff/ no plants may .e planted at the 7ocation or its %ollen *rap e2cept: 8a9 GM @otton grown p"rs"ant to this licenceF 8"9 grasses #grass past"res$F 8/9 cereals #cereal crops$F 8d9 plants agreed to in writing .y the +eg"lator-
Trans ortation of t1e GM&s= Pollen Tra
lants and Plant Material
44+ S".Aect to the condition immediately .elow in respect of transportation/ the GMOs/ %ollen *rap plants and %lant Material m"st .e transported in accordance with the OG*+ G"idelines for the *ransport of GMOs #M"ne 2001$ iss"ed .y the +eg"lator4&+ Hvery container "sed to transport the GMOs/ %ollen *rap plants and %lant Material m"st .e la.elled: #a$ to indicate that it contains GM @ottonF and #.$ with telephone contact n"m.ers for the licence holder and instr"ctions to contact the licence holder in the event that the container is .ro(en or misdirected4'+ ;arvested GMOs/ %ollen *rap plants and %lant Material my .e transported to a ginning facility in a @otton mod"le that is: 8a9 completely enclosed within 2 layers or tarpa"lin #>do".le wrapped in tarpa"lin4$F
A--endi2 )
7i/en/e /#nditi#ns
!04
8"9 completely enclosed within a layer of tarpa"lin inside a layer of shade cloth #>do".le wrapped in tarpa"lin and shade cloth4$F or 8/9 contained within a @otton mod"le in an enclosed chainB.ed tr"c( specifically designed for the p"rposes of transporting @otton mod"les4)+ Ginned seed from the GMOs or %ollen *rap plants may only .e transported to the e2tent necessary to store it/ e2port it/ Destroy it .y ."rning it or relocate it to a facility certified .y the +eg"lator to physical containment level 2 #%@2$48+ *he licence holder m"st have in place acco"nting proced"res to verify whether the same ,"antity of GMOs/ %ollen *rap plants and %lant that is sent is delivered- +o"tes/ methods and proced"res "sed for transportation in accordance with this licence m"st .e doc"mented-
+ontingenc0 Plans
4*+ 9ithin 60 days of the date of the commencement of this licence/ a written @ontingency %lan m"st .e s".mitted to the +eg"lator detailing meas"res to .e ta(en in the event of the "nintended presence of the GMOs/ %ollen *rap plants or %lant Material/ o"tside an area that m"st .e inspected&0+ *he @ontingency %lan m"st incl"de details of proced"res to: #a$ ens"re the +eg"lator is notified immediately if the licence holder .ecomes aware of the eventF #.$ destroy any of the GMOs/ %ollen *rap plants and %lantF and #c$ inspect and Destroy any &ol"nteer plants that may e2ist as a res"lt of the event&!+ *he @ontingency %lan m"st .e implemented in the event that the "nintended presence of the GMOs/ %ollen *rap plants or %lant Material is discovered o"tside an area that m"st .e inspected-
+om liance Management Plan

&2+ %rior to growing the GMOs/ a written @ompliance Management %lan m"st .e provided to the +eg"lator- *he @ompliance Management %lan m"st descri.e in detail how the licence holder intends to ens"re compliance with these conditions and doc"ment that compliance-
Re orting
&3+ *he licence holder m"st provide the +eg"lator with a written report within C0 days of each anniversary of this licence/ in accordance with any G"idelines iss"ed .y the +eg"lator in relation to ann"al reporting- *his report m"st incl"de information on any adverse impacts on h"man health and safety or the environment/ ca"sed as a res"lt of the GMOs/ %ollen *rap plants or %lant Material-
A--endi2 )
7i/en/e /#nditi#ns
!0&
Researc1 re72irements
&4+ *he licence holder m"st/ in cons"ltation with the OG*+/ develop an agreed research program to collect information regarding: #a$ the levels of e2pression of the insecticidal and her.icide tolerance genes in GM @otton tiss"es "nder A"stralian field conditionsF #.$ the effect of GM @otton on nonBtarget organisms "nder A"stralian field conditionsF #c$ the potential for the introd"ced proteins to acc"m"late in the soil "nder A"stralian field conditionsF and #d$ the efficacy of gene flow containment meas"res for any release sites in e2cess of one hectare/ within the associated +esearch one/ and for any release site for which a pollen trap is not re,"ired/ d"e to having no other cotton crops or pop"lations within 400 m-
Testing met1odolog0
&&+ *he licence holder m"st provide a written instr"ment to the +eg"lator descri.ing an e2perimental method that is capa.le of relia.ly detecting the presence of the GMOs and the presence of the genetic modifications descri.ed in this licence #at Attachment '$ in a recipient organism- *he instr"ment m"st .e provided within 60 days of planting the GMOs-
GM&s= Pollen Tra
lants and Plant Material m2st not 8e cons2med
&'+ *he licence holder m"st ens"re that the GMOs/ %ollen *rap plants and prod"cts derived from these plants are not cons"med .y h"mans or "sed as stoc(feed-
A--endi2 )
7i/en/e /#nditi#ns
!0'
APP$'DI, D )$GI()ATI%$ R$>-IR$M$'T( *&R A(($((I'G D$A)I'G( I'%&)%I'G I'T$'TI&'A) R$)$A($(

($+TI&' 9 T/$ R$G-)ATI&' &* G$'$ T$+/'&)&G. I' A-(TRA)IA
30)+ The Gene Technology Act 2000 #the Act$ too( effect on 21 M"ne 2001- *he Act/ s"pported .y the Gene Technology Regulations 2001/ an interBgovernmental agreement and corresponding legislation that is .eing enacted in each State and *erritory/ "nderpins A"stralia4s nationally consistent reg"latory system for gene technology- 5ts o.Aective is to protect the health and safety of people/ and the environment/ .y identifying ris(s posed .y or as a res"lt of gene technology/ and managing those ris(s .y reg"lating certain dealings with genetically modified organisms #GMOs$- *he reg"latory system replaces the former vol"ntary system overseen .y the Genetic Manip"lation Advisory @ommittee #GMA@$308+ *he Act esta.lishes a stat"tory officer/ the Gene *echnology +eg"lator #the +eg"lator$/ to administer the legislation and ma(e decisions "nder the legislation30*+ *he +eg"lator is s"pported .y the Office of the Gene *echnology +eg"lator #OG*+$/ a A"stralian Government reg"latory agency located within the ;ealth and Ageing portfolio3!0+ *he Act prohi.its persons from dealing with GMOs "nless the dealing is e2empt/ a Notifia.le 7ow +is( Dealing/ on the +egister of GMOs/ or licensed .y the +eg"lator #see section 61 of the Act$3!!+ *he re,"irements "nder the legislation for cons"ltation and for considering and assessing licence applications and preparing ris( assessment and ris( management plans #+A+M%s$ are disc"ssed in detail in Division 4/ %art 0 of the Act and s"mmarised .elow3!2+ Detailed information a.o"t the national reg"latory system and the gene technology legislation is also availa.le from the OG*+ we.site #www.ogtr.gov.au$-
($+TI&' 2 T/$ )I+$'+$ APP)I+ATI&'

3!3+ 7icence applications for dealings involving the intentional release #D5+$ of a genetically modified organism into the environment m"st .e s".mitted in accordance with the re,"irements of section 40 of the Act- As re,"ired .y Sched"le 4/ %art 2 of the +eg"lations/ the application m"st incl"de information a.o"t: !3)( the parent organismF !38( the GMOsF !3*( the proposed dealing with the GMOsF !40( interaction .etween the GMOs and the environmentF !4!( ris(s the GMOs may pose to the health and safety of peopleF !42( ris( managementF
A--endi2 8
7egislati6e .e:0i.ements $#. assessing dealings in6#l6ing intenti#nal .eleases
!0)
!43( previo"s assessments of approvalsF and !44( the s"ita.ility of the applicant3!4+ *he application m"st also contain:
!4&( additional information re,"ired for a GMO that is: 0( a plantF !( 2( 3( 4( &( '( )( 8( *( a microBorganism #not living in or on animals and not a live vaccine$F a microBorganism that lives in or on animalsF a live vaccine for "se in animalsF a verte.rate animalF an a,"atic organismF an inverte.rate animalF to .e "sed for .iological controlF to .e "sed for .ioremediationF and intended to .e "sed as food for h"man or verte.rate animal cons"mptionF
!4'( s"pporting information from the 5nstit"tional 'iosafety @ommittee3!&+ A preliminary screening of an application is "nderta(en .y OG*+ staff to determine whether it complies with the Act and the +eg"lations/ .y containing the re,"ired information- 5f this information is provided in the application/ the +eg"lator may then accept the application for formal consideration- Section 46 of the Act provides that the +eg"lator is not re,"ired to consider an application if the application does not contain the re,"ired information3!'+ After accepting an application for consideration/ the +eg"lator m"st decide to iss"e/ or ref"se to iss"e/ a licence- *he decision m"st .e ta(en following an e2tensive cons"ltation and eval"ation process/ as detailed in sections 6B8 of this Appendi2+eg"lation = of the +eg"lations prescri.e a period of 1I0 wor(ing days within which this decision m"st .e ta(en- *his period does not incl"de wee(ends or p".lic holidays in the A"stralian @apital *erritory- Also/ this period does not incl"de any days in which the +eg"lator is "na.le to progress the application .eca"se information so"ght from the applicant in relation to the application has not .een received-
($+TI&' 3 T/$ I'ITIA) +&'(-)TATI&' PR&+$(($(

3!)+ 5n accordance with section 00 of the Act/ the +eg"lator m"st see( advice in preparing a +A+M% from prescri.ed agencies: !4)( State and *erritory GovernmentsF !48( the Gene *echnology *echnical Advisory @ommittee #G**A@$F !4*( prescri.ed A"stralian Government agencies #+eg"lation C of the Gene Technology Regulations 2001 refers$F
A--endi2 8
!08
!&0( the A"stralian Government Minister for Hnvironment and ;eritageF and !&!( relevant local co"ncil#s$ where the release is proposed3!8+ Section 4C of the Act re,"ires that if the +eg"lator is satisfied that at least one of the dealings proposed to .e a"thorised .y the licence may pose significant ris(s to the health and safety of people or to the environment/ the +eg"lator m"st p".lish a notice #in national and regional news papers/ in the Gazette and on the OG*+ we.site$ in respect of the application/ inviting written s".missions on whether the licence sho"ld .e iss"ed3!*+ As a meas"re over and a.ove those re,"ired "nder the Act/ in order to promote the openness and transparency of the reg"latory system/ the +eg"lator may ta(e other steps- !or e2ample/ receipt of applications is notified to the p".lic .y posting a notice of each application4s receipt on the OG*+ we.site and directly advising those on the OG*+ mailing list- @opies of applications are availa.le on re,"est from the OG*+-
($+TI&' 4 T/$ $%A)-ATI&' PR&+$(($(

320+ *he ris( assessment process is carried o"t in accordance with the Act and Regulations/ "sing the +is( Analysis !ramewor( #the !ramewor($ developed .y the +eg"lator #availa.le on the OG*+ we.site$- 5t also ta(es into acco"nt the g"idelines and ris( assessment strategies "sed .y related agencies .oth in A"stralia and overseas*he !ramewor( was developed in cons"ltation with the States and *erritories/ A"stralian Government agencies/ G**A@ and the p".lic- 5ts p"rpose is to provide general g"idance to applicants and eval"ators and other sta(eholders in identifying and assessing the ris(s posed .y GMOs and in determining the meas"res necessary to manage any s"ch ris(s32!+ 5n "nderta(ing a ris( assessment/ the following are considered and analysed:
!&2( the data presented in the proponent4s applicationF !&3( data provided previo"sly to GMA@/ the interim OG*+ or the OG*+ in respect of previo"s releases of relevant GMOsF !&4( s".missions or advice from States and *erritories/ A"stralian Government agencies and the A"stralian Government Minister for Hnvironment and ;eritage and the p".licF !&&( advice from G**A@F !&'( information from other national reg"latory agenciesF and !&)( c"rrent scientific (nowledge and the scientific literat"re322+ 5n considering this information and preparing the +A+M%/ the following specific matters are ta(en into acco"nt/ as set o"t in section 4C and re,"ired .y section 01 of the Act: !&8( the ris(s posed to h"man health and safety or to the environmentF
A--endi2 8
!0*
!&*( the properties of the organism to which the dealings relate .efore it .ecame a GMOF !'0( the effect/ or the e2pected effect/ of the genetic modification that has occ"rred on the properties of the organismF !'!( provisions for limiting the dissemination or persistence of the GMO or its genetic material in the environmentF !'2( the potential for spread or persistence of the GMO or its genetic material in the environmentF !'3( the e2tent or scale of the proposed dealingsF !'4( any li(ely impacts of the proposed dealings on the health and safety of people323+ 5n accordance with +eg"lation 10 of the +eg"lations/ the following are also ta(en into acco"nt: !'&( any previo"s assessment/ in A"stralia or overseas/ in relation to allowing or approving dealings with the GMOF !''( the potential of the GMO concerned to: !0( .e harmf"l to other organismsF !!( adversely affect any ecosystemsF !2( transfer genetic material to another organismF !3( spread/ or persist/ in the environmentF !4( have/ in comparison to related organisms/ a selective advantage in the environmentF and !&( .e to2ic/ allergenic or pathogenic to other organisms!')( the short and long term when ta(ing these factors into acco"nt-
($+TI&' : *-RT/$R +&'(-)TATI&'

324+ ;aving prepared a ris( assessment and a ris( management plan/ the +eg"lator m"st/ "nder section 02 of the Act/ see( comment from sta(eholders/ incl"ding those o"tlined in section 6 and the p".lic32&+ All iss"es relating to the protection of h"man health and safety and the environment raised in written s".missions on an application or a ris( assessment and a ris( management plan are considered caref"lly/ and weighed against the .ody of c"rrent scientific information/ in reaching the concl"sions set o"t in a final +A+M%Section 08 of the Act re,"ires that these .e ta(en into acco"nt in ma(ing a decision on whether or not to iss"e a licence for the proposed release32'+ @omments received in written s".missions on this +A+M% are very important in shaping the final +A+M% and in informing the +eg"lator4s decision on an application- A s"mmary of p".lic s".missions and an indication of where s"ch iss"es have .een ta(en into acco"nt are provided in an Appendi2 to the final +A+M%!!0
A--endi2 8
32)+ 5t is important to note that the legislation re,"ires the +eg"lator to .ase the licence decision on whether ris(s posed .y the dealings are a.le to .e managed so as to rotect 12man 1ealt1 and safet0 and t1e enAironment- Matters in s".missions that do not address these iss"es and)or concern .roader iss"es o"tside the o.Aective of the legislation will not .e considered in the assessment process- 5n most instances/ as determined in the e2tensive cons"ltation process that led to the development of the legislation/ they fall within the responsi.ilities of other a"thorities-
($+TI&' ; D$+I(I&' &' )I+$'+$

328+ ;aving ta(en the re,"ired steps for assessment of a licence application/ the +eg"lator m"st decide whether to iss"e or ref"se a licence #section 00 of the Act$*he +eg"lator m"st not iss"e the licence "nless satisfied that any ris(s posed .y the dealings proposed to .e a"thorised .y the licence are a.le to .e managed in s"ch a way as to protect the health and safety of people and the environment32*+ *he +eg"lator m"st also .e satisfied/ "nder section 0I of the Act/ that the applicant is a s"ita.le person to hold the licence- Section 0= o"tlines matters the +eg"lator m"st consider in deciding whether a person or company is s"ita.le to hold a licence eg-: !'8( any relevant convictionsF !'*( any relevant revocations or s"spensions of a licences or permitsF and !)0( the capacity of the person or company to meet the conditions of the licence330+ *he +eg"lator caref"lly considers all of this information which is s"pplied in a declaration signed .y licence applicants33!+ *he Monitoring and @ompliance Section of the OG*+ compiles compliance histories of applicants/ considering all previo"s approvals to deal with GMOs "nder the Act and the previo"s vol"ntary system- *hese histories as well as other information s"ch as followB"p actions from a"dits may .e ta(en into acco"nt- *he a.ility of an organisation to provide reso"rces to ade,"ately meet monitoring and compliance re,"irements may also .e ta(en into acco"nt332+ 5f a licence is iss"ed/ the +eg"lator may impose licence conditions #section 82 of the Act$- !or e2ample/ conditions may .e imposed to: !)!( limit the scope of the dealingsF !)2( re,"ire doc"mentation and recordB(eepingF !)3( re,"ire a level of containmentF !)4( specify waste disposal methodsF !)&( manage ris(s posed to the health and safety of people/ or to the environmentF !)'( re,"ire data collection/ incl"ding st"dies to .e cond"ctedF !))( limit the geographic area in which the dealings may occ"rF !)8( re,"ire contingency planning in respect of "nintended effects of the dealingsF and
A--endi2 8 7egislati6e .e:0i.ements $#. assessing dealings in6#l6ing intenti#nal .eleases
!!!
!)*( limit the dissemination or persistence of the GMO or its genetic material in the environment333+ 5t is also re,"ired as a condition of a licence that the licence holder inform any person covered .y the licence of any condition of the licence which applies to them #section 86 of the Act$- Access to the site of a dealing m"st also .e provided to persons a"thorised .y the +eg"lator for the p"rpose of a"diting and monitoring the dealing and compliance with other licence conditions #section 84 of the Act$- 5t is a condition of any licence that the licence holder inform the +eg"lator of: !80( any new information as to any ris(s to the health and safety of people/ or to the environment/ associated with the dealings a"thorised .y the licenceF !8!( any contraventions of the licence .y a person covered .y the licenceF and !82( any "nintended effects of the dealings a"thorised .y the licence334+ 5t sho"ld .e noted that/ as well as imposing licence conditions/ the +eg"lator has additional options for ris( management- *he +eg"lator has the legislative capacity to enforce compliance with licence conditions/ and indeed/ to direct a licence holder to ta(e any steps the +eg"lator deems necessary to protect the health and safety of people or the environment- *he OG*+ also independently monitors trial sites to determine whether the licence holder is complying with the licence conditions/ or whether there are any "nforseen pro.lems-
A--endi2 8
!!2
APP$'DI, E (-MMAR. &* P-#)I+ (-#MI((I&'( &' T/$ RI(5 A(($((M$'T A'D RI(5 MA'AG$M$'T P)A'
(28mission from: A: agric"lt"ral)ind"stry organisationF I: individ"al Iss2es raised/consideration: A : appendi2F AP%MA: iss"es dealt with .y A%&MAF +1: chapterF $': environmental ris(sF *(: feed safetyF /: h"man health and safetyF /R: her.icide resistanceF IR: insecticide resistanceF 6: weediness-
(28< 'o< 1
T0 e
(2mmar0 of iss2es raised
Iss2e
+onsideration of iss2e
Hffects of cotton wastes Qfrom GM cottonR in animal feeds have not .een tested e2perimentally- *heir effects on health of po"ltry and dairy animals have not .een tested*he effects of h"man cons"mption of animals or prod"cts from animals fed QGMR cotton resid"es has not .een tested/ or at least no p".lished evidence is availa.le*he effects of QGMR cotton on soil microorganisms have not .een scientifically e2amined- THval"ationU is no s".stit"te for e2perimentations5 .elieve this application is for commercial Qp"rposeR and not QforR environmental or health testing5 do not .elieve eno"gh testing for health and environmental effects have .een done to warrant proceeding with testing for commercial via.ility-
!S
@h 2/ App 6/ I
App 2/ 6
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App 6
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@h 1/ App 2BI
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App 2B8
*he introd"ction of genetically modified insecticidal cotton of this type #5NGA+DE and 'ollgardE 55$ has shown a dramatic red"ction in their pesticide "se re,"irements/ compared to conventional cottonStac(ing Qcom.iningR of different genes in
A%&MA
OSA
A%&MA/
App 8
A--endi2 *
0mma.1 #$ -0"li/ s0"missi#ns
!!3
commercial crops is seen as one of the .est meas"res to improve efficacy of the plants as well as to red"ce the ris( of resistance developing to any one to2inAfter 40 years of cotton growing and processing thro"gho"t the northwestern regions of New So"th 9ales cotton has not .een a.le to esta.lish itself as a weed*he li(elihood of the trial cotton containing the cry1Fa gene or cry1Ac gene and mar(er gene conferring tolerance to gl"fosinate ammoni"m her.icide .ecoming a weed is a.sol"tely minimalstrongly s"pports the proposed field trial of the GM cottons-
5+
App 4
App 4
None
Noted
R$*$R$'+$(
Ama.ileB@"evas/ @-!-/ @hic"rel/ M- #1CC6$- ;ori<ontal gene transfer- A"erican %cientist D9: 662B641AN !A #1CCC$- !"ll assessment report and reg"latory impact statement/ A641: Oil and linters derived from insect resistant cotton- A641/ A"stralia New ealand !ood A"thority/ @an.erra/ A"stralia- pp 1B0CAN !A #2001a$- !inal assessment report- Application A6I2: Oil derived from gl"fosinateB ammoni"m tolerant canola lines *opas 1C)2 and *40 AND Oil derived from gl"fosinateB ammoni"m tolerant and pollination controlled canola lines MS1/ MS=/ +!1/ +!2 and +!600)02/ pp 1B==AN !A #2001.$- !inal assessment report- Application A6I2: Oil derived from gl"fosinateB ammoni"m tolerant canola lines *opas 1C)2 and *40 AND Oil derived from gl"fosinateB ammoni"m tolerant and pollination controlled canola lines MS1/ MS=/ +!1/ +!2 and +!600)02/ pp 1B==AN !A #2001c$- !inal assessment report- Application A6=0: !ood from insectBprotected and gl"fosinate ammoni"mBtolerant D'*41= corn- A"stralia New ealand !ood A"thority/ AN !A #2002$- Draft assessment report #!"ll assessment B S-10$ Application A468: Oil and linters derived from insectBprotected cotton containing event 10C=0- !"ll assessment B S-10 Application A468/ A"stralia New ealand !ood A"thority/ @an.erra/ A"stralia- pp 1BI=-
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!!4
Ao(i/ S-/ Syono/ J- #1CCC$- ;ori<ontal gene transfer and m"tation: Ngrol genes in the genome of :icotiana glauca- 1roceedings o( the :ational Acade"y o( %ciences o( the >nited %tates o( A"erica E;: 1622CB16264Astwood/ M-D-/ 7each/ M-N-/ !"chs/ +-7- #1CC8$- Sta.ility of food allergens to digestion in vitro- :ature Biotechnology 94: 128CB12I6A"stralian @otton @ooperative +esearch @entre #2002$- 9HHDpa( B A g"ide for integrated management of weeds in cotton- A"stralian @otton @ooperative +esearch @entre/ Narra.ri/ NS9'ar.era/ %-9- #1CC0$- *o2icity)pathogenecity testing of Bacillus thuringiensis strain HG I=28 following ac"te oral challenge in rats- 55*+5 %roAect N"m.er 70=0I4/ 55* +esearch 5nstit"te/ @hicago 57'ar(er/ +-!-/ 5dler/ J-'-/ *hompson/ D-&-/ Jemp/ M-D- #1C=6$- N"cleotide se,"ence of the *B DNA region from the Agro'acteriu" tu"e(aciens octopine *i plasmid p*i10C00- 1lant .olecular Biology 2: 660B600'eever/ D-H-/ Jemp/ @-!- #2000$- Safety iss"es associated with the DNA in animal feed derived from genetically modified crops- A review of scientific and reg"latory proced"res:utrition A'stracts and Reviews9 %eries B) /ivestock Feeds and Feeding C0: 1I0B1=2'er.erich/ S-A-/ +eam/ M-H-/ Mac(son/ *-7- #1CC8$- Safety assessment of insectBprotected cotton: the composition of the cottonseed is e,"ivalent to conventional cottonseed- ?ournal o( Agricultural and Food !he"istry 49: 680B6I1'ergelson/ M-/ %"rrington/ @-'-/ 9ichmann/ G- #1CC=$- %romisc"ity in transgenic plants:ature 3E:: 20'ernstein/ 5-7-/ 'ernstein/ M-A-/ Miller/ M-/ *ier<ieva/ S-/ 'ernstein/ D-5-/ 7"mm"s/ -/ Selgrade/ M-J-/ Doerfler/ D-7-/ Seligy/ &-7- #1CCC$- 5mm"ne responses in !arm wor(ers after e2pos"re to Bacillus thuringiensis pesticides- 7nviron"ental ,ealth 1erspectives 90C: 0I0B 0=2'ertolla/ !-/ Simonet/ %- #1CCC$- ;ori<ontal gene transfers in the environment: nat"ral transformation as a p"tative process for gene transfers .etween transgenic plants and microorganisms- Research in .icro'iology 9:0 #8$: 6I0B6=4'et</ !-S-/ ;ammond/ '-G-/ !"chs/ +-7- #2000$- Safety and advantages of Bacillus thuringiensisBprotected plants to control insect pests- Regulatory To4icology and 1har"acology 32: 108B1I6'evan/ M- #1C=4$- 'inary Agro'acteriu" vectors for plant transformation- :ucleic Acids Research 92: =I11B=I21-
A--endi2 *
!!&
'ietlot/ ;-/ @arey/ %-+-/ Japlan/ ;-/ 7essard/ *-/ %o<sgay/ - #1C=C$- !acile preparation and characterisation of the to2in from Bacillus thuringiensis var- kurstaki- Bioche"istry 2;0: =IB C1'remmer/ M-N-/ 7eist/ J-;- #1CC8$- Statement on the lac( of allergenic potential of %A*B protein and gl"fosinate tolerant crops containing %A*Bprotein- 3np".lished- ;oechst Schering AgrHvo Gm.; 5nternal report C8-0601)A088C0/ 'rimecom.e/ M-M-/ De 7eiA/ !-A-/ 7ynch/ M-M- #2001$- *he effect of root e2"dates on rhi<osphere micro.ial pop"lations- -n X*he rhi<osphere: .iochemistry and organic s".stances at the soilBplant interfaceX/ + %inton/ &aranini/ % Nannipieri/ eds- Marcel De((er/ 5nc-/ New Nor(/ 3SA- pp C0B140'ryngelsson/ *-/ G"stafson/ M-/ Green/ '-/ 7ind/ @- #1C==$- 3pta(e of host DNA .y the parasitic f"ng"s 1las"odiophora 'rassicae- 1hysiological and .olecular 1lant 1athology 33: 186B1I1'"hariwalla/ ;-/ Mithen/ +- #1CC0$- @loning of a Brassica repetitive DNA element from resting spores of 1las"odiophora 'rassicae- 1hysiological and .olecular 1lant 1athology 4C: C0B101@anadian !ood 5nspection Agency #1CC0$- Decision Doc"ment DDC0B04: Determination of environmental safety of %lant Genetic Systems 5nc- #%GS$ novel hy.ridi<ation system for canola #Brassica napus 7-$- pp 1B=http:))www-inspection-gc-ca)english)plaveg)p.o)dd)ddC004e-shtml @anadian !ood 5nspection Agency #1CC8$- S"pplement to Decision Doc"ment DDC8B11 S"ppl: Determination of 7ivestoc( !eed Safety of AgrHvo @anada 5nc-Vs Gl"fosinate Ammoni"mB*olerant @anola 7ine ;@N2=- decision docu"ent DDE;399 (2 l< @anadian !ood 5nspection Agency #1CC=a$- Decision Doc"ment #DDC=B2=$: determination of the safety of AgrHvo @anada 5ncVs gl"fosinateBammoni"m her.icideBtolerant Brassica rapa canola line ;@+B1- http:))www-inspection-gc-ca)english)plaveg)p.o)dd)ddC=2=e-shtml @anadian !ood 5nspection Agency #1CC=.$- Decision Doc"ment C=B26: Determination of Hnvironmental Safety of De(al. Genetics @orporationVs H"ropean @orn 'orer #H@'$ +esistant @orn #&ea "ays 7-$ 7ine D'*41=@arter/ M-N-/ 7igget/ M-%- #1CC4$- Ac"te oral to2icity and infectivity)pathogenicity to rats of HG I=41- ;+@ St"dy +eport n"m.er H@O 8)C4206=/ ;"ntingdon +esearch @entre 7td-/ ;"ntington @am.ridgeshire Hngland@astillo/ A-+-/ Gallardo/ M-+-/ Maciel/ M-/ Giordano/ M-M-/ @onti/ G-A-/ Gaggiotti/ M-@-/ :"aino/ O-/ Gianni/ @-/ ;artnell/ G-!- #2001a$- Hffect of feeding dairy cows with cottonseeds containing 'ollgard#+$ and +o"nd"p +eady#+$ genes or control nonBtransgenic cottonseeds on feed inta(e/ mil( yield and mil( composition- 5nternational Animal Agric"lt"re and !ood
A--endi2 *
!!'
Science @onference- http:))www-asas-org)Aas)Aointa.s)Antmttoc-htm ?ournal o( 8airy %cience D4 #S"pplement 1$: 416@astillo/ A-+-/ Gallardo/ M-+-/ Maciel/ M-/ Giordano/ M-M-/ @onti/ G-A-/ Gaggiotti/ M-@-/ :"aino/ O-/ Gianni/ @-/ ;artnell/ G-!- #2001.$- Hffect of feeding dairy cows with either 'ollgardE/ 'ollgard 55E/ +o"nd"p+eadyE or control cottonseeds on feed inta(e/ mil( yield and mil( composition- http:))www-adsa-org)Aointa.s)Antmttoc-htm ?ournal o( 8airy %cience D4 #S"pplement 1$: 416@ham.ers/ M-A-/ Melen/ A-/ Gil.ert/ M-%-/ Many/ @-S-/ Mohnson/ *-'-/ GawronB'"r(e/ @- #1CC1$5solation and characteri<ation of a novel insecticidal crystal protein gene from 'acill"s th"ringiensis s".sp- ai<awai- ?ournal o( Bacteriology 9C3: 6C88B6CI8@homa/ @-*-/ Japlan/ ;- #1CC0$- !olding and "nfolding of the proto2in from Bacillus thuringiensis: evidence that the to2ic moiety is present in an active conformationBioche"istry 2E: 10CI1B10CII@howdh"ry/ H-;-/ Mi(ami/ O-/ M"rata/ ;-/ S"ltana/ %-/ Shimada/ N-/ Noshio(a/ M-/ G"r"ge/ J-S-/ Namamoto/ S-/ Miya<a(i/ S-/ Namana(a/ N-/ Na(aAima/ N- #2004$- !ate of mai<e intrinsic and recom.inant genes in calves fed genetically modified mai<e 't11- ?ournal o( Food 1rotection ;C: 680B6I0@hristensen/ A-;-/ Sharroc(/ +-A-/ :"ail/ %-;- #1CC2$- Mai<e poly".i,"itin genes: str"ct"re/ thermal pert"r.ation of e2pression and transcript splicing/ and promoter activity following transfer to protoplasts .y electroporation- 1lant .olecular Biology 9D: 8I0B8=C@oghlan/ A- #2000$- So far so good: for the moment/ the gene genie is staying in its .ottle:ew %cientist 2239: 4@onner/ A-M-/ Glare/ *-+-/ Nap/ M-%- #2006$- *he release of genetically modified crops into the environment- %art 55- Overview of ecological ris( assessment- 1lant ?ournal 33: 1CB48@raven/ 7- A-/ Stewart/ M- M-/ 'rown/ A- ;- D-/ and Grace/ M- %- #1CC4$- @hallenging the f"t"reF the A"stralian wild species of Gossypiu"- 5n X1roceedings o( the 1st @orld !otton Research !on(erenceA/ pp- 2I=B2=1@ric(more/ N-/ eigler/ D-+-/ Schnepf/ H-/ &an +ie/ M-/ 7erecl"s/ D-/ 'a"m/ M-/ 'ravo/ A-/ and Dean/ D-;- Bacillus thuringiensis to2in nomenclat"re- #2002$ Availa.le from: http:))www-.iols-s"s2-ac-"()home)NeilY@ric(more)'t) Davies/ M-H- #1C=8$- AminoglycosideBaminocyclitol anti.iotics and their modifying en<ymes-n XAnti.iotics in la.oratory medicineX/ & 7orian/ ed Hd- 2- 9illiams and 9il(ins/ Haston/ MD- 3SA- pp IC0B=0CDe &ries/ M-/ Meier/ %-/ 9ac(ernagel/ 9- #2001$- *he nat"ral transformation of the soil .acteria 1seudo"onas stutzeri and Acineto'acter sp- .y transgenic plant DNA strictly depends on homologo"s se,"ences in the recipient cells- F7.% .icro'iology /etters 9E:: 211B210!!)
A--endi2 *
De &ries/ M-/ 9ac(ernagel/ 9- #1CC=$- Detection of npt-- #(anamycin resistance$ genes in genomes of transgenic plants .y mar(erBresc"e transformation- .olecular and General Genetics 2:C: 808B816D"lmage/ ;-*-a-A-J- #1C=2$- Distri."tion of 'acill"s th"ringiensis in nat"re- -n XMicro.ial and &iral %esticidesX/ H J"rsta(/ ed- Marcel De((er/ New Nor(- pp 20CB26IHastic(/ +- #2002$- Hval"ation of the potential weediness of transgenic cotton in northern A"stralia- *echnical '"lletin no- 600/ Northern *erritory Government and A"stralian @otton @ooperative +esearch @entre/ pp 1B1II- http:))cotton-pi-csiro-a")Assets)%D!!iles)*'6001-pdf Hnglish/ 7-/ Slatin/ S-7- #1CC2$- Mode of action of deltaBendoto2ins from Bacillus thuringiensis: a comparison with other .acterial to2ins- -nsect Bioche"istry and .olecular Biology 22: 1BIH%A #2001$- 'iopesticides registration action doc"ment: Bacillus thuringiensis plantB incorporated protectants- 3S H%A/ 55A1B55@10Chttp:))www-epa-gov)pesticides).iopesticides)pips).tY.rad-htm H%A #1CCI$- %hosphinothricin Acetyltransferase and the genetic material necessary for its prod"ction in all plantsF e2emption from the re,"irement of a tolerance on all raw agric"lt"ral commodities- Federal Register ;2: 1II1IB1II20H%A #2000$- 'iopesticide fact sheet: Bacillus thuringiensis s".sp- kurstaki @ry1Ac deltaB endoto2in and its controlling se,"ences as e2pressed in cotton #008440$ 5ss"ed: 4)00- pp 1B12http:))www-epa-gov)pesticides).iopesticides)factsheets)fs008440t-htm H"ropean Scientific @ommittee on %lants #1CC=$- Opinion of the Scientific @ommittee on %lants regarding the gl"fosinate tolerant/ hy.rid rape derived from genetically modified parental lines #MS= 2 +!6$ notified .y %lant Genetic Systems #notification @)')C8)01$- *he H"ropean @ommission/ http:))e"ropa-e"-int)comm)food)fs)sc)scp)o"t0CYen-html H"ropean Scientific @ommittee on %lants #2000$- Opinion of the Scientific @ommittee on %lants regarding s".mision for placing on the mar(et of gl"fosinate tolerant oilseed rape transformation event 7i.erator %;OH 8)A@ notified .y the Agrevo company Qnow Aventis @ropScienceR #Notification @)DH)C=)8$- *he H"ropean @ommission/ pp 1B10http:))e"ropa-e"-int !AO and 9;O Safety aspects of genetically modified foods of plant origin- #2000$ Availa.le from: http:))www-who-int)fsf)GMfood)!AOB9;OY@ons"ltationYreportY2000-pdf !arrell/ *-/ +o.erts/ G- #2002$- S"rvey of cotton vol"nteers north of latit"de 22Z so"thA"stralian @otton @+@ and @S5+O %lant 5nd"stry/ Narra.ri!DA #1CC0$- Gl"fosinate tolerant canola #*opas 1C)2$- 3nited States !ood and Dr"g Administration-/ http:))www-cfsan-fda-gov)Slrd).iocon-html[lis
A--endi2 *
!!8

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Risk Assessment and Risk Management Plan

DIR 048/2003 Risk Assessment and Risk Management Plan

A"stralia New ealand !ood A"thority #now !SAN $

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DIR 048/2003 Risk Assessment and Risk Management Plan

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DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

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Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

Office of the Gene Technology Regulator

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DIR 044/2003 Risk assessment and Risk management Plan

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