NUTRITIONAL STUDIES OF CHARA CORALLINA

Uday Bhan Singh1*, R.K. Thakur2, Ravi h !"r#a$, A.S. A%LU&ALIA1

*

e-mail: ubsday@gmail.com

Laboratory of Algal Biology and Diversity, Department of Botany, Panjab University, Chandigarh-16
#

1!, "ndia 1!, "ndia !, "ndia

Department of $oology, Panjab University, Chandigarh-16 Department of Botany, University of &ajasthan, 'aip(r-% #

ABSTRA'T ( )hy i*+,*h"#i*a- ana-y i +. /h" 0+nd 1a/"r u00+r/ing gr+1/h +. Chara corallina and +. /h+ " gr+1n in di.."r"n/ *u-/ur" #"dia in /h" -a2+ra/+ry *+ndi/i+n hav" 2""n *arri"d +u/. Th" 0hy i*+,*h"#i*a- 0ara#"/"r +. /h" 1a/"r +. Na0-i .+r" / and ).N. 3"hra B+/ani*a- 4ard"n h+1 /ha/ Chara corallina r"5uir" -" a#+un/ +. nu/ri"n/ and

/h" -"v"- +. 0+--u/i+n i 5ui/" -+1 a i "vid"n/ .r+# /h" da/a and /andard . Th" 1a/"r i *-"ar "n+ugh /+ a--+1 0"n"/ra/i+n +. vi i2i-i/y /+ i/ 2+//+#. Na0-i .+r" / and ).N. 3"hra B+/ani*a- 4ard"n -a*k ri*h div"r i/y 1hi*h #ay 2" a//ri2u/"d /+ +#" *h"#i*a- r"-"a "d 2y Chara corallina. In -a2+ra/+ry *u-/ur", i/ i .+und /ha/ Chara corallina *an 2" gr+1n in a-- /hr"" *u-/ur" #"dia, viz., A--"n and Arn+n6 #"diu# , 'hu,17 #"diu# and B4,11 #"diu# 2u/ gr+1/h i ""n /+ 2" /h" 2" / in A--"n and Arn+n6 *u-/ur" #"diu# in

*+#0ari +n 1i/h /h" +/h"r /1+ *u-/ur" #"dia. K"y 1+rd ( Chara, Physi)o)hemi)al parameters, Algae, *ater poll(tion+ INTRODU'TION

Chara)ean algae are )alled stone,orts or brittle,orts d(e to e-tensive e-tra)ell(lar )al)ifi)ation+ .he presen)e of Charophy)eae /stone,orts0 is often asso)iated ,ith )lean and rather hard ,ater ,ith a lo, phosphoro(s /P0 )on)entration /1ig+ 10+

Figur" 1. Sh+1ing /h" gr+1/h +. Chara corallina in Ba/ani*a- 4ard"n 1orsberg reported that sol(ble rea)tive phosphor(s /2&P0 )on)entration as lo, as 13 4g P L51 inhibited the plant gro,th and the sensitivity to P e-plained the absen)e of )harophytes in very e(trophi) ,aters and their disappearan)e from poll(ted lo)alities 617+ Contrary to earlier findings, later st(dies have sho,n that the plant )an gro, ,ell in ,ater ,ith a high P )on)entration, for e-ample 1+ mg P L51 6#7 and +8 mg P L51 6%7 and the disappearan)e of the plant is )a(sed primarily by shading thro(gh phytoplan9ton and other algae 6!7+ Dense Chara beds in hard ,ater may a)t as n(trient sin9s in several ,ays, s()h as n(trient in)orporation in plant biomass, n(trient ,ithdra,al from de)omposing detrit(s, release of allelophati)

)ompo(nds, red()ed sediment res(spension, slo, de)omposition rate on plant senes)en)e, and the )o-pre)ipitation of P ,ith )al)ite 637+ Charophytes thrive and may dominate the vegetation in (npoll(ted fresh and bra)9ish ,aters 667+ Charophytes may also indire)tly affe)t n(trient )y)ling in la9es+ Utili:ation of bi)arbonate is a))ompanied by pre)ipitation of )al)ite d(ring periods of intensive photosynthesis, favoring immobili:ation of P by binding in the )rystal str()t(re or sorption on sedimenting mineral parti)les+ Charophytes are able to deliver o-ygen to the sediment, th(s potentially enhan)ing nitrifi)ation;denitrifi)ation pro)esses and preventing ironbo(nd sediment phosphor(s from being released to the overlying ,ater+ 1(rthermore, dense Chara meado,s restri)t sediment res(spension, )onse<(ently blo)9ing an important internal so(r)e of n(trients to plan9toni) algae+ =oreover, the plant )an be heavily )al)ified-more than 3 > /as CaC?%0 of the total plant biomass dry ,eight has been reported 6@,87+ Aeeping this in vie,, the present investigation has been (nderta9en to assess the ,ater <(ality of fresh,ater ponds s(pporting vegetation of Chara corallina, and also s(itability of best gro,th medi(m to gro, Chara corallina, for its e-ploitation as a so(r)e for )arbon se<(estration programs+ 3ATERIALS AND 3ET%ODS '+--"*/i+n i/" "n the present st(dy, t,o sample )olle)tion sites ,ere sele)ted in /@6 !6B!3+C6BBD, % !!B 1+1CBBE0 Chandigarh+ Both sites sho,ed prof(se gro,th of Chara corallina+ /10 Botani)al garden pondF "nside Panjab University botani)al garden, ,ater re)harged every ,ee9 ,ith gro(nd ,ater+ Almost )rystal )lear ,ater maintained and presen)e of Chara corallina. Eo addition of ,aste,ater+ /#0 Eepli forest pondF Eat(ral and )onserved pond, s(rro(nded by forest )over, no

anthropogeni) a)tivities allo,ed+ Gro,th of Chara corallina has been observed+ Eo addition of ,aste,ater+ )hy i*+,*h"#i*a- ana-y i Physi)o-)hemi)al parameters of the ,ater ,ere analy:ed (sing standard methods APHA 6C7+ *ater samples in tripli)ates ,ere pro)(red from the P+E+ =ehra Botani)al Garden Chandigarh, and Eapli forest, Chandigarh+ Parameters li9e temperat(re, pH, fi-ation of dissolved o-ygen, se))hi-transparen)y, and free )arbon dio-ide ,ere determined on the spot+ 1or rest of the parameters, #+3 L of pond ,ater ,as bro(ght to the laboratory+ "t ,as filtered and analy:ed for total al9alinity, total hardness, )hlorides, s(lphate, nitrates, and phosphates+ .otal dissolved solids /.D20 by (sing D-=er)9Bs .D2 meterI *ater temperat(re by (sing )entigrade thermometer and pH ,as re)orded ,ith BDH paper on the spot and later by pH meter in the laboratory+ .he data ,as statisti)ally analy:ed (sing soft,are 2P22 version 18, to )al)(late average and standard deviation+ E /i#a/i+n +. gr+1/h ra/" 1resh and dry ,eight ,as estimated a))ording to the method given by "nternational Atomi) Dnergy Agen)y 61 7+ .he media sele)ted for the )(lt(re of Chara corallina are Ch(-1 medi(m 6117, Allen and ArnonBs medi(m 61#7, and BG-11 medi(m 61%7+ .he api)al gro,ing tips of the plant material ,ere )(t at appro-imately 1 )m ea)h and ino)(lated in the medi(m ,ith (tmost )are (nder asepti) )onditions+ ?bservations ,ere re)orded for gro,th in terms of length, fresh ,eight and dry ,eight in order to estimate their gro,th rates after every fo(r days, (pto 1# days+ RESULTS AND DIS'USSION

Physi)o-)hemi)al )hara)teristi)s of ,ater in P+E+ =ehra Botani)al Garden and Eapli forest ,ere investigated /.able 10+ Ta2-". 1. )hy i*+,*h"#i*a- 0ara#"/"r +. 1a/"r u00+r/ing gr+1/h +. Chara corallina in ).N. 3"hra B+/ani*a- 4ard"n and Na0-i F+r" /, 'handigarh.
S.N+. 1 # % ! 3 6 @ 8 C 1 11 1# 1% 1! 13 )ara#"/"r 2e))hi-transparen)y /)m0 .otal Dissolved 2olid/mg;L0 pH .emperat(re /air0/ C0 .emperat(re /,ater0/ C0 1ree C?# /mg;L0 Carbonate - C?%#K /mg;L0 Al9alinity /mg;L0 .otal Hardness /mg;L0 Ca#K /mg;L0 =g#K /mg;L0 Cl- /mg;L0 Dissolved ?# /mg;L0 .otal Phosphate P?!#- /mg;L0 Eitrate - E?%- /mg;L0 B+/ani*a- 4ard"n @1+%J +C #16+@J%+% @+3J + % #3+%J +1@ #6+1J + % #1+@J + 8 Absent C1+@J + % %!8+%J6+ 1 1+@J +!3 #!6+6J3+8C 6!+ J%+3 # +8J1+1 + #3J + # +1#%J + ! Na0-i F+r" / 1#!+%J#+% # 1+@J6+ @+@J + 6 #3+8J + 8 #6+3J + 8 %8+ J1+13 Absent 1 %+6J#+C6 %66+@J#+! 11 +CJ +%1 #33+8J#+6 63+ J#+6 # +@J#+! + %6J + % +#1%J + 6

?bservations of se))hi-transparen)y of t,o sites indi)ate that ,ater <(ality of Eapli forest ,ater body is )leaner as visibility of se))hi dis) is more in this /.able 10+.otal dissolved solids /mg;l0 and pH ,as fo(nd to be broadly in an identi)al range+ 1ree C? # ,as re)orded to be higher in Eapli forest ,ater body+ =ost of the parameters /al9alinity,hardness and )al)i(m0 ,ere fo(nd to be higher in P+E+ =ehra Botani)al Garden ,ater+ 1e, of the parameters ,ere signifi)ant in having (niform val(es in both the habitats /.able 10+Comparing the observed val(es ,ith the standard val(es of potable ,ater, the ,ater in the P+E+ =ehra Botani)al Garden and Eapli forest is non-poll(ted+ Ho,ever, it s(pported the gro,th of Chara corallina+ .his also indi)ates that Chara ,o(ld prefer )lean ,ater for its establishment and f(rther gro,th+ .hese ,ater bodies la)9 ri)h biodiversity, may be d(e to non-poll(ted and poor levels of nitrogen and phosphor(s+

Comparing vario(s physi)o)hemi)al parameters of st(died ,ater samples ,ith Eational and "nternational 2tandards, .able # 61!,13,167, reveals that all the parameters analy:ed ,ere ,ith the permissible limits of drin9ing ,ater standards+ Ta2-" 2. S/andard .+r drinking 1a/"r
Parameters *H? pH 6+3-8+3 .otal Al9alinity 1# .otal hardness 3 Chloride #3 Eitrate 3 -1 Phosphate +1-1 B?D 3+ D? 3+ .D2 3 All parameters are e-pressed in mgL-1 e-)ept pH "2" 6+3-8+3 % #3 -1 3 -1 +3-1 %+ "C=& @-8+3 % #3 -1 # -1 3

Based on )ertain parameters, Central Poll(tion Control Board /.able %0 has )lassified fresh ,ater into five )lasses /A, B, C, D and D0 s(itable for vario(s p(rposes+ ?n the basis of this )lassifi)ation, the ,ater at ponds )o(ld be )ategori:ed as LAM+ .he val(es of ,ater <(ality )learly revealed that the ,ater )an be (sed for drin9ing p(rposes after disinfe)tion+ Ta2-" $. &a/"r 5ua-i/y 'ri/"ria .+r vari+u u "
D" igna/"d,B" /,U " '-a +. 1a/"r 'ri/"ria .otal Coliforms ?rganism =PE;1 Drinking &a/"r S+ur*" 1i/h+u/ *+nv"n/i+na- /r"a/#"n/ 2u/ a./"r di in."*/i+n A Dissolved ?-ygen 6mg;L or more ml shall be 3 or less

pH bet,een 6+3 and 8+3

Bio)hemi)al ?-ygen Demand 3 days # NC #mg;L or less Ou/d++r 2a/hing 8Organi "d9 B .otal Coliforms ?rganism =PE;1 less ml shall be 3 or

pH bet,een 6+3 and 8+3 Dissolved ?-ygen 3mg;L or more

Bio)hemi)al ?-ygen Demand 3 days # NC %mg;L or less .otal Coliforms ?rganism =PE;1 ml shall be 3 Drinking 1a/"r +ur*" a./"r *+nv"n/i+na- /r"a/#"n/ and di in."*/i+n Bio)hemi)al ?-ygen Demand 3 days # NC %mg;L or less )r+0aga/i+n +. &i-d -i." and D Fi h"ri" 1ree Ammonia /as E0 1+# mg;L or less pH bet,,n 6+ to 8+3 pH bet,een 6+3 to 8+3 Dissolved ?-ygen !mg;L or more ' pH bet,een 6 to C Dissolved ?-ygen !mg;L or more less or

Dle)tri)al Cond()tivity at #3NC mi)ro mhos;)m Irriga/i+n, Indu /ria- '++-ing, E '+n/r+--"d &a /" di 0+ a2odi(m absorption &atio =a-+ #6 =a-+##3

Boron =a-+ #mg;L B"-+1,E N+/ 3""/ing A, B, ', D : E 'ri/"ria Central poll(tion )ontrol board /httpF;;)p)b+ni)+in;*aterOP(alityOCriteria+php0

As stated earlier, 1 )m long api)al portions of Chara corallina ,ere gro,n in % different media, ea)h medi(m being ino)(lated ,ith ten samples ea)h /1ig+ #0+

Figur" 1. Sh+1ing /h" gr+1/h +. Chara corallina in +-id agar #"diu# % repli)ates of ea)h medi(m ,as maintained to minimi:e the error+ "n these samples some profo(ndly visible observations )o(ld be made+ After ! days of )(lt(re, the older basal plant portion of the samples in Ch(-1 medi(m be)ame pale in )oloration /lost green )olor0 ,hile those gro,n in Allen and ArnonBs and BG-11 media retained their green )oloration /.able !0+ As the )(lt(re pro)eeded to the 8th day after )(lt(re /.able 30, most of the samples in Ch(-1 medi(m ,ere pale yello, in )olor, be)ame soft and had rotten baseI s(rvival ,as lo, in Ch(-1 medi(m b(t those ,hi)h s(rvived, attained proper length+ "n BG-11 medi(m, samples be)ame greenish-bro,n+ 1e, of them ,ere seen to have deteriorated ,ith rotten basal portion+ B(t in Allen and ArnonBs medi(m, all samples s(rvived and ,ere l(sh green in )olor+ &hi:oids of abo(t +%- +3 )m ,ere also seen to have developed on all the samples+ Ta2-". ;. '+#0ari +n +. /h" gr+1/h ra/" +. Chara corallina a./"r ; day +. *u-/ur" in di.."r"n/ *u-/ur" #"dia /hr+ugh .r" h and dry 1"igh/ and di.."r"n*" in -"ng/h.
)ara#"/"r Fr" h 1"igh/ 8g9 Dry 1"igh/ 8g9 L"ng/h8*#9 Surviva- +u/

3"diu# 'hu 17 #"diu# B4 11 #"diu# A--"n and Arn+n6 #"diu#

R"0-i*a/" + 3C + 33 + 6# + !C + %! + 3# + ! + 3! + %@

3"an + 3C

R"0-i*a/" + 1 + 1 + 1# + 1# + 11 + 1# + 1% + 1% + 16

3"an + 11

R"0-i*a/" 1#+1 1%+# 1#+6 1%+# 1%+@ 1%+ 1!+ 1%+! 1#+8

3"an 1#+6%

+. 17 1 1 1 1 1 1 1 1 1

+ !3

+ 1#

1%+%

+ !!

+ 1!

1%+#@

Ta2-" <. '+#0ari +n +. /h" gr+1/h ra/" +. Chara corallina a./"r = day +. *u-/ur" in di.."r"n/ *u-/ur" #"dia /hr+ugh .r" h and dry 1"igh/ and di.."r"n*" in -"ng/h.
)ara#"/"r Fr" h 1"igh/ 8g9 &epli)ates =ean Dry 1"igh/ 8g9 &epli)ates =ean L"ng/h8*#9 &epli)ates =ean Surviva+u/ +. 17 3"diu# + !3 'hu 17 #"diu# B4 11 #"diu# A--"n and Arn+n6 #"diu# +18 + 8! +1@@ +1@C +# 1 +1!C +1!@ + 13 + 8 + 18 + 1@ + 18 + 16 + 1! 13+8 1 +3 1%+! 16+@ 1!+6 13+! 1%+#% 1 8 1 1 1 1 + C +1!3 + C% + @ + 11 8+% 1!+% 16+% 1%+ 6 C C + 1 + 13

+1@6

+ 1@

13+6

After the 1#th day of )(lt(re /.able 60, s(rvival of sample ,as lo, in Ch(-1 medi(m+ "n Allen and ArnonBs medi(m, basal portion be)ame yello,ish-green li9e old stem and initiation of ne, ape- from nodes ,as seen+ 1(rther, all samples be)ame fertile and rhi:oids of abo(t 1+3-#+3

)m ,ere seen to arise from all samples+ Ho,ever, in BG-11 medi(m, the old basal body of plant got deteriorated b(t the (pper portions still retained their green )oloration+ Ta2-". >. '+#0ari +n +. /h" gr+1/h ra/" +. Chara corallina a./"r 12 day +. *u-/ur" in di.."r"n/ *u-/ur" #"dia /hr+ugh .r" h and dry 1"igh/ and di.."r"n*" in -"ng/h.
)ara#"/"r 3"diu# + !1 'hu 17 #"diu# B4 11 #"diu# A--"n and Arn+n6 #"diu# + @! + C3 +11% +1#% + 68 + 38 + C! + 8 + 1 + 1! + 16 + 13 + 13 + 11 11+% 11+# 1#+6 1%+6 11+6 1%+% 11+@ 1 1 1 1 1 1 + C + @3 + 6C + + 1 + 1 8 + C 11+C 1!+6 1%+# 1%+%% Fr" h 1"igh/ 8g9 R"0-i*a/" 3"an Dry 1"igh/ 8g9 R"0-i*a/" 3"an L"ng/h8*#9 R"0-i*a/" 3"an Surviva- +u/ +. 17 8 C C

+ 8%

+ 13

1#+8%

'ON'LUSIONS Con)l(sions )an be made from these observations that the samples gro,n in Allen and ArnonBs medi(m had the ma-im(m s(rvival and gro,th rate /.ables !-60+ 1(rther, rhi:oids ,ere seen to arise from the plant body only in the Allen and ArnonBs medi(m+ =a-im(m deterioration ,as seen in plant gro,n in Ch(-1 medi(m ,hile BG-11 medi(m )o(ld s(pport the gro,th of the algae moderately+ As stated earlier, after the 8th day of )(lt(re, no f(rther in)rease in gro,th )o(ld be seen in all media /.ables 3-60+.his may be d(e to the reason that Chara corallina re<(ires hard s(rfa)e to an)hor the plant body thro(gh rhi:oids, so after some time the deterioration of plant body starts+ D(e to lo, n(tritional re<(irements and fast gro,th rate, Chara )horallina )an be e-ploit as a possible )andidate for )arbon se<(estration programs+

Physi)o-)hemi)al parameters of ,ater s(pporting gro,th of Chara corallina also indi)ates that Chara ,o(ld prefer )lean ,ater for its establishment and f(rther gro,th+ .hese ,ater bodies la)9 ri)h biodiversity, may be d(e to non-poll(ted and poor levels of nitrogen and phosphor(s+ Chara corallina ,ith the lo, poll(tion )learly indi)ates that these ma)rophytes prefer ,ater ,ith signifi)antly different n(trient )on)entrations for their gro,th, and )an be (sed as indi)ators of poll(tion stat(s <(ality of ,ater+

A*kn+1-"dg"#"n/ .he a(thors are than9f(l to the Chairperson, Department of Botany, Panjab University, Chandigarh for providing ne)essary resear)h fa)ilities, University Grants Commission, Ee, Delhi for 2AP-D&2-"" grant and to the Council of Scientific and Industrial Research , Ee, Delhi /U+B+2ingh0 for providing finan)ial assistan)e in the form of '(nior &esear)h 1ello,ship and 2enior &esear)h 1ello,ship+

REFEREN'ES 617 1orsberg, C+ 1C6!+ Phosphor(s, a ma-im(m fa)tor in the gro,th of Chara)eae+ Eat(re+ # 1F31@-318+ 6#7 Blindo,, "+ 1C88+ Phosphor(s to-i)ity in Chara+ A<(at+ Bot+ %#F%C%-%C3+ 6%7 A(fel, L+, and .+ ?:ime9+ 1CC!+ Can Chara )ontrol phosphor(s )y)ling in La9e (9najno /Poland0Q Hydrobiol+ #@3;#@6F#@@-#8%+ 6!7 Blindo,, "+ 1CC#+ De)line of )harophytes d(ring e(trophi)ationF Comparison ,ith angiosperms+ 1resh,ater Biol+ #8FC-1!+ 637 A(fel, L+, and "+ A(fel+ # #+ Chara beds a)ting as n(trient sin9s in shallo, la9es-A revie,+

A<(at+ Bot+ @#F#!C-#6 +

667 2tephen, '+, Lambert, and '+ Anthony+ # 11+ Davy *ater <(ality as a threat to a<(ati) plantsF dis)riminating bet,een the effe)ts of nitrate, phosphate, boron and heavy metals on )harophytes+ Ee, Phytologist+ 18CF1 31-1 3C+ 6@7 H(t)hinson, G+D+ 1C@3+ A treatise on limnology, Rol+ """, Limnologi)al botany, 'ohn *iley S 2ons, Ee, Tor9+ 687 ArUli9o,s9a, '+ 1CC@+ D(trophi)ation pro)esses in a shallo,, ma)rophyte-dominated la9espe)ies differentiation, biomass and the distrib(tion of s(bmerged ma)rophytes in La9e (9najno /Poland0+ Hydrobiol+ %!#;%!%F!11-!16+ 6C7 APHA, A**A, and *D1+ # 3+ 2tandard methods for the e-amination of ,ater and ,aste

,ater /#1st ed+0, Ee, Tor9, *ashington, DCF jointly prepared and p(blished by Ameri)an P(bli) Health Asso)iation, Ameri)an *ater *or9s Asso)iation and *ater Dnvironment 1ederation+ 61 7 "nternational Atomi) Dnergy Agen)y+ # "ADA-%C#+ &eferen)e sheetF 1-!+ 6117 Ch(, 2+P+ 1C!#+ .he infl(en)e of mineral )omposition of the medi(m on the gro,th of plan9toni) algae+ "+ =ethods and )(lt(re media+ '+ D)o+ % F#8!-%#3+ 61#7 Allen, =+ B+ and D+ "+ Arnon+ 1C33+ 2t(dies on nitrogen-fi-ing bl(e-green algae+ ", Gro,th and nitrogen-fi-ation by Anabaena cylindrica Lemm+ Plant Physiol+ % F%66-%@#+ 61%7 2tainer, &+T+, A(nisa,a, &+, =andel, =+ and G+C+ Ba:ira+ 1C@1+ P(rifi)ation and properties of (ni)ell(lar bl(e-green algae /order Chro)o))ales0+ Ba)teriol+ &ev+ %3F1@1-# 3+ 61!7 *H? /1CC#0+ "nternational standards for drin9ing ,ater+ *orld Health ?rganisation, Geneva+ 6137 "C=& /1C@30+ Drin9ing ,ater standards+ "ndian Co(n)il of =edi)al &esear)h, Ee, Delhi+ 3+ .ra)e, minor and major elements in algae+

6167 "2" /1C@%0+ "ndian sandard methods for sampling and test /physi)al and )hemi)al0 for ,ater (sed in ind(stry+ "ndian 2tandard "nstit(te, =ana9 Bha,an, Ee, Delhi+