Background

Background

Bacillus thuringiensis Gram+, Soil bacterium, secretes -endotoxins with insecticidal property Belongs to Kingdom Eubacteria Phylum Firmicutes Class Bacilli Order Bacillales Family Bacillaceae Genus Bacillus Species thuringiensis Discovered by Japanese biologist Shigetane Ishiwatari in 1902 Important reservoir of Cry toxins to produce insect resistant crops

Background

Background

(Sandhu, 2010)

Background

- endotoxin

Insecticidal crystal protein secreted by many Bt strains during sporulation Encoded by Cry gene located on plasmid Toxic to Lepidoptera, Diptera, Coleoptera, Hymenoptera, and Nematodes Domain III: Determines insect specificity Domain II: Binds membrane receptor Domain I: Makes transmembrane lytic

Molecular structure

Background

Background

Components of Gene Construct

Promoter Selectable marker Scorable marker Gene of interest Terminating sequence

Promoter

A sequence of bases in a nucleic acid strand, that serves as a signal to start transcription e.g 35S,35S+sh1-i1, Vicilin

Gene of interest Selectable marker

cry1A(a), cry1A(b) etc

Neomycin phophotransferase (nptII) Hygromycin phophotransferase (hpt) Gentamycin acetyl transferase Streptomycin phophotransferase Phosphinothricin acetyl transferase lux (Luciferase) lacZ (-galactosidase) Green florescencent protein (GFP)

(bar)

Scorable marker

Terminator sequences

Background

Creation of a Bt crop

Why does India need Bt food

Need for Bt food in India

Insect pest damage adds to food insecurity

Need for Bt food in India

Insect pest damage adds to food insecurity

Need for Bt food in India

Insect pest damage adds to food insecurity

Need for Bt food in India

Insect pest damage adds to food insecurity

Need for Bt food in India

Insect pest damage adds to food insecurity

State of food insecurity in rural India

Reported jointly by WFP and MSSRF

Need for Bt food in India

Climate change may encourage insect pests

Petzoldt and Seaman in Climate Change Effects on Insects and Pathogens Climate change and agriculture: Promoting practical and profitable responses

Need for Bt food in India

Climate change may encourage insect pests Increased survival

(Frazier et al, 2006)

Need for Bt food in India

Climate change may encourage insect pests Increased survival

Need for Bt food in India

Climate change may encourage insect pests Increased survival

Need for Bt food in India

Environmental, ecological and health hazard of insecticides

Need for Bt food in India

Environmental, ecological and health hazard of insecticides

Need for Bt food in India

Insecticide resistance development

Detoxification by esterases in B tabaci caused resistance against organosulfates and pyrethroids
(Ahmed et al, 1999)

Insecticide resistance to cotton insect pests had been static despite significantly reduced application of insecticides in India
(Jadhav, 1999)

Resistance to deltamethrin and fenvalrate, parathion methyl was developed in field populations of Pink Bollworm (Pectinophora gossypiella)
(Li et al, 1997)

Need for Bt food in India

Target non-specificity of insecticides

Toxicity-: to natural enemies Biomagnification: Most organochlorines (banned in US in 1970s and 80s) were found to magnify in birds of prey thinning of egg shells and breakage during incubation Hormoligosis: Sublethal dose of insecticides can stimulate the growth of insects. Found in: Two-spotted spider mite (Tetranychus urticae) Western corn rootworm (Diabrotica virgifera) Brown planthopper (Nilaparvata lugens) Over 500 insect spp. have developed resistance to at least one insecticide -

Need for Bt food in India

High expense on insecticides

Despite extensive plant breeding efforts, over 10 billion USD is spent each year worldwide on management and chemical control of pests
(Khush, 2001)

India spends 630 million USD on pesticides annually of which 380 million USD are spent solely on FSB and sucking pests
(Reddy and Zehr, 2004)

Present scenario of Bt

Present scenario of BT

Bt Research

Private undertakings in Bt research

Present scenario of BT

Bt Research

Present scenario of BT

Bt Research
Institute IARI, N.Delhi

Plants Brinjal Tomato Cauliflower Cabbage

Transgenes Bt.cry1A(b) Bt.cry1A(b) Bt.cry1A(b) Bt.cry1A(b) ACCsynthase Bt.cry1A(b)

Aim /target Resistant to lepidopteron Resistant to lepidopteron Plutella scylostella (DBM) Plutella scylostella To control fruit ripening Resistant to lepidopteron

CPRI(Simla)

Tomato Potato

Crop Cabbage Cauliflower Okra

Event name / genes cry1Ac cry1Ac cry1Ac

Trait Insect resistance Insect resistance Insect resistance

Possible commercialisation 2015 2015 2015

R&D in vegetables and fruits in world, 2008

R&D in vegetables and fruits in world, 2008

R&D in vegetables and fruits in world, 2008

Present scenario of BT

Bt Research Status of lab and field trials in India, 2008

Present scenario of BT

Bt Research

Status of lab and field trials in India, 2008

ISAAA, 2008

Present scenario of BT

Bt Adoption and Commercialization

Present scenario of BT

Bt Adoption and Commercialization

Panopio and Mercado in BIOLIFE, Nov2010-Jan2011 issue

James, 2009

Present scenario of BT

Bt Adoption and Commercialization

Bt food in Indian context

Bt food in Indian context

Pros Boon to small scale farmers and agrial economy

98% mortality in shoots and 100% in fruits of Bt brinjal average increase of 116166% marketable fruits over non-Bt brinjal Projected net economic benefit range from Rs 16,299 to Rs 19,744 per acre to farmers and national benefit to India will be 400 million USD per year
(Executive summary, ISAAA, 2009)

Reduced tractor fuel usage as a result of reduced number of sprays

(DBT, GoI)

Farmers in Gujarat, Maharashtra, A.P, T.N were significantly benefitted from Bt cotton cultivation (IIM-A, Gandhi and Namboodiri, 2006) During 1996-2008, In US, 7 billion USD economic benefits from Bt corn
(Susckiw, 2010)

In 2008, global net economic benefits to biotech crop farmers was 9.2 billion USD (4.7 billion in developing, 4.5 billion in developed countries) During 1996-2008, these benefits were 26.1 billion USD for developing and 25.8 billion USD for developed nations
(ISAAA, 2009)

Bt food in Indian context

Pros Food security

If not all GMs but definitely Bt crops show enhanced yield

(Gurien and Sherman, 2009)

Bt food in Indian context

Pros Environment friendly

Significant reduction in pesticides, saving on fossil fuels, decreased GHG emission due to no/reduced spraying

During 1996-2008, cummulative reduction in pesticide usage was 356 million Kg active ingredient , which meant: 1. 8.4% saving on pesticides 2. 16.1% reduction in associated environmental impact of pesticides use measured by Environmental Impact Quotient

In 2008 alone, the reduction was 34.6 million Kg a.i, which meant: 1. 9.6% saving on pesticide 2. 18.2% reduction in environmental impact In 2008, saving of 1.22 billion Kg CO2 equivalent to saving from about half a million reduction in no.of cars on roads!

(ISAAA, 2009)

In 1998, 8.3 million pound a.i. was cut by planting Bt crops in US

(USDA, 1998)

Bt food in Indian context

Pros Environment friendly

Bt food in Indian context

Cons-Bt technology too expensive for resource poor farmers

Rakesh Tuli, National Botanical Research Institute

Bringing Bt food to market is costly and most of them are patented raises seed cost many fold Small scales farmers of developing world cant afford Widens gap between rich and poor countries
Whitman, CSA discovery guides, April 2000

Bt food in Indian context

Cons-Corporate control over Indian agriculture

MNC monopoly is a serious issue Beneficiaries of Biotechnology giants work to meet their own ends Strong intellectual property rights make the technology slave to MNCs and inaccessible to the real Target, the Farmer

Indigenous research should be encouraged to resolve MNC monopoly Large funding to Public sector R&D Cost reduction programmes and loan systems should be introduced

BT food in general context

Concerns

BT food in general context

Concerns- Antibiotic resistance due to marker genes

Antibiotic marker genes present in Bt transgene is suspected to get transferred our gut micro-organisms to cause them survive oral antibiotic doses But in human stomach all food first passes through acidic environment where all foreign DNA is digested Still research is being carried on to preclude a rare occurrence of gene transfer, if any Use of safer markers
1. Green flourescent protein as visual marker 2. - glucoronidase gene 3. Phosphomannose isomerase (PMI) and xylose isomerase (xylA) genes 4. rol genes Removal of markers 1. Co-transformation 2. Site specific recombination 3. Intrachromosomal recombination 4. Use of transposases Modulation of marker gene expression

BT food in general context

Concerns- Development of Bt resistance in insects

http://www.biotechknowle dge.monsanto.com

Sandhu, 2010

BT food in general context

Concerns- Allerginicity and toxicity

Source: FDA

BT food in general context

Concerns- Allerginicity and toxicity

Bt toxin has been used for more than 50 years as a biological insecticide
(Musser and Shelton ,2003; Carriere et al , 2003; Qaim and Zilberman, 2003)

Show specific activity against narrow group of insects and have no or little direct effect on non-target organisms (Perlak
et al , 1990; Carriere et al , 2003; Qaim and Zilberman, 2003)

The Cry1Ac endo-toxin when cooked breaks down into common amino acids in the digestive system, which are part of the normal diet and are neither toxic nor allergic The Cry1Ac endo-toxin works only in alkaline medium. The human stomach is acidic, the digestive process will not be affected by the introduction of the Cry1Ac toxin (Centre for
Environmental Education, Ministry of Environment and Forests, GoI)

BT food in general context

Concerns- Environmental hazard

BT food in general context

Concerns- Environmental hazard

Loss of biodiversity and weediness

There has been no significant risk of gene transfer or capture in Bt Corn or potato in the US (EPA, USA) Bt crop should never be grown in centre of diversity S. melongena does not cross naturally with other species or wild relatives to produce fertile offspring Over 90% self-pollinated Bt brinjal transgene will not confer any fitness advantage to the wild species because no major insects have been found feeding on them. Solanum pollens are sticky and do not travel long distances even if they become airborne as observed in Bt brinjal pollen flow studies.

GEAC expert committee (2007)

BT food in general context

Concerns- Environmental hazard

Soil contamination
Bt protein were below detectable levels during study Bioassays and ELISA showed no detectable Bt protein in any of the microflora and fauna samples tested No differences were reported in cultivable bacterial and fungal populations, collembola and earthworm populations and soil nematode populations in Bt and non-Bt brinjal fields
(GEAC Minutes of Second Expert Committee, 2007; Mahyco 2008b)

BT food in general context

Concerns- Environmental hazard

Harm to non-target organisms

A Nature article suggested that Bt pollen is toxic High mortality rates were reported in Monarch Butterfly larvae by feeding on milkweed adjacent to Bt Corn
(Nature, Vol 399, No 6733, p 214, May 1999) Bt and the Monarch Butterfly: Update by Dr. Douglas Powell http://www.agcare.org/AGCareUpdate.htm#Monarch)

The study was not conducted in natural field conditions and was flawed
USDA, EPA

There is a strong network to assure the safety of Bt crops

Role and jurisdiction of competent authorities

Role and jurisdiction of competent authorities

Role and jurisdiction of competent authorities

Data and information typically required by regulatory authorities

Feature of the GMO Parent organism Donor organism Altered phenotype Introduced genetic material Altered phenotype Genetic modification Detection technique Dietary intake Nutritional data Toxicological data Allergenicity Horizontal gene transfer Example data sets and information History of safe use as a food Known pathogenicity and previous history of safe use as a food, potential allergenic properties The encoded protein and its function Description of the vector, description of genetic elements, sequence of the vector, the transformation process, sequence of flanking regions at insertion Description, morphology compared to parental organism, any intended levels of nutrient alteration, variation in concentration of nutrients in different environments, potential for modification of the modified phenotype (for example after processing) Southern analysis, Northern analysis, measure of expression of the transgene such as Westerns, immunoblotting PCR, Biochemical assay How the food will be processed and consumed Key metabolite concentrations, comparison to nutrient levels in other foods Sequence comparison to databases, similarity to know toxins, levels of exposure, heat stability of introduced protein, simulated digestion studies, acute oral toxicity studies, animal feeding studies Results of immune tests, similarity of protein to know allergens, resistance of protein to heat and digestion Assessment of interaction with gut microflora

Little about Bt Brinjal

Leucinodes orbonelis larva feeds inside the shoots and fruits of the crop it attacks

It plagues most of African and Asian agriculture

Bt brinjal is armed with Bt toxin against FSB Bacillus thuringiensis cry1Ac gene
cry1Ac,CaMV35S,npt II

Agrobacteriummediated transformation

Resistance against lepidopteron insects like the Brinjal Fruit and Sho orbonalis) and Fruit Borer (Helicoverpa armigera)

Sathimoorthy et al (2006)

Bt Brinjal at our Department

Large scale field trials of Bt Brinjal hybrids at our department

Conducted during Kharif, 2007 and Kharif, 2008 seasons 15 entries comprised Bt and non-Bt varieties and a national check Pusa hybrid.6 Observations were made on: 1. Number and weight of healthy and damaged fruits 2. Economic benefits 3. Effect on target insect, non-target pests and beneficial pests Seedlings were provided by MAHYCO seeds and trials were monitored by Institutional Bio-safety Committee (IBSC) on 19.92008 and 15.11.2008 Bt brinjal hybrids showed better performance than non-Bt and national check on all attributes (Evaluation report on Large Scale Field Trial of Bt-Brinjal
Hybrids, Kharif, 2007 and 2008)

Conclusions