bt foods in india geetika pau, ldh

8/6/2019 BT Foods in India Geetika PAU, LDH

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Background


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Background

Bacillus thuringiensis

Gram+, Soil bacterium,secretes -endotoxins withinsecticidal property

Belongs to Kingdom Eubacteria

Phylum Firmicutes Class Bacilli Order Bacillales Family Bacillaceae Genus

Bacillus

Speciesthuringiensis

Discovered by Japanesebiologist ShigetaneIshiwatari in 1902

Important reservoir of Cry

toxins to produce insectresistant cro s
http://en.wikipedia.org/wiki/Bacteriahttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Bacillihttp://en.wikipedia.org/wiki/Bacillaleshttp://en.wikipedia.org/wiki/Bacillaceaehttp://en.wikipedia.org/wiki/Bacillushttp://en.wikipedia.org/wiki/Bacillushttp://en.wikipedia.org/wiki/Bacillaceaehttp://en.wikipedia.org/wiki/Bacillaleshttp://en.wikipedia.org/wiki/Bacillihttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Bacteria


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Background


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Background

(Sandhu, 2010)


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Background

- endotoxin

Molecular structure

Insecticidal crystalprotein secreted bymany Bt strains duringsporulation

Encoded by Crygene

located on plasmid

Toxic to Lepidoptera,Diptera, Coleoptera,Hymenoptera, andNematodes

Domain III: Determinesinsect specificity

Domain II: Bindsmembrane receptor

Domain I: Makestransmembrane l tic


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Background


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PromoterScorable

marker

Gene of

interest

Selectable

marker

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 cry1A(a), cry1A(b) etcSelectable marker Neomycin phophotransferase (nptII)

Hygromycin phophotransferase (hpt)Gentamycin acetyl transferaseStreptomycin phophotransferasePhosphinothricin acetyl transferase

(bar)

Scorable marker lux(Luciferase)lacZ (-galactosidase)Green florescencent protein (GFP)

Terminator sequences

Background

Components of Gene Construct


11/68Creation of a Bt crop

Background


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Why does India need Bt food


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Need for Bt food in IndiaInsect pest damage adds to food insecurity


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N d f Bt f d i I di


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Need for Bt food in India

Insect pest damage adds to food insecurity

State of food insecurity in rural IndiaReported jointly by WFP and MSSRF


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N d f Bt f d i I di


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Need for Bt food in IndiaClimate change may encourage insect pests Increased survival

(Frazier et al, 2006)



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Need for Bt food in IndiaClimate change may encourage insect pests Increasedsurvival


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Climate change may encourage insect pests Increased

survival


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Need for Bt food in IndiaEnvironmental, ecological and health hazard of insecticides


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Need for Bt food in IndiaEnvironmental, ecological and health hazard of insecticides

N d f B f d i I di


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Detoxification by esterases in B tabacicaused resistance againstorganosulfates and pyrethroids

(Ahmed et al, 1999)

Insecticide resistance to cotton insect pests had been staticdespite significantly reduced application of insecticides in India

(Jadhav, 1999)

Resistance to deltamethrin and fenvalrate, parathion methyl wasdeveloped in field populations of Pink Bollworm (Pectinophoragossypiella)(Li et al, 1997)

Need for Bt food in IndiaInsecticide resistance development

N d f Bt f d i I di


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Toxicity-: to natural enemies

Biomagnification: Mostorganochlorines (banned in US in1970s and 80s) were found tomagnify in birds of prey thinningof egg shells and breakage duringincubation

Hormoligosis: Sublethal dose ofinsecticides can stimulate thegrowth 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 oneinsecticide -

Need for Bt food in IndiaTarget non-specificity of insecticides



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Despite extensive plant breeding efforts, over 10 billion USD isspent each year worldwide on management and chemicalcontrol of pests

(Khush, 2001)

India spends 630 million USD on pesticides annually of which 380million USD are spent solely on FSB and sucking pests

(Reddy and Zehr, 2004)


High expense on insecticides


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Present scenario of Bt

Present scenario of BT


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Bt Research

Private undertakings in Bt research



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Bt Research



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Institute Plants Transgenes Aim /target

IARI, N.Delhi Brinjal Bt.cry1A(b) Resistant to lepidopteron

Tomato Bt.cry1A(b) Resistant to lepidopteron

Cauliflower Bt.cry1A(b) Plutella scylostella (DBM

Cabbage Bt.cry1A(b) Plutella scylostella

Tomato ACCsynthase To control fruit ripeningCPRI(Simla) Potato Bt.cry1A(b) Resistant to lepidopteron


Bt Research

Crop Event name /genes

Trait Possiblecommercialisation

Cabbage cry1Ac Insect resistance 2015

Cauliflower cry1Ac Insect resistance 2015

Okra cry1Ac Insect resistance 2015

R&D in vegetables and fruits in world 2008


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R&D in vegetables and fruits in world, 2008



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Bt Adoption and Commercialization



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Panopio and Mercado in BIOLIFE, Nov2010-Jan2011 issue

James, 2009



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Bt food in Indian context


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t oo n n an contextPros Boon to small scale farmers and agrial economy


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Pros Boon to small scale farmers and agrial economy

98% mortality in shoots and 100% in fruits of Bt brinjal average increase of 116-166% marketable fruits over non-Bt brinjal

Projected net economic benefit range from Rs 16,299 to Rs 19,744 per acre tofarmers 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 Btcotton 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 25.8billion USD for developed nations

(ISAAA, 2009)


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If not all GMs but definitely Bt crops show

enhanced yield (Gurien and Sherman, 2009)

Pros Food security



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

During 1996-2008, cummulative reduction in pesticide usage was 356million 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)

Pros Environment friendly



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Pros Environment friendly

B f d i I di


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Bt food in Indian contextCons-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 raisesseed cost many fold

Small scales farmers of developing world cant afford

Widens gap between rich and poor countriesWhitman, CSA discovery guides, April 2000

Bt f d i I di t t


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MNC monopoly is a serious issue

Beneficiaries of Biotechnology giants work to meet their own ends

Strong intellectual property rights make the technology slave toMNCs and inaccessible to the real Target, the Farmer

Indigenous research should be encouraged to resolve MNCmonopoly

Large funding to Public sector R&D

Cost reduction programmes and loan systems should be

introduced

Bt food in Indian contextCons-Corporate control over Indian agriculture

BT food in general context


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Concerns



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Antibiotic marker genes present in Bt transgene is suspected to gettransferred our gut micro-organisms to cause them survive oralantibiotic doses

But in human stomach all food first passes through acidic environmentwhere all foreign DNA is digested

Still research is being carried on to preclude a rare occurrence of genetransfer, if any

Use of safer markers1.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

Concerns- Antibiotic resistance due to marker genes



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Concerns- Development of Bt resistance in insects

Sandhu, 2010

http://www.biotechknowle

dge.monsanto.com



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Concerns- Allerginicity and toxicity

Source: FDA



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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 orlittle direct effect on non-target organisms (Perlaket al, 1990; Carriere et al, 2003; Qaim and Zilberman, 2003)

The Cry1Ac endo-toxin when cooked breaks down into common aminoacids in the digestive system, which are part of the normal diet andare 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 forEnvironmental Education, Ministry of Environment and Forests, GoI)

Concerns- Allerginicity and toxicity



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Concerns- Environmental hazard


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BT food in general contextC E i t l h d


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Soil contamination

Bt protein were below detectable levels during study

Bioassays and ELISA showed no detectable Bt protein in any of themicroflora and fauna samples tested

No differences were reported in cultivable bacterial and fungal populations,collembola and earthworm populations and soil nematode populations inBt and non-Bt brinjal fields

(GEAC Minutes of Second Expert Committee, 2007; Mahyco 2008b)

BT food in general contextC E i t l h d


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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 onmilkweed 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
http://www.agcare.org/AGCareUpdate.htmhttp://www.agcare.org/AGCareUpdate.htm


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


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Role and jurisdiction of competent authorities


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Role and jurisdiction of competent authorities


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Data and information typically required by regulatory authorities


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Feature of the GMO Example data sets and information

Parent organismDonor organism

ltered phenotype

Introduced genetic material

ltered phenotype

Genetic modification

Detection technique

Dietary intakeutritional data

Toxicological data

llergenicity

Horizontal gene transfer

History of safe use as a foodKnown pathogenicity and previous history of safe use as a food, potentialallergenic properties

The encoded protein and its functionDescription of the vector, description of genetic elements, sequence of the vector,the transformation process, sequence of flanking regions at insertionDescription, morphology compared to parental organism, any intended levels ofnutrient alteration, variation in concentration of nutrients in differentenvironments, potential for modification of the modified phenotype (for exampleafter processing)

Southern analysis, Northern analysis, measure of expression of the transgene suchas Westerns, immunoblottingPCR, Biochemical assayHow the food will be processed and consumedKey metabolite concentrations, comparison to nutrient levels in other foodsSequence comparison to databases, similarity to know toxins, levels of exposure,heat stability of introduced protein, simulated digestion studies, acute oral toxicity

studies, animal feeding studiesResults of immune tests, similarity of protein to know allergens, resistance ofprotein to heat and digestionAssessment of interaction with gut microflora

Little about Bt Brinjal


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Little about Bt Brinjal

Leucinodes orbonelis larva feeds inside the shoots and fruits


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Leucinodes orbonelis larva feeds inside the shoots and fruitsof the crop it attacks

It plagues most of African and Asian agriculture


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p g g

Bt brinjal is armed with Bt toxin against FSB


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cry1Ac gene

Sathimoorthy et al(2006)

Resistance against lepidopteron insects like the Brinjal Fruit and Sh

orbonalis) and Fruit Borer (Helicoverpa armigera)

Agrobacterium-mediatedtransformation

Bt brinjal is armed with Bt toxin against FSB

Bacillus thuringiensis

cry1Ac,CaMV35S,npt II


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Bt Brinjal at our Department

l fi ld i l f i j l h b id


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Large scale field trials of Bt Brinjal hybrids at ourdepartment

Conducted during Kharif, 2007 and Kharif, 2008 seasons

15 entries comprised Bt and non-Bt varieties and a national checkPusa 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 monitoredby Institutional Bio-safety Committee (IBSC) on 19.92008 and15.11.2008

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


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Conclusions

bt foods in india geetika pau, ldh

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