Bt Eggplant

Bharat CharMahyco Research Centre

Maharashtra Hybrid Seeds Company Ltd.Jalna, India

Eggplant (Solanum melongena L.)

• Cultivated on 510,000 ha in India• 8,200,000 metric tons production • Mainly cultivated on small family farms. • Source of cash income for resource-poor

farmers.• Grown as Kharif (wet season) and Rabi

crop

Concerns associated with cultivation

• Eggplant fruit and shoot borer (EFSB) is the most destructive pest

• Losses range from 50-70%• 25 to 80 sprays undertaken for effective control.• Chemical control measures are undertaken but are

not effective due to larval behaviour• Human health concerns due to

– Pesticide exposure to farm workers – Pesticide residues on the produce

Primary damagePrimary damage

Fruit plugged with excreta

Shoot damage

Fruit damage

Larvae enters main stem and makes

tunnel which results in either

stunting or withering of plants.

Healthy stem and Damaged stem

Stem borer damage

Stem borer damage(Euzophera perticella)

Eggplant pests

Eggplant Fruit and Shoot Borer adult

Larvae (different instars)

Helicoverpa armigeraLarva (Fruit borer)

India partners: UAS Dharwad, TNAU, IIVR

Bangladesh: BARI, East-West Seeds

Philippines: UPLB

PBS-BBI

Baseline information on ESFB susceptibility to Cry1Ac protein

Genetic diversity among different EFSB populations

Mahyco hybrid fruit characteristics

• Variegated (Green+Purple+White), thorny

• Green long • Variegated

(Purple+White), thorny• Purple round (Large) • Green round (small)

• Purple+white• Shining

reddish purple• Black

Regulatory bodies in India

• Institutional Biosafety Committee (IBSC)

• Review Committee on Genetic Manipulation (RCGM) – Monitoring and Evaluation Committee (MEC) – (Dept of Biotechnology)

• Genetic Engineering Approval Committee (GEAC) – (Min of Env Forests)

Development of Bt brinjal at Mahyco

2000 Brinjal transformation initiated at Mahyco

• Gene: cry1Ac (B.t.k. strain HD73), nptII, CaMV 35S promoter,construct from Monsanto

• Transformed by Agrobacterium tumefacians-mediated method

2000-01 Lab and greenhouse evaluation for efficacy

2002 Pollen flow studies

Backcrossing program into different hybrid parents initiated

Studies conducted in 2003-04

• Acute oral toxicity studies in rats (Intox, Pune)

• Mucous membrane irritation test in female rabbit (Intox, Pune)

• Primary skin irritation test in rabbit (Intox, Pune)

• RCGM multilocation field trials-11 Locations, five hybrids (MHB-4, -9, -10, -80 and -99)Effects on non-target and beneficial insects

• ICAR first year trials with five hybrids under the All India Coordinated Vegetable Improvement Program

Studies conducted in 2005

• Subchronic oral toxicity study in Sprague Dawley rats (Intox, Pune)

• Assessment of allergenicity of protein extract using Brown Norway Rats (Rallis, Bangalore)

• Effects as a dietary feed ingredient on common carp (Cyprinus carpio) (Central Institute of Fisheries Education, Mumbai)

Studies conducted in 2005

• IRM workshop and recommendations• RCGM trials for three new hybrids

(MHB-11, -39, -112) • ICAR second year trials for five

hybrids• ICAR first year trials for three new

hybrids

Additional studies requested by RCGM in 2005(Completed in 2006)• Chemical fingerprinting of Bt and non-Bt eggplant

(alkaloids) (Indian Institute of Chemical Technology, Hyderabad)

• Subchronic feeding studies using New Zealand white rabbit (Advinus Theraputic, Bangalore)

• Effect on performance and health of broiler chickens (Central Avian Research Institute, Izatnagar)

• Subchronic feeding studies in goats (AdvinusTheraputic, Bangalore)

• Feeding studies in lactating dairy cows (G. B. Pant University of Agriculture and Technology)

• Socioeconomic studies

Other studies completed

• Germination and weediness studies• Aggressiveness studies• Soil micro-biota studies (two years)• Substantial equivalence studies• Protein expression studies on all hybrids• Baseline susceptibility studies (two years

with 29 populations)• Protein detection in cooked fruits• Molecular characterization and line ID

Gene stability and integrity

• Behaves as single gene dominant trait

• Single copy insert• Line ID and zygosity assays

developed

Protein expression and quantification

• Crop analyzed from 30-180 d.a.t• Leaf, shoot, calyx, fruit, stem and root

tissues sampled• Protein levels highest in shoots and

fruit• Stable expression through out the

growing period.

Mean Cry1Ac concentration in tissues of MHB-112grown in two locations in North India.

Days after transplanting

20 40 60 80 100 120 140 160 180 200

Con

cent

ratio

n of

Cry

1Ac

(ppm

)

0.1

1

10

100

Terminal LeafShootStemFlowerFruitRoot

MIC95 = 0.059 ppm

2005

Insect Resistance Management (IRM) 2005-2006

(In consultation with TNAU, UAS Dharwad, Mahyco, Cornell Univ., Univ. of Philippines, BARI and East West Seeds)

• A structured refuge in which 5% of the plants in the field will be non-Bt

• Development of guidelines for crop management and post-release monitoring

Bt crop

Results of agronomic evaluation

• All Bt eggplant hybrids have higher percentage of marketable yield as compared to their non-Bt counterpart, local and commercial checks.

• Bt hybrids yield significantly higher (50%) than their non-Bt counterparts, local and commercial checks.

• The incidence of EFSB is significantly lower in the Bt hybrids as compared to their non-Bt counterparts, local and commercial checks.

• Stem tunneling was also reduced in Bt hybrids• Observations of non-target pests and beneficial

insects show no significant differences between the Bt hybrids, their non-Bt counterpart, local and commercial checks.

Conclusions

• The target pest is controlled by Bt eggplant

• Biosafety studies conducted to date show no significant differences between Bt and non-Bt eggplant

Present Status

• Based on the data presented we have requested GEAC to permit Large Scale Trials of Bt eggplant

• Written queries submitted to GEAC by NGOs and other groups have been responded to

• Awaiting GEAC decision on LST approval

Thanks