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Biotech research at tamil nadu agricultural university 2011


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  • 1. GenesisA manpower development programme in Biotechnology initiated during 1988 through PG-Biotechnology programme with the support of DBT, GoI. Plant Molecular Biology and BiotechnologyMajor activities Education Research
  • 2. EducationUnder Graduate Education B. Tech. (Biotechnology) B. Tech. (Bioinformatics)Post Graduate Education M.Sc. (Biotechnology) M.Tech (Biotech & Business Management) PhD (Biotechnology)
  • 3. Major Areas of Biotech Research Isolation of genes conferring agronomic traits  Novel genes to overcome IPR issues Genetic transformation of crops for improved agronomic traits  Wider choice of genes for incorporation into crops Marker Assisted Breeding for improving agronomic traits in crop plants  Improves the efficiency with which breeders can select plants with desirable combinations of genes
  • 4. Major Target TraitsAbiotic stress resistance Drought resistance Submergence tolerance Salt toleranceBiotic stress resistance Insect resistance Disease resistanceNutritional quality
  • 5. Abiotic Stress Resistance
  • 6. • Duration– 100 -105 days• Season– September-October (NE Monsoon)• Yield (from 40 OFTs) – 3900 kg/ha (rainfed) – 6600 kg/ha (semi dry)• Grain type – Long slender white rice• Increase in yield over PMK (R) 3 - 14.40 %• Suitable for cultivation under rainfed condition in Ramnad and Sivagangai Districts• Moderately resistant to sheath rot and blast diseases
  • 7. Submergence Tolerance About 3 lakh ha paddy is affected frequently by submergence in cauvery delta zone Necessitates the development of submergence tolerant rice varieties Marker Assisted Introgression of Sub1 locus conferring tolerance to flooding into elite rice varieties of Tamil Nadu
  • 8. Submergence Tolerance FR13A Sub1 locus was introgressed into a widely grown Asian rice cultivar “Swarna” using marker-assisted selection Submergence tolerant versions of CR1009, Savithri, Samba Mahsuri and IR 64 are already available for cultivation Submergence tolerant versions of CO 43 and White Ponni are being developed to target flooding prone areas White Ponni Co43 FR13A Recovery after 13 days of submergence
  • 9. Salt ToleranceIntrogression of salT locus conferring tolerance against salinity stress into elite rice varieties of Tamil NaduA major QTL called salT located on chromosome 1 of a salt tolerant variety “FL 478” controlling salinity tolerance will be targeted for introgressionDevelopment of salinity tolerant versions of ADT 36 and White Ponni White Ponni FL 478 Control 80mM 150 mM Control 80 mM 150 mM 10 days after treatment
  • 10. Genetic engineering of sugarcane for abioticstress toleranceAtDREB1a gene from Arabidopsis thalianaBcZAT12 gene from Brassica carinataCo86032 transformed with AtDREB1a20 transgenic lines obtainedCoC671 transformed with BcZAT1211 transgenic lines obtained
  • 11. Cotton Varieties with Drought Toleranceand Extra Long Staple FibreObjectives To characterize cotton germplasm for drought tolerance using biochemical, physio-morphological traits and molecular markers To identify DNA markers linked to drought tolerance and fibre quality traitsDevelopment of mapping populations in progress MCU5 X KC3 MCU5 X TCH1218 KC3 X Suvin and TCH1218 X Suvin
  • 12. Insect & Disease Resistance
  • 13. Development of DNA markers linked toplant-hopper resistance in riceA Mapping population involving from IR50 and Rathu-Heenathi (a genotype resistant to BPH) developed and now in F8 stage Two SSR markers viz., RM3180 (18.22 Mb) and 25 RM2453 (20.19 Mb) showed significant association with BPH resistance in the Bulked Segregant Analysis at F7 stage
  • 14. Developing bacterial blight resistant riceXa genes conferring resistance to bacterial blight are routinely transferred to elite rice cultivars through maker assisted breedingPyramiding resistance genes Xa4,xa5, xa13 and Xa21 into high yielding adopted rice varieties (ADT 43 and ADT 47) through MAS using IRBB60-1-5 (resistant) as donor is in progress30 genotypes showing higher level of resistance were identified with three genes (xa5, xa13 and Xa21) in homozygous state. Resistance reaction of susceptible (A) and pyramided (B) lines against Xoo isolates A B
  • 15. Development of gall midge, blast andbacterial blight resistant rice varietiesGall midge resistant version of Co43 pyramided with GM1 and GM4 developed through marker assisted breedingBacterial blight resistant version of ASD16 pyramided with xa13 and Xa21 developed through markers assisted selectionGall midge resistant Co43 and bacterial blight resistant ASD16 will be crossed with a DRR line (BPT5204 X Tetap) and VRP1 for incorporation of blast resistant genes Pikh and Pi9 respectively Silver shoot in ratoon crop
  • 16. Bt Brinjal• Four elite genotypes, CO2, MDU1, KKM1 and PLR1 were converted into Bt versions by introgression of cry1Ac gene from Mahyco’s elite event through back-cross breeding programme• All Bt genotypes had significantly higher fruit yield as compared to their non-Bt counterparts.• Bt brinjal offer adequate level of resistance to BFSB and show potential for significantly higher marketable yield Co2 MDU1 KKM1 PLR1
  • 17. Indigenous Bt genesLocal isolates of Bacillus thuringiensis collected and evaluated against major lepidopteran pestsNovel Bt genes isolated from promising isolatesProteins from cloned Bt genes were tested by bioassayA novel chimeric cry2A gene madePromising Bt genes (belonging to cry2A group) being introduced in crops cry2A for leaf folder in rice
  • 18. New Bt gene discoveryS. No Gene Year Country1 cry2Aa 1989 USA2 cry2Ab 1989 USA3 cry2Ac 1991 USA4 cry2Ad 1999 Korea5 cry2Ae 2003 USA6 cry2Af 2007 Australia7 cry2Ag 2008 China8 cry2Ah 2008 China9 cry2Ai 2009 TNAU, India
  • 19. Bt CottonInsect Resistant CottonStandardization of regeneration and transformation protocols for Coker genotypes Regeneration ofAgrobacterium-mediated transformed plants Transformation of Coker genotypes with a chimeric cry2A geneIn planta transformation of MCU5, MCU12 and MCU13
  • 20. Bt BrinjalIndigenously isolated, synthetic cry2A genes to control fruit and shoot borerTarget genotypes: CO2, Elavambadi LocalTransgenic plants generated using Agrobacterium method Regeneration of plants from cotyledons Regenerated of plants Transgenic plant
  • 21. Engineering resistance against bananabunchy top virusThe replication of virus is targeted through RNA interference approachRNAi vector targeting the BBTV Rep gene constructedAgrobacterium- mediated transformation of hill banana with developed RNAi construct to impart BBTV resistance
  • 22. Engineering resistance against banana bunchy topvirusMore than 75 putative transgenic hill banana plants were generated and established in transgenic greenhouse.Bioassay results showed that BBTV resistance in all eight transgenic hill banana lines tested
  • 23. Engineering resistance in papayaagainst the papaya ring spot virusViral coat protein gene and replicase genes are suppressed through RNA interference approach to contain virus replicationRNAi vector targeting these genes madeBiolistic and Agrobacterium- mediated transformation of Co7 papaya with the RNAi construct is on-going
  • 24. Genetic engineering of Cassava forresistant to Cassava Mosaic DiseaseRNAi construct targeting the ICMV and SLCMV Replicase geneAgrobacterium-mediated transformation of cassava with RNAi construct is on-going
  • 25. Nutritional Quality
  • 26. Golden Rice IPP• Vitamin A deficiency is prevalent in the developing world Geranylgeranyl diphosphate – responsible for the deaths of Phytoene synthase Maize psy two million children annually Phytoene – deficiency is a leading cause of Phytoene desaturase blindness Erwinia crtl• β-carotene expressed in many ξ-carotene desaturase plants but not in cereal grains Lycopene-beta-cyclase• Introduction of correct β -carotene metabolic steps into rice (vitamin A precursor) endosperm to facilitate β- carotene synthesis• ASD16 and ADT43 converted
  • 27. Iron riceFerritin is a ubiquitous protein that stores iron and releases it in a controlled ADT43 fashionOver-expression of Avecennia marina ferritin in ASD16 rice using an endosperm- specific promoter
  • 28. Low phytate maizeTwenty nine maize inbred lines were selected based on the percent of phytate phosphorus content in total phosphorus content in the maize kernels, as low and high and were included in the crossing programmeSSR Polymorphism survey among the low and high phytate maize inbreds for the lpa1 (10 SSR markers) and lpa2 (10 SSR markers) regions in chromosome1 is under progressContent (mg/g) Reported Variability Observed variability lpa1 / lpa2 mutants ( Raboy et al.2002) in UMI inbreds Low High Low High lpa1 lpa2 Phytate P 2.50 3.50 1.12 4.30 1.10 2.60 Total P 3.50 4.50 2.04 5.07 4.60 4.70
  • 29. Phytase maizeExpression of fungal phytase in maize grain is attempted with a view to make more micronutrients bioavailable to the poultry birdsPhytase gene from Aspergillus niger isolated, expressed in E. coli and characterizedGene construct made to express the phytase in maize endosperm in tissue specific mannerGenetic transformation resulted in development of 26 events Somatic embryos from Callus on shooting Shoots on rooting Immature embryos media medium
  • 30. Tissue cultureMicro-propagation of vegetatively propagated cropsSecondary metabolite production in medicinal plants through suspension culture