Biotech research at tamil nadu agricultural university 2011
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Transcript of Biotech research at tamil nadu agricultural university 2011
GenesisA manpower development programme in Biotechnology
initiated during 1988 through PG-Biotechnology programme with the support of DBT, GoI.
Plant Molecular Biology and Biotechnology
Major activitiesEducationResearch
EducationUnder Graduate Education
B. Tech. (Biotechnology)B. Tech. (Bioinformatics)
Post Graduate EducationM.Sc. (Biotechnology)M.Tech (Biotech & Business
Management)PhD (Biotechnology)
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
Major Target TraitsAbiotic stress resistance
Drought resistanceSubmergence toleranceSalt tolerance
Biotic stress resistanceInsect resistanceDisease resistance
Nutritional quality
Abiotic Stress Resistance
• 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
Submergence ToleranceAbout 3 lakh ha paddy is affected frequently by submergence
in cauvery delta zoneNecessitates the development of submergence tolerant rice
varietiesMarker Assisted Introgression of Sub1 locus conferring
tolerance to flooding into elite rice varieties of Tamil Nadu
Submergence ToleranceFR13A Sub1 locus was introgressed into a widely grown Asian
rice cultivar “Swarna” using marker-assisted selectionSubmergence tolerant versions of CR1009, Savithri, Samba
Mahsuri and IR 64 are already available for cultivationSubmergence 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
Salt ToleranceIntrogression of salT locus conferring tolerance against salinity
stress into elite rice varieties of Tamil NaduA 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 478White Ponni FL 478
Control 80mM 150 mM Control 80 mM 150 mM 10 days after treatment10 days after treatment
Genetic engineering of sugarcane for abiotic stress toleranceAtDREB1a gene from Arabidopsis thalianaBcZAT12 gene from Brassica carinata
Co86032 transformed with AtDREB1a 20 transgenic lines obtainedCoC671 transformed with BcZAT12 11 transgenic lines obtained
Cotton Varieties with Drought Tolerance and Extra Long Staple FibreObjectives
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 progressMCU5 X KC3 MCU5 X TCH1218 KC3 X Suvin and TCH1218 X Suvin
Insect & Disease Resistance
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
Developing bacterial blight resistant riceXa 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.
A BResistance reaction of susceptible (A) and
pyramided (B) lines against Xoo isolates
Development of gall midge, blast and bacterial blight resistant rice varietiesGall 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
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
Indigenous Bt genesLocal isolates of Bacillus thuringiensis collected and
evaluated against major lepidopteran pestsNovel Bt genes isolated from promising isolatesProteins from cloned Bt genes were tested by
bioassayA novel chimeric cry2A gene madePromising Bt genes (belonging to
cry2A group) being introduced in crops
cry2A for leaf folder in rice
S. No Gene Year Country
1 cry2Aa 1989 USA
2 cry2Ab 1989 USA
3 cry2Ac 1991 USA
4 cry2Ad 1999 Korea
5 cry2Ae 2003 USA
6 cry2Af 2007 Australia
7 cry2Ag 2008 China
8 cry2Ah 2008 China
9 cry2Ai 2009 TNAU, India
New Bt gene discovery
http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/intro.html
Bt CottonInsect Resistant CottonStandardization of regeneration and
transformation protocols for Coker genotypes
Agrobacterium-mediated Transformation of Coker genotypes with a chimeric cry2A gene
In planta transformation of MCU5, MCU12 and MCU13
Regeneration of transformed plants
Bt BrinjalIndigenously isolated, synthetic cry2A genes to
control fruit and shoot borerTarget genotypes: CO2, Elavambadi LocalTransgenic plants generated using Agrobacterium
method
Regeneration of plants from cotyledons
Regenerated of plants Transgenic plant
Engineering resistance against banana bunchy top virusThe replication of virus is targeted
through RNA interference approachRNAi vector targeting the BBTV Rep
gene constructedAgrobacterium- mediated
transformation of hill banana with developed RNAi construct to impart BBTV resistance
Engineering resistance against banana bunchy top virusMore 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
Engineering resistance in papaya against the papaya ring spot virusViral coat protein gene and replicase genes are suppressed
through RNA interference approach to contain virus replication
RNAi vector targeting these genes madeBiolistic and Agrobacterium- mediated transformation of Co7
papaya with the RNAi construct is on-going
Genetic engineering of Cassava for resistant to Cassava Mosaic Disease
RNAi construct targeting the ICMV and SLCMV Replicase gene
Agrobacterium-mediated transformation of cassava with RNAi construct is on-going
Nutritional Quality
Golden Rice• Vitamin A deficiency is prevalent
in the developing world – responsible for the deaths of
two million children annually – deficiency is a leading cause of
blindness• β-carotene expressed in many
plants but not in cereal grains• Introduction of correct metabolic
steps into rice endosperm to facilitate β-carotene synthesis
• ASD16 and ADT43 converted into GR lines
IPP
Geranylgeranyl diphosphate
Phytoene
-carotene(vitamin A precursor)
Phytoene synthase
Phytoene desaturase
Lycopene-beta-cyclase
ξ-carotene desaturase
Maize psy
Erwinia crtl
Iron riceFerritin is a ubiquitous
protein that stores iron and releases it in a controlled fashion
Over-expression of Avecennia marina ferritin in rice using an endosperm-specific promoter
ASD16
ADT43
Low phytate maizeTwenty 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 progress
Content (mg/g) Reported Variability ( Raboy et al.2002)
Observed variability in UMI inbreds
lpa1 / lpa2 mutants
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
Phytase maizeExpression 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
Shoots on rooting medium
Callus on shooting media
Somatic embryos from Immature embryos
Tissue cultureMicro-propagation of vegetatively propagated
cropsSecondary metabolite production in medicinal
plants through suspension culture