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Genetic Transformation for Drought Resistance in Cotton

KrishiKosh

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Title Genetic Transformation for Drought Resistance in Cotton
 
Creator Prashanth Sangannavar
 
Contributor I.S. Katageri
 
Subject Genetics & Plant Breeding
 
Description Different concentrations and combinations of growth regulators supplemented to MS
medium were tried to study the callogenesis from cotyledon and hypocotyl explants of
Coker-312 cotton. The media combination of 0.1 mg/l 2, 4-D and 0.5 mg/l kinetin resulted in
early callus initiation (10days), high per cent callus induction (99%) and high amount of fresh
callus weight (0.88g) with cream friable calli. Irrespective of growth regulators hypocotyl, is
better explant than cotyledon for callus initiation, per cent callus induction and fresh callus
proliferation. Higher callus induction (94%) was observed in 3% glucose. Reduction of 2, 4-
D (0.01mg/l) and kinetin (0.1mg/l) resulted in highest per cent of embryogenesis (71%).
Embryo maturation from torpedo embryo’s to plantlets was highest (95%) in media devoid of
growth regulators. Incubation in basal MS medium for 4 weeks in in vitro culture condition
followed by 1 week incubation in growth chamber in soil and peat mixture resulted in
establishment of higher number of plants (95%).
Agrobacterium strain LB-4404 carrying pCambia with AtDREB1a and pBinAR with
BcZAF12 transcriptional factors were used for genetic transformation. Colonization for 10
minutes followed by 24 hours co-cultivation resulted in explants free of Agrobacterium
contamination. Cefotaxime at 1000 mg/l showed complete elimination of excess
Agrobacterium from explant surface. Pre-culture of explants for 48 hours prior to
transformation resulted in highest number of kanamycin resistant calli (29). Colonization of
Agrobacterium with vacuum infiltration for 30 minutes resulted in kanamycin resistant calli
(28).
Analysis of the putative transformants for the presence and expression of gene
revealed that the transformation efficiency was 2% and 0.6% respectively for AtDREB1a and
BcZAF12 transcriptional factors in transformation via somatic embryogenesis in Coker-312,
and transformation efficiency of 1.4% and 2.5% respectively for AtDREB1a and BcZAF12 in
transformation via in planta in Sahana was recorded. RT-PCR and dot blot confirmed
expression and integration of genes for transcriptional factors in plants.
 
Date 2016-07-26T09:52:15Z
2016-07-26T09:52:15Z
2013
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/69829
 
Format application/pdf
 
Publisher UAS Dharwad