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Studies on in vitro regeneration and genetic transformation in chilli (capsicum annuum L.)

KrishiKosh

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Title Studies on in vitro regeneration and genetic transformation in chilli (capsicum annuum L.)
 
Creator S.B.Channappagoudar
 
Contributor S.A.Patil
 
Subject Genetics Plant Bredding
 
Description In vitro regeneration and Agrobacterium-mediated genetic transformation was
studied in popular local cultivar of chilli viz., Byadagi Kaddi. Different explants (cotyledon,
hypocotyls and cotyledonary nodal region) were cultured on MS medium supplemented with
various levels of BAP (2–12 mg/l) and TDZ (0.1-1.0 mg/l) for adventitious shoot induction.
Frequency of adventitious shoot induction was significantly more in cotyledonary nodal
region explant than cotyledon and hypocotyls explants. Adventitious shoot induction
response was high on medium with 0.5 mg/l TDZ. Addition of 1 mg/l of GA3 to this medium
resulted in higher frequency of shoot elongation. The elongated shoots were rooted on MS
medium with 0.5 mg/l IAA and 0.1 mg/l BAP.
Different concentrations and combinations of growth regulators were tried to study
callus induction and regeneration from cotyledon, hypocotyls and cotyledonary nodal region
explants. The media combination of 0.5 mg/l 2, 4-D and 0.5 mg/l kinetin resulted in higher
quantity of callus. Elimination of 2, 4-D from this media induced somatic embryos. Complete
regeneration was also observed on the same media but with very low frequency (10%).
Shoot apical meristems were co-cultivated with Agrobacterium strain EHA105
carrying pBinBt3 construct having cry1Ac gene. The explants pre-cultured for 48 hours,
infected with agrobacterial suspension having 0.5 × 108 cells/ml for 10 min along with
acetosyringene @ 200 μM for 48 hours yielded the transformants under in vitro conditions.
Analysis of the putative transformants for the presence and expression of cry gene revealed
that the transformation efficiency was 0.5 per cent. In two step RT-PCR amplicon of 1000 bp
revealed the expression of transformed gene.
 
Date 2016-07-22T15:10:53Z
2016-07-22T15:10:53Z
2007
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/69375
 
Format application/pdf
 
Publisher UAS Dharwad