GENETIC TRANSFORMATION FOR FRUIT BORER RESISTANCE USING cry1Ab GENE IN TOMATO (Lycopersicum esculentum MILL.)
Loading...
Date
2003
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
University of Agricultural Science, Dharwad
Abstract
Genotypic response for in vitro callus induction and regeneration was studied in
six-tomato genotypes viz., Pusaruby, Megha (LI5), PKMl, Dharwad local, SI40 and
SI42. Different concentrations and combinations of 2,4-D, kinetin and lAA were used to
induce callus and plantlet regeneration. Highly significant differences among genotypes
and hormonal concentrations were noticed for callus induction and regeneration. A 2,4-D
concentration of 2 mg/1 was found to induce highest frequency of good callus among
genotypes. Different calli quality parameters were significantly influenced by the
genotype, explant and 2,4-D concentration in the medium.
Highest regeneration frequency from callus was obtained in MS basal
supplemented with 3 mg/1 of kinetin and 0.3 mg/I of lAA. Megha found to be highly
responsive for regeneration through callus. However, kinetin @ 5mg/l induced highest
frequency of direct plantlet regeneration in cotyledonary leaf and hypocotyl explants
which responded differently across genotypes. Sufficient and healthy i/i vitro root
induction was achieved on MS basal supplemented with NAA (0.1 mg/1) and lAA
(0.3rag/l). A plantlet survival of more than 95% was recorded during hardening process.
Agrobacterium EHA 105 carrying pCAMBIA 1301 with uidA (GUS) and
hygromycin selectable gene and pBinBtl with cryl Ab along with nptll gene, both driven
by CaMV 35S promoter were used for transformation. Different parameters of
transformation process were optimized using i« gene. Cefotaxime @ 300 mg/1 was used
to eliminate Agrobacterium. A repeatable average transformation frequency of 2.32%
was recorded in Megha. PCR assay using nptll specific primers and insect bioassay
studies confirmed the transgenic status of Megha. RT-PCR analysis indicated the
expression of transgene in tomato leaves.
Description
Keywords
null
Citation
No. of references120