GENETIC TRANSFORMATION STUDIES IN BELL PEPPER (Capsicum annuum L.)

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dc.contributor.advisorSRIVASTAVA, D.K.
dc.contributor.authorVERMA, SWATI
dc.date.accessioned2017-01-19T16:20:20Z
dc.date.available2017-01-19T16:20:20Z
dc.date.issued2009
dc.description.abstractABSTRACT The research work was conducted to standardize a protocol for genetic transformation in bell pepper. Plant regeneration studies were carried out using two types of explants viz hypocotyl and cotyledon. The cotyledon explants showed high frequency of shoot regeneration (80.95%) on MS medium supplemented with 6.0 mg/l BA and 0.3 mg/l IAA as compared to hypocotyl (only 8%) on MS medium supplemented with 4.5 mg/l BA and 0.1 mg/l IAA. The shoots regenerated after 25-30 days. The regenerated shoots were in the form of profuse rosette, which elongated to about 2-2.5 cm on MS medium supplemented with 2.25 mg/l BA and 2 mg/l GA3. MS medium containing 0.25 mg/l IAA was found to be best for root regeneration (50%). Regenerated plantlets were acclimatized successfully. During kanamycin sensitivity test, kanamycin concentration as low as 20 mg/l was found to be inhibitory to the growth of bell pepper cells. For genetic transformation, disarmed Agrobacterium tumefaciens LBA 4404 strain containing a reporter -glucuronidase (gus) gene in binary vector (pBI 121) system along with kanamycin resistance gene (npt-II) for selection in both bacteria and plant was used for co-cultivation experiment to transfer gus and npt-II genes in bell pepper cells (cotyledon and hypocotyl explants). After co-cultivation only the transformed cells were able to grow on selective shoot regeneration medium which formed callus and regenerated shoots, whereas non transformed cells/explants died on the selective medium. The elongation of the putative transgenic shoots was achieved on selective elongation medium. Root regeneration in the putative elongated transgenic shoots was obtained on selective root regeneration medium. Out of the total 15 putative transgenic shoots developed, 5 were randomly selected for PCR analysis using designed primers for npt-II and gus genes. Only two shoots/plantlets showed amplified gene separately indicating the presence of npt-II and gus gene(s) in the transgenic shoots/plantlets. Expression of the gus gene was confirmed by GUS assay. The PCR positive shoots/plantlets have shown the expression of gus gene as well. A protocol for genetic transformation in bell pepper has been standardized.en_US
dc.identifier.other47119
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/97184
dc.language.isoenen_US
dc.subBiotechnology and Molecular Biology
dc.titleGENETIC TRANSFORMATION STUDIES IN BELL PEPPER (Capsicum annuum L.)en_US
dc.typeThesisen_US
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