Journal of Biotechnology & Biomaterials
Open Access

Abstract

Alternaria leaf spot is an economical disease of sunflower causing serious yield losses and reduced oil content. Sources of resistance to this pathogen are limited in the cultivated germplasm of sunflower. In the present study, attempts have been made to transform sunflower using the defensin gene TVD1 . TVD1 gene extracted from Tephrosia villosa has potent antifugal activity and confers protection againest several pathogens including Alternaria leaf spot in sunflower. Incorporation of TVD1 gene into sunflower is achieved by Agrobacterium mediated genetic transformation method. The Agrobacterium containing PCAM 2300 vector with TVD1 genes are cocultivated with sunflower cotyledons from mature seeds. (split seeds through embryonal axis). The explants were kept for shoot regeneration(2BI supplemented with BAP 0.5mg/l and 2ip 1mg/l) and selection. The TVD1 and gene has npt gene for selection. The transformed tissues showed resistance to kanamycin in medium and remained green while the non-transformed tissue were bleached on kanamycin medium. Following shoot regeneration they were transferred for rooting. The rooted shoots were acclimatizated and rootless microshoots were grafted and finally transferred to green house. DNA was isolated from the putative transformed plants and PCR analysis was done with primers specific to the gene of interest and also the selectable marker and positives for both TVD1 and npt were identified.

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