+ All Categories
Home > Documents > Genetic transformation of sunflower using TVD 1 for ...lternaria leaf spot is an economical disease...

Genetic transformation of sunflower using TVD 1 for ...lternaria leaf spot is an economical disease...

Date post: 09-Jul-2020
Category:
Upload: others
View: 0 times
Download: 0 times
Share this document with a friend
1
BIOTECHNOLOGY-2011 579 21-23 March 2011 World Congress on Biotechnology A lternaria 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. e Agrobacterium containing PCAM 2300 vector with TVD1 genes are cocultivated with sunflower cotyledons from mature seeds. (split seeds through embryonal axis). e explants were kept for shoot regeneration(2BI supplemented with BAP 0.5mg/l and 2ip 1mg/l) and selection. e TVD1 and gene has npt gene for selection. e 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. e rooted shoots were acclimatizated and rootless microshoots were graſted 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. Genetic transformation of sunflower using TVD 1 for resistance against Alternaria leaf spot disease Ch.Sirisha, M.Sravani, D.Sri Rami reddy and N.M.Yugandhar Andhra University, India doi:10.4172/2155-952X.1000001
Transcript
Page 1: Genetic transformation of sunflower using TVD 1 for ...lternaria leaf spot is an economical disease of sunflower causing serious yield losses and reduced oil content. Sources of resistance

BIOTECHNOLOGY-2011

579

21-2

3 M

arch

2011

World Congress on

Biotechnology

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.

Genetic transformation of sunflower using TVD 1 for resistance against Alternaria leaf spot diseaseCh.Sirisha, M.Sravani, D.Sri Rami reddy and N.M.YugandharAndhra University, India

doi:10.4172/2155-952X.1000001

Recommended