Optimization of explants for Agrobacterium mediated gene transfer in Eleusine coracana (PES-400)
| dc.contributor.advisor | Lohani, Pushpa | |
| dc.contributor.author | Raingam, Masochon | |
| dc.date.accessioned | 2018-08-23T07:28:25Z | |
| dc.date.available | 2018-08-23T07:28:25Z | |
| dc.date.issued | 2017-08 | |
| dc.description.abstract | Finger millet is the primary food source for millions of people living in tropical dry lands.It posses many desirable properties in terms of adaptability and nutrition. Finger millet can become a hub for fishing out vital genes involved in adaptation to biotic and abiotic stress which can be used for improvement and development of superior crop varieties. For genetic improvement of this important crop, development of efficient protocol for transformation is necessary. In this study, an effort was made to optimize the explants for Agrobacterium mediated gene transfer. Several experimental factors influencing the transformation efficiency were evaluated for four explants; seed, coleoptile, mesocotyl and callus. Among different hormone concentrations and combinations studied, the best callus induction rate for all the explants was observed in 3(2,4 D)+1.5 NAA while the size of the callus was biggest in 2 mg/l NAA .The best proliferation was observed in 1mg/l NAA, 1.5mg /l NAA, 2mg/l NAA and 1mg/l 2,4 D + 1mg/l NAA. Antibiotic sensitivity test for kanamycin showed callus induction and survival in presence of 400 mg/l kanamycin. Hygromycin above 20 mg/l was found to be lethal for finger millet explants. A good Callus induction of explants was observed in presence of 200mg/l-500mg/l cefotaxime. Cefotaxime enhanced the formation of shoot at the concentration of 200 and 250mg/l. Agrobacterium strain LBA4404 harboring pCAMBIA1301 and optimized callus induction media(3mg/l 2,4 D+1.5 mg/l NAA) and antibiotics (300mg/l cefotaxime and 25 mg/l hygromycin for selection) and were used for further optimization of acetosyringone concentration, infection time, co-cultivation time. Infection time of 30 minutes, acetosyringone concentration of 200µM and co-cultivation for three days were found to be optimum for Agrobacterium mediated transformation. Therefore, it can be concluded that callus is the best target explants for Agrobacterium transformation with the efficiency of 15.2 %. | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810067075 | |
| dc.keywords | optimization, explants, Agrobacterium, gene transfer, Eleusine coracana | en_US |
| dc.language.iso | en | en_US |
| dc.pages | 110 | en_US |
| dc.publisher | G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand) | en_US |
| dc.research.problem | Eleusine coracana | en_US |
| dc.sub | Agricultural Biotechnology | en_US |
| dc.subject | null | en_US |
| dc.theme | Molecular Biology and Biotechnology | en_US |
| dc.these.type | M.Sc | en_US |
| dc.title | Optimization of explants for Agrobacterium mediated gene transfer in Eleusine coracana (PES-400) | en_US |
| dc.type | Thesis | en_US |