Effect of antibiotics on shoot regeneration and suppression of Agrobacterium growth in apple rootstock
| dc.contributor.advisor | MODGIL, MANJU | |
| dc.contributor.author | CHAUHAN, ARJUN | |
| dc.date.accessioned | 2017-02-06T09:37:36Z | |
| dc.date.available | 2017-02-06T09:37:36Z | |
| dc.date.issued | 2010 | |
| dc.description.abstract | Among 24 combinations of BA/TDZ and NAA/IAA, 1 mg/l of IAA and 4.0 mg/l of BA proved better and resulted in direct shoot regeneration (22.13%) of leaf explants. Antibiotics viz. cefotaxime, carbenicillin, timentin, augmentin and rifampicin were used to see their effect on callus/shoot regeneration, growth of bacteria in suspension cultures & for suppressing Agrobacterium tumefaciens growth during genetic transformation studies in apple rootstock MM111 using strain LBA4404 harboring the chitinase gene of rice within the vector pCAMBIA chi 11 with hygromycin phosphotransferase (hpt) as a selectable marker. All the antibiotics increased the callus induction with the increase in their concentration upto certain extent and declined at higher concentration. However, browning of leaf explants increased with the increase in concentration. Maximum callusing (85 – 93%) was observed in 300 mg/l Carb., 200 mg/l Aug. & 100-200 mg/l Tim., but no browning of leaf explants occurred only on Tim. Cefo. & Rif. showed reduced callus formation. It was also found that cfu/ml and OD values at 540 nm showed decrease with the increase in concentration of antibiotics added in bacterial suspension culture. Among all the antibiotics, Tim. & Aug. comparatively showed rapid decrease in OD values as the concentration increased of. No colony was observed in Tim. at 200 – 500 mg/l and Carb. & Aug. at 500 mg/l. Tim.(200 mg/l) and Aug.(300 mg/l) completely controlled the bacteria after co-cultivation even after 1st blot. At 500 mg/l, Cefo. showed the Agrobacterium growth in 25.07% of explants, whereas Carb. and Rif. showed no growth after 1st blot. But these antibiotics showed deleterious effects on the leaf explants at this concentration. Therefore, timentin(100-200 mg/l) and augmentin(300 mg/l) can be recommended for suppression of Agrobacterium growth during transformation experiments in apple rootstock MM111. | en_US |
| dc.identifier.other | 47332 | |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810000302 | |
| dc.keywords | apple rootstock MM111,Agrobacterium tumefaciens, | en_US |
| dc.language.iso | en | en_US |
| dc.publisher | UHF,NAUNI,SOLAN | en_US |
| dc.sub | Biotechnology | en_US |
| dc.subject | biological phenomena, farmyard manure, cotton, land resources, productivity, sowing, gossypium hirsutum, organic fertilizers, yields, nitrogen | en_US |
| dc.theme | APPLES, GENETICS,MOLECULAR BIOLOGY | en_US |
| dc.these.type | M.Sc | en_US |
| dc.title | Effect of antibiotics on shoot regeneration and suppression of Agrobacterium growth in apple rootstock | en_US |
| dc.type | Thesis | en_US |