Effect of antibiotics on shoot regeneration and suppression of Agrobacterium growth in apple rootstock
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Date
2010
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Publisher
UHF,NAUNI,SOLAN
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.
Description
Keywords
biological phenomena, farmyard manure, cotton, land resources, productivity, sowing, gossypium hirsutum, organic fertilizers, yields, nitrogen