• Diksha R. Jawale
• Sanjay S. Moharekar
• Shubhangi S. Moharekar

In the present study, Agrobacterium was isolated from rhizosphere soil obtained from Botanical Garden of New Arts Commerce and Science College, Ahmednagar by using selective YEM medium. On the basis of morphological, Biochemical and Gram characters and comparing them with Bergy’s Manual of Determinative Biology, it was confirmed as Agrobacterium. Isolate was further characterized for presence of Ti plasmid. An Alternative Boiling of Miniprep method is used for Plasmid isolation which is very efficient method and gives results in short time and gives reproducible isolation of Ti plasmid. Sensitivity to different antibiotics viz: Cefotaxime, Streptomycin and Penicillin was done by disc diffusion method and it showed high sensitivity towards Cefotaxime, moderate sensitive towards Streptomycin, and slight resistant towards Penicillin. Effect of various temperatures (25oC, 35oC and 40oC) on growth of Agrobacterium and its plasmid was checked and it was found that high temperature (40oC) affected growth of Agrobacterium but had no effect on plasmid. To our knowledge it was first attempt to investigate the stability of the Ti plasmid at these temperatures which reveals that Ti plasmid of this isolate is stable even at 40⁰C. Further, the Restriction Fragment Length Polymorphism (RFLP) analysis showed no fragment patterns on agarose gel for EcoRI, HindIII and BamHI which clearly indicates that there are no restriction sites for these endonucleases.

Keywords: Agrobacterium, Ti-plasmid, Antibiotic sensitivity, Temperature optimization, RFLP analysis

"Investigative study of Agrobacterium isolated from Rhizosphere soil", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.9, Issue 2, page no.b262-b270, February-2022, Available :http://www.jetir.org/papers/JETIR2202131.pdf

"Investigative study of Agrobacterium isolated from Rhizosphere soil", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.9, Issue 2, page no. ppb262-b270, February-2022, Available at : http://www.jetir.org/papers/JETIR2202131.pdf