PJB-2019-528
Isolation, characterization and evolution of wild virulent strains of Agrobacterium for their potential transformation through use of potato discs
Muhammad Rafique Rind, Aneela Yasmin, Saboohi Raza and Wajid Ali Jatoi
Abstract
The present study was aimed to investigate the Crown galls of the trees and plants caused by Agrobacterium. For the purpose, the Crown galls were collected from Botanical and Horticultural Gardens of Sindh Agriculture University, Tandojam, Pakistan and examined for Agrobacterium species. In all 90 Crown galls were examined for Agrobacterium species. The Crown galls extracts were obtained and streaked on PYGA (Peptone, Yeast extract, Glycerol, Agar) and Luria–Bertani (LB) media supplemented with rifampicin (30 µl L-1) antibiotic to inhibit the growth of other bacterial organisms and the plates were incubated at 28°C for 48 hours. Only thirty (33.333%) wild isolates of the Agrobacterium tumefaciens were isolated from Crown galls. However, nineteen (19) isolates out of 30 were confirmed as Agrobacterium tumefaciens. Among these 19 isolates, At224 was found to be the most virulent strain which showed 80% transformation efficiency (TE) while other three isolates i-e At213-TE: 70%; At222-TE: 70% and At230-TE:70% were induced solid green cell masses/tumors/ galls on the potato discs vigorously. Of the tested isolates, the isolate At224 (Mangifera indica) was the most virulent strain that caused 80% transformation as compared to other isolates. Furthermore, when comparison was made among isolates recovered from Crown galls and that of controlled for phenotypical characteristics, the majority of isolates showed similar characteristics as demonstrated in the controlled (At 2441- EHA101). Moreover, Ti plasmid was found visible in by using 0.8% in 50 ml of TAE buffer solution. Thus Ti plasmid that helped in for the characterization of wild Agrobacterium by molecular system
To Cite this article:
Rind, M.R., A. Yasmin, S. Raza and W.A. Jatoi. 2020. Isolation, characterization and evolution of wild virulent strains of Agrobacterium for their potential transformation through use of potato discs. Pak. J. Bot., 52(6): DOI: http://dx.doi.org/10.30848/PJB2020-6(34)
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