Wild-type Agrobacterium rhizogenes-mediated gene transfer in plants: Agrobacterium virulence and selection of transformants

Agrobacterium rhizogenes ATCC 15834 wild type strain was transformed with the binary vector pBI121 using the heat shock method. The transformed Agrobacterium was then tested for virulence through tobacco leaf explant transformation. Compared to the non-transformed Agrobacterium, the transformed Agrobacterium showed reduced virulence, producing signifi cantly lower number of hairy roots in tobacco leaf explants. Although the transformed Agrobacterium showed reduced virulence, it was able to transfer the T-DNA of the binary vector into the plant genome, resulting in stable GUS expression in the generated hairy roots. This indicated that in addition to the transfer DNA (T-DNA) from its root inducing (Ri) plasmid, the transformed Agrobacterium is also capable of transferring the binary vector T-DNA and allowing the integration of a foreign gene. Results also showed that hairy root generation effi ciency of the transformed Agrobacterium varied with the concentration of the selection agent (kanamycin). Hairy root generation effi ciency (hairy roots·explant-1) progressively increased with decreasing concentrations of kanamycin; and the effi ciency was highest in the absence of kanamycin. Generated hairy roots showed very strong to tiny GUS expression even those that grew under the highest concentration of the kanamycin (50 mg·L-1). This indicated that co-transformation and effi cient transgene expression does not always occur. Research Article Wild-type Agrobacterium rhizogenesmediated gene transfer in plants: Agrobacterium virulence and selection of transformants Mohammad M Rana1,2, Muhammad Abdullah1, Ferdinand L Shamalla1 and Shu Wei1* 1State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Ave W Hefei, Anhui 230036, China 2Bangladesh Tea Research Institute, Srimangal-3210, Moulvibazar, Bangladesh *Address for Correspondence: Shu Wei, State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Ave W, Hefei, Anhui, 230036, China, Tel & Fax: +86-551-65783941; Email: [email protected] Submitted: 27 May 2017 Approved: 09 June 2017 Published: 12 June 2017 Copyright: 2017 Rana MM, et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.