Transgenic Expression of Stress Tolerance Gene in Sugarcane (Saccharum officinarum L.)

Abstract

Abstract

Agrobacterium tumefaciens LBA4404 mediated transformation of sugarcane variety 31 with two plasmids of pB1121 constructs, one containing PsCBL gene and the other containing PsCIPK gene was studied for stress tolerance capability induction. Calli of the variety Isd 31 were infected with both plasmid containing strains using three different infection times of 60, 90 and 120 min in acetosyringone infection medium. In GUS (β-glucuronidase) assay, randomly selected calli from 120 min infection time for PsCBL gene showed 100% of callus GUS positive, and for PsCIPK gene, it was 98%. The expression of these two genes was confirmed by the expression of GUS reporter gene present in the construct. Transformed calli from Isd 31 showed lower levels of regeneration response. Regenerated plantlets derived from transformed calli of sugarcane varieties Isd 31 were used as plant material for DNA isolation. The GUS gene transformed in the plant genome was detected by amplifying through two sets of primer: Primer set 1 amplifies 600 bp DNA segment from GUS gene and Primer set 2 amplifies 960 bp DNA segment from GUS gene. The transformed plantlets produced bands of the expected size 600 bp (primer set 1) and 960 bp (primer set 2) for the GUS fragment at the same position. No band was produced in DNA isolated from the non-transformed sugarcane control plants. The stably attached transgenic plants showed stress tolerance behaviors.

Keywords: Agrobacterium, sugarcane, callus, GUS assay, PCR

Cite this Article

Md Munan Shaik, Mst Monira Khaton, Md Amzad Hossain et al. Transgenic Expression of Stress Tolerance Gene in Sugarcane (Saccharum officinarum L.). Research & Reviews: Journal of Agricultural Science and Technology. 2017; 6(2): 32–39p.