RT Journal T1 TRANSFORMATION OF THE OSC3H52 GENE PROMOTER IN TISSUE CULTURE OF SUGARCANE A1 M. S. Khan1 A1 W. Ahmad A1 M. Z. Ahmad A1 M. Ullah A1 A. Jan JF Journal of Animal and Plant Sciences JO JAPS SN 1018-7081 VO 31 IS 1 SP 156 OP 165 YR 2020 FD 2020/08/26 DO DOI https://doi.org/10.36899/JAPS.2021.1.0203 AB

Sugarcane growth is negatively affected by abiotic stresses. Genetic engineering of sugarcane with abiotic stress-responsive genes has been the method of choice to confer abiotic stress tolerance. In the present work, we optimized callus induction and transformation of the sugarcane variety CP-77-400 with the promoter region of the OsC3H52 gene to analyze its regulatory function under drought and salt stress. Calli were induced using different callus induction media (CIM). The promoter region of the OsC3H52 gene that belongs to the zinc finger protein family was cloned into two different expression vectors i.e. pBI221 and pGreenII0129, which were subsequently transformed to sugarcane calli through agrobacterium and biolistic transformation methods. Among various callus induction media, CIM3 (5 mg L-1 2,4-D + 10% coconut water) showed high callus induction (93%). Biolistic transformation using recombinant pGreenII0129 at 8.5µg/µl concentration resulted in higher efficiency of transformation (28%). While the agrobacterium mediated transformation using recombinant pBI221 plasmid gave 13% transformation efficiency. Amongst the various concentrations of acetosyringone used in agrobacterium mediated transformation, 100 µM showed higher efficiencies (13%) of transformation. Selection of the transformed calli was performed at 25 mg L-1 Hygromycin and 25 mg L-1 Hygromycin + 300 mg L-1 cefotaxime for 15 days, both for biolistic and agrobacterium mediated transformation, respectively. Transformed calli were then successfully confirmed through PCR and GUS expression. The regulatory role of the OsC3H52 promoter in transgenic calli was further evaluated under drought and salt stress. Quantitative real-time PCR analysis of the GUS reporter gene showed high expression in transgenic calli under drought and salt stress conditions.

K1 Abiotic stress, Callus induction, Gene Transformation, GUS Analysis, Saccharum officinarum PB Pakistan Agricultural Scientists Forum LK https://thejaps.org.pk/AbstractView.aspx?mid=AG-19-0319