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EFFECT OF BACTERIAL CULTURE DENSITY AND ACETOSYRINGONE CONCENTRATION ON AGROBACTERIUM MEDIATED TRANSFORMATION IN WHEAT

HAMID RASHID¹, ASMA AFZAL², M. HAROON KHAN¹, ZUBEDA CHAUDHRY³ AND SALMAN A. MALIK²

Abstract: In the present study high efficiency Agrobacterium tumefaciens mediated transformation system of wheat (Triticum aestivum L.) cv. Inqilab-91 was developed by exploiting bacterial culture density and acetosyringone concentration. Agrobacterium strain EHA101 harboring binary vector pIG121Hm, containing gene for GUS activity and hygromycin resistance was used in transformation experiments. Different optical densities (0, 0.25, 0.5, 0.75 and 1.0) of bacterial suspension and different concentrations (0, 50, 100μM) of acetosyringone were evaluated in transformation studies. Maximum transformation efficiency (4.16%) was obtained with bacterial suspension of O.D._600nm = 0.5. Contamination could not be controlled when O.D._600nm = 0.75 and 1 was used. In case of concentration of acetosyringone, 100μM was found to be better for transformation where transformation efficiency was 15.62%. Transformation results were confirmed with GUS analysis.

¹Department of Bioinformatics, Mohammad Ali Jinnah University, Islamabad, Pakistan.

²Department of Biochemistry, Quaid-i-Azam University, Islamabad, Pakistan

³Department of Botany, Hazara University, Mansehra, Pakistan.