1Deptt. of Molecular Biology and Biotechnology, CCS Haryana Agricultural University, Hisar-125004, India
Department of Molecular Biology and Biotechnology, M.P. University of Agriculture & Technology, Udaipur-313 001, India
*Corresponding author e-mail: vinodsaharan@gmail.com
Online published on 28 March, 2013.
High frequency stable GUS gene expression has been achieved in three chickpea varieties. Standardized regeneration protocol from epicotyl explant has been used for regeneration of transformed GUSexpressing shoots of chickpea. Agrobacterium tumefaciensstrain EHA 105 containing vector pCAMBIA 2301 and strain LB 4404 harboring pBI121 were used in present study. Infection time (15 min.), co-cultivation duration (72 hrs.), acetosyringone (200 μM) were found to be optimum for transient and stable GUS gene expression via pCAMBIA vector. Maximum percentage of stable GUS gene expression was recorded 33.3% in microshoots of Pratap-1, followed by 19.0% in microshoots of GNG-469. Efforts have been made to perform PCR for regenerated microshoots to confirmed stable GUS and npt-II gene integration in transformed microshoots. Present protocols developed for popular local varieties of chickpea can be used for transfers of genes for varietal improvement programme.
Agrobacterium tumefaciens, Epicotyl explants, GUS gene