Analysis and optimization of DNA delivery into chickpea (Cicer arietinum L.) seedlings by Agrobacterium tumefacience

Abstract

The main purpose of this study was to develop a non-tissue culture based Agrobacterium mediated transformation method for chickpea. The influences of several factors were investigated on the transfer
of -glucuronidase (GUS) gene into chickpea (Cicer arietinum) seedlings during the early stages of Agrobacterium-mediated gene transfer, including cocultivation period in liquid induction medium (2, 8,
16 and 24 h), strains of Agrobacterium tumefaciens (C58C1, EHA105, KYRT1) containing the plasmid pTJK136, developmental stage (16 h imbibed and 40 h germinated), microwounding, vacuum infiltration
(200, 400, 600 mmHg for 20 and 40 min) and genotype (5 different). The number of GUS-expressing foci was counted to evaluate the gene transfer process. The KYRT1/pTJK136 strain of A. tumefaciens was
significantly more effective for transformation than the  58C1/pTJK136 and EHA105/pTJK136 strains. The highest transient GUS activity was obtained from 16  h imbibed seedlings of cv.Uzunlu wounded
with a needle and co-cultivated in liquid induction medium for 24 h with the KYRT1 strain (226 GUS foci/per explant).