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

AKBULUT M., Yucel M., Oktem H. A.

AFRICAN JOURNAL OF BIOTECHNOLOGY, vol.7, no.8, pp.1011-1017, 2008 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 7 Issue: 8
  • Publication Date: 2008
  • Journal Name: AFRICAN JOURNAL OF BIOTECHNOLOGY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.1011-1017
  • Keywords: chickpea, transformation, Agrobacterium, vacuum infiltration, transient expression, MEDIATED GENE-TRANSFER, GENERATING TRANSGENIC PLANTS, LENS-CULINARIS MEDIK, TISSUE-CULTURE, TRANSFORMATION SYSTEM, ARABIDOPSIS-THALIANA, VACUUM INFILTRATION, BETA-GLUCURONIDASE, GRAIN LEGUMES, EXPRESSION
  • Middle East Technical University Affiliated: Yes

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 C58C1/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).