ISSN 2756-3707
International Journal of Histology and Cytology ISSN 2756-3707 Vol. 7 (2), pp. 001-007, February, 2020. © International Scholars Journals
Full Length Research Paper
Analysis and optimization of DNA delivery into chickpea (Cicer arietinum L.) seedlings by Agrobacterium tumefacience
Mikail Akbulut1*, Meral Yücel2, Hüseyin Avni Öktem2
1Department of Biology, Erciyes University, 38039, Kayseri, Turkey.
2Department of Biology, Plant Biotechnology R&D Laboratories, Middle East Technical University, 06531 Ankara, Turkey.
Accepted 17 November, 2019
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 β-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).
Key words: Chickpea, transformation, Agrobacterium, vacuum infiltration, transient expression.