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Full Length Research Paper
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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.
*Corresponding author.
E-mail:
akbulut@erciyes.edu.tr.
Accepted 17 March, 2008 |
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Abstract |
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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. |
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