Abstract
Morphologically normal and fertile transgenic chickpea plants have been regenerated through a standardized transformation protocols. This protocol is based on the infection of apical meristem explants (AME) with Agrobacterium strain EHA105. The stain, carrying pCAMBIA2301 vector contained β-glucuronidase (uidA) gene and neomycin phosphotransferase (nptII) genes. Different explants of chickpea and Agrobacterium specific conditions were standardized with the help of transient β-glucuronidase (uidA) gene expression to further optimize the transformation protocol. Pre-conditiong of the explants, vacuum infiltration and presence of acetosyringone significantly enhanced the frequency of gus expression. Positive transformants with nptII and gus genes were confirmed by PCR and histochemical gus analysis. An overall successful chickpea transformation frequency of 1.2 was achieved. This high efficiency and easy to use method may provide opportunities for the development of transgenic lines with different useful genes in chickpea in near future.
Acknowledgements
The authors are thankful to Dr. L. Sahoo, IIT-Guwahati for providing the Agrobacterium strain EHA105 having plasmid pCAMBIA2301. We also acknowledge the help provided by Dr. Bansa Singh, IIPR, Kanpur, in histological studies. The senior research fellowship awarded to J. Srivastava by the Council of Scientific and Industrial Research, New Delhi, is gratefully acknowledged.
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© 2017 Institute of Botany, Slovak Academy of Sciences
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- Cellular and Molecular Biology
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- Accumulation and physiological response of cadmium in Hydrocharis dubia
- Botany
- Development of an efficient Agrobacterium mediated transformation system for chickpea (Cicer arietinum)
- Botany
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