Robust Genetic Transformation System to Obtain Non-Chimeric Transgenic Chickpea
- 1Queensland University of Technology, Australia
- 2Centre for Tropical Crops and Biocommodities, Institute for Future Environments, Queensland University of Technology, Australia
- 3Agriculture & Food, Commonwealth Scientific and Industrial Research Organisation, Australia
Chickpea transformation is an important component for the genetic improvement of this crop through modern biotechnological approaches. However, recalcitrant tissue culture and occasional chimerism encountered during transformation hinder efficient generation of transgenic chickpeas. Two key parameters, namely micro-injury and light emitting diode (LED)-based lighting were used to increase transformation efficiency. Early PCR confirmation of positive in vitro transgenic shoots, together with efficient grafting and an extended acclimatisation procedure contributed to the rapid generation of transgenic plants. High intensity LED light facilitate chickpea plants to complete their life cycle within 9 weeks thus enabling up to two generations of stable transgenic chickpea lines within 8 months. The method was validated with several genes from different sources, either as single or multi-gene cassettes. Stable transgenic chickpea lines containing GUS (uidA), stress tolerance (AtBAG4 and TlBAG), as well as Fe-biofortification (OsNAS2 and CaNAS2) genes have been successfully produced.
Keywords: Agrobacterium, Transgenic Chickpea, Chimeric Chickpea, Micro-Injury of In Vitro Explants, LED light, Legume transformation
Received: 24 Nov 2018;
Accepted: 04 Apr 2019.
Edited by:Roger Deal, Emory University, United States
Reviewed by:Rohini Garg, Shiv Nadar University, India
Sergio J. Ochatt, INRA UMR1347 Agroécologie, France
Copyright: © 2019 Das Bhowmik, Cheng, LONG, Tan, Hoang, Karbaschi, Williams, Higgins and Mundree. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Prof. Sagadevan G. Mundree, Centre for Tropical Crops and Biocommodities, Institute for Future Environments, Queensland University of Technology, Brisbane, 4000, Queensland, Australia, firstname.lastname@example.org