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Development of an Agrobacterium-delivered codon-optimized CRISPR/Cas9 system for chickpea genome editing by Santosh Kumar Gupta; Niraj Kumar Vishwakarma; Paheli Malakar; Poonam Vanspati; Nilesh Kumar Sharma; Debasis Chattopadhyay is a Agricultural and Biological Sciences article available to read on EtoBox.

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Chickpea is considered recalcitrant to in vitro tissue culture amongst all edible legumes. The clustered, regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-based genome editing in chickpea can remove the bottleneck of limited genetic variation in this cash crop, which is rich in nutrients and protein. However, generating stable mutant lines using CRISPR/Cas9 requires efficient and highly reproducible transformation protocols. As an attempt to solve this problem, we developed a modified and optimized protocol for chickpea transformation. This study transformed the single cotyledon half-embryo explants using CaMV35S promoter to drive two marker genes (β-glucuronidase gene; GUS and green fluorescent protein; GFP) through binary vectors pBI101.2 and modified pGWB2, respectively. These vectors were delivered in the explants through three different strains of Agrobacterium tumefaciens, viz., GV3101, EHA105, and LBA4404. We found better efficiency with the strain GV3101 (17.56%) compared with two other strains, i.e., 8.54 and 5.43%, respectively. We recorded better regeneration frequencies in plant tissue culture for the constructs GUS and GFP, i.e.

Who reads Development of an Agrobacterium-delivered codon-optimized CRISPR/Cas9 system for chickpea genome editing?

Author

Santosh Kumar Gupta; Niraj Kumar Vishwakarma; Paheli Malakar; Poonam Vanspati; Nilesh Kumar Sharma; Debasis Chattopadhyay

Publisher

Springer Science and Business Media LLC

Published

2023

Language

EN

Field

Agricultural and Biological Sciences (Life Sciences)