Callus induction and efficient in vitro plant regeneration protocol for Chickpea

Abstract

The development of an efficient and consistent callus-mediated in vitro regeneration protocol is crucial for biotechnological approaches aimed at improving chickpea, an economically important crop legume. In this study, we assess the effectiveness of callus-mediated regeneration in different chickpea genotypes. Through in vitro screening of explants, we identified the Indian cultivar Pusa 240 as a favourable genotype with higher efficiency of somatic embryogenesis and in vitro plant regeneration. Building upon this finding, we have successfully established two distinct protocols for chickpea callus-mediated somatic embryogenesis, utilizing leaf and hypocotyl explants obtained from the Pusa 240 genotype. These protocols achieved plant regeneration efficiencies of 27% using leaf explants and 46.6 − 66% using hypocotyl explants. Extensive literature review and comparative analysis underscored the superiority of our current protocol. Subsequently, the regenerated plants were successfully acclimatized and transferred to the greenhouse, exhibiting normal phenotypic growth. This detailed regeneration method will provide a valuable resource for chickpea genetic transformations and the generation of large mutant populations where embryogenesis via callus formation is required. The protocol presented here establishes a powerful tool for studying the functional genomics of chickpea plants and lays the foundation for future advancements in this field.