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    Callus induction and efficient in vitro plant regeneration protocol for Chickpea
    (Springer Nature Publishing AG, 2024) Jangid, Vinod Kumar; Senthil-Kumar, Muthappa; Chandran, Divya; Sinharoy, Senjuti
    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.
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    Genotype independent regeneration and agrobacterium-mediated genetic transformation of sweet potato (Ipomoea batatas L.)
    (Bangladesh Association for Plant Tissue Culture and Biotechnology, 2013) Shekhar, Shubhendu; Agrawal, Lalit; Buragohain, Alak Kumar; Datta, Asis; Chakraborty, Subhra; Chakraborty, Niranjan
    Development of an efficient genotype independent regeneration and genetic transformation system in sweet potato continues to be of great interest. Agrobacterium‐mediated genetic transformation protocol was established in two different cultivars of sweet potato using Agrobacterium strain EHA105 harbouring binary plasmid pBI121 containing GUS and nptII genes. The internodal stem segments from 30‐day‐old micropropogated plants were used as explant with different combinations of media and hormones. MS and LS media with various concentrations of growth regulators proved to be non‐responsive and the infecundity was severe with the addition of cytokinins. Nonetheless, MS with 2,4‐D and TDZ gave a good percentage of callusing but with low differentiation. In different concentrations of NAA, significant amount of callusing was observed but percentage of rooting remained low in both the genotypes. Gamborg’s B5 supplemented with NAA proved to be the most suitable media and hormone combination, which yielded shoot formation after 8 ‐ 10 weeks with a regenera‐ tion efficiency of 40 ‐ 70%. Stable integration of transgene was confirmed by PCR analysis. Furthermore, qRT‐PCR analysis was performed to assess the transcript accumulation in addition to the GUS enzymatic assay in the transgenic lines.