Agrobacterium-mediated rapid and efficient development of transgenics using shoot apex explants in two elite Indica rice cultivars

Sundararajan, Sathish and Nayeem, Safia and Sivakumar, Hari Priya and Ramalingam, Sathishkumar (2023) Agrobacterium-mediated rapid and efficient development of transgenics using shoot apex explants in two elite Indica rice cultivars. CEREAL RESEARCH COMMUNICATIONS, 51 (4). pp. 917-929. ISSN 0133-3720 (print), 1788-9170 (online)

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Abstract

An efficient multiple shoot induction, plant regeneration, and subsequent genetic transformation system for two locally adapted recalcitrant indica rice cultivars ASD16 and ADT43 using shoot apex explants were established. Three different plant growth regulators (PGRs) were tested for their efficacy in forming multiple shoots from shoot apices. Thidiazuron (TDZ) (5.0 mg/L) induced the highest frequencies of multiple shoot induction compared to Benzyl Amino Purine (BAP) and Kinetin (Kin). Shoot apices cultured on MS basal medium devoid of plant growth regulators formed single shoots. All tested PGRs produced multiple shoots at concentrations higher than 3.0 mg/L. Indole-3-Acetic Acid (IAA) and Indole-3-Butyric Acid (IBA) were supplemented to media containing TDZ (5.0 mg/L) for assessing the root induction efficiencies and it was found that media devoid of growth hormones were sufficient to produce better-rooting frequencies comparatively. Among the two tested cultivars, ASD16 responded with better shooting frequencies than ADT43. Further, genetic transformation experiments were carried out with the shoot apex explants of both cultivars. The putative transgenic plants were screened and confirmed by transient GUS expression, PCR, and Southern hybridization in both T0 and T1 generations, respectively. This rapid and efficient regeneration system amenable to Agrobacterium-mediated genetic transformation from shoot apices may be used for genetic manipulation of recalcitrant indica varieties for varietal crop improvement strategies.

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