Optimization of Rice (Oryza sativa L.) ‘Tubtim Chumphae', for Callus Induction, Proliferation and Plantlet Regeneration

Worasitikulya Taratima, Kongtong Plaikhuntod, Chanpen Wichachai and Pitakpong Maneerattanarungroj

Background and Objective: Tissue culture techniques are now used to increase rice yield and quality. This technology offers wide-ranging plant improvements including development, preservation and genetic variability. A rapid and reliable protocol for callus production and plantlet regeneration of Thai rice (Oryza sativa L.) cultivar Tubtim Chumphae was investigated. Materials and Methods: Murashige and skoog (MS) medium supplemented with various concentrations of 2,4-D (0, 1, 1.5, 2 and 2.5 mg L^–1) and BAP (0, 0.1 and 0.5 mg L^–1) was used for callus induction from mature seed. High callus formation was recorded in MS medium containing 2.5 mg L^–1 2,4-D alone and medium with 2 mg L^–1 2,4-D in combination with 0.5 mg L^–1 BAP, giving callus induction at 96.42 and 92.86%, respectively. The effect of chitosan on callus growth was determined using an MS medium supplemented with 2 mg L^–12,4-D and 0.5 mg L^–1 BAP, combined with chitosan concentrations of 0, 25, 50 and 100 ppm. Results: Results indicated that 50 ppm chitosan treatment was the most suitable for callus growth. Three weeks old calli were cultured on MS medium comprising 0.5 mg L^–1 NAA with various concentrations of BAP (0, 1, 2, 3, 4 and 5 mg L^–1). The highest regeneration performance was obtained in 0.5 mg L^–1 NAA with 3 mg L^–1 BAP, with new plantlets completely regenerated within 4 weeks. Conclusion: These research results can be used as a starting point for in vitro rice propagation based on demand and provide valuable important information that can be used in combination with other agronomic traits in rice improvement or breeding programs to increase the adaptability of tolerant cultivars under abiotic stress.

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