Plant Tissue Cult. & Biotech. 36(1): 107-116, 2026 (June)

General

High Frequency Direct Organogenesis Protocol for Mass Multiplication of Centrosema pubescens Benth

Kamrun Nahar, Md. Abul Kashem, Tapash Kumar Bhowmik and Md. Mahbubur Rahman

Laboratory of Plant Tissue Culture and Biotechnology, Department of Botany, University of Chittagong, Chattogram-4331, Bangladesh

Key words: Centrosema, Direct regeneration, Callus induction, PGRs, Acclimatization

Abastract

This study was aimed to develop a suitable protocol for rapid and mass propagation of Centrosema pubescens Benth. Fresh seeds from field grown plants were aseptically cultured on full and half-strength MS media in combination with or without PGRs (plant growth regulators). Cent percent seed germination was achieved on half-strength MS medium without any PGRs and MS medium in combination with 1.0 mg/l BAP. The shoot apices and nodal segments from in vitro grown seedlings were aseptically cultured for direct organogenesis. The MS medium supplemented with 2.0 mg/l BAP + 1.0 mg/l IAA produced maximum number of multiple shoot buds (4.44 ± 0.12) per shoot apices segment. On the other hand, 93% of nodal segments gave response for induction of multiple shoot buds when cultured them on MS medium supplemented with 2.0 mg/1 BAP + 1.0 mg/1 NAA. The highest percentage of leaf segments (86%) produced brownish friable callus on MS + 1.0 mg/l 2, 4-D + 0.5 mg/I BAP. It was reported that brownish friable callus did not undergo any kind of differentiation. Maximum root induction was 5.20 ± 0.10 per elongated shoot and the highest average length 3.07 ± 0.26 cm was observed on half-strength MS medium fortified with 1.0 mg/l IAA. The strong and stout rooted plantlets were successfully hardened with 78% of survival rate. Further research could be initiated to produce secondary metabolites from the friable callus of this plant species to fulfill the demands of the pharmaceutical industries.

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