An Improved Method for In Vitro Large Scale Propagation of Piper betle L

Abstract

A protocol for large-scale propagation of Piper betle cvs Desawari and Desi Bangla was developed through axillary shoot proliferation. Due to systemic infection as well as high phenol content the crop was very difficult to establish in aseptic condition. But a mixture of 5 mg l⁻¹ each of chloramphenicol and oxytetracyclin and 100 mg l⁻¹ each of citric acid and ascorbic acid used in MS medium for 2 days helped in establishment. After 48 h, the explants were transferred to the antibiotic free medium having PVP and ascorbic acid (100 mg l⁻¹ each), citric acid (50 mg l⁻¹), and glutathione (20 mg l⁻¹). Regular subculturing of the explants into liquid medium, use of antioxidants and incubation of the cultures in the dark for initial 7–10 days played a crucial role for keeping them fresh and green. Maximum numbers of axillary shoots were obtained with 2 mg l⁻¹ BA and 0.2 mg l⁻¹ NAA as growth supplements. The plants were rooted in 0.25 mg l⁻¹ IBA and hardened in the soil. Phenolic compound analysis showed almost the same results in tissue-raised and in vivo grown plants in Desawari.