Env Exp Biol (2014) 12: 167–178

Direct organogenesis, phytochemical screening and assessment of genetic stability in clonally raised Chlorophytum borivilianum

Abstract

Keeping in mind the immense commercial and pharmacological importance of the rare aphrodisiac herb safed musli (Chlorophytum borivilianum Sant et Fern), the present investigation describes the development of a rapid and efficient protocol for direct plant regeneration from leaf explants of two populations of C. borivilianum collected from different agroclimatic zones of India. The induction of shoot initials was achieved on both Murashige and Skoog’s (MS) and Gamborg’s basal media supplemented with varying levels of 6-benzyladenine (1.11 to 8.90 μM) and thidiazuron (1.14 to 9.08 μM). MS basal medium supplemented with 4.54 μM thidiazuron was found ideal for shoot induction and gave rise to numerous shoot primordia within three weeks without any callus phase. Seperation of shoot clumps and their subsequent transfer to MS basal medium supplemented with 4.54 μM thidiazuron resulted in differentiation of more than 90% of the shoot initials into well developed shoots. Healthy shoots were rooted on half strength MS supplemented with varying concentrations of indole-3-acetic acid (2.28 to 11.42 μM), indole-3-butyric acid (1.97 to 9.80 μM) and naphthalene acetic acid (2.7 to 10.8 μM). Regenerated plantlets were transferred to the field with more than 80% success. Comparative cytological, molecular and phytochemical screening of the donor populations and their regenerates revealed genetic and phytochemical stability. The fingerprinting patterns generated in our study using high performance thin layer chromatography can be used effectively for quality screening and checking adulteration among different populations and cultivars of C. borivilianum. The micropropagation strategy may be effectively utilized for mass propagation as well germplasm conservation of this rare medicinal herb.