In vitro multiple shoot induction through axillary bud of Cocculus hirsutus (L.) Diels: A threatened medicinal plant

Abstract

In this study, we describe a method for rapid in vitro multiplication for conservation of a threatened medicinal plant, Cocculus hirsutus (Menispermaceae) via enhanced axillary bud proliferation from nodal explants collected from young shoots of a mature elite plant. The physiological effect of different plant growth regulators such as 6-Benzylaminopurine (BAP), kinetin (Kn), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) on in vitro multiplication was studied. Multiple shoots were induced in vitro from the nodal segments of stem on Murashige and Skoog’s (MS) medium containing BAP or Kn alone or in combination. Maximum number of shoots (45 ± 0.69 shoots per explant) was observed on the medium containing BAP (0.5 mg/L) along with additives like adenine sulphate (50.0 mg/L) and glutamine (150 mg/L). The shoots were best elongated on medium containing 0.25 mg/L BAP along with additives. Elongated shoots were rooted on ¼ MS medium supplemented with 0.5 mg/L IBA. The rooted in vitro raised plantlets were acclimatized in growth chamber. The method described here can be successfully employed for large-scale multiplication, long term in vitro conservation and for further investigation of medicinally active constituents of C. hirsutus. This is the first report on in vitro multiplication of C. hirsutus.

Key words: Cocculus hirsutus, in vitro multiplication, threatened, nodal explants.