Regeneration

Murraya koenigii (L.) Spreng, commonly known locally as “curry patta” or “mitha neem” in India, is a valuable medicinal plant known for its biochemical and aromatic properties. This paper presents a protocol for the rapid and high frequency shoot regeneration from nodal and inter-node explants from matured plant and epicotyl, cotyledons, cotyledonary node (embryonic axis), juvenile leaf, hypocotyl and root segment of in vitro derived seedling via axillary and adventitious shoot formation of M. koenigii. Adventitious regeneration, which is a pre-requisite in most genetic transformation studies using Agrobacterium and ballistics, needs to be developed as a protocol for micropropagation of M. koenigii. Cytokinins benzylaminopurine (BAP), Kinetin (Kin), adenine sulphate (ADS) and indole-3-acetic acid (IAA) were used for multiple shoot induction. Addition of an auxin along with cytokinin improved the shoot production capacity. Shoot buds could be initiated from all the explants tested, with epicotyl explants producing the highest average number of shoots/explant. N6-benzyle adenine (BA), kinetin, adenine sulphate (ADS) and indole-3-acetic acid (IAA) in combination were the most effective PGRs for shoot induction.

Tissue culture of medicinal plants is widely used to produce clones and has been found in the conservation of threatened plants species face risk of extinction. Vajradanti is an ethno-medicinally important plant showing pharmacological effects such as anti-microbial, anti-helminthic, anti-fertility, anti-oxidant, anti-diabetic, anti-arthritic, hepato-protective, diuretic, cytoprotective, antidiarrhoeal, analgesic, anti-leukemic, anti-inflammatory and hypoglycemic properties. Micropropagation in most of the plant species involve proliferation of explants using MS medium. There are scanty reports on tissue culture and regeneration of Vajradanti. However, no effective in vitro regeneration protocol has been developed for this medicinally important plant. Development of high frequency regeneration pathway was undertaken using shoot tip and nodal segments of Barleria prionitis on MS medium supplemented with auxin and cytokinins alone and in various combinations.