Effect of Various Sterilizing agents on the Micropropagation of Endangered Alpine Medicinal Plant Trillium govanianum Wall. ex-Royle.

Abstract

The use of higher altitude grown medicinal plants as a direct source of explant material to produce ‘clean’ in vitro plantlets, presents a major challenge. The microorganisms (fungi or bacteria or endophytes) that usually live within plant tissues without showing symptoms of disease. In most cases, these endophytes exhibit a symbiotic relationship with the host plant, but in tissue culture procedures, these endophytes overgrow in established cultures, thus hindering the practice of in vitro establishment. In our research we use the plant Trillium govanianum from the family melantheiaceae. Surface sterilization trials were conducted on the various plants including leaf, rhizome stem and roots using as the explants for the in vitro propagation. This study aims to obtain an optimal explant surface sterilization protocol for Trillium govanianum tissue culture that is free from contamination, browning, and explant death. Initial sterilization steps i.e., explants under running tap water time duration should be at least one hour for underground plant parts and 40- 45 minutes for other plants parts. Nine types of chemicals are used as a sterilizing agent along with different concentration with different exposure of the time. The only part of the plant that can grow roots & shoots is rhizome bud; the other plant parts show no response. Therefore, sterilizing agents such as streptomycin (antibiotic), Bavistin (Fungicide), Mercuric chloride, 70% ethanol, sodium hypochlorite shows the best response. Therefore, this study helps in maintaining healthy in vitro cultures of this valuable and endangered medicinal herb for its mass propagation and conservation aspects.