ex situ conservation of trichoglottis tenera (lindl.) a threatened, and endangered orchid of western ghats using asymbiotic seed germination technique

Research Article
*M. Bastin and R. Jeyachandran
DOI: 
xxx-xxxx-xxx
Subject: 
science
KeyWords: 
Trichoglottis tenera, Seed germination, Asymbiotic seed culture, BAP, NAA TDZ.
Abstract: 

Orchids have been used as traditional healing system of medicine as well as in the treatment of a number of diseases in many parts of the country and in the world. Orchids are a group of extremely interesting plants now recognized as economically important crops having floricultural, commercial and therapeutic values. They are well represented in India, distributed in Himalayan, NE Indian, Western Ghats (WGs) and Eastern Ghats regions. Trichoglottis tenera is an endangered epiphytic orchid of Western Ghats. Orchid seeds are dust-like structures and difficult to use directly in the field for mass propagation of orchid seed germination. We have used ten different media for orchid seed germination namly; we investigated seed germination in the ten basic inorganic media which were a). full and half strength Knudson-C medium6- benzylaminopurine (BAP- 0.2 – 2.0 mg/l), α-Naphthalene acetic acid (NAA-0.1 -1.0 mg/l), Thidiazuron (TDZ 0.2 – 2.0 mg/l) coconut water (15% ml/l) and banana powder (0.4 % mg/l). b). Full and half strength Murashige & Skoog (MS) medium (1962) full strength and half strength of MS medium has been fortified with different combinations of plant growth regulators i.e., 6-Benzylaminopurine (BAP 0.2 – 2.0 mg/l and 0.1-1.0 mg/l), α-Naphthalene acetic acid (NAA 2.0 - 0.2 mg/l and 0.2-2.5mg/l), Kinetin (1.0 – 1.0 mg/l), Thidiazuron (0.2 – 2.0 mg/l), coconut water (15% ml/l) and tomato powder (4.0% and 0.4%). c). Full and half strength Oat meal agar medium (OMA) full strength of banana agar medium (BAM) was supplemented with different types of plant growth regulators (PGRs) i.e., 6-Benzylaminopurine (BAP 2.0 – 0.2 mg/l and 0.2-1.0 mg/l), α-Naphthalene acetic acid (NAA 0.2 – 2.5 mg/l and 2.0 -0.2 mg/l), Kinetin (1.0 – 0.1 mg/l), Gibberellic acid (GA3 0.2 – 2.0 mg/l) and different concentrations of hormonal additives like Tryptone powder (0.1 – 1.0 mg/l) banana powder (0.4%) coconut water (15% ml/l).

d). Full and half strength Banana agar medium (BAM) which were contains 6-Benzylaminopurine (BAP 0.1 – 1.0 mg/l and 0.2 – 2.0 mg/l), α-Naphthalene acetic acid (NAA 2.0 – 0.2 mg/l), Kinetin (1.0 – 0.1 mg/l) and Thidiazuron (0.1 – 1.0 mg/l) different strength of hormonal additives like Tryptone powder (0.1 – 1.0 mg/l) banana powder (0.1 – 1.0 mg/l and coconut water (15% ml/l). e). Full and half strength R medium which were contain 6-Benzylaminopurine (BAP 0.2 – 2.0 mg/l and 0.1 – 1.0 mg/l) α-Naphthalene acetic acid (NAA 2.0 – 0.2 mg/l and 2.0 -0.2 mg/l), Thidiazuron (0.1 – 1.0 mg/l and 0.2 – 2.0 mg/l), and different concentrations of additives of tomato powder (0.4% mg/l) and coconut water (15% ml/l). The rooting media were prepared and supplemented with plant growth regulators (IBA -0.2 mg/l), and IAA (0.3 mg/l). A maximum seed germination of 90% percentage was observed after 60-70 days on half strength MS medium with banana powder. 90% of seed germination was observed on half strength MS medium with commercially purchased oats. Significant seed germination was observed on half strength MS medium with corn flour. Optimum seed germination was observed on half strength MS medium and banana powder. The well developed shoots were removed from their conical flask and transferred in to subculture medium, and supplemented with BAP (0.2 – 0.5 mg/l), NAA (0.3 – 0.7 mg/l). The rooting media also media prepared and supplemented with plant growth regulators (IBA -0.2 mg/l) and IAA (0.3 mg/l). The results of these experiments showed that the studied species can be effectively propagated by in vitro seed germination of orchids with the aim of ex situ biodiversity conservation.