In vitro symbiotic seed germination of South Indian endemic orchid Coelogyne nervosa |
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| Institution: | 1. Root and Soil Biology Laboratory, Department of Botany, Bharathiar University, Coimbatore 641 046, Tamil Nadu, India;2. Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore 641 046, Tamil Nadu, India;3. Department of Life Sciences, Manipur University, Canchipur, Imphal 795 003, India;1. Beijing Botanical Gardens, Wofosi Road Xiangshan, Haidian District, Beijing 100093, China;2. Botany Department, National Museum of Natural Science, No. 1, Kuan-Chien Road, Taichung, Taiwan;1. Departamento de Micología, Real Jardín Botánico, CSIC, Plaza de Murillo 2, E-28014 Madrid, Spain;2. Laboratorio de Micología Experimental, Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina;1. Centro Regional de Investigaciones Científicas y Transferencia Tecnológica, CRILAR- CONICET, Entre Ríos y Mendoza, 5301 Anillaco, La Rioja, Argentina;2. Instituto de Botánica Darwinion, Labardén 200, San Isidro, B1642HYD, Buenos Aires, Argentina;1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 151, Malianwa North Road, Haidian District, Beijing 100193, China;2. Guangxi Botanical Garden of Medicinal Plant, Guangxi Key Laboratory of Medicinal Resources Conservation and Genetic Improvement, Nanning 530023, China;3. Botanical Garden of Xishuangbanna South Medicine, Yunnan 666100, China;1. Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand;2. Pharmaceutical Chemistry and Natural Products Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand;3. Natural Products for Ageing and Chronic Diseases Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand;4. Department of Food and Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand |
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| Abstract: | Study on the dependence of orchids on fungi for seed germination and seedling development provides a mean for understanding the role of fungi in the orchid development process. The epiphytic orchid Coelogyne nervosa endemic to south India is exploited in an unsustainable manner for its therapeutic value. So a protocol for symbiotic seed germination was established for C. nervosa. We isolated a fungus by plating mycorrhizal root discs of the terrestrial orchid Eulophia epidendreae and identified it as Epulorhiza sp., by sequencing the internal transcribed spacer (ITS) regions of the ribosomal RNA gene. Germination of C. nervosa seeds was higher when inoculated with Epulorhiza sp. Uninoculated seeds of C. nervosa ceased to develop soon after the initiation of germination, and the embryo failed to rupture the seed testa. The isolated fungal hyphae entered the germinating seeds either through the pores in-between the integuments, or through the rhizoids. After the fungal establishment (peloton formation) in embryonic cells, the embryo transformed into a protocorm and after 45 days, 66% of the germinated seeds were transformed into protocorms. Nevertheless, promeristem formation occurred only after fungal association. Sixty-three percent of the protocorms developed their first leaf by 90 days and 62% of these produced a second leaf by 120 days after fungal inoculation. All the seedlings in green leaf stage produced roots and contained fungal pelotons. Our results suggest that the Epulorhiza sp. could be successfully used in the in vitro production of C. nervosa for their reintroduction into its natural environment. |
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| Keywords: | Mycorrhiza Pelotons Protocorm Seedling development |
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