Priming abiotic factors for optimal hybrid Cymbidium (Orchidaceae) PLB and callus induction,plantlet formation,and their subsequent cytogenetic stability analysis |
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Authors: | Jaime A Teixeira da Silva Ming-Tsair Chan Sanjaya Ming-Liang Chai Michio Tanaka |
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Institution: | 1. Department of Horticultural Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan;2. Institute of BioAgricultural Sciences, Academia Sinica, Taipei 11529, Taiwan;3. Zhejiang University, Department of Horticulture, College of Agriculture and Biotechnology, Huajiachi Campus, Hangzhou 310029, People''s Republic of China |
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Abstract: | Abiotic factors affect the induction of PLBs and callus in hybrid Cymbidium Twilight Moon ‘Day Light’. The initiation and proliferation of new PLBs and callus could be achieved on NAA and kinetin, supplemented at 0.1 mg l−1 each, respectively, both within 45–60 days. Bacto agar was found to be the most suitable solidifying agent for PLB induction, although a higher shoot fresh weight was obtained on Gelrite; a pH 5.3 was optimal while pH 4.5 caused 100% explant necrosis; coconut water, when supplied at 10–20% (v/v) resulted in a significant increase in the number of PLBs formed per PLB segment (23.1 versus 14.6 in controls) while a massive (almost four-fold) increase in fresh top weight occurred when PLB explants were placed in liquid culture, as a result of hyperhydricity; Fe-EDTA (1 mg l−1) and activated charcoal (1 g l−1) stimulated total fresh weight and PLB formation in the presence of PGRs; PLB formation decreased but total fresh shoot weight increased with the addition of niacin or myo-inositol, both vitamins. Dark-grown PLB-induced plants were etiolated and had longer internodes and higher fresh weight than light-grown control plants at 45 μmol m−2 s−1; at 15 μmol m−2 s−1 shoots were slightly etiolated, fragile, and PLB formation was scarce. RAPD and mtDNA analysis of all resultant PLBs, callus or plants showed them to be genetically identical, with comparable chlorophyll contents. Despite the detection of cytological variation between different plant parts, little variation resulted from abiotic factor treatment. |
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Keywords: | AC activated charcoal NAA α-naphthaleneacetic acid PGR plant growth regulator PLB protocorm-like body TDZ N-phenyl-N&prime -1 2 3-thiadiazol-5-yl-urea or thidiazol VW Vacin and Went |
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