Obtaining plant Miscanthus sacchariflorus (Maxim.) Hack and Miscanthus sinensis Andersson in vitro culture by indirect morphogenesis
Keywords:Miscanthus, callus, biotechnological methods, seed, nutrient medium
Purpose. To obtain Miscanthus sacchariflorus (Maxim.) Hack and Miscanthus sinensis Andersson in vitro culture by indirect morphogenesis.
Methods. Biotechnological procedures, mathematical and statistical analyses.
Results. Composition of nutrient medium was developed intended for induction of callusogenesis from Miscanthus seeds with a poor germination and viability of seedlings – Murashige and Skoog (MS) medium was modified for the amount of macroelements (half-dose) that was supplemented with amino acids (300 mg/l of glutamic acid, 50 mg/l of aspartic acid, 5 mg/l of tyrosine, 3 mg/l of arginine, 2 mg/l of hydroxyproline) and plant growth regulators [2,5 mg/l of 2.4D (2.4-Dichlorophenoxyacetic acid), 0,6 mg/l of BAP (6-Benzyl-aminopurine) and 0,3 mg/l of ABA (Abscisic acid)]. Composition of nutrient medium was developed for regeneration of microplants from callus – agar MS medium was modified for the amount of macroelements (half-dose) supplemented with vitamins: 10 mg/l of thiaminum, 1,0 mg/l of pyridoxine, 1,0 mg/l of nicotinic acid (by White), 1,0 mg/l of ascorbic acid, 250 mg/l of glutamic acid, 2,0 mg/l of BAP, 0,3 mg/l of NAA (Naphthaleneacetic acid). On this medium, 100% regeneration of M. sacchariflorus (Maxim.) Hack and 50% regeneration of M. sinensis Andersson was obtained. Due to media modification aimed at initiating callusogenesis and microplants regeneration, reproduction factor of M. sinensis was increased 20 times at the average, M. sacchariflorus – 35–40 times.
Conclusions. Plants of M. sacchariflorus (Maxim.) Hack and M. sinensis Andersson were obtained in vitro culture by initiation of callusogenes and microplants regeneration from the Miscanthus seeds with poor germination and viability on nutrient media of certain composition.
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