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Method of propagation, stimulation of rhizomes growth <em>in vitro</em> culture and adaptation in the open ground for the genus <em>Miscanthus </em>representatives
2017
Гонтаренко, С. М | Лашук, С. О
Purpose. To develop a method of propagation, stimulation of rhizomes growth in vitro culture for the genus Miscanthus representatives and their adaptation in the open field without the use of greenhouse complexes for acclimatization and completion of growing. Methods. Biotechnological procedures, mathematical and statistical analyses. Results. Prescription of nutrient medium was developed for explants inoculation, sprouts propagation, rhizomes growth stimulation in vitro. Such sterile explants as seeds, buds to be removed from rhizomes, parts of stems with bud were placed on modified media with mineral portion by Murashige and Skoog (MS) that contained 0,5–1 dose of macroelements and one dose of microelements, vitamins (10 mg/l of thiaminum, 1,0 mg/l of pyridoxine, 1,0 mg/l of nicotinic acid and 1,0 mg/l of ascorbic acid) supplemented with amino acids (250 mg/l of glutamic acid, 3 mg/l of tyrosine, 3 mg/l of arginine, 2 mg/l of hydroxyproline), plant growth regulators [0,5–1,0 mg/l of GA (gibberelline acid), 0,2 mg/l of 6-BAP (6-Benzylaminopurine, 0,1 mg/l of NAA (α-naphtylacetic acid)] in different variations. After seed germination, buds emerging and sprouts formation 1–2 cm in height, for propagation purpose they were passivated on the medium of other composition that differed from previous one by the content and ratio of growth regulators, especially by a high concentration of cytokinins [6-BAP (0,4–0,5 mg/l), kinetin (0,5 mg/l), adenine (0,5 mg/k)] in different variations in presence of GA (0,2 mg/l). In order to stimulate rhizomes growth, microclones were transferred on media with other composition and ratio growth regulators (6-BAP (0,2–0,3 mg/l) + GA (0,5–1,0 mg/l) or 6-BAP (0,2–0,3 mg/l) + GA (0,5–1,0 mg/l) + NAA (0,1 mg/l), in other words, with a high content of gibberellins. After the formation of rhizomes 10–15 cm in length, miscanthus plants were planted out in the open ground. Stimulation of rhizomes initiation and elongation on appropriate nutrient media before Miscanthus giganteus, M. sacchariflorus and M. sinensis planting in vivo resulted in 100% adaptation and 100% survival of plants in the winter period without the use of greenhouse complexes. Conclusions. The method of miscanthus propagation in vitro and adaptation in the open ground was developed that included stimulation of rhizomes growth and favoured the increase of their length on media supplemented with gibberelline that guaranteed 100% preservation of microplants to be propagated from in vitro culture during adaptation in the open ground and acclimatization in winter.
Show more [+] Less [-]Clonal micropropagation of peppermint (Mentha piperita L.) varieties of Ukrainian breeding
2016
Таланкова-Середа, Т. Є | Коломієць, Ю. В | Григорюк, І. П
Purpose. Developing technology for clonal micropropagation of peppermint (Mentha piperita L.) plants of Ukrainian breeding based on the complex of methods of isolated tissue and organ culture in vitro. Methods. During the experiment, such methods as isolated tissue and organ culture in vitro, clonal micropropagation, detached scion grafting, chemotherapy with adding of virucide Ribavirin to the nutrient medium, biometric and statistical ones were used. Results. The stepped procedure of sterilization that we have developed allows to receive 88–100% of sterile explants. For M. piperita L. introduction into culture and clonal micropropagation, Murashige and Skoog (MS) nutrient medium appeared to be optimal supplemented with 6-benzylaminopurine (0.75 mg/l), adenine (0.05 mg/l), indole-3-acetic acid (IAA) (0.05 mg/l) and gibberellic acid (0.5 mg/l) on which the reproduction ratio on the 28th day ranged between 1:7 and 1:15. For recovery of plants from viral infection, virucide Ribavirin at concentration of 10 mg/l was added to the nutrient medium. The proposed nutrient medium for rhizogenesis, that contained IAA (0.5 mg/l) and indole butyric acid (IBA) (0.5 mg/l), allows to obtain the frequency of rhizogenesis up to 84–100%. Regenerated plants were adapted to the conditions in vivo on substrate peat : universal soil : perlite : sand in the ratio 2:1:1:1. The survival rate for peppermint varieties amounted to 96–100%. Conclusions. Biotechnological scheme was developed that permits to get healthy, purebred planting material and intensively propagate plants for supplying breeding programs of the Experimental Station for Medicinal Plants of the Institute of Agroecology and Environmental Management of the National Academy of Agrarian Sciences of Ukraine, among which such varieties as ‘Lebedyna pisnia’ and ‘Ukrainska pertseva’ were selected as the most promising for clonal micropropagation.
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