Micropropagation of Chrysanthemum leiophyllum (Asteraceae)
Abstract
The East Asian species Chrysanthemum leiophyllum is a source of adaptive traits in the breeding of Ch. × morifolium. The specifics of introduction of Ch. leiophyllum into an in vitro culture and its micropropagation were studied. Stem segments with axillary buds were used as explants. A technique for sterilization of stems to obtain viable explants was developed. Shorter and vitrified shoots, with the callus tissue developing in their basal part, formed on Murashige and Skoog (MS) medium supplemented with 0,5 mg/L 6-benzylaminopurine (BAP) and 30 g/L sucrose. A decrease in the BAP level to 0.2 mg/L promoted shoot growth. A two-fold decrease in the sucrose concentration raised the multiplication coefficient of Ch. leiophyllum shoots to 10,3, while a two-fold decrease in the macro-salts concentration raised the multiplication coefficient to11.7. It was found that concentration of macro-salts has an effect on shoot development. The medium with half-strength MS macrosalts in combination with 0.2 mg/L BAP was optimal for shoot propagation. Adventitious buds formed in the basal part of shoots. Roots developed intensively in the medium with half-strength MS macrosalts and in the presence of 0,5 mg/L indoleacetic acid (IAA) and 30 g/L sucrose. Growth of roots was influenced mostly by plant growth regulators (PGR); the optimal PGR at the rooting stage was IAA. The efficiency of adaptation of plants to soil conditions constituted 98 %. The plants obtained in vitro and transplanted to soil grew normally. The ground-hugging rosettes of leaves developed by the end of the first year on the open soil; in the second year, the plants entered the generative phase. The chromosome number was 2n = 36 both in the in vitro obtained plants and in the original plant.
Key words: Chrysanthemum leiophyllum, in vitro culture, organogenesis,phytohormones, chromosome number.
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