In Vitro Storage of Stevia rebaudiana Bertoni under Slow Growth Conditions and Mass Multiplication after Storage

Abstract

Author(s): Ely Zayova, Trendafil Nedev, Ludmila Dimitrova

Stevia rebaudiana Bertoni (Asteraceae) is a valuable medicinal plant widely used in the pharmaceutical and food industry. An efficient protocol for in vitro storage of S. rebaudiana plants under slow growth conditions was established. Slow growth cultures were obtained on half strength Murashige and Skoog basal medium (½ MS) supplemented with the osmotic agent mannitol (0; 1; 2; 3 and 4% w/v) for three and six months without any subculture. The 3% mannitol reduced the growth of S. rebaudiana plants in terms of plant height, root length and number of roots formed under low light intensity. The all studied growth parameters decreased significantly with the increase of the storage period. Cultures stored at 22±1 °C did not survive more than six months. The conserved plants were retrieved on the ½ MS basal medium. The plants storaged for three months multiplied more rapidly on propagation medium with 1 mg L-1 6-benzylaminopurine (BAP) than these conserved for six months. The micropropagated plants were successfully in vitro rooted and ex vitro acclimatizated. They showed very good growth and development under field conditions.

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