Micropropagation of Medicago truncatula Gaertn. cv. Jemalong and Medicago truncatula ssp. Narbonensis
2001
Neves, L.O. | Tomaz, L. | Fevereiro, M.P.S.
A simple and reliable method was established for the maintenance of a permanent stock of several Medicago truncatula genotypes selected from a general seed stock by their in vitro culture amenability and embryogenic capacity. In the first step, multiple shoots were induced from the cotyledon axillary meristem meristem of pre-germinated seeds in Murashige and Skoog medium supplemented with cytokinins (9.3 micromolar zeatin, 22.2 micromolar benzylaminopurine or 4.5 micromolar thidiazuron). In the second step, the induced shoots were allowed to develop in growth-regulator-free medium. Benzylaminopurine at 22.2 micromolar supported the best combination of shoot quality and number of shoots produced. Rooting of microshoots depended on the cytokinin used for shoot induction and was faster for zeatin-treated shoots. In this work a propagation system was devised where the addition of growth regulators was restricted to the induction phase therefore reducing the risks of epigenetic and somaclonal variation.
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