Adventitious shoot regeneration in Lotus glaber (Mill)

SANSBERRO, PEDRO A.; ESPASANDIN, FABIANA DANIELA; LUNA, CLAUDIA V; MROGINSKY, LUIS A.

Chascomús, Buenos Aires, Argentina

Taller; 2º taller interdisciplario sobre aspectos genéticos, moleculares y ecofisiológicos de Lotus spp. y sus simbiontes; 2004

Unidad de Biotecnología 1. IIB-INTECh (UNSAM-CONICET).

In order to get a suitable protocol of regeneration for the genetic transformation of Lotus glaber Mill. several experiments from different explants were conduced. Roots, cotyledons and leaves from seedlings grown in vitro were cut into pieces and used as source of explants. Subsequently, they were cultured on tubes of Murashige and Skoog (1962) semisolid medium supplemented with different combinations of auxins, cytokinins. After 45 days of culture, de novo shoot organogenesis was noticed from explanted tested. The type of explants markedly influenced and the growth of the regenerated shoots. The regeneration frequencies were higher with leaf and cotyledons explants while the number of shoot formed per responsive explant was greater with leaf and roots. The regenerated shoot from the best induction treatment were transferred to a fresh medium of similar chemical composition and without plant growth regulators for 30 days; in wich, the in vitro rooting was stimulated. In conclusion, our study provides a practical technique for efficient plantets production of Lotus glaber. The procedure described here for the direct shoot organogenesis from various explants, and subsequently plantlets regeneration, facilitates the rapid propagation of this species. It will also be of use cryoconservation and genetic breeding aimed at improving the abiotic stress tolerance.