Authors: Duoue, Ana; Pires, Ana; Santos, Dulce; Fevereiro, Pedro

Source: In Vitro Cellular and Development Biology - Plant, Volume 42, Number 3, May 2006 , pp. 270-273(4)

Publisher: Springer

Abstract:

Plants were suecessfully régenerated via somatic embryos from 3-yr-old cell suspension cultures of Medicago Truncatula Gaertin. cv. Jemalong line M9-10a. The cultures were originally initiated from callus induced in well-expanded leaflets of 30 d in vitro-grown plants, Suspension cultures were established in stirred-liquid Murashige and Skoog (MS) basal salts and vitamins supplemented with 2.3 μM 2.4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin (Kin) and subeultured weekly. Somatic embryogenesis induction step was conducted in liquid MS medium containing 0.45 μM 2,4-D and 0.91 μM zeatin (Zea), during 1,2, and 3wk after subculture. Induced and non-induced cultures were transferred to solid embryo proliferation medium [EPM-MS basal salts and vitamins solidified with 0.2% (w/v) gelrite]. Somatic embryos developed until the late torpedo/dicotyledonary stages. We found that the best condition for the development of somatic embryos was achieved when suspension cultures were not subjected to the induction step. Induction of 1 and 2 wk led to a decrease in the recovery of somatic embryos and the 3-wk treatment resulted in no differentiation of somatic embryos. Plant regeneration was obtained in all conditions (except for 3wk induction) when embryos were transferred to an embryo conversion medium [ECM, similar to EPM but solidified with 0.7% (w/v) agar]. Embryo conversion rates were 54.5±1.6, 52.5±18.5, and 41.6±8.4% for 0, 1, and 2 wk induction treatments, respectively. These plants were successfully transferred to the greenhouse where they matured and produced seeds.

Keywords: embryogenic competence; cell suspension culture; Medicago Truncatula; line M9-10a

Document Type: Research article

DOI: http://dx.doi.org/10.1079/IVP2006775

Affiliations: 1: Email: sduque@itqb.unl.pt

Publication date: 2006-05-01

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