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1 May 2006 EFFICIENT SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM LONG-TERM CELL SUSPENSION CULTURES OF MEDICAGO TRUNCATULA CV. JEMALONG
ANA SOFIA DUQUE, ANA SOFIA PIRES, DULCE METELO DOS SANTOS, PEDRO FEVEREIRO
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Abstract

Plants were successfully regenerated via somatic embryos from 3-yr-old cell suspension cultures of Medicago truncatula Gaertn. 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 subcultured 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 3 wk 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 3 wk 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.

ANA SOFIA DUQUE, ANA SOFIA PIRES, DULCE METELO DOS SANTOS, and PEDRO FEVEREIRO "EFFICIENT SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM LONG-TERM CELL SUSPENSION CULTURES OF MEDICAGO TRUNCATULA CV. JEMALONG," In Vitro Cellular and Developmental Biology - Plant 42(3), 270-273, (1 May 2006). https://doi.org/10.1079/IVP2006775
Received: 22 August 2005; Accepted: 1 March 2006; Published: 1 May 2006
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KEYWORDS
cell suspension culture
embryogenic competence
line M9-10a
Medicago truncatula
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