Plant regeneration from proroplasts of alfalfa (Medicago sativa) via somatic embryogenesis

Abstract

Alfalfa is one of the most frequently studied species from the production of tissue culturederived embryos point of view. In this study, five alfalfa cultivars were analyzed with reference to their ability to regenerate plants from protoplast cultures via somatic embryogenesis. Plant regeneration from leafderived protoplasts isolated from the cultivar Rangelander was achieved using a protocol defined for alfalfa cell suspension-derived embryogenesis. Because of its high efficiency, this procedure is recommended for protoplast electroporation-mediated genetic transformation of alfalfa.

Protoplast production and isolation from Etlingera elatior

“…For this experiment, in vitro leaves of E. elatior were incubated in the enzymatic solution E. A higher yield of protoplasts was obtained for leaves incubated in the shaking system for 15 hours (22.0 x 10 5 protoplasts g -1 of fresh tissue) (Table 3). Expressive results were obtained by Monteiro et al (2003) when they isolated protoplasts from the alfalfa Medicago sativa using the system of continuous shaking (35 rpm) in the dark.…”

Gene Transfer in Legumes

“…Atanasov and Brown (1984) reported a system for successful regeneration of plants from protoplasts from either leaf mesophyll or cotyledon-derived cell suspension cultures, which divided and formed colonies on Kao medium, followed by somatic embryo formation on a high auxin/low cytokinin medium. Monteiro et al (2003) also reported plant regeneration from leaf-derived protoplast cultures of several alfalfa cultivars via somatic embryogenesis. Because of its high efficiency, this procedure can be used for electroporation-mediated protoplast transformation of alfalfa.…”

Date Palm Cell and Protoplast Culture

“…It was reported that in the purple coneflower, Echinacea angustifolia, the yield of protoplasts released also increased when the cellulase concentration was increased up to 2% (w/v), and the lower cellulase concentration (less than 1%) could not lead to liberation of enough protoplast (Zhu et al 2005). Cellulase at a concentration higher than 2% may cause overdigestion of plant material (Koster et al 2003;Monteiro et al 2003). Several factors influence protoplast release, including the extent of cell-wall thickening, temperature, duration of enzyme incubation, pH of enzyme solution (Sinha et al 2003), agitation, the nature of osmoticum and plasmolysis prior to enzyme digestion of source tissues, manual or enzymatic removal of the epidermis and conditioning of the donor material or its culture on media containing osmoticum (Davey et al 2000).…”