An efficient protocol for plant regeneration from protoplasts of hydroxyproline(HYP)resistant cell line of Onoblychis viciaefolia was established. In SH medium supplemented with 1 mg/L 2, 4-dichlorophenoxy-acetic acid...An efficient protocol for plant regeneration from protoplasts of hydroxyproline(HYP)resistant cell line of Onoblychis viciaefolia was established. In SH medium supplemented with 1 mg/L 2, 4-dichlorophenoxy-acetic acid (2,4-D), 0.5 mg/L kinetin (KT) and 0.2 mg/L naphthalene acetic acid (NAA), the division frequency of protoplastderived cells reached uP to over 60 %, and microcalli were obtained in 5-6 wk. Upon transferring them on agar solidified MS medium plus 2 mg/L indole-3-acetic acid (IAA), shoots were induced. After cultivating them on MS medium with or without IAA, roots were regenerated.Chromosome number of all protoplast-regenerated plants examined were normal (2n=28). The protoplast-derived calli and plants grew vigorously on the medium containing 10 mmol/L HYP.展开更多
A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic sig...A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic signal molecules and multiple natural extracts from susceptible plants were used to pretreat the primary vigorous Agrobacterium(PVA) cells for approximately 16 h; (b) the gyratory magnetic field condition was used during cocultivation; (c) the cocultivating period and selecting condition were modified; (d) the recipient cells were at exuberant metabolism and active division while infected with Agrobacterium. Both neomycin phosphotransferase and nopaline synthase assays demonstrated the expression of NPT II and NOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850::1103neo, in transformed wheat cell colonies by adopting the techniques of dot blot, ndPAGE or high voltage paper electrophoresis. Integration of the foreign genes into wheat genome was confirmed by Southern blot hybridization. Moreover, a relatively rational method was described for the estimation of transformation frequencies from cultured cell levels.展开更多
文摘An efficient protocol for plant regeneration from protoplasts of hydroxyproline(HYP)resistant cell line of Onoblychis viciaefolia was established. In SH medium supplemented with 1 mg/L 2, 4-dichlorophenoxy-acetic acid (2,4-D), 0.5 mg/L kinetin (KT) and 0.2 mg/L naphthalene acetic acid (NAA), the division frequency of protoplastderived cells reached uP to over 60 %, and microcalli were obtained in 5-6 wk. Upon transferring them on agar solidified MS medium plus 2 mg/L indole-3-acetic acid (IAA), shoots were induced. After cultivating them on MS medium with or without IAA, roots were regenerated.Chromosome number of all protoplast-regenerated plants examined were normal (2n=28). The protoplast-derived calli and plants grew vigorously on the medium containing 10 mmol/L HYP.
文摘A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic signal molecules and multiple natural extracts from susceptible plants were used to pretreat the primary vigorous Agrobacterium(PVA) cells for approximately 16 h; (b) the gyratory magnetic field condition was used during cocultivation; (c) the cocultivating period and selecting condition were modified; (d) the recipient cells were at exuberant metabolism and active division while infected with Agrobacterium. Both neomycin phosphotransferase and nopaline synthase assays demonstrated the expression of NPT II and NOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850::1103neo, in transformed wheat cell colonies by adopting the techniques of dot blot, ndPAGE or high voltage paper electrophoresis. Integration of the foreign genes into wheat genome was confirmed by Southern blot hybridization. Moreover, a relatively rational method was described for the estimation of transformation frequencies from cultured cell levels.
