In this paper, we reported firstly the transgenic plants of Orychophragmus violateus in the world. Excised cotyledon and hypocotyls of Orychophragmus violaceus were used as explants for genetic transformation. After 2...In this paper, we reported firstly the transgenic plants of Orychophragmus violateus in the world. Excised cotyledon and hypocotyls of Orychophragmus violaceus were used as explants for genetic transformation. After 2-3 days of co-cultivation with Agrobacterium tumefaciens strain A208SE(PROA93),the hypocotyls and cotyledon were transferred onto selection medium containing 25 mg/L Km and 250 mg/L Ap. 8 weeks later, shoots emerged,then the shoots were excised and transferred onto root medium containing 25 mg/L Km and 100 mg/L Cef. The roots were formed within 4-5 weeks.The whole plants were transplanted into pots and grew well. The frequency of plant regeneration of hypocotyls was about 30%,and that of cotyledon was 51%.The regenerated plants showed high enzymatic activities ofglucuronidase and neomycin phosphotransferase II. Southern blot analysis confirmed that NPT II gene had been stably integrated into the chromosomal genome of Orychophragmus violaceus .the transformation frequency of hypocotyls was 10%,and that of cotyledon was 5.5%.展开更多
Transgenic plants were obtained by PEG-mediated tranfer of foreign gene into cotyledon protoplasts of Orychophragums violaceus. Systematic study was carricd out on PEG-mediatcd transformation of cotyledon protoplast u...Transgenic plants were obtained by PEG-mediated tranfer of foreign gene into cotyledon protoplasts of Orychophragums violaceus. Systematic study was carricd out on PEG-mediatcd transformation of cotyledon protoplast using transient expression system, which showed 25-30 μg of pasmid, 15% PEG and a pH value of 8.0 as the optimal parameters contributing to the highest expression level. Using these parameters, cotyledon protoplasts were isolated, treated with bacterial plasmid DNA (pBI222 with HPT as selective marker) and PEG, and cultured at a density of 5×10 4/ml.After 10-15 days,they were selected by adding 25 μg/ml hygromycine. One month later, a few calli were observed, which were then transferred onto a solid medium with 50-100 μg/ml hygromycine for proliferation. Later they were transferred successively onto differentiation and rooting media and finally hygromycineresistant whole plants were obtaincd. The plants grew well in pots and a regeneration rate of 5 ×10(-5) was achieved. Then,excised leaves of the transgenic plants were used as explants for Southern blot analysis, which confirmed the stable integration of HPT gene into the chromosomal genome of Orychophragmus violaceus The transformation frequency was 10-5.展开更多
Beneficial interactions between microorganisms and plants, particularly in the rhizosphere, are a research area of global interest. Four cadmium(Cd)-tolerant bacterial strains were isolated from heavy metal-contaminat...Beneficial interactions between microorganisms and plants, particularly in the rhizosphere, are a research area of global interest. Four cadmium(Cd)-tolerant bacterial strains were isolated from heavy metal-contaminated sludge and their effects on Cd mobility in soil and the root elongation and Cd accumulation of Orychophragmus violaceus were explored to identify the capability of metalresistant rhizobacteria for promoting the growth of O. violaceus roots on Cd-contaminated soils. The isolated strains, namely, Bacillus subtilis, B. cereus, B. megaterium, and Pseudomonas aeruginosa, significantly enhanced the plant Cd accumulation. The Cd concentrations in the roots and shoots were increased by up to 2.29- and 2.86-fold, respectively, by inoculation of B. megaterium, as compared with the uninoculated control. The bacterial strains displayed different effects on the shoot biomass. Compared with the uninoculated plants, the shoot biomass of the inoculated plants was slightly increased by B. megaterium and significantly decreased by the other strains. B. megaterium was identified as the best candidate for enhancing Cd accumulation in O. violaceus. Thus, this study provides novel insight into the development of plant-microbe systems for phytoremediation.展开更多
文摘In this paper, we reported firstly the transgenic plants of Orychophragmus violateus in the world. Excised cotyledon and hypocotyls of Orychophragmus violaceus were used as explants for genetic transformation. After 2-3 days of co-cultivation with Agrobacterium tumefaciens strain A208SE(PROA93),the hypocotyls and cotyledon were transferred onto selection medium containing 25 mg/L Km and 250 mg/L Ap. 8 weeks later, shoots emerged,then the shoots were excised and transferred onto root medium containing 25 mg/L Km and 100 mg/L Cef. The roots were formed within 4-5 weeks.The whole plants were transplanted into pots and grew well. The frequency of plant regeneration of hypocotyls was about 30%,and that of cotyledon was 51%.The regenerated plants showed high enzymatic activities ofglucuronidase and neomycin phosphotransferase II. Southern blot analysis confirmed that NPT II gene had been stably integrated into the chromosomal genome of Orychophragmus violaceus .the transformation frequency of hypocotyls was 10%,and that of cotyledon was 5.5%.
文摘Transgenic plants were obtained by PEG-mediated tranfer of foreign gene into cotyledon protoplasts of Orychophragums violaceus. Systematic study was carricd out on PEG-mediatcd transformation of cotyledon protoplast using transient expression system, which showed 25-30 μg of pasmid, 15% PEG and a pH value of 8.0 as the optimal parameters contributing to the highest expression level. Using these parameters, cotyledon protoplasts were isolated, treated with bacterial plasmid DNA (pBI222 with HPT as selective marker) and PEG, and cultured at a density of 5×10 4/ml.After 10-15 days,they were selected by adding 25 μg/ml hygromycine. One month later, a few calli were observed, which were then transferred onto a solid medium with 50-100 μg/ml hygromycine for proliferation. Later they were transferred successively onto differentiation and rooting media and finally hygromycineresistant whole plants were obtaincd. The plants grew well in pots and a regeneration rate of 5 ×10(-5) was achieved. Then,excised leaves of the transgenic plants were used as explants for Southern blot analysis, which confirmed the stable integration of HPT gene into the chromosomal genome of Orychophragmus violaceus The transformation frequency was 10-5.
基金Supported by the National Natural Science Foundation of China(Nos.40771203 and 40871243)the Shanghai Key Laboratory of Bio-Energy Crops,China(No.10DZ2271800)the Shanghai Leading Academic Discipline Project,China(No.S30109)
文摘Beneficial interactions between microorganisms and plants, particularly in the rhizosphere, are a research area of global interest. Four cadmium(Cd)-tolerant bacterial strains were isolated from heavy metal-contaminated sludge and their effects on Cd mobility in soil and the root elongation and Cd accumulation of Orychophragmus violaceus were explored to identify the capability of metalresistant rhizobacteria for promoting the growth of O. violaceus roots on Cd-contaminated soils. The isolated strains, namely, Bacillus subtilis, B. cereus, B. megaterium, and Pseudomonas aeruginosa, significantly enhanced the plant Cd accumulation. The Cd concentrations in the roots and shoots were increased by up to 2.29- and 2.86-fold, respectively, by inoculation of B. megaterium, as compared with the uninoculated control. The bacterial strains displayed different effects on the shoot biomass. Compared with the uninoculated plants, the shoot biomass of the inoculated plants was slightly increased by B. megaterium and significantly decreased by the other strains. B. megaterium was identified as the best candidate for enhancing Cd accumulation in O. violaceus. Thus, this study provides novel insight into the development of plant-microbe systems for phytoremediation.
