以基因枪法转化日本结缕草获得转基因植株

Transgenic Zoysia japonica plants obtained by biolistic bombardment transformation.

将含有DREB1A和gus基因的双基因载体质粒包裹在金粉上,以基因枪轰击日本结缕草胚性愈伤组织,以gus基因的瞬时表达研究日本结缕草基因枪转化参数.结果表明,在1 100 psi压力下6 cm的轰击距离、每皿轰击2次的转化效果最好,而金粉和质粒用量对转化效率影响不显著;经过筛选和再生培养,检测到gus基因的稳定表达,并获得日本结缕草潮霉素抗性株系,PCR-Southern杂交证实外源DREB1A基因已整合到日本结缕草基因组中.

Embryogenic calli induced from zoysiagrass particles coated with double-gene vector plasmid （Zoysia japonica Steud. ） seeds were bombarded with gold containing DREB1 A and gus gene. Physical bombardment parameters were evaluated by transient expression of gus gene, and the best result was obtained by a target distance of 6 cm and twice bombardments under the pressure of 1 100 psi, while the concentration of gold particle and plasmid had no significant effect on the transformation efficiency. Stable gus expression was observed in hygromycin-resistant calli, while regenerants survived hygromycin selection and showed gusactivity. PCR-Southem blot analysis further indicated that the foreign gene DREB1A had been integrated into the genome of recovered zoysiagrass plants.