Transgenic Zoysia japonica plants obtained by biolistic bombardment transformation
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摘要: 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.Abstract: Embryogenic calli induced from zoysiagrass(Zoysia japonica Steud.) seeds were bombarded with gold particles coated with double-gene vector plasmid containing DREB1A 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 gus-activity.PCR-Southern blot analysis further indicated that the foreign gene DREB1A had been integrated into the genome of recovered zoysiagrass plants.
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