根癌农杆菌介导的S6PDH基因转化香蕉研究

研究以香蕉栽培品种"天宝蕉"(Musaspp.cv.Tianbao)横切薄片(Thin cross-sections,TCSs)为材料,采用根癌农杆菌介导的方法,进行S6PDH基因转化香蕉的研究。结果表明,在横切薄片继代增殖培养基M4中添加5%~7%(V/V)的椰汁明显增强了香蕉芽苗的生长势;GUS基因瞬时表达检测表明,长势旺盛的香蕉芽苗(直径为7~8 mm)适宜作为香蕉遗传转化的受体材料,横切薄片厚度以2 mm左右为佳;采用两步法进行抗性芽的筛选得到37个抗性芽苗,生根移栽后获得31株成活苗;目的基因S6PDH和报告基因GUS的PCR检测表明其中4株是转基因植株。该研究为将蔷薇科山梨醇代谢途径引入香蕉以提高其耐渗透胁迫的能力奠定了重要的基础。

Agrobacterium-mediated transformation of Musa spp.cv.Tianbao with cDNA encoding S6PDH(NADP-dependent sorbitol-6-phosphate dehy-drogenase) from Prunus salicina var.cordata

Musa spp.cv.Tianbao was transformed with cDNA encoding S6PDH(NADP-dependent sorbitol-6-phosphate dehydrogenase)isolated from Prunus salicina var.cordata by an Agrobacteriummediated thin cross-sections(TCSs) transformation system.The condition of the buds was effectively improved when the TCSs were transferred onto the medium M4 adding 5%-7%(V/ V) coconut water.And the highest GUS transient expression occurred while 2 mm thin TCSs from the healthy and strong buds were used as the recepted material.Total 37 putative transformants were selected via the two-step method and 31 putative transformants survived after transplanting.Finally,four transgenic lines were conformed by PCR analysis of S6PDH gene and GUS gene.Sorbitol synthesis pathway which was unique to the Rosaceae plants had been introduced into Musa spp.cv.Tianbao,laying the groundwork to increase its tolerance to environmental stress.