农杆菌介导的枳壳转化系统建立研究初报

以枳壳为对象 ,进行了农杆菌介导的上胚轴转化系统建立研究 ,得到了携带柑桔衰退病病毒外壳蛋白基因的枳壳转化植株。研究表明 :枳壳各类外植体中 ,上胚轴是较好的转化材料 ;上胚轴出芽部位主要在形态学上端。以卡那霉素为选择剂 ,外植体水平面放置时 ,选择剂的剂量是 50～ 70mg/ L。转化研究中发现感染液中乙酰丁香酮 10 0μmol/ L的加入促进转化外植体 Gus表达阳性率的提高。延迟 1a的选择培养 ,有助于 Gus阳性芽的增多。所得 Gus阳性无性系经 Southern blot杂交检测 ,证明外源基因已稳定整合到了植物基因组中 ,所得 Gus阳性植株确为携带柑桔衰退病病毒外壳蛋白基因的转化植株

Preliminary study on establishment of transfor-mation system in Poncirus trifoliata Raf.

WT5”BZ]Transformation and regeneration of Poncirus trifoliata Raf. plants were studied by Agrobacterium mediated. Transgenic plants with CTV cp have been obtained. Of the various explants, epicotyls are suitable for transformation reseach. Shoot is formed on basal end of the epicotyls segment. As a selection agent, the concentration of kanmaycin is 50～70 mg/L. The frequeney of explants with Gus positive was promoted when 100 μmol/L acetosyingone was added in Agrobacterium suspension liquid. It is useful for Gus positive shoot producing when selectable cultivation is postponed for 1 day. DNA fragment from plasmid pGA482GG, which was cut by NcoⅠE, was used as probe, foreign gene is proved going to P. trifoliata by Southern blot. WT5”HZ]