农杆菌介导西瓜转葡聚糖酶及几丁质酶双基因

Transformation of glucanase and chitinase genes to watermelon mediated by Agrobacterium tumefaciens

采用根癌农杆菌(Agrobacteriumtumefaciens)介导法将含有葡聚糖酶基因和几丁质酶基因表达载体遗传转化西瓜(Citrulluslanatus),遗传转化体系为以子叶为外植体经过组织培养获得再生植株。结果表明,Kan抗性芽的分化率随着共培养时间的延长先上升后下降,GUS阳性芽的百分率逐渐升高,共培养时间30h后,GUS阳性芽的百分率趋于平稳,同时,通过对候选转基因植株中GUS基因染色鉴定、卡那基因、葡聚糖酶基因及几丁质酶基因特异引物PCR检测,表明葡聚糖酶和几丁质酶基因已成功导入西瓜植株中,转基因植株的表现在进一步研究中。

A constitute expression vector containing a rice chitinase gene and an alfalfa glucanase gene was transferred into watermelon mediated by Agrobacterium tumefaciens. Planflets were regenerated in vitro by resistance selection on MS medium containing various concentrations of kanamycin, h was shown that the regeneration rate of shoots of anfi-kanamycin was firstly increased and then decreased depended on co-cuhure duration. Furthermore, the positive rate of shoots for GUS staining was increased gradually depended on co-culture duration, and was stable after 30 h of co-culture duration. Meanwhile, GUS staining and PCR-aided detection indicated that both chitinase and glucanase genes had been successfully transferred into watermelon.