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利用hrpZPsg12转化玉米,以期得到对玉米病害具有广谱抗性的新种质材料,为抗病育种提供新的种质资源。以植物表达载体pCAMBIA3301为基础载体,采用PCR法从克隆载体pGM-hrpZPsg12克隆来自丁香假单胞菌大豆致病变种(P.syringae pv.glycinea)的hrp ZPsg12基因,两端分别引入XhoⅠ酶切位点,构建了1个具有卡那霉素和除草剂草丁膦抗性标记的植物表达载体pCAMBIA3301-hrpZPsg12,并通过热激法转入到大肠杆菌DH5α中。采用花粉管通道法将构建好的植物表达载体的重组质粒导入玉米优良自交系“综31”中。结果表明:对得到的575株转化植株经PCR检测有45株呈阳性,阳性转化率为7.83%。
Transformation of maize with hrpZPsg12 in order to obtain new germplasm materials with broad spectrum resistance to maize diseases and provide new germplasm resources for disease resistance breeding. Using the plant expression vector pCAMBIA3301 as the basic vector, the hrp ZPsg12 gene from P. syringae pv.glycinea was cloned by PCR from the cloning vector pGM-hrpZPsg12, A plant expression vector pCAMBIA3301-hrpZPsg12 with kanamycin and herbicide glufosinate resistance was constructed and transformed into E. coli DH5α by heat shock. The recombinant plasmid of the constructed plant expression vector was introduced into the maize inbred line “Comprehensive 31” by using the pollen tube channel method. The results showed that 45 of the 575 transformed plants were positive by PCR and the positive conversion rate was 7.83%.