CAJ | 학술논문

稻瘟病是水稻生产中的主要病害，抗病品种选育和利用是控制稻瘟病最有效措施。研究稻瘟病菌与水稻品种互作机制是培育抗病品种的基础。GFP（Green fluorescent protein）基因因其具有片段小、易与多种不同蛋白质N端或C端融合等特点，广泛应用于病菌与寄主植物互作研究中。研究以pBGgHg作为转化载体，根癌农杆菌C58C1作为转化介体，转化稻瘟病菌的强致病菌株py1022。研究发现，250μg·mL-1潮霉素B能完全抑制稻瘟病菌生长，利用根癌农杆菌介导稻瘟病菌遗传转化的最佳条件为：稻瘟病菌分生孢子浓度为1×105个·mL-1，共培养时间为4 d，共培养AS浓度为200μmol·mL-1，共培养温度为28℃。随机挑取8个转化子分别进行PCR扩增后测序、荧光显微镜下观察、致病力及稳定性测定，结果表明GFP成功转化到稻瘟病菌中，可为稻瘟病菌与水稻品种的互作机制研究提供技术支撑。
도온병시수도생산중적주요병해，항병품충선육화이용시공제도온병최유효조시。연구도온병균여수도품충호작궤제시배육항병품충적기출。GFP（Green fluorescent protein）기인인기구유편단소、역여다충불동단백질N단혹C단융합등특점，엄범응용우병균여기주식물호작연구중。연구이pBGgHg작위전화재체，근암농간균C58C1작위전화개체，전화도온병균적강치병균주py1022。연구발현，250μg·mL-1조매소B능완전억제도온병균생장，이용근암농간균개도도온병균유전전화적최가조건위：도온병균분생포자농도위1×105개·mL-1，공배양시간위4 d，공배양AS농도위200μmol·mL-1，공배양온도위28℃。수궤도취8개전화자분별진행PCR확증후측서、형광현미경하관찰、치병력급은정성측정，결과표명GFP성공전화도도온병균중，가위도온병균여수도품충적호작궤제연구제공기술지탱。
Rice blast is one of the main devastating diseases in rice production, breeding and utilization of resistant varieties are the most economical and effective measures to control of rice blast disease. Studying the interaction mechanism between Magnaporthe grisea and rice varieties is the base of breeding resistant varieties. GFP ( Green fluorescent protein ) gene fragment is so smal that it can fuse with different protein N terminal or C terminal easily, and it can be used in the interaction study between pathogen and host. In this study, we developed an Agrobacterium tumefaciens-mediated transformation system for Magnaporthe grisea by using a high pathogenic isolate py1022 and the Agrobacterium tumefaciens strain C58C1 carrying plasmid pBGgHg. The results showed that 250 μg·mL-1 hygromycin B could completely inhibit the growth of Magnaporthe grisea. The optimal genetic transformation system were as fol owed:the conidia concentration of Magnaporthe grisea was 1×105 conidia·mL-1, the total culture time for Agrobacterium tumefaciens-mediated rice blast fungus transformation was 4 d, co-culture with AS concentration was 200 μmol·mL-1, co-culture temperature was 28 ℃. We randomly chosed eight clones and verified the GFP transformation by PCR amplifying with the hph primers and sequencing, observed mycelia and conidia under fluorescence microscopy, determinated pathogenicity and genetic stability, the results showed that the GFP had been successful y transformed into the rice blast fungus, and it can provide powerful tool for the study of interaction mechanism between Magnaporthe grisea and rice varieties .