【作者】 张玉娟； 阙友雄； 张木清； 许莉萍； 邓祖湖； 林彦铨； 陈如凯；

【Author】 ZHANG Yu-juan QUE You-Xiong~(**) ZHANG Mu-qing~(**) XU Li-Ping DENG Zu-hu LIN Yan-quan CHEN Ru-Kai (Key Lab of Sugarcane Genetic Improvement,Ministry of Agriculture,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)

【机构】 福建农林大学/农业部甘蔗遗传改良重点开放实验室；

【摘要】 甘蔗梢腐病是甘蔗生产中一种常见真菌病害,该病由镰刀菌[Gibberella fujikuroi(Saw.)Wholle.]引起,是甘蔗生长中期最主要的一种真菌性病害。近年来,梢腐病在我国蔗区的发生呈逐渐加重的趋势,对我国甘蔗生产和糖业可持续性发展构成严重威胁。因此,甘蔗梢腐病菌的分离、培养和鉴定以及其病原快速检测体系的建立具有重要意义。本研究采集感染梢腐病的甘蔗品种福农28上的病斑叶片,接种培养获得病原菌菌落,经电子显微镜和扫描电镜进行形态学观察后,分离病原菌单孢,培养后获得相应菌丝体,提取基因组DNA,而后利用ITS-PCR、ATP-PCR和Effd-PCR进行分子鉴定。多重序列比对结果表明,甘蔗梢腐病菌ITS序列与Gibberella moniliformis(甘蔗梢腐病菌的有性态)具有100%同源性,与Gibberella fujikuroi具有99%的同源性,而Effd片段与Fusarium fujikuro的同源性也高达92%。同时,基于甘蔗梢腐病菌ITS-PCR和Effd-PCR的鉴定结果与电子显微镜和扫描电镜的鉴定结果高度符合,其中根据ITS序列设计的PCR检测特异性引物,可用于甘蔗梢腐病的快速检测。相比较而言,ATP较难区分甘蔗梢腐病菌以及其他近缘真菌,ATP片段与F.oxysporum相比仅有79%的同源性。更多还原

【Abstract】 Pokkah Boeng Disease of Sugarcaene caused by Gibberella fujikuroi(Saw.)Wholle causes serious damage to sugarcane and is a worldwide disease.Research on the isolation,culture and identification of the corresponding pathogen should be helpful to the establishment of the fast identification system and aid to sugarcane Pokkah Boeng disease resistance breeding.In this paper,we cut and inoculation-culture the spot leaves from sugarcane cultivar FN28 infected by Gibberella fujikuroi(Saw.)Wholle culture,and the electron microscope and scanning electron microscopy for morphological observation of the corresponding fungi colonies was conducted.Then,the separation of single-spore pathogens,the extraction of its DNA and the molecular evaluation by ITS-PCR,ATP-PCR and Effd-PCR were conducted.The results showed that the ITS sequence amplified from the above pathogen above was 100%homoly with that of Gibberella moniliformis,99%homoly with that of Gibberella fujikuroi,which demonstrated that the outcome of ITS-PCR and Effd-PCR identification existed a high degree of compliance with the observation through electron microscope and scanning electron microscope. From above,a rapid detection system of sugarcane Pokkah Boeng was established and the specific PCR primers were designed according to the ITS sequence.Comparatively speaking,ATP-PCR is relatively more difficult to distinguish between sugarcane Gibberella fujikuroi(Saw.)Wholle and its relative fungi, ATP sequence of sugarcane Gibberella fujikuroi(Saw.)Wholle was only 79%homology with F.oxysporum.更多还原