基于PCR和巢式PCR技术的甘蔗黑穗病早期检测

Early Stage Molecular Diagnosis Based on PCR and Nested PCR Assays for Sugarcane Smut Disease

为了探索甘蔗黑穗病早期检测的可能性,应用本实验室已建立的甘蔗黑穗病菌PCR和巢式PCR检测技术,对经人工接种甘蔗黑穗病菌冬孢子的植蔗叶片进行检测,并调查采样植株的黑穗病实际发生情况。结果表明,黑穗病实际发生率80%(16/20),PCR检测阳性率35%(7/20),巢式PCR检测阳性率70%(14/20),PCR和巢式PCR检测为阳性的样品,其植株黑穗病发生率几乎为100%。这一结果表明了甘蔗黑穗病菌PCR和巢式PCR可用于甘蔗黑穗病早期检测,但巢式PCR检测结果与黑穗病的实际发生情况比较吻合,更适合于甘蔗黑穗病早期检测。另外,太幼小的甘蔗株(三叶期前),PCR和巢式PCR均不适合于甘蔗黑穗病早期检测。

In order to explore the possibility of early stage detection for sugarcane smut disease, caused by the basidiomycetous fungus, Ustilago scitaminea, samples of the youngest expanding leaf upon growing point from plants derived from seedeane inoculated with U. scitamina teliospores by artificial inoculation method, were detected with PCR or nested PCR assays of U. scitamina developed by our laboratory, and actual smut occurrence of sampling plants was surveyed. The results showed that the eventual smut incidence in the field, PCR and nested PCR detection positive rate was 80% （16/20）, 35% （7/20） and 70% （14/20）, respectively, and positive samples of PCR or nested PCR detection could nearly 100% produce whips of smut in the field. It indicated that the PCR or nested PCR technique could be used for the early stage detection of sugarcane smut, but the nested PCR technique was more suitable than PCR technique, because the results of nested PCR assays appeared to correspond quite well with the eventual percentage of seedlings were very young, before 3-leaf stage, PCR or detection of sugarcane smut. whips produced in the field. In addition, if sugarcane nested PCR assays were not suitable for the early stage