Detection of Latent Infection of Wheat Leaves Caused by Blumeria graminis f.sp tritici Using Nested PCR
Detection of Latent Infection of Wheat Leaves Caused by Blumeria graminis f.sp tritici Using Nested PCR
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巢式PCR检测小麦叶片由小麦白粉菌专化型(Blumeria graminis f.sp tritici)引起的潜伏感染
DOI：
10.1111/j.1439-0434.2009.01594.x
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发表时间：
2010-04-01
影响因子：
1.5
通讯作者：
Zhou, Yilin
中科院分区：
文献类型：
--
作者：
Zeng, Xiaowei;Luo, Yong;Zhou, Yilin
由小麦白粉菌引起的小麦白粉病。小黑麦(Tritici，BGT)是一种危害世界的重要病害。小麦苗期潜伏侵染的检测是估计田间疫情初始接种潜力的关键。为了改进常规方法，本研究建立了一种检测BGT潜伏侵染小麦叶片的套式聚合酶链式反应方法。设计了套式聚合酶链式反应的外引和内引两对DNA引物，并利用BGT和其他小麦真菌检测了它们对BGT的特异性。敏感性试验表明，套式聚合酶链式反应最低可检出0.1fg模板DNA，灵敏度约为标准聚合酶链式反应的1万倍。人工接种试验结果表明，套式聚合酶链式反应检测小麦幼苗潜伏侵染的时间比标准聚合酶链式反应提前2天。套式聚合酶链式反应测定的小麦幼苗潜伏侵染率与常规培养法测定的潜伏侵染率呈线性相关(R2=0.66，P=0.0023)。套式聚合酶链式反应检测的潜伏感染率高于常规方法。本研究为有效估计田间小麦白粉病流行的初始接种潜力提供了一种准确的方法。
Wheat powdery mildew caused by Blumeria graminis f.sp. tritici (Bgt) is an important and destructive disease worldwide. Detection of latent infection of wheat seedlings is critical to estimate initial inoculum potential of epidemics in the fields. To improve the conventional method, a nested PCR approach had been established in this study to detect latent infections of wheat leaves caused by Bgt. The DNA primer sets including external and internal primer pairs for the nested PCR were designed followed by testing their specificities to Bgt by using Bgt and other fungal species of wheat. Sensitivity test demonstrated that the nested PCR could detect as low as 0.1 fg template DNA and about 10,000 times more sensitive than the standard PCR. Results of artificial inoculation experiments showed that the nested PCR assay can detect a low level of latent infection of wheat seedlings 2 days earlier than did standard PCR. The incidences of latent infection of wheat seedlings determined by the nested PCR linearly correlated with those by the conventional incubation method (r2 = 0.66, P = 0.0023). The incidences of latent infection detected with nested PCR were higher than that with the conventional method. This study provides an accurate method to efficiently estimate the initial inoculum potential of wheat powdery mildew epidemics in the fields.