Presymptomatic and quantitative detection of Mycosphaerella graminicola development in wheat using a real-time PCR assay
Presymptomatic and quantitative detection of Mycosphaerella graminicola development in wheat using a real-time PCR assay
复制标题
DOI：
10.1111/j.1574-6968.2006.00393.x
复制
发表时间：
2006-09-01
影响因子：
2.1
通讯作者：
Verreet, Joseph-Alexander
中科院分区：
文献类型：
--
作者：
Guo, Jian-Rong;Schnieder, Frank;Verreet, Joseph-Alexander
对稻瘟病菌进行症状前、准确的诊断，对病情预测和及时使用杀菌剂具有重要意义。为了建立一种灵敏的聚合酶链式反应方法，设计了四对特异的引物。它们比已知的三对特定的引物更具特异性。其中三种方法在常规的聚合酶链式反应中可以检测到低至0.5pg的禾谷分枝杆菌DNA。利用所设计的引物对ST-rRNA F/R对接种和自然侵染的小麦植株进行了实时定量PCR检测，结果表明，接种植株在接种后即刻即可检测到稻瘟病菌DNA，并在接种后8d出现明显症状前呈稳定上升趋势。快速生长期发生在接种后6~16d。田间观察了流行期间顶层3个叶层的病情发展。与病情指数呈极显著正相关(R=0.8986)，与每叶拟线虫数呈极显著正相关(R=0.9227)。这表明，实时荧光定量聚合酶链式反应技术是一种准确、无症状、准确检测田间麦草分枝杆菌生长发育的可靠方法。
Presymptomatic and accurate diagnosis of Mycosphaerella graminicola leaf blotch is desirable for the disease prediction and the timely application of fungicides. To develop a sensitive PCR assay, four specific primer pairs were designed. They were more specific than three known specific primer pairs. Three of them could detect as little as 0.5 pg M. graminicola DNA in a conventional PCR. A real-time PCR assay was applied for monitoring the disease progression in both inoculated and naturally infected wheat plants using the primer pair ST-rRNA F/R. In inoculated plants, M. graminicola DNA could be detected immediately after inoculation and a steady increase was detected before visible symptoms appeared at 8 days. The rapid growth period took place between 6 and 16 days postinoculation. In the field, the disease progression in the top three leaf layers was followed during the epidemic period. The results were significantly correlated to the disease indices (R=0.8986) and also to the number of pycnidia per leaf (R=0.9227). These suggest that the real-time PCR assay is a reliable approach for the presymptomatic and accurate detection of M. graminicola development in the field.