Rapid quantification of plant-powdery mildew interactions by qPCR and conidiospore counts.
Rapid quantification of plant-powdery mildew interactions by qPCR and conidiospore counts.
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DOI：
10.1186/1746-4811-8-35
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发表时间：
2012-08-31
期刊：
影响因子：
5.1
通讯作者：
Panstruga R
中科院分区：
文献类型：
--
作者：
Weßling R;Panstruga R
白粉病为研究植物寄主与专性生物营养真菌病原体之间的相互作用提供了一个有价值的病理系统。在植物与白粉病相互作用的定量评估的基础上，特别是在超敏感和/或抗性植物突变体的背景下，已经有了许多发现。然而，目前可用的方法来评分白粉病真菌的致病成功是费力的，因此不太适合中到高通量分析。在这里，我们提出了两种新的方案，允许快速定量评估白粉病的发展。一种方法依赖于基于定量聚合酶链反应（qPCR）的真菌生物量评估，而另一种方法依赖于真菌分生孢子的定量。我们使用白粉病病原体goovinomyces orontii在一组超敏感和抗性拟南芥突变体上验证了这两种技术，发现两者分别覆盖了1到2个（qPCR）和4到5个（分生孢子定量）数量级的广泛动态范围。这两种方法产生可重复的结果，并且无需专门设备即可轻松执行。qPCR和孢子计数检测可快速、可重复地定量白粉病的发病机制。我们的方法是在感染的后期进行的，并准确地辨别突变表型。因此，该方法补充了目前使用的白粉病定量方法，并克服了它们的一些局限性。此外，它们可以很容易地适应其他植物白粉病的病理系统。
The powdery mildew disease represents a valuable patho-system to study the interaction between plant hosts and obligate biotrophic fungal pathogens. Numerous discoveries have been made on the basis of the quantitative evaluation of plant-powdery mildew interactions, especially in the context of hyper-susceptible and/or resistant plant mutants. However, the presently available methods to score the pathogenic success of powdery mildew fungi are laborious and thus not well suited for medium- to high-throughput analysis. Here we present two new protocols that allow the rapid quantitative assessment of powdery mildew disease development. One procedure depends on quantitative polymerase chain reaction (qPCR)-based evaluation of fungal biomass, while the other relies on the quantification of fungal conidiospores. We validated both techniques using the powdery mildew pathogen Golovinomyces orontii on a set of hyper-susceptible and resistant Arabidopsis thaliana mutants and found that both cover a wide dynamic range of one to two (qPCR) and four to five (quantification of conidia) orders of magnitude, respectively. The two approaches yield reproducible results and are easy to perform without specialized equipment. The qPCR and spore count assays rapidly and reproducibly quantify powdery mildew pathogenesis. Our methods are performed at later stages of infection and discern mutant phenotypes accurately. The assays therefore complement currently used procedures of powdery mildew quantification and can overcome some of their limitations. In addition, they can easily be adapted to other plant-powdery mildew patho-systems.
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影响因子：
30.8
作者：
Consonni, Chiara;Humphry, Matthew E.;Panstruga, Ralph
通讯作者：
Panstruga, Ralph
DOI：
10.1199/tab.0115
发表时间：
2008-01-01
期刊：
The arabidopsis book
影响因子：
--
作者：
Micali, Cristina;Gollner, Katharina;Panstruga, Ralph
通讯作者：
Panstruga, Ralph
影响因子：
6.5
作者：
Gachon, C;Saindrenan, P
通讯作者：
Saindrenan, P
影响因子：
11.6
作者：
Stuttmann, Johannes;Hubberten, Hans-Michael;Parker, Jane E.
通讯作者：
Parker, Jane E.
影响因子：
11.6
作者：
Vogel, JP;Raab, TK;Somerville, SC
通讯作者：
Somerville, SC