Real-time PCR monitoring of fungal development in Arabidopsis thaliana infected by Alternaria brassicicola and Botrytis cinerea
Real-time PCR monitoring of fungal development in Arabidopsis thaliana infected by Alternaria brassicicola and Botrytis cinerea
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DOI：
10.1016/j.plaphy.2004.04.001
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发表时间：
2004-05-01
影响因子：
6.5
通讯作者：
Saindrenan, P
中科院分区：
文献类型：
--
作者：
Gachon, C;Saindrenan, P
可靠的病害严重度评估方法在植物病原体相互作用的研究中至关重要，无论是对于田间病害诊断还是评估植物或病原体菌株的表型差异。目前，用于真菌疾病诊断的大多数测定依赖于症状的视觉评估、病变直径测量或孢子计数。然而，这些测试是乏味的，并且通常不能区分稍微不同的阻力水平。此外，它们不太适合在感染的早期阶段，在肉眼可见的症状之前或孢子形成之前评估真菌的发育。在这里，我们描述了一种致病性测定的基础上的相对定量的真菌和植物DNA在受感染的拟南芥叶片通过实时定量PCR。我们表明，它可以定量监测的真菌Alternaria brassicicicola和灰葡萄孢的生长在一个敏感和可靠的方式。尽管高度敏感，但该测试也表现出高稳健性，这对于显著区分显示出略微不同抗性水平的品系至关重要。因此，它允许从感染的最初阶段评估真菌发育，并为目前描述的用于对植物突变株系或真菌菌株进行表型分型的测定提供了快速且非常实用的替代方案。(C)2004年，Elsevier SAS。All rights reserved.
Reliable methods for disease severity assessment are of crucial importance in the study of plant pathogen interactions, either for disease diagnostic on the field or to assess phenotypical differences in plants or pathogen strains. Currently, most of the assays used in fungal disease diagnostic rely on visual assessment of the symptoms, lesion diameter measurement or spore counting. However, these tests are tedious and often cannot discriminate between slightly different levels of resistance. Besides, they are not well suited to assess fungal development in the early phases of the infection, before macroscopical symptoms are visible or before sporulation. We describe here a pathogenicity assay based on the relative quantification of fungal and plant DNA in infected Arabidopsis thaliana leaves by means of real-time quantitative PCR. We show that it allows to monitor quantitatively the growth of the fungi Alternaria brassicicola and Botrytis cinerea in a sensitive and reliable way. Although highly sensitive, this test also exhibits a high robustness, which is crucial to significantly discriminate between lines displaying slightly different levels of resistance. Therefore, it allows to assess fungal development from the very first stages of infection and provides a fast and very practical alternative to currently described assays for phenotyping either plant mutant lines or fungal strains. (C) 2004 Elsevier SAS. All rights reserved.