Quantification of Magnaporthe grisea during infection of rice plants using real-time polymerase chain reaction and northern blot/phosphoimaging analyses
Quantification of Magnaporthe grisea during infection of rice plants using real-time polymerase chain reaction and northern blot/phosphoimaging analyses
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DOI：
10.1094/phyto.2002.92.8.870
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发表时间：
2002-08-01
期刊：
影响因子：
3.2
通讯作者：
Yang, YN
中科院分区：
文献类型：
--
作者：
Qi, M;Yang, YN
稻瘟病，由稻瘟病菌引起。是世界范围内严重的水稻真菌病害。目前，对真菌致病性和宿主抗性的评估主要基于疾病等级或对blast病变数量和大小的测量。然而。这些方法只能提供直观的估计，而不能准确测量真菌在水稻植株中的生长情况。在本研究中，采用基于dna的实时聚合酶链反应（PCR）和基于rna的非免疫印迹/荧光成像分析来定量稻瘟病菌。两种方法都很灵敏。具体。可准确测定稻瘟病菌的相对生长量和绝对生物量。实时荧光定量PCR分析表明，接种后4和6 d，感病品种（M201和Wells）幼苗叶片中稻瘟病菌的生长速度是抗性品种（Drew）的约46 ~ 80倍。实时PCR检测结果与northern blot/phosphoimaging分析结果一致。然而。在大多数情况下，实时PCR方法更快、更方便。因此，该方法是测定稻瘟病菌在植株内的致病性和抗性的有效工具。
Rice blast, caused by Magnaporthe grisea. is a serious fungal disease of rice worldwide. Currently, evaluation of the fungal pathogenicity and host resistance is mainly based on a disease rating or measurement of blast lesion number and size. However. these methods only provide visual estimation rather than accurate measurement of fungal growth in rice plants. In this study, DNA-based real-time polymerase chain reaction (PCR) and RNA-based nor-them blot/phosphoimaging analyses were evaluated to quantify M. grisea. Both methods were sensitive. specific. and reproducible and could accurately measure the relative growth and absolute biomass of M. grisea. The real-time PCR analysis showed that the growth of M. grisea in seedling leaves of susceptible cultivars (M201 and Wells) was approximate to46 to 80 times higher than that of a resistant cultivar (Drew) at 4 and 6 days after inoculation. The data obtained from the real-time PCR assays also were consistent with that from northern blot/phosphoimaging analysis. However. the real-time PCR approach was much faster and more convenient in most cases. Therefore, it is an excellent tool for in planta quantification of M. grisea and can be used for reliable assessment of fungal pathogenicity and host resistance.