| 王 翀1, 张小菊1, 张祥林1, 张 伟1, 张 瑜1, 李亚伟1, 孙燕飞2*.实时荧光PCR检测瓜炭疽病菌[J].植物保护,2016,42(2):129-135. |
| 实时荧光PCR检测瓜炭疽病菌 |
| Detection of Colletotrichum orbiculare by the real-time PCR |
| 投稿时间：2015-01-07 修订日期：2015-04-06 |
| DOI： |
| 中文关键词: 实时荧光PCR 检测 瓜炭疽病菌 |
| 英文关键词:real-time PCR detection Colletotrichum orbiculare |
| 基金项目:国家质量监督检验检疫总局科研计划项目(2013IK287, 2011IK168); 国家科技部质检公益性行业科研专项(201310091) |
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| 中文摘要: |
| 采用实时荧光PCR技术建立了瓜炭疽病菌(Colletotrichum orbiculare)的检测方法。根据瓜炭疽病菌甘油醛-3-磷酸脱氢酶(GAPDH)基因和谷氨酰胺合成酶(GS)基因序列, 设计了该病菌特异性引物和TaqMan探针, 并对所设计的引物和探针的反应条件进行了优化。采用本试验建立的实时荧光PCR方法对瓜上的其他菌株及近似菌株进行检测, 可将瓜炭疽病菌与其他病原菌区分开。灵敏度试验表明, 25 μL体系中只要有39.6 pg的核酸量就可以被检测到, 检测灵敏度达到1.584 pg/μL, 比普通PCR检测灵敏度高100倍。同时对田间采集的病株和未知样品进行的检测证明了引物和TaqMan探针的特异性。 |
| 英文摘要: |
| The detection of Colletotrichum orbiculare by real-time PCR was established.The specific primers and TaqMan probes were designed according to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene and glutamine synthase (GS) gene sequences in C. orbiculare, and the reaction conditions were optimized. C. orbiculare could be detected from other strains in melon and other similar strains by real-time PCR. The sensitivity test indicated that 39.6 pg DNA could be detected in 25 μL reaction system. The real-time PCR sensitivity could reach to 1.584 pg/μL, 100 times more sensitive than ordinary PCR. The primers and Taqman probe were shown to be specific by detecting infected plants sampled in the field and unknown samples. |
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