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| 以Ypt1基因序列为靶标的辣椒疫病菌快速分子检测 | |
| 引用本文： | 李毛毛,赵伟,汪涛,谷雨,高智谋,戚仁德.以Ypt1基因序列为靶标的辣椒疫病菌快速分子检测[J].植物病理学报,2014,44(5):546-551. |
| 作者姓名： | 李毛毛 赵伟 汪涛 谷雨 高智谋 戚仁德 |
| 作者单位： | 安徽农业大学植物保护学院,合肥 230036; 安徽省农业科学院植物保护与农产品质量安全研究所,合肥 230031 |
| 基金项目： | 国家公益性行业(农业)科研专项(201303018);安徽省农科院院长青年创新基金项目(11B1111) |
| 摘 要： | |
| 收稿时间： | 2014-01-28 |
Rapid molecular detection of Phytophthora capsici based on its Ypt1 gene |
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| LI Mao-mao,ZHAO Wei,WANG Tao,GU Yu,GAO Zhi-mou,QI Ren-de.Rapid molecular detection of Phytophthora capsici based on its Ypt1 gene[J].Acta Phytopathologica Sinica,2014,44(5):546-551. | |
| Authors: | LI Mao-mao ZHAO Wei WANG Tao GU Yu GAO Zhi-mou QI Ren-de |
| Institution: | School of Plant Protection, Anhui Agricultural University,Hefei 230036, China; Institute of Plant Protection and Agro-products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China |
| Abstract: | PCR primers were designed based on the sequence of Ras-related protein gene (Ypt1) of P. capsici. According to the multiple sequence alignment, Ypt1 has the sufficiently polymorphic intron region for the development of P. capsici-specific primers (PcYpt1F/PcYpt1R). One primer pair was developed which can amplify one P. capsici-specific fragment of 156 bp. Using the primer pair, the P. capsici infected plants and soils were detected. Additionally, Ypt1 has an appropriate region for the development of Phytophthora genus-specific primers (Ypt1F/Ypt1R), which can amplify a fragment of about 540 bp from 14 different Phytophthora specices and a fragment of about 350 bp in Pythium species, with no amplification from fungal species. By PCR optimization using P. capsici genomic DNA, the detection sensitivities of 10 pg and 10 fg DNA were achieved in standard PCR (PcYpt1F/PcYpt1R) and nested PCR (Ypt1F/Ypt1R and PcYpt1F/PcYpt1R), respectively. The developed primers were proved to be efficient in detection of Phytophthora pathogens from diseased plant tissues and residues in soils. |
| Keywords: | Phytophthora capsici molecular detection Ypt1 gene |
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