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| 利用基因YKT6鉴定疫霉属的不同种 | |
| 引用本文： | 赵伟,迟元凯,段劲生,陈晴晴,Myat Phyu Khine,汪涛,戚仁德.利用基因YKT6鉴定疫霉属的不同种[J].植物病理学报,2019,49(1):43-55. |
| 作者姓名： | 赵伟 迟元凯 段劲生 陈晴晴 Myat Phyu Khine 汪涛 戚仁德 |
| 作者单位： | 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; 农业部合肥作物有害生物科学观测实验站,合肥 230031; 缅甸教育部研究与创新部生物技术研究系,皎克西 05151 |
| 摘 要： | 利用分子标记或对特异位点的碱基序列进行分析是植物病原物分子检测的基础,可以在属和种的水平上对物种进行区分和鉴定。对疫霉属的不同种已有一系列的分子检测方法。SNARE蛋白相关基因YKT6拥有保守的侧翼编码区,适于设计疫霉属特异性的PCR引物,同时其内含子所具有的多态性可开发出几乎所有疫霉种的分子标记。利用疫霉属特异性引物对P-YKT6-F/P-YKT6-R可在31个疫霉种中特异地扩增出一条约600 bp的条带,而在腐霉或其他真菌中不能扩增出该条带。利用大豆疫霉的引物对Ps-YKT6-F/ Ps-YKT6-R和辣椒疫霉的引物对Pc-YKT6-F/Pc-YKT6-R,能分别从大豆疫霉菌株和辣椒疫霉菌株中扩增出一条399 bp和282 bp的条带,常规PCR和巢式PCR的灵敏度分别达到100 pg和10 fg。利用这些引物也可从土壤和病组织中检测到目标病原菌。此外,利用上述特异性引物开发出了大豆疫霉和辣椒疫霉的实时定量PCR检测方法。基于YKT6基因的分子标记和检测方法可用于疫霉种的调查检测和法定定量检测。 |
| 关 键 词： | 分子标记 辣椒疫霉 大豆疫霉 YKT6 |
| 收稿时间： | 2018-01-23 |
Identification of Phytophthora Species by Specific Primers Derived from YKT6 Gene |
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| ZHAO Wei,CHI Yuan-kai,DUAN Jin-sheng,CHEN Qing-qing,MYAT Phyu Khine,WANG Tao,QI Ren-de.Identification of Phytophthora Species by Specific Primers Derived from YKT6 Gene[J].Acta Phytopathologica Sinica,2019,49(1):43-55. | |
| Authors: | ZHAO Wei CHI Yuan-kai DUAN Jin-sheng CHEN Qing-qing MYAT Phyu Khine WANG Tao QI Ren-de |
| Institution: | Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China; Scientific Observing and Experimental Station of Crop Pests in Hefei, Ministry of Agriculture, Hefei 230031, China; Biotechnology Research Department, Department of Research and Innovation, Ministry of Education, Kyaukse 05151, Myanmar |
| Abstract: | A series of molecular methods have been developed for the detection of Phytophthora species. Development of molecular markers and sequence analysis of specific loci has been proved to be the most accurate technique for identification at generic and species level. The SNARE-related gene YKT6 possesses conserved flanking coding regions, which are appropriate for the design of Phytophthora genus-specific PCR primers, with sufficient polymorphism for the development of molecular markers for almost all Phytophthora species. The Phytophthora genus-specific primer set P-YKT6-F/P-YKT6-R were able to specifically amplify a single product of about 600 bp in 31 different Phytophthora species. With no product from Pythium spp. or fungi tested. The specific primers sets Ps-YKT6-F/Ps-YKT6-R or Pc-YKT6-F/Pc-YKT6-R generated specific products of 399 bp and 282 bp from isolates of P. sojae and P. capsici, respectively. For both species, the sensitivity achieved was 100 pg for stan-dard PCR and 10 fg for nested PCR. These primers also proved to be efficient in detecting pathogens from soil and diseased plants. In addition, real-time quantitative PCR assays coupled with the species-specific primers were developed to detect and quantify the two species. The results demonstrated that the YKT6-based molecular marker and assays have clear potential for detection and quantification of Phytophthora spp. in a range of contexts from pathogen surveys to statutory testing. |
| Keywords: | molecular marker Phytophthora capsici P sojae YKT6 |
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