|
|||||
|
|
| 一步法RT-PCR和两步法RT-PCR对马铃薯病毒诊断研究的比较 | |
| 引用本文： | 陈阳婷,宁红,张敏.一步法RT-PCR和两步法RT-PCR对马铃薯病毒诊断研究的比较[J].中国农学通报,2010,26(11):298-302. |
| 作者姓名： | 陈阳婷 宁红 张敏 |
| 作者单位： | 1. 成都市农林科学院园艺研究所,成都,610027 2. 四川省植物检疫站,成都,610041 3. 四川农业大学,四川雅安,625014 |
| 摘 要： | 本研究采用试剂盒和Total试剂两种方法从马铃薯叶片和茎秆中提取总RNA，同时设计了一步法RT-PCR和两步法RT-PCR的反应体系，依据马铃薯X病毒、马铃薯Y病毒、马铃薯S病毒、马铃薯A病毒以及马铃薯卷叶病毒的基因组RNA保守序列设计特异性引物对，并运用两种RT-PCR反应体系对染病的马铃薯进行病毒检测，结果与报道的这五种马铃薯病毒核苷酸序列进行BLAST比对, 表明扩增的五种马铃薯病毒靶带的序列与靶序列完全一致。两种诊断方法对马铃薯病毒核酸进行浓度检测，结果证明，两步法RT-PCR具有较高灵敏性。但这两种方法均可快速、简便、有效地诊断马铃薯病毒。 |
| 关 键 词： | 芹菜 芹菜 沼液 品质 干物质含量 |
| 收稿时间： | 2010-01-07 |
| 修稿时间： | 2010-02-27 |
Research on the Detection of Potato Viruses by One-step RT-PCR and Two-step RT-PCR |
|
| Chen Yangting,Ning Hong,Zhang Min.Research on the Detection of Potato Viruses by One-step RT-PCR and Two-step RT-PCR[J].Chinese Agricultural Science Bulletin,2010,26(11):298-302. | |
| Authors: | Chen Yangting Ning Hong Zhang Min |
| Institution: | 1Institue of Horticultural Science, Chengdu Academy of Agricultural and Forestry Sciences, Chengdu 610027; 2Plant Quarantine Station of Sichuan Agricultural Department, Chengdu 610041; 3Sichuan Agricultural University, Ya’an Sichuan 625014 |
| Abstract: | In order to establish a technique using RT-PCR to detect potato viruses, which was rapid, accurate and sensitive, the RNAprep Plant Kit and RNAiso Plus RNA-release methods were developed for extracting potato viruses （PVX, PVY, PVS, PVA, PLRV） from potato stems and leaves. The primers for the viruses were designed. The one step RT-PCR and two step RT-PCR were used to detect the five virus and the amplification fragment were 620 bp（PVX）, 550 bp（PVY）, 435 bp（PVS）, 300 bp（PVA） and 222 bp（PLRV）. The plasmids were extracted and sequenced, compared with those in NCBI BLAST; the result showed that the sequences of amplification products were correct. Serial dilutions were used to detect the sensitivity of one and two-step RT-PCR testing system. One-step RT-PCR testing system was also can used to detect many potato viruses with the optimized conditions but the sensitivity was inferior to two-step RT-PCR. |
| Keywords: | potato virus RNA-release methods one-step RT-PCR two-step RT-PCR |
| 本文献已被 维普 万方数据 等数据库收录！ | |
| 点击此处可从《中国农学通报》浏览原始摘要信息 | |
| 点击此处可从《中国农学通报》下载免费的PDF全文 | |