Development of one-step multiplex RT-PCR assay for rapid simultaneous detection of five RNA viruses and Acidovorax citrulli in major cucurbitaceous crops in China
文献类型：期刊论文
| 作者 | Zhao, Zhenxing; Xiang, Jun; Tian, Qian; Zhao, Wenjun; Zhou, Tao; Zhao, Lixia ; Zhang, Yongjiang
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| 刊名 | ARCHIVES OF MICROBIOLOGY
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| 出版日期 | 2022 |
| 卷号 | 204期号:12页码:696-1-12 |
| ISSN号 | 0302-8933 |
| 关键词 | One-step mRT-PCR Detection Cucurbitaceous crops Virus Acidovorax citrulli |
| 英文摘要 | Cucurbitaceous fruits and vegetables are important crops. Viral and bacterial diseases cause substantial economic losses to cucurbit crops globally. For rapid detection of these pathogens and improved disease control, a one-step multiplex reverse-transcription polymerase chain reaction (mRT-PCR) system was created. This method allowed for the concurrent detection of Tobacco mosaic virus (TMV), Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), Cucumber green mottle mosaic virus (CGMMV), Cucumber mosaic virus (CMV), and Acidovorax citrulli. Five pairs of specific primers were created according to the conserved regions around the coat protein (CP) genes of each virus, and one pair was based on the A. citrulli internal transcribed spacer (ITS). To limit false negatives, one pair of primers, created based on the Transcriptional elongation factor 1-alpha (EF1-alpha) from the major cucurbitaceous crop species, was put into the mRT-PCR reaction system. Primer concentrations, annealing temperature, extension time, and amplification cycles were optimized. Anticipated fragments of 152 bp (TMV), 205 bp (ZYMV), 318 bp (WMV), 419 bp (CGMMV), 529 bp (CMV), 662 bp (A. citrulli), and 821 bp (EF1-alpha) were amplified by the multiplex RT-PCR system, and their origin was established via DNA sequencing. This method was successfully used to examine field-collected seed samples of cucurbitaceous crops from China. The results demonstrated that the one-step mRT-PCR technique is a quick, efficient, and sensitive assay for the concurrent detection of six pathogens of cucurbits. It provides a method for monitoring and preventing these diseases. |
| 源URL | [https://ir.rcees.ac.cn/handle/311016/48484]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 作者单位 | 1.Nanjing Agricultural University 2.China Agricultural University 3.Chinese Academy of Sciences 4.Research Center for Eco-Environmental Sciences (RCEES) 5.Ministry of Agriculture & Rural Affairs 6.Chinese Academy of Inspection & Quarantine |
| 推荐引用方式 GB/T 7714 | Zhao, Zhenxing,Xiang, Jun,Tian, Qian,et al. Development of one-step multiplex RT-PCR assay for rapid simultaneous detection of five RNA viruses and Acidovorax citrulli in major cucurbitaceous crops in China[J]. ARCHIVES OF MICROBIOLOGY,2022,204(12):696-1-12. |
| APA | Zhao, Zhenxing.,Xiang, Jun.,Tian, Qian.,Zhao, Wenjun.,Zhou, Tao.,...&Zhang, Yongjiang.(2022).Development of one-step multiplex RT-PCR assay for rapid simultaneous detection of five RNA viruses and Acidovorax citrulli in major cucurbitaceous crops in China.ARCHIVES OF MICROBIOLOGY,204(12),696-1-12. |
| MLA | Zhao, Zhenxing,et al."Development of one-step multiplex RT-PCR assay for rapid simultaneous detection of five RNA viruses and Acidovorax citrulli in major cucurbitaceous crops in China".ARCHIVES OF MICROBIOLOGY 204.12(2022):696-1-12. |
入库方式： OAI收割
来源：生态环境研究中心
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