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首页> 中文期刊> 《农业科学学报（英文版）》 >Combining simplified DNA extraction technology and recombinase polymerase amplification assay for rapid and equipment-free detection of citrus pathogen Phytophthora parasitica

Combining simplified DNA extraction technology and recombinase polymerase amplification assay for rapid and equipment-free detection of citrus pathogen Phytophthora parasitica

         
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摘要

Foot and root rot caused by Phytophthora parasitica is a substantial threat to citrus cultivation,affecting both yield and quality.Thus,rapid and accurate detection of P.parasitica plays an important role in disease management.The aim of this study was to develop a simple diagnostic method to detect P.parasitica infection by combining recombinase polymerase amplification and lateral flow strips(LF-RPA).To establish the LF-RPA assay of P.parasitica,the primers and probe designed based on the Ypt1 gene were tested for specificity to P.parasitica,which showed no cross-reactivity with DNAs of other related oomycete species.The LF-RPA assay detected the amount of genomic DNA of P.parasitica which was as low as 1 pg.To make the LF-RPA assay useful in low-resource settings,four simplified DNA extraction methods were compared,after which the LF-RPA assay was applied,with no specialized equipment,to analyze a diverse range of citrus tissues by using a simplified PEG-NaOH method for DNA extraction.This method was successful in detecting P.parasitica in infected plant samples within 30 min.Combining the LF-RPA assay and a simplified DNA extraction method could be a potential detection test for P.parasitica,especially in areas with limited resources.

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  • 来源
    《农业科学学报（英文版）》 |2021年第10期|2696-2705|共10页
  • 作者

    CHEN Wei-yu; YU Jia; XU Heng; LU Xin-yu; DAI Ting-ting; TIAN Yue-e; SHEN Dan-yu; DOU Dao-long;

  • 作者单位

    Nanjing Plant Protection and Quarantine Station Nanjing 210036 P.R.China;

    Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 P.R.China;

    College of Chemical and Biological Engineering Taiyuan University of Science and Technology Taiyuan 030024 P.R.China;

    Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 P.R.China;

    Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 P.R.China;

    College of Forestry Nanjing Forestry University Nanjing 210037 P.R.China;

    Department of Plant Protection Henan University of Science and Technology Luoyang 471000 P.R.China;

    Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 P.R.China;

    Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 P.R.China;

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