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Image_2_Development and application of crude sap-based recombinase polymerase amplification assay for the detection and occurrence of grapevine geminivirus A in Indian grapevine cultivars.jpeg

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  • معلومة اضافية
    • الموضوع:
      2023
    • Collection:
      Frontiers: Figshare
    • نبذة مختصرة :
      Geminiviruses are known to infect several fields and horticultural crops around the globe. Grapevine geminivirus A (GGVA) was reported in the United States in 2017, and since then, it has been reported in several countries. The complete genome recovered through high-throughput sequencing (HTS)-based virome analysis in Indian grapevine cultivars had all of the six open reading frames (ORFs) and a conserved nonanucleotide sequence 5′-TAATATTAC-3′ similar to all other geminiviruses. Recombinase polymerase amplification (RPA), an isothermal amplification technique, was developed for the detection of GGVA in grapevine samples employing crude sap lysed in 0.5 M NaOH solution and compared with purified DNA/cDNA as a template. One of the key advantages of this assay is that it does not require any purification or isolation of the viral DNA and can be performed in a wide range of temperatures (18°C–46°C) and periods (10–40 min), which makes it a rapid and cost-effective method for the detection of GGVA in grapevine. The developed assay has a sensitivity up to 0.1 fg μl -1 using crude plant sap as a template and detected GGVA in several grapevine cultivars of a major grapevine-growing area. Because of its simplicity and rapidity, it can be replicated for other DNA viruses infecting grapevine and will be a very useful technique for certification and surveillance in different grapevine-growing regions of the country.
    • Relation:
      https://figshare.com/articles/figure/Image_2_Development_and_application_of_crude_sap-based_recombinase_polymerase_amplification_assay_for_the_detection_and_occurrence_of_grapevine_geminivirus_A_in_Indian_grapevine_cultivars_jpeg/22239232
    • الرقم المعرف:
      10.3389/fpls.2023.1151471.s002
    • الدخول الالكتروني :
      https://doi.org/10.3389/fpls.2023.1151471.s002
      https://figshare.com/articles/figure/Image_2_Development_and_application_of_crude_sap-based_recombinase_polymerase_amplification_assay_for_the_detection_and_occurrence_of_grapevine_geminivirus_A_in_Indian_grapevine_cultivars_jpeg/22239232
    • Rights:
      CC BY 4.0
    • الرقم المعرف:
      edsbas.5E6D798E