Pineapple mealybug wilt associated virus-2 （PMWaV-2） is an important pathogen causing Mealybug wilt of pineapple （MWP）. We established a realtime quantitative RT-PCR （RT-qPCR） method with TaqMan probe based on s...Pineapple mealybug wilt associated virus-2 （PMWaV-2） is an important pathogen causing Mealybug wilt of pineapple （MWP）. We established a realtime quantitative RT-PCR （RT-qPCR） method with TaqMan probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was higldy sensitive to positive sample, but had no fluorescence signal to health sample and blank control. The sensitivity of RTqPCR was about 100 times higher than general PCR. Reproducibihty test revealed that the coefficients of variation between intra-and inter-assay were beth within 1.85%, indicating it was a quantitative detection method for PMWaV-2 with simple operation, strong specificity, high sensitivity, and reliable reproducibility.展开更多
[ Objective ] The paper aimed to establish a real-time fluorescent quantitative PCR （qPCR） detection method for Pineapple mealybug wilt associated vi- rus-3 （ PMWaV3 ）. [ Method] Specific TaqMan probe and primers ...[ Objective ] The paper aimed to establish a real-time fluorescent quantitative PCR （qPCR） detection method for Pineapple mealybug wilt associated vi- rus-3 （ PMWaV3 ）. [ Method] Specific TaqMan probe and primers were designed and synthesized according to the conserved sequence of coat protein（CP） gene of PMWaV-3, and the standard curve was established after optimizing the amplification condition of qPCR. [ Result] The results showed that the method was specific for the detection of PMWaV-3, and the sensitivity of the present method was about 10 times higher compared to general RT-PCR ; the variation coefficients of intra- assay and inter-assay were less than 1.73, respectively. [ Conclusion] The qPCR is an easy, fast and reliable method for quantitative detection of PMWaV-3.展开更多
为掌握海南琼海地区菠萝凋萎病(Mealybug Wilt Disease of Pineapple,MWP)发生概况的基础数据,基于2018年以来琼海菠萝主产乡镇MWP田间显症率数据,从菠萝生育期、果园土壤类型和季节3方面分析了MWP田间发生特点。结果显示,MWP总体显症率...为掌握海南琼海地区菠萝凋萎病(Mealybug Wilt Disease of Pineapple,MWP)发生概况的基础数据,基于2018年以来琼海菠萝主产乡镇MWP田间显症率数据,从菠萝生育期、果园土壤类型和季节3方面分析了MWP田间发生特点。结果显示,MWP总体显症率约30%;全生育期MWP显症率呈现先高后低再渐升的趋势,即育苗期>大果期>小果期>花期>催花期>返青期>速生期;从果园土壤类型来看,MWP显症率沙土>砂壤土>壤土;从季节上看,MWP显症率呈夏季<秋季<冬春季的特点,水肥管理水平较高的果园显症率较管理不到位的低,且存在带毒种苗在产区间调运传播可能。琼海地区MWP发生程度是种苗、生育期、土壤、季节和管理等因子综合作用的结果,而非单一因子决定,鉴于此提出相关防控措施建议。展开更多
基金Supported by Applied Research and Industrialization Projects of Key Science and Technology Plan of Hainan Province(ZDXM20130031)Special Fund for Agro-scientific Research in the Public Interest(201203021)+3 种基金Scientific Operation Fund of Hainan Province(QCY[2013]131)Natural Science Foundation of Hainan Province(QK[2013]32)Key Research and Development Project of Hainan Province(ZDYF2016035)Special Program for Agricultural Science and Technology Innovation of Hainan Academy of Agricultural Sciences(CXZX201410)
文摘Pineapple mealybug wilt associated virus-2 （PMWaV-2） is an important pathogen causing Mealybug wilt of pineapple （MWP）. We established a realtime quantitative RT-PCR （RT-qPCR） method with TaqMan probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was higldy sensitive to positive sample, but had no fluorescence signal to health sample and blank control. The sensitivity of RTqPCR was about 100 times higher than general PCR. Reproducibihty test revealed that the coefficients of variation between intra-and inter-assay were beth within 1.85%, indicating it was a quantitative detection method for PMWaV-2 with simple operation, strong specificity, high sensitivity, and reliable reproducibility.
基金Supported by Applied Research and Industrialization Projects of Key Science and Technology Plan of Hainan Province(ZDXM20130031)Special Fund for Agro-scientific Research in the Public Interest(201203021)+1 种基金Scientific Operation Fund of Hainan Province(QCY[2013]131)Natural Science Foundation of Hainan Province(QK[2013]32)
文摘[ Objective ] The paper aimed to establish a real-time fluorescent quantitative PCR （qPCR） detection method for Pineapple mealybug wilt associated vi- rus-3 （ PMWaV3 ）. [ Method] Specific TaqMan probe and primers were designed and synthesized according to the conserved sequence of coat protein（CP） gene of PMWaV-3, and the standard curve was established after optimizing the amplification condition of qPCR. [ Result] The results showed that the method was specific for the detection of PMWaV-3, and the sensitivity of the present method was about 10 times higher compared to general RT-PCR ; the variation coefficients of intra- assay and inter-assay were less than 1.73, respectively. [ Conclusion] The qPCR is an easy, fast and reliable method for quantitative detection of PMWaV-3.
文摘为掌握海南琼海地区菠萝凋萎病(Mealybug Wilt Disease of Pineapple,MWP)发生概况的基础数据,基于2018年以来琼海菠萝主产乡镇MWP田间显症率数据,从菠萝生育期、果园土壤类型和季节3方面分析了MWP田间发生特点。结果显示,MWP总体显症率约30%;全生育期MWP显症率呈现先高后低再渐升的趋势,即育苗期>大果期>小果期>花期>催花期>返青期>速生期;从果园土壤类型来看,MWP显症率沙土>砂壤土>壤土;从季节上看,MWP显症率呈夏季<秋季<冬春季的特点,水肥管理水平较高的果园显症率较管理不到位的低,且存在带毒种苗在产区间调运传播可能。琼海地区MWP发生程度是种苗、生育期、土壤、季节和管理等因子综合作用的结果,而非单一因子决定,鉴于此提出相关防控措施建议。
