A sensitive SYBR Green RT-qPCR method for grapevine virus E and its application for virus detection

来源 :农业科学学报(英文) | 被引量 : 0次 | 上传用户:zq19900303
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
To develop a rapid and high-sensitivity method for detection of grapevine virus E(GVE),a SYBR Green based real-time fluorescence quantitative RT-PCR method(RT-qPCR)was established.This method could be used to detect GVE specifically,and the sensitivity was about 100 times greater than conventional RT-PCR.An excellent linear correlation(R2=0.997)and a high amplification efficiency(E=97.5%)were obtained from the standard curve of this method.Reproducibility tests revealed that the coefficients of variation in the intra-and inter-assay results were 0.31-1.03%and 0.82-2.62%,respectively,indicating a good reproducibility.The RT-qPCR method could be used to detect GVE in a wide range of grapevine sample types.The detection rates of RT-qPCR for nearly all sample types from different positions and seasons were higher than conventional RT-PCR.The detection rates in spring,summer,autumn and winter increased gradually.Samples in autumn and winter were best for detection,and the detection rates of most samples were 80-100%,which were 10 to 40%higher than conventional RT-PCR.In general,old petioles and branches were the best tissues for GVE detection.The detection rates of these samples in each season were all 100%,which were 20 to 40%higher than conventional RT-PCR.The second highest rates were in the old leaf,with detection rates for RT-qPCR of 80-100%in all seasons,which were 20 to 40%higher than conventional RT-PCR.GVE could be difficultly detected in young leaves by conventional RT-PCR,and the detection rates were only 0-50%,while by RT-qPCR the rates could increase to 0-80%.A total of 33 out of 363 samples(belonging to 68 cultivars)from 20 regions in China were detected to be positive by RT-qPCR(9.1%),which was more than twice the rate of the conventional RT-PCR(3.9%).
其他文献
社会网络分析的应用拓宽了医疗与公共卫生的研究范式和实验场域,促进了流行病学、医疗与公共卫生服务、卫生系统等研究的纵深发展.公共卫生突发事件及流行病防疫加速了社会网
地源热泵是利用浅层地热能对建筑物进行供暖制冷的高效节能环保型空调,其中垂直土壤埋管系统由于节约用地、换热性能好、不消耗地下水,在我国得到了最广泛的应用。地源热泵的
热计量仪表是用于供暖系统中计算和实时显示介质通过热交换回路过程中热能变化量的装置。热计量仪表不仅要计算和显示换热过程中的热能变化量,还要方便人们的监测与数据保存。热计量系统采用超低功耗单片机MSP430F149为控制器,在以LabVIEW为平台实现监测软件的设计。本文对热计量计算方法及原理进行研究,选择了合适的热计量算法。研究温度测量、流量测量及热量计算的原理及方法,选择合适的的传感器及相关计量仪
活性炭是一种孔隙发达、比表面积巨大、吸附性能强的功能性的碳材料,广泛应用于溶剂与废气回收、空气净化、水处理等工业。随着社会的发展以及人们对生活环境的重视,活性炭的需求量呈逐年上升趋势。提高活性碳生产技术、研究活性炭生产设备的全自动化控制,从而提高生产效率、节约生产成本成为活性炭生产中的一个严峻的问题。通过对活性炭混捏成型生产工艺的研究,结合厂家的要求,本论文设计了一套针对活性炭生产过程中混捏成型过
杏鲍菇工厂化生产的目标是获得高品质、高产量的优质商品菇。杏鲍菇生长环境因子的调控理论的研究对品质与产量提高有积极的意义。光照是影响杏鲍菇生长发育的关键因子,目前
断裂构造的定量预测一直是地质工作中的一个难点。断裂构造影响了矿井的安全生产和煤矿开采效率。本文在前人研究煤矿断裂构造预测的基础上,收集北阳庄煤矿相关资料,研究相关区
浅部煤炭开采资源的逐渐枯竭使得深部开采的问题逐渐凸显,煤系地层中普遍存在的砂岩裂隙水是影响矿井正常生产的重要因素,而砂岩裂隙水的富水性与裂隙的发育与分布密不可分,开采
新建铁路南钦线D2K93+000~D2K95+000段经过横岭井煤矿采空区及稔子坪、大垌煤矿采空区边缘地带,线路附近已有明显的地表塌陷。为了确保南钦线在建设及将来运行中的安全,需要对
绕组变形是诱发电力变压器多种故障和事故的直接原因,因此及时检测绕组变形是预防变压器故障的重要措施之一。相比传统的电量检测法,振动检测法在检测绕组变形故障上有其明显优
拉力试验机是通过拉伸试验来检测金属材料、非金属材料的强度、刚度、弹性、塑性等机械性能的基本设备之一,拉力试验机所测定的试验数据是从事工程设计和保证产品安全工作的依