我国是第一大苹果生产国.苹果茎沟病毒(apple stem grooving virus,ASGV)在我国各苹果产区分布广泛,侵染果树导致树势变弱,生长量减少,果实品质下降,严重威胁我国苹果产业的可持续发展.病毒检测是果树病毒病防控的重要前提.本研究根据苹果茎沟病毒外壳蛋白基因保守序列,设计了引物和探针,建立了微滴数字PCR(droplet digital PCR,ddPCR)检测方法.所建立的检测方法能特异性检出苹果茎沟病毒,检测重复性好,灵敏度比qRT-PCR法高10倍,可适用于田间样品及组培苗检测.该方法的建立为苹果茎沟病毒的检测提供了重要的技术手段.
Establishment and application of droplet digital RT-PCR detection method for apple stem grooving virus
China is the largest apple producer in the world.Apple stem grooving virus(ASGV),which impairs the growth and yield of apple trees,is widely distributed in apple producing areas of China.Virus detection is crucial for the prevention and control of apple viral diseases.In this research,we designed primers and probes according to the conserved regions of ASGV coat protein gene.A droplet digital PCR(ddPCR)for ASGV de-tection was established and assessed.The results showed that the ddPCR could detect ASGV specifically with high repeatability.The sensitivity of ddPCR is 10 times higher than that of qPCR.The ddPCR established in this research could be applied for virus detection in field samples of orchard and virus-free seedlings.
apple stem grooving virus(ASGV)droplet digital reverse transcription-polymerase chain reaction(ddRT-PCR)detection
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