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为了开发苹果锈果类病毒(ASSVd)快速、准确的RT-PCR检测方法,以感染苹果锈果类病毒的田间苹果枝条为试材,对苹果锈果类病毒RT-PCR反应体系和程序进行选择和优化,利用优化的检测体系对保定市曲阳县苹果园苗木及3~5年生母本树苹果锈果病的发生情况进行检测,验证该体系的可靠性。结果表明:自行设计引物ASSVd QCxin3特异性最好,优化体系灵敏度达到能够检测3.75 ng新鲜样本中的病毒,可准确、灵敏检测苹果锈果类病毒田间样本;曲阳县苹果园48株苗木中携带苹果锈果类病毒13株,带毒率为27.1%;18株母本树中携带苹果锈果类病毒5株,带毒率为27.8%。
In order to develop a rapid and accurate RT-PCR detection method for apple rust-causing virus (ASSVd), the field apple shoots infected with apple rust-causing virus were used as materials to select the RT-PCR reaction system and program of apple rust- And optimize the use of optimized detection system in Quyang County, Baoding apple orchard seedlings and 3 to 5-year-old female tree apple rust test results detected the system to verify the reliability of the system. The results showed that ASSVd QCxin3 was the best one, and the sensitivity of optimized system was able to detect the virus in fresh sample of 3.75 ng, which could accurately and sensitively detect field samples of apple rust-causing virus. 48 apple trees in apple orchard of Quyang County carried apple There were 13 rust fruit viruses with a rate of 27.1%. Five strains of apple rust-causing fruit viruses were carried in 18 mother trees with a virulence rate of 27.8%.