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期 刊 ：植物保护学报 2012年

关键词：苹果花叶病毒；RNA提取；RT-PCR；分布；

Keywords:Apple mosaic virus, RNA extraction, RT-PCR, distribution,

摘 要 ：为了开发苹果花叶病毒(Apple mosaic virus,ApMV)田间样本RT-PCR检测方法和揭示我国ApMV的发生情况,以田间染病苹果组织为材料,对ApMV RT-PCR检测体系中总RNA提取方法、反应体系及程序进行了选择和优化,并利用优化的检测方法对我国苹果主产区13个省的23个市(县)ApMV发生情况进行了检测。以RNA提取改良法提取的田间样本总RNA为模板,RT-PCR优化体系的灵敏度达到能够检测15 μg田间样本组织中的ApMV,且在果树整个生长发育时期能够检测1年生枝条树皮组织的带毒情况;ApMV在我国普遍发生,13个省的23个市(县)均检测到了ApMV,采集的327个样本带毒率为80.1%,其中未显花叶症状样本205个,ApMV阳性率为68.3%;在23个地区中,只有河北张家口、河南三门峡和陕西白水样本带毒率低于50%。

Abstract:In order to develop RT-PCR method for detection of Apple mosaic virus (ApMV) in field and make clear the occurrence of ApMV in China, RT-PCR detection system, including RNA-extraction method, reaction system and program of PCR, was optimized using the apple tissue infected by ApMV. The occurrence of ApMV in 13 provinces (23 cities/counties) of China was clarified by the modified RT-PCR assay. The result showed that the sensitivity of the optimized RT-PCR method could reach as less as 15 μg fresh sample using the total RNA extracted by the improved RNA-extraction method as template. In addition, the modified RT-PCR assay could detect ApMV using the bark tissue of one-year old branch as material during the whole growth period. In China, ApMV was detected in all of the 23 cities (counties). The ApMV positive rate of the 327 samples reached 80.1% and the positive rate of the 205 symptomless samples was 68.3%. The positive rate was relative lower in three areas, including Zhangjiakou City of Hebei, Sanmenxia City of Henan and Baishui City of Shaanxi, which was lower than 50%.

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