苹果花叶病毒的RT-PCR检测及其在我国苹果产区的分布

The RT-PCR detection and distribution of ApMV in apple producing area of China

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15μg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%。

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 Ixg 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 （coun- ties）. The ApMV positive rate of the 327 samples reached 80.1% and the positive rate of the 205 symp- tomless 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%.