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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 an RT-PCR method for the detection of apple mosaic virus (ApMV) in field and to reveal the occurrence of ApMV in China, the methods of total RNA extraction from ApMV RT-PCR system System and program were selected and optimized. The detection of ApMV in 23 cities (counties) in 13 provinces of main apple producing areas in China was carried out by using optimized detection methods. The total RNA extracted from the field samples extracted by modified RNA extraction method was used as a template. The sensitivity of RT-PCR optimized system was able to detect ApMV in 15 μg of field samples and to detect the virulence of bark in 1-year-old branches throughout the growth and development of fruit trees ApMV was detected in 23 provinces (counties) in 13 provinces in China. The infected rate of ApMV was 80.1% in 327 samples, of which 205 were not detected, and the positive rate of ApMV was 68.3 %; In 23 regions, only zhangjiakou, henan, sanmenxia and shaanxi white water sample rate of less than 50%.