摘要： 为了调查天津地区种植梨的已知病毒病的发病情况，以雪花梨叶片为材料，采用改良的CTAB法对梨的嫩叶进行总RNA提取，并通过RT-PCR对天津3个地区70株不同品种的梨树所带的潜隐性病毒进行鉴定，包括苹果褪绿叶斑病毒（ACLSV）、苹果锈果类病毒（ASSVd）、苹果茎痘病毒（ASPV）、苹果茎沟病毒（ASGV）、苹果凹果类病毒（ADFVd）。结果表明，改良方法1-巯基乙醇法是提取梨叶片总RNA较为理想的方法，可获得的RNA杂质含量较少，总RNA得率较高，平均质量分数可达73.880 μg/g，获得的总RNA符合后续试验要求；利用RT-PCR法在70个检测样品中，上述5种病毒病的平均发生率依次为47.1%，11.4%，10.0%，8.6%，1.4%，复合侵染率平均为28.6%，种植年份越高的植株带毒率越高，不同品种对病毒的敏感性也不相同。研究结果表明，天津地区梨病毒病发病普遍，其中苹果褪绿叶斑病毒是主要带毒种类，而且具有一定比例的多种病毒复合侵染现象。梨树种植管理水平低，地区间梨树引种缺乏规范的检测监控均可导致梨病毒病的发生和扩散，这对于天津梨产业的健康发展十分不利。

关键词: 总RNA提取, 梨病毒病, 类病毒, 分子检测

Abstract: In order to investigate the incidence of known viral diseases of pear in Tianjin Region,the total RNA of snow pear leaves was extracted by improved CTAB method. And the latent virus species of the 70 pear trees from three regions of Tianjin were detected through RT-PCR identification,including Apple cholortotic leaf spot virus (ACLSV), Apple scar skin viroid (ASSVd), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple dimple fruit viroid (ADFVd). The results showed that the improved CTAB-mercaptoethanol method was a kind of comparatively ideal method for total RNA extraction from snow pear,the RNA had less impurity, the rate of total RNA extrate was higher, and the mean concentration was 73.880 μg/g. The obtained RNA meted the requirements for the next analysis. By RT-PCR technique the infection rates of the 70 pear trees with the above five viruses were 47.1%,11.4%,10.0%,8.6%,and 1.4%,respectively,and the mixed infection rate was 28.6% on average. The higher the age of pear trees, the higher the virus carrying rate. And the sensitivity of different varieties to viruses was different. The results showed that the pear virus disease in Tianjin was common, in which apple chlorotic leaf spot virus was the main type of virus, and had a certain proportion of multiple virus complex infection. The low level of pear planting management and the lack of standardized detection and monitoring of pear introduction among regions could lead to the occurrence and spread of pear virus disease. It was very unfavorable for the healthy development of Tianjin pear industry.

Key words: Total RNA extraction, Pear viral disease, Viroid, Molecular detection

中图分类号: 

• S661.2