阿克苏地区枣树花叶伴随病毒A(JuMaVA)的分子检测  

Molecular detection of jujube Mosaic-associated Virus A in Aksu

作　　者：王权 刘保军 张国新 宋单波 白剑宇 WANG Quan;LIU Baojun;ZHANG Guoxin;SONG Danbo;BAI Jianyu(College of Agriculture,Kimjiamng Agricultural University,Urimqi 830042,Xmjiang,China;Research Institute of Economic Forestry,Xinjiang Academy of Forestry Sciences,Urumqi 830063 Xinjiang,China)

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]新疆林科院经济林研究所,乌鲁木齐830063

出　　处：《果树学报》2022年第3期449-455,共7页Journal of Fruit Science

基　　金：新疆维吾尔自治区科技援疆计划项目(2018D02026)。

摘　　要：【目的】明确枣树花叶伴随病毒A的田间侵染动态,探索出病害防治的关键时间节点。【方法】以枣树花叶伴随病毒A全基因组序列为靶标设计序列特异性引物,建立枣树花叶伴随病毒A分子检测技术,并应用于枣树花叶伴随病毒A田间侵染动态的追踪检测与分析。【结果】该检测技术对枣树花叶伴随病毒A检测的最低质量浓度为4.168 pg·μL^(-1),可在叶片显症之前检测到该病毒。通过田间侵染动态的追踪检测与分析,明确了5月中下旬是防治该枣树花叶病毒病的关键时间节点。【结论】建立的分子检测技术具有可操作性,能够快速准确对枣树花叶伴随病毒A田间侵染动态进行追踪检测。利用已建立的分子检测技术开展了枣树花叶伴随病毒A田间侵染动态的追踪检测,初步明确了枣树花叶伴随病毒A的田间侵染动态,明确了枣树叶片带毒率随着月份的增加而增加。其中5—6月份检出率较低,7—8月份的检出率明显高于5—6月份,这与病害前期隐症、7月份叶片显症、8月份进入显症高峰期的田间调查结果相一致,但枣树花叶病毒A与枣树花叶和果实畸形的关系还需进一步实验验证。【Objective】Jujube Mosaic-associated Virus A(JuMaVA) was a novel member of the genus Badnavirus in the Caulimoviridae, infesting Chinese jujube tree grown in Aksu, Xinjiang, China. The occurrence regularity of the jujube mosaic-associated virus A in the field was not clear, so it was very difficult to provide proper prevention and control technology guidance for local jujube farmers. Growers usually ignore the prevention and control of the disease in the early stage due to the lack of obvious symptom of mosaic and malformation of jujube leaves. Therefore, it was very important to develop the early diagnosis and monitoring of jujube virus disease and explore the key time point of disease control using the molecular detection technology.【Methods】A pair of highly specific primers(JCD-F)/(JCDR) were designed using Primer 5.0 software based on the genome sequence of Chinese jujube mosaicassociated virus A which had been cloned from jujube leaves by high throughput sequencing. The specificity of primers were tested by Blast using Blastn in NCBI, and 7 positive samples of plant virus diseases were used as primer-specific verification materials, including jujube witches’ broom, clove virus, fig virus, apple mosaic virus, tomato yellow leaf curl virus, alfalfa virus and watermelon melon virus collected from different fields and orchards in 2019 to 2020 by plant pathology laboratory of Xinjiang Academy of Forestry in different areas in Xinjiang. The total DNA of samples for the specificity and sensitivity verification of primers were extracted according to the instructions of the plant DNA extraction kit, and the molecular detection technology was established by optimizing the reaction system. The samples of jujube leaves with typical mosaic and malformation were collected from Chinese jujube orchards in different growth periods(from May to September) in 2020. The total DNA of 360 diseased leaves samples of jujube were extracted for specificity detection tests. The molecular detection technology was applied to det

关 键 词：枣树花叶伴随病毒 枣树DNA病毒 特异性PCR引物分子检测 侵染动态 

分 类 号：S665.1[农业科学—果树学] S436.65[农业科学—园艺学]