朱志强,集贤,兰青阔,赵新,张小栓,张平.鲜食葡萄贮运期黑斑病害致病病原菌分离鉴定[J].包装工程,2020,41(21):31-37.
ZHU Zhi-qiang,JI Xian,LAN Qing-kuo,ZHAO Xin,ZHANG Xiao-shuan,ZHANG Ping.Isolation and Identification of Pathogens Causing Grape Blackspot during Storage and Transportation[J].Packaging Engineering,2020,41(21):31-37. |
| 鲜食葡萄贮运期黑斑病害致病病原菌分离鉴定 |
| Isolation and Identification of Pathogens Causing Grape Blackspot during Storage and Transportation |
| 投稿时间：2020-04-24 |
| DOI：10.19554/j.cnki.1001-3563.2020.21.005 |
| 中文关键词: 黑斑病 致病病原菌 分离鉴定 ITS序列分析 链格孢霉 |
| 英文关键词: black spot pathogen isolation and identification ITS sequences analysis Alternaria |
| 基金项目:天津市青年科研人员创新研究与实验项目(201920);天津市自然科学基金(17JCQNJC14700);国家葡萄产业技术体系(CARS-29-jg-1) |
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| 中文摘要: |
| 目的 通过研究鲜食葡萄黑斑病害的致病病原菌分离和鉴定，为鲜食葡萄采后病害防控措施开发提供理论支撑。方法 选用贮藏后的马奶等4个鲜食葡萄品种为研究对象，用组织分离法对引起黑斑病害的病原菌进行分离纯化，在PDA培养基上，观察拟菌株形态以及菌丝和孢子形态特征，采用有伤和无伤活体接种健康果实，并观察其发病特征，应用ITS通用引物对病原菌DNA进行扩增并测序。结果 分离出3种优势致病菌(A，B和C)，检出频率分别为75.0%，32.0%和42.11%。有伤和无伤活体接种均具有致病性，且有伤大于无伤。对比《真菌鉴定手册》，初步确定拟菌株A为链格孢霉，拟菌株B为灰葡萄孢霉，拟菌株C为拟茎点霉，采用rDNA-ITS序列分析法证实拟菌株A与KJ489375.1、GU190188.1的序列相似度达到99%，结合两者结果确定主要致病病原菌为链格孢霉属小孢子种。结论 明确引起鲜食葡萄贮运期黑斑病害的主要致病菌为链格孢霉属小孢子种。 |
| 英文摘要: |
| The work aims to provide theoretical support for development of prevention and control measures for post-harvest disease of grape by isolation and identification of pathogenic bacteria of fresh grape black spot disease. The article used four table grapes (MANAI grape etc.) during storage as testing material to separate and purify pathogenic bacteria which caused disease of black spot disease. The colonial morphology on PDA medium and microscopic characteristics of spores and hyphae morphology of every quasi strain was analyzed. The epidemiological characteristics on berry inoculated with and without wound. ITS universal primers were applied to amplify the genomic DNAs of pathogenic bacteria and sequence the fragments. Three priori pathogenic pseudo strains (identified as A, B and C) were isolated and the frequency of them was detected as 75.0%, 32.0% and 42.11% in sequence. Grape, inoculated with and without wound with three pseudo strains, were pathogenic, and there were higher pathogenicity in those with wound than those without wound. By comparison with the fungus identification manual, the article preliminarily judged that pseudo strain A, B and C was Alternaria, Botrytis cinereal and Phomopsis, respectively. The sequence analysis of rDNA-ITS confirmed that the sequence similarity between strain A and KJ489375.1 and GU190188.1 was 99%. The main pathogenic fungus was confirmed to be Alternariaalternata. The main pathogen causing the black spot disease in fresh grape during storage and transportation is Alternaria alternata. |
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