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福建省柑橘褐斑病菌的分离、鉴定和系统进化分析

引用本文：李亚,吕佳芮,朱玲,鲁国东,王宗华,刘国坤.福建省柑橘褐斑病菌的分离、鉴定和系统进化分析.植物保护学报,2019,46(6):1243-1251

DOI：10.13802/j.cnki.zwbhxb.2019.2018205

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作者	单位	E-mail
李亚	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002
吕佳芮	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002
朱玲	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002
鲁国东	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002
王宗华	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002
刘国坤	福建农林大学, 生物农药与化学生物学教育部重点实验室, 福州 350002	liuguok@126.com

中文摘要:为明确在福建省南平市的橘柚和三明市的温州蜜橘上发现的疑似柑橘褐斑病的病原菌种类，采用组织分离法获得纯化菌株，通过回接法验证菌株的致病性，利用形态学特征对病原菌进行初步鉴定，并采用最大似然法以多聚半乳糖醛酸酶基因endoPG为靶标对本研究以及国内外已报道的链格孢菌株构建系统发育树，分析其遗传多样性。结果表明，从病组织中共分离获得26株纯培养菌株，经形态学鉴定均为链格孢菌Alternaria spp.。利用分生孢子液接种橘柚离体嫩叶发现，有22株菌株能侵染橘柚叶片并产生与田间相似的褐斑病症状，确认该病害为链格孢引起的柑橘褐斑病。系统发育树分析结果显示，分离所得的26株菌株均聚在已报道的4个柑橘链格孢进化分支Clade1~Clade4中，其中21株菌株聚在国内特有的分支Clade4中，有3株菌株和1株菌株分别聚在国内外兼有的分支Clade3和Clade1中，1株菌株聚在国外特有的分支Clade2中，表明在福建省采集的这些柑橘褐斑病菌均为链格孢菌，且遗传多样性较丰富。

中文关键词:柑橘褐斑病链格孢菌病原菌分离鉴定系统进化分析

Isolation, identification and phylogenetic analysis of citrus brown spot pathogens in Fujian Province

Author Name	Affiliation	E-mail
Li Ya	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China
Lü Jiarui	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China
Zhu Ling	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China
Lu Guodong	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China
Wang Zonghua	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China
Liu Guokun	Key Laboratory of Biopesticides and Chemical Biology, Ministry of Education Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian Province, China	liuguok@126.com

Abstract:To define the pathogens causing suspected citrus brown spot disease on tangelo at Nanping and Mikan at Sanming in Fujian Province, the pathogen fungi were isolated out from infected tissues, the pathogenicity of isolates to citrus leaves were verified, the morphology features of isolates were analyzed to identify the fungi, and the phylogenetic tree of Alternaria isolates from this study and those reported was constructed based on endopolygalacturonase gene by using maximum likelihood method to analyze the fungus genetic diversity. The results showed that 26 pure fungal cultures were morphologically identified as Alternaria spp. By inoculating tangelo tender leaves with conidial suspension, 22 isolates could infect citrus and caused brown spot symptom similar to that discovered in the orchard, which further confirmed this disease as citrus brown spot induced by Alternaria. The phylogenetic tree displayed, 26 Fujian isolates clustered in all four reported citrus Alternaria clades (Clade1-Clade4). Among them, 21 isolates clustered in Clade4 that only reported in China; three isolates and one isolate belonged to Clade3 and Clade1, respectively, which were reported both at home and abroad; one isolate located in Clade2 that only reported at abroad. These results suggested that the citrus brown spot pathogens collected from Fujian Province were Alternaria strains and had higher level of genetic diversity.

keywords:citrus brown spot Alternaria spp. pathogen isolation identification phylogenetic analysis

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