Analysis of Pathogen Species and Their Genetic Relationship of Apple Tree canker

doi:10.6048/j.issn.1001-4330.2022.03.020

Abstract

Abstract:

【Objective】 Based on the observation of the characteristics of pathogen culture, combined with molecular biology technology, to determine the species and related relationship of apple tree rot fungus in Aksu area in the hope of providing theoretical basis for effective control of apple canker. 【Methods】 The pure culture of Apple Tree Valsa Canker was obtained by Koch's Rule. The morphology and culture characters were observed and recorded. The DNA of the strain was extracted by improved CTAB method and then sequenced. After sequencing, the blast homology was compared and analyzed on NCBI website. Then the β - tubulin and EF1 - α sequences were analyzed by adjacency method and after that, phylogenetic tree was constructed. 【Results】 The optimum temperature for mycelium growth was 20-30℃, the optimum PH value was 5-6, and the optimum carbon and nitrogen sources were glucose and peptone; A total of 11 strains were isolated. The strain could be inoculated on healthy apple branches and the same strains could be isolated. It turned out that each strain was closely related to Valsa Mali, Valsa ceratosperma. 【Conclusion】 The apple tree rotting bacteria in Aksu area are V. Mali (asexual C. Mali) and V. ceratosperma (asexual C. sacculus), among which, V. Mali is the main pathogenic species. There are different types of culture characters on PDA.

Key words: apple tree; valsa canker; β-tubulin; EF1-α; culture traits

摘要：

【目的】研究苹果树腐烂病菌的种类及其亲缘关系,为有效防治苹果树腐烂病提供理论依据。【方法】以阿克苏地区不同区域苹果树腐烂病病样为材料,采用常规组织分离法和枝条烫伤接种法分离和鉴定菌株,记录形态学特征和发病情况;利用分子生物学方法获得β-tubulin与EF1-α序列,在NCBI网站对比分析Blast同源性,采用邻接法构建系统发育树分析同源性。【结果】菌丝生长最适温度 20~30℃,最适pH值为5~6,最适碳氮源分别是葡萄糖和蛋白胨;共分离出11个菌株,将菌株接种到健康苹果枝条上均可发病且能分离出相同的菌株,各菌株分别与Valsa mali和Valsa ceratosperma具有较近的亲缘关系。【结论】阿克苏地区苹果树腐烂病菌为黑腐皮壳属V. mali(无性型C. mali)和V. ceratosperma(无性型C. sacculus),其中V. mali为阿克苏地区的主要致病种,在PDA上存在不同类型的培养性状。

关键词: 苹果树, 腐烂病菌, β-tubulin, EF1-α, 培养性状

CLC Number:

S436.611

LI Yapeng, ZHANG Wangbin, YI Zibo. Analysis of Pathogen Species and Their Genetic Relationship of Apple Tree canker[J]. Xinjiang Agricultural Sciences, 2022, 59(3): 700-706.

李亚鹏, 张王斌, 仪子博. 苹果树腐烂病病原种类及其亲缘关系分析[J]. 新疆农业科学, 2022, 59(3): 700-706.

Figures/Tables 7

References 20

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编号 Number	寄主 Host	采集地点 Collection location	采集时间 Acquisition time
HQP₁	苹果	红旗坡一分厂	2018.5.16
HQP₂	苹果	红旗坡一分厂	2018.5.16
HQP₃	苹果	红旗坡一分厂	2018.5.16
YLC₁	苹果	第一师5团7连	2018.5.17
YLC₂	苹果	第一师5团7连	2018.5.17
YLC₃	苹果	第一师5团7连	2018.5.17

编号 Number	寄主 Host	采集地点 Collection location	采集时间 Acquisition time
HQP₁	苹果	红旗坡一分厂	2018.5.16
HQP₂	苹果	红旗坡一分厂	2018.5.16
HQP₃	苹果	红旗坡一分厂	2018.5.16
YLC₁	苹果	第一师5团7连	2018.5.17
YLC₂	苹果	第一师5团7连	2018.5.17
YLC₃	苹果	第一师5团7连	2018.5.17

菌株类别 Strain classification	菌落颜色 Colony color (3 d)	菌落颜色 Colony color (15 d)	菌落边缘 Edge of the colony	菌丝气生性 Mycelial aerogenesis	菌落直径 Diameter of the colony (2 d)(mm)	平均生长速率 Verage growth rate (mm/d)	菌落产孢情况 Sporulation of bacterial colonies
淡黄色菌株 Pale yellow strain	淡黄色	黄色、黄褐色	放射状	无	55.12±1.26	26.69±1.32	6株产孢第7~8 d
乳白色菌株 Milky white strain	乳白色	黄褐色	羽毛状	无	44.98±2.81	23.16±1.75	2株产孢第7~8 d

菌株类别 Strain classification	菌落颜色 Colony color (3 d)	菌落颜色 Colony color (15 d)	菌落边缘 Edge of the colony	菌丝气生性 Mycelial aerogenesis	菌落直径 Diameter of the colony (2 d)(mm)	平均生长速率 Verage growth rate (mm/d)	菌落产孢情况 Sporulation of bacterial colonies
淡黄色菌株 Pale yellow strain	淡黄色	黄色、黄褐色	放射状	无	55.12±1.26	26.69±1.32	6株产孢第7~8 d
乳白色菌株 Milky white strain	乳白色	黄褐色	羽毛状	无	44.98±2.81	23.16±1.75	2株产孢第7~8 d

编号 Number	寄主 Host	菌株类别 Strain classification	病原菌 Pathogenic bacteria
HQP2-1	苹果	淡黄色	V.ceratosperma
HQP2-2	苹果	淡黄色	V.ceratosperma
HQP2-3	苹果	淡黄色	V.ceratosperma
HQP2-4	苹果	淡黄色	V.ceratosperma
YLC3-1	苹果	淡黄色	V.mali
YLC3-2	苹果	淡黄色	V.mali
HQP3-1	苹果	乳白色	V.mali
YLC1-1	苹果	乳白色	V.mali
YLC1-2	苹果	乳白色	V.mali
YLC2-1	苹果	乳白色	V.mali