新疆山桃腐烂病病原菌鉴定

doi:10.6048/j.issn.1001-4330.2024.12.024

新疆农业科学 ›› 2024, Vol. 61 ›› Issue (12): 3089-3096.DOI: 10.6048/j.issn.1001-4330.2024.12.024

• 草业·植物保护·畜牧兽医 • 上一篇下一篇

新疆山桃腐烂病病原菌鉴定

唐丽¹(), 李春艳², 贾文浩¹, 刘振亚¹, 李亚鹏¹, 但红侠¹, 张王斌¹()

1.塔里木大学/南疆特色果树高效优质栽培与深加工技术国家地方联合工程实验室/南疆农业有害生物综合治理兵团重点实验室/农业农村部阿拉尔作物有害生物科学观测实验站,新疆阿拉尔 843300
2.新疆应用职业技术学院,新疆奎屯 833200

收稿日期:2024-05-28 出版日期:2024-12-20 发布日期:2025-01-16
通信作者: 张王斌(1974-),男,陕西澄城人,教授,硕士生导师,研究方向为植物病理学,(E-mail)zwbzky@163.com
作者简介:唐丽(1993-),女,四川德阳人,硕士研究生,研究方向为植物病理学,(E-mail)susuzaizai@163.com
基金资助:
“新疆枣园林果树木腐烂病菌遗传多样性及致病力分化研究”(31660034)

Identification of the pathogen of Prunus davidiana canker in Xinjiang

TANG Li¹(), LI Chunyan², JIA Wenhao¹, LIU Zhenya¹, LI Yapeng¹, DAN Hongxia¹, ZHANG Wangbin¹()

1. The National Local Joint Engineering Lab of High Efficiency and Superior-Quality Cultivation and Fruit Deep Processing Technology of Characteristic Fruit Tress in Southern Xinjiang/Key Lab of Xinjiang Production and Construction Corps in Comprehensive Agricultural Pest Management in Southern Xinjiang/Scientific Observing/Experimental Station of Crops Pests in Aral, Ministry of Agriculture and Rural Affairs, Tarim University, Aral Xinjiang 843300, China
2. Xinjiang Applied Vocational and Technical College, Kuitun Xinjiang 833200, China

Received:2024-05-28 Published:2024-12-20 Online:2025-01-16
Supported by:
Genetic Diversity and Pathogenicity Differentiation of Fruit Trees in Xinjiang(31660034)

摘要/Abstract

摘要：

【目的】 鉴定在新疆奎屯市(44°26'35″N,84°54'24″E)和阿拉尔市(40°32'30″N,81°17'35″E)的山桃腐烂枝条病症。【方法】 从山桃腐烂枝条上分离出2株山桃腐烂病病原菌(KTST和TDST),测定分析田间症状、无性形态、致病性,并结合ITS、RPB2、Tef1-α多基因序列联合鉴定分析病原菌。【结果】 KTST和TDST的子座结构、孔口的数量、腔室形状及孢子类型均符合壳囊孢属(Cytospora)特征,KTST与TDST分离株与GenBank中已有的Cytospora leucostoma、Cytospora chrysosperma菌株聚为一个独立的分支。将腐烂病病原菌接种在离体的健康山桃枝上,引起腐烂病症状,验证其致病性。【结论】 新疆奎屯市和阿拉尔市的山桃腐烂病致病菌为Cytospora chrysosperma。

关键词: 山桃腐烂病; 形态特征; 致病性测定; 多基因联合分析

Abstract:

【Objective】 There is a typical symptom about canker disease in Prunus davidiana from Kuitun City (44°26'35″N, 84°54'24″ E) and Aral City (40°32'30″N, 81°17'35″E), Xinjiang.With this in mind, this research aim to identify the real cause.【Methods】 Pathogens (KTST and TDST) were isolated from the susceptible Prunus davidiana branches.【Results】 Based on morphological characteristics, combined with ITS, RPB2 and Tef1-α multi-genesequence analysis, Cytospora leucostoma and Cytospora chrysosperma were isolates from KTST and TDST.【Conclusion】 The Koch postulates was verified by inoculating the pathogen of canker disease on the healthy branches in vitro to cause canker disease symptoms.

Key words: Valsa canker; morphological characteristics; pathogenicity determination; multigene analysis

中图分类号:

S436.629

唐丽, 李春艳, 贾文浩, 刘振亚, 李亚鹏, 但红侠, 张王斌. 新疆山桃腐烂病病原菌鉴定[J]. 新疆农业科学, 2024, 61(12): 3089-3096.

TANG Li, LI Chunyan, JIA Wenhao, LIU Zhenya, LI Yapeng, DAN Hongxia, ZHANG Wangbin. Identification of the pathogen of Prunus davidiana canker in Xinjiang[J]. Xinjiang Agricultural Sciences, 2024, 61(12): 3089-3096.

图/表 9

表1 供试材料

Tab.1 experimental material

分离株 Strain	寄主 Host plant	寄主植物科 Host plant family	寄主植物属 Host plant genus	采集地点 Collection site	地理位置 Geographic position	海拔 Elevation(m)
KTST	山桃 Prunus davidiana	蔷薇科 Rosaceae Juss.	桃属 Amygdalus L.	新疆奎屯市	44°26'35″N,84°54'24″E	470
TDST	山桃 Prunus davidiana	蔷薇科 Rosaceae Juss.	桃属 Amygdalus L.	新疆阿拉尔市	40°32'30″N,81°17'35″E	1 010

表2 引物序列及反应程序

Tab.2 Primers sequence and reaction procedure

基因名称 Gene	引物名称 Primer	引物序列 Primer sequence	反应程序 Reaction program
核糖体基因转录间隔区 ITS	ITS1	5’-CCGTAGGTGAACCTGCGG-3’	95℃预变性5 min,95℃变性1 min,48℃退火1 min,72℃延伸1 min,72℃延伸10 min,35个循环
核糖体基因转录间隔区 ITS	ITS4	5’-TCCTCCGCTTATTGATATGC-3’
延伸因子 EF1-a	EF1-512F	5’-ATGTGCAAGGCCGGTTTCGC-3’	95℃预变性5 min,95℃变性1.5 min,56℃退火1 min,72℃延伸1.5 min,72℃延伸10 min,35个循环
延伸因子 EF1-a	EF1-783R	5’-TACGAGTCCTTCTGGCCCAT-3’
RBP2	RBP2-5F	5’-GATCGATCACGATGATCATCTTTCGG-3’	95℃预变性5 min,95℃变性1.5 min,56℃退火1 min,72℃延伸1.5 min,72℃延伸10 min,35个循环
RBP2	RBP2-7Cr	5’-CCCATAGGCTTGTCTTAGCCCAT-3’

图1 KTST形态特征注:A:田间发病症状;B,C:分生孢子器横切面;D:无性型子座;E,F:分生孢子器纵切面;G:分生孢子梗;H:分生孢子;I:菌落形态(3d和30d)。标尺:B,C,F=500 μm ;G-H=5 μm

Fig.1 Morphological characteristics of KTST Note:A: Symptoms of field disease; B, C: Transverse section of conidia; D: Asexual substratum; E, F: Longitudinal section of conidia; G: Conidiospore stalk; H: Conidium; I: Colony morphology (3 d and 30 d).Gauge: B,C,F=500 μm; G-H=5 μm

图2 TDST形态特征注:A:田间发病症状;B,C:分生孢子器横切面;D:无性型子座;E:分生孢子器纵切面;G:分生孢子梗;H:分生孢子;F,I:菌落形态(3 d和30 d)。标尺:B-E=500μm;G-H=5μm

Fig.2 Morphological characteristics of TDST Notes:A: Symptoms of field disease; B, C: Transverse section of conidia; D: Asexual substratum; E: Longitudinal section of conidia; G: Conidiospore stalk; H: Conidium; F, I: Colony morphology (3 d and 30 d).Gauge: B,C,F=500 μm; G-H=5 μm

表3 分离株KTST和TDST的致病性变化

Tab.3 Changes of KTST and TDST for pathogenicity determination

分离株 Strain	发病率 Morbidity (%)	显症时间 Disease time (h)	病斑扩展速度 Rate of lesion expansion (cm/d)	产孢时间 Sporulation time(d)	分生孢子角颜色 Conidia color	病斑颜色 Lesion color
KTST	100	48	0.56	13	橘黄色	褐色
TDST	100	48	0.79	15	橘黄色	褐色
CK	0	0	0	-	-	-

图3 山桃枝条接种注:A:KTST接种5 d;B:KTST接种14 d;C:TDST接种7 d

Fig.3 Inoculation of Prunus davidiana branches Notes:A: KTST vaccination for 5 days; B: 14 days after KTST inoculation; C: TDST was inoculated for 7 days

图4 ITS基因系统发育树

Fig.4 Phylogenetic tree of ITS gene

图5 RBP2基因系统发育树

Fig.5 Phylogenetic tree of RBP2 gene

图6 Tef1-α基因系统发育树

Fig.6 Phylogenetic tree of Tef1-α gene

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新疆山桃腐烂病病原菌鉴定

Identification of the pathogen of Prunus davidiana canker in Xinjiang

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