新疆马铃薯黑痣病病原菌的分离及定

doi:10.6048/j.issn.1001-4330.2024.07.024

新疆农业科学 ›› 2024, Vol. 61 ›› Issue (7): 1766-1771.DOI: 10.6048/j.issn.1001-4330.2024.07.024

新疆马铃薯黑痣病病原菌的分离及定

洪飞¹^,²(), 贾丰莲¹, 刘易², 吴燕², 杨茹薇²(), 孙慧², 李广悦¹()

1.中国农业科学院植物保护研究所,北京 100193
2.新疆农业科学院综合试验场,乌鲁木齐 830000

收稿日期:2024-01-05 出版日期:2024-07-20 发布日期:2024-09-04
通信作者: 李广悦(1983-),男,河北沧州人,研究员,博士生导师,研究方向为植物病害生物防治,(E-mail)liguangyue@caas.cn；
杨茹薇(1984-),女,宁夏灵武人,硕士,正高级农艺师,研究方向为植物保护,(E-mail)617950493@qq.com
作者简介:沈洪飞(1989-),男,云南昆明人,助理研究员,研究方向为马铃薯育种及病害防治,(E-mail)hongfeiscience@163.com
基金资助:
新疆维吾尔自治区区域协同创新专项(科技援疆计划)(2021E02024);新疆维吾尔自治区自然科学基金(地州科学基金)项目(2022D01F69);国家马铃薯产业技术体系(CARS-09-ES36);新疆维吾尔自治区重点研发计划(厅厅联动)(2022B02044)

Isolation and identification of the pathogen for potato black scurf in Xinjiang

SHEN Hongfei¹^,²(), JIA Fenglian¹, LIU Yi², WU Yan², YANG Ruwei²(), SUN Hui², LI Guangyue¹()

1. Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2. Xinjiang Academy of Agricultural Sciences Comprehensive Test Farm, Urumqi 830000, China

Received:2024-01-05 Published:2024-07-20 Online:2024-09-04
Supported by:
Regional Collaborative Project of Xinjiang Uyghur Autonomous Region(S & T Assisting Xinjiang Project)(2021E02024);Natural Science Foundation of Xinjiang Autonomous Region(2022D01F69);China Agriculture Research System- -National Potato Industry Technology System(CARS-09-ES36);Key R&D Program of Xinjiang Autonomous Region (Linkage between departments and departments)(2022B02044)

摘要/Abstract

摘要：

【目的】分离鉴定引起新疆马铃薯黑痣病病原菌,为筛选防治该病药剂奠定基础。【方法】从新疆吉木萨尔县、奇台县、乌鲁木齐市达坂城区和昭苏县等马铃薯种植区采集马铃薯块茎,通过组织分离法从带有马铃薯黑痣病的块茎分离病原菌,利用电子显微镜观察病原菌的形态特征,进行对峙培养和ITS序列分析,鉴定融合群。【结果】从新疆吉木萨尔县、奇台县、乌鲁木齐市达坂城区和昭苏县分离的4株病原菌均能使马铃薯块茎出现马铃薯黑痣病症状,分离菌株均为AG3融合群。【结论】AG3融合群是新疆马铃薯黑痣病的主要致病群。

关键词: 马铃薯黑痣病; 分离鉴定; 融合群

Abstract:

【Objective】To isolate and identify the pathogen causing potato black scurf in Xinjiang in the hope of providing a foundation for screening of control agents and establishment of control technology. 【Methods】Potato tubers were collected from Jimsar, Qitai, Dabancheng and Zhaosu in Xinjiang, and Rhizoctonia solaniwere isolated from tubers with potato black scurf by tissue separation method. Anastomosis groups of Rhizoctonia solaniwere identified by paired culture and ITS sequence analysis. 【Results】The four strains isolated from Jimsar, Qitai, Dabancheng and Zhaosu were all able to cause the symptoms of potato black scurf on potato tubers. The isolated strains were Anastomosis groups AG3. 【Conclusion】Anastomosis Groups AG3 is the main pathogenic group of potato black scurf in Xinjiang.

Key words: potato black scurf; isolation and identification; anastomosis groups

中图分类号:

S436.32

洪飞, 贾丰莲, 刘易, 吴燕, 杨茹薇, 孙慧, 李广悦. 新疆马铃薯黑痣病病原菌的分离及定[J]. 新疆农业科学, 2024, 61(7): 1766-1771.

SHEN Hongfei, JIA Fenglian, LIU Yi, WU Yan, YANG Ruwei, SUN Hui, LI Guangyue. Isolation and identification of the pathogen for potato black scurf in Xinjiang[J]. Xinjiang Agricultural Sciences, 2024, 61(7): 1766-1771.

图/表 5

表1 病原菌鉴定引物

Tab.1 Primers for pathogen identification

引物名称 Primer name	引物序列(5'-3') Primer sequence(5'-3')
ITS1	TCCGTAGGTGAACCTGCGG
ITS4	TCCTCCGCTTATTGATATGC
AG-2-1-F	CAAAGGCAATGGGTTATTGGAC
AG-2-1-R	CCTGATTTGAGATCAGATCATAAAG
Rs1F2	TTGGTTGTAGCTGGTCTATTT
AG-3PR2	ACACTGAGATCCAGCTAATA

图1 病薯样品中分离出病原菌在不同时间点的形态特征的变化注:A~C为菌丝生长特征;E为72 h菌落形态,F为96 h菌落形态,G为168 h菌落形态

Fig.1 Changes of morphological characteristics of pathogen isolated from potato sample infected with R.solani Note: A-C is the hyphal growth characteristic; E is the colony of pathogen incubatedon in PDA media for 72 hours, F is the colony of pathogen incubatedon in PDA media for 96 hours, and G is the colony of pathogen incubatedon in PDA media for 168 hours

图2 分离菌株的致病性验证注:CK为不接带菌土壤,收获的块茎;XJ1-4为回接带分离菌株毒土,收获的块茎

Fig.2 Verification of pathogenicity of the isolated strains Note: CK is harvested tuber from soil without tuber; XJ1-4 was a tieback strain isolated from tuber and harvested tuber

图3 特异性引物PCR扩增的凝胶电泳图注:A图为原始病原菌的ITS区通用引物ITS1/IT4的PCR扩增的电泳图,M为Star Marker D2000 PlusⅡ,1~4为新疆马铃薯分离的菌株XJ1-4;B图为回接后重新分离的病原菌的PCR扩增验证的电泳图,1~4是通用引物ITS1/IT4;5-8是特异性引物AG-2-1-F/R,9-12是特异性引物Rs1F2/AG-3PR2

Fig.3 Gel electrophoresis of the PCR amplification with specific primer Note: A is the electrophoretic PCR amplification of universal primer ITS1/IT4 in ITS region of the original pathogen, M is Star Marker D2000 PlusⅡ, 1-4 is strain XJ1-4 isolated from Xinjiang potato.The B diagram is an electrophoretic diagram of PCR amplification verification of pathogens reisolated after tieback.1-4 are universal primers ITS1/IT4.5-8 is the specific primer AG-2-1-F/R, and 9-12 is the specific primer Rs1F2/AG-3PR2

图4 马铃薯黑痣病病原菌与标准菌株AG3-PT的菌丝融合

Fig.4 Hyphal fusion reaction of the Rhizoctonia solani isolated from potato with standard strain AG3-PT

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新疆马铃薯黑痣病病原菌的分离及定

Isolation and identification of the pathogen for potato black scurf in Xinjiang

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