雪腐微座孢引起的小麦雪腐病病原鉴定及生物学特性分析

doi:10.6048/j.issn.1001-4330.2024.05.018

新疆农业科学 ›› 2024, Vol. 61 ›› Issue (5): 1201-1208.DOI: 10.6048/j.issn.1001-4330.2024.05.018

雪腐微座孢引起的小麦雪腐病病原鉴定及生物学特性分析

王莉¹(), 周小云²^,³, 鄢蓉¹, 张军高²^,³, 李进²^,³, 梁晶²^,³, 龚静云²^,³, 杜雨¹, 马德英¹, 雷斌²^,³()

1.新疆农业大学农学院, 乌鲁木齐 830052
2.新疆农业科学院核技术生物技术研究所/科技成果转化中心/农药试制中心, 乌鲁木齐 830091
3.新疆作物化学调控工程技术研究中心,乌鲁木齐 830091

收稿日期:2023-10-15 出版日期:2024-05-20 发布日期:2024-07-09
通信作者: 雷斌(1973-),男,四川巴中人,研究员,博士,硕士生导师,研究方向为作物根腐类病害防控及绿色农药研发,(E-mail)leib668@xaas.ac.cn
作者简介:王莉(1998-),女,河南周口人,硕士研究生,研究方向为小麦病害防控,(E-mail)1669655475@qq.com
基金资助:
中央引导地方项目“‘一带一路’国际绿色农药研制及应用技术创新基地建设”(ZYYD2023B06);省部共建课题“干旱半干旱地区旱地小麦提质增效关键技术研究”(SBGJXTZXKF-9);新疆维吾尔自治区乡村振兴产业发展科技行动项目“春小麦增密抗逆增产关键技术引进与示范”(2022NC078);新疆维吾尔自治区科技特派员农村科技创业行动项目“春小麦高产创建关键技术集成与示范”(2022BKZ022)

Pathogen identification and biological characteristics analysis of wheat snow rot caused by Microdochium nivale

WANG Li¹(), ZHOU Xiaoyun²^,³, YAN Rong¹, ZHANG Jungao²^,³, LI Jin²^,³, LIANG Jing²^,³, GONG Jingyun²^,³, DU Yu¹, MA Deying¹, LEI Bin²^,³()

1. College of Agriculture, Xinjiang Agricultural University, Urumqi 830052, China
2. Scientific and Technological Achievements Transformation Center /Pesticide Trial Production Center / Institute of Nuclear Technology and Biotechnology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
3. Xinjiang Crop Chemical Control Engineering Technology Research Center, Urumqi 830091, China

Received:2023-10-15 Published:2024-05-20 Online:2024-07-09
Correspondence author: LEI Bin (1973 -), male, from Bazhong, Sichuan, researcher, Ph.D., research direction: pesticide research and crop chemical regulation technology, (E-mail)leib668@xaas.ac.cn
Supported by:
The central government guides local projects " Construction of 'Belt and Road' international green pesticide development and application technology innovation base"(ZYYD2023B06);Provincial and ministerial joint projects "Study on Key techniques of improving quality and efficiency of dryland wheat in arid and semi-arid areas"(SBGJXTZXKF-9);Agriculture,Rural Areas and Farmers Backbone Talent Training Project of Xinjiang Uygur Autonomous Region"Introduction and demonstration of key technologies for increasing density, stress resistance, and yield of spring wheat"(2022NC078);Xinjiang Uygur Autonomous Region Science and Technology Commissioner Rural Science and Technology Entrepreneurship Action Project "Integration and Demonstration of Key Technologies for High Yield Creation of Spring Wheat"(2022BKZ022)

摘要/Abstract

摘要：

【目的】研究新疆伊犁哈萨克自治州(简称伊犁州)冬小麦主产区小麦雪腐病病原菌种类、生物学特性,为农药研制及该病害防控提供科学依据。【方法】通过组织分离法,分离冬小麦雪腐病病原菌并接种到健康小麦上验证其致病性,结合生物学特征及ITS基因序列分析鉴定病原菌种类,采用生长速率法研究病原菌生物学特性。【结果】分离获得的菌株YLXF-1、YLXF-2及YLXF-3均可侵染健康小麦,且发病症状和田间症状相同;病原菌为Microdochium nivale。病原菌在5~35℃、pH值3~12、不同光照条件及供试培养基上均可生长,且大部分供试碳、氮源均可利用,具有较强的环境适应性。病原菌在20℃、pH值8、12 h光暗交替培养和PDA培养基上菌丝生长较快,利用率较好的碳源、氮源分别为蔗糖和牛肉膏。【结论】新疆伊犁州冬小麦主产区小麦雪腐病病原菌为M.nivale,病原菌菌丝最适生长温度为20℃,pH值8,光照为12 h光暗交替,培养基为PDA,最佳碳源为蔗糖,最佳氮源为牛肉膏。

关键词: 小麦; 雪腐病; 病原鉴定; 生物学特性

Abstract:

【Objective】The study aims to investigate the species and biological characteristics of the causal agents of wheat snow mold disease in the main wheat-producing areas of Yili Kazak Autonomous Prefecture (Yili Prefecture), Xinjiang Uygur Autonomous Region, China, in order to provide scientific basis for the development of pesticides and the prevention and control of the disease. 【Methods】The pathogen of winter wheat snow mold disease was isolated by tissue culture method and inoculated onto healthy wheat plants to verify its pathogenicity. The species of the pathogen was identified by combining its biological characteristics and ITS gene sequence analysis. The biological characteristics of the pathogen were studied by the method of growth rate. 【Results】The isolates YLXF-1, YLXF-2, and YLXF-3 could infect healthy wheat and exhibit the same symptoms of disease in the field. Morphological and molecular biological identification results indicated that the pathogen was Microdochium nivale. The pathogen could grow at temperatures of 5-35°C, pH values of 3-12, under different light conditions, and on various test media, and could utilize most of the tested carbon and nitrogen sources, indicating strong environmental adaptability. The pathogen had faster mycelial growth at 20°C, pH 8, and 12 h of light-dark alternation culture on PDA medium, and utilized sucrose and beef extract as the better carbon and nitrogen sources, respectively.【Conclusion】The pathogen of wheat snow mold disease in the main production area of winter wheat in Yili, Xinjiang is M. nivale. The optimal growth temperature of the pathogen mycelium is 20°C, pH value is 8, light is 12 h of light and darkness alternation, culture medium is PDA, the best carbon source is sucrose, and the best nitrogen source is beef extract.

Key words: wheat; snow rot; pathogen identification; biological characteristics

中图分类号:

S513

王莉, 周小云, 鄢蓉, 张军高, 李进, 梁晶, 龚静云, 杜雨, 马德英, 雷斌. 雪腐微座孢引起的小麦雪腐病病原鉴定及生物学特性分析[J]. 新疆农业科学, 2024, 61(5): 1201-1208.

WANG Li, ZHOU Xiaoyun, YAN Rong, ZHANG Jungao, LI Jin, LIANG Jing, GONG Jingyun, DU Yu, MA Deying, LEI Bin. Pathogen identification and biological characteristics analysis of wheat snow rot caused by Microdochium nivale[J]. Xinjiang Agricultural Sciences, 2024, 61(5): 1201-1208.

图/表 8

表1 采样地点及坐标

Tab.1 Sampling locations and coordinates

采样地点 Sampling site		GPS坐标GPS coordinates
采样地点 Sampling site		北纬 Northern latitude(°)	东经 East longitude(°)	海拔 Altitude(m)
察布查尔锡伯自治县 Qapqal Xibe Autonomous County	琼博拉镇	43.581 403	80.962 036	1 105.22
	海努克镇	43.688 163	81.290 940	833.23
	阔洪奇乡X715	43.745 650	91.308 734	691.00
巩留县 Gongliu County	阿尕尔森镇X736	43.478 332	82.405 158	776.36
	阔什阿尕尔羊场	43.468 804	82.342 651	790.46
	巩留镇健康路	43.419 643	84.201 696	793.37

图1 小麦雪腐病田间发病症状注:A为田间症状;B为采集病株

Fig.1 The symptoms of the field of wheat snow rot disease Note: A is field symptom; B is for the collection of disease strains

表2 小麦雪腐病病样中病原菌分离频率的比较

Tab.2 Comparison of pathogen isolation frequency in wheat snow rot disease samples

真菌种属 Genus of fungi	菌株数量 Number of strains(株)	分离频率 Isolating frequency(%)
微座孢属Microdochium	24	41.38
镰刀菌属Fusarium	10	17.24
链格孢属Alternaria	19	32.86
其它属Other Genus	5	8.62

图2 病原菌的致病力比较注:A:叶片接种无菌PDA(CK);B:叶片接种菌株YLXF-1菌饼;C:叶片接种菌株YLXF-2菌饼;D:叶片接种菌株YLXF-3菌饼;E:土壤接种无菌培养物(CK);F:土壤接种菌株YLXF-2固体培养物

Fig.2 Comparison of pathogenicity of pathogenic bacteria Note: A: leaf inoculated sterile PDA (CK); B: leaf inoculated strain YLXF-1 cake; C: leaf inoculated strain YLXF-2 cake; D: leaf inoculated strain YLXF-3 cake; E: soil inoculated sterile culture (CK); F: soil inoculated strain YLXF-2 solid culture

图3 病原菌菌落形态特征注:A:培养7d菌落正面形态;B:培养7d菌落背面形态;C:培养25d菌落正面形态;D:分生孢子

Fig.3 Morphological characteristics of pathogenic bacteria colonies Note: A: Positive form of colony cultured for 7 days; B: Morphology of colony backside after 7 days of culture; C: Positive colony morphology after 25 days of culture; D: Conidia

图4 基于ITS基因扩增片段

Fig.4 ITS gene based amplification fragment

图5 基于ITS基因序列构建的系统发育树

Fig.5 Phylogenetic tree constructed through method based on combined ITS gene sequences

图6 不同培养条件下雪腐微座孢菌丝生长的比较注:A~F依次为温度、pH值、光照、培养基、碳源、氮源。不同小写字母表示在P<0.05水平差异显著

Fig.6 Comparison of mycelial growth of M. nivale under different culture conditions Note: A-F is temperature, pH, light, medium, carbon source, nitrogen source.Different small letters mean significant differences (P<0.05)

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雪腐微座孢引起的小麦雪腐病病原鉴定及生物学特性分析

Pathogen identification and biological characteristics analysis of wheat snow rot caused by Microdochium nivale

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