云南高原粳稻白叶枯病菌的抗药性室内鉴定及其rpfC基因序列分析

杨雅云1,张恩来1,阿新祥1,汤翠凤1,张斐斐1,陈丹1,董超1,吴坤1,2,徐福荣1,周英3,祁春学4,戴陆园1,*

doi:10.3969/j.issn.1001-7216.2014.06.013

中国水稻科学 >

2014 , Vol. 28 >Issue 6: 665 - 674

DOI: https://doi.org/10.3969/j.issn.1001-7216.2014.06.013

研究报告

云南高原粳稻白叶枯病菌的抗药性室内鉴定及其rpfC基因序列分析

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¹云南省农业科学院生物技术与种质资源研究所/云南省农业生物技术重点实验室/农业部西南作物基因资源与种质创制重点实验室，昆明650223；²云南农业大学农学与生物技术学院，昆明650201；³西双版纳州农业科学研究所，云南景洪 666100； ⁴西双版纳州勐海县西家乡农业综合服务中心，云南勐海 666200；

收稿日期: 2013-11-25

修回日期: 2014-07-17

网络出版日期: 2014-11-10

基金资助

国家自然科学基金资助项目(31160274)；云南省农业科学院专项(YAAS2013JC009)；云南省自然科学基金资助项目( 2013FZ147)。

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Laboratory Identification of Resistance to Pesticides and rpfC Gene Sequence Analysis of Xanthomonas oryzae pv. oryzae in japonica Rice from Yunnan Plateau

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¹ Institute of Biotechnology and Germplasm Resources/ Yunnan Provincial Key Laboratory of Agricultural Biotechnology/Key Laboratory of Southwestern Crop Gene Resource and Germplasm Enhancement, Ministry of Agriculture, Yunnan Academy of Agricultural Sciences,Kunming 650223, China; ²College of Agriculture and Biotechnology, Yunnan Agricultural University,Kunming 650201, China; ³Institute of Agricultural Science of Xishuangbanna, Jinghong 666100, China; ⁴Agricultural Comprehensive Service Center， Xiding Township, Menghai County, Xishuangbanna Precture, Menghai 666200, China;

Received date: 2013-11-25

Revised date: 2014-07-17

Online published: 2014-11-10

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摘要

为了探索云南省高原粳稻上十种不同致病型的白叶枯病菌对噻枯唑、叶枯灵和新植霉素三种农药的抗药性及其机制，分别用含有不同浓度农药的NA培养基进行白叶枯病菌的室内抗药性筛选，并设计与菌株抗药性密切相关的rpfC基因特异引物，对抗药性不同的菌株进行扩增、测序、基因和氨基酸序列比对分析。结果表明，噻枯唑对参试的所有菌株的最小抑菌浓度（MIC）为40~180 mg/L，而叶枯灵为10~100 mg/L，没有发现对新植霉素产生抗性的菌株；病原菌的致病型与对农药的敏感性相关，致病性强的菌株，其抗药性较强。将致病型和抗药性不同的10个菌株的rpfC基因序列与GenBank中登录号为X97865.1的基因序列比对，序列同源性为92%～98%，而RpfC蛋白序列同源性差异较大（8.3%至99%）。致病性和抗药性最强的Ⅵ型菌株200131的RpfC蛋白序列的六个功能域完整，致病性和抗药性最弱的0型菌株DHL1的RpfC蛋白序列的信号接收区域REC已经消失。7个致病性和抗药性中等的菌株中，Ⅳ型菌株⑤较为特殊，其RpfC蛋白序列已经不能形成功能域。

关键词： 云南高原粳稻; 白叶枯病菌; 致病型; 抗药性; rpfC基因分析

本文引用格式

杨雅云1,张恩来1,阿新祥1,汤翠凤1,张斐斐1,陈丹1,董超1,吴坤1,2,徐福荣1,周英3,祁春学4,戴陆园1,* . 云南高原粳稻白叶枯病菌的抗药性室内鉴定及其rpfC基因序列分析[J]. 中国水稻科学, 2014 , 28(6) : 665 -674 . DOI: 10.3969/j.issn.1001-7216.2014.06.013

Abstract

In order to reveal pesticide resistance and its mechanisms of Xoo strains belonging to 10 pathogenic types in japonica rice from Yunnan plateau, the indoor resistance identification of Xoo strains to pesticides were conducted in NA medium containing pesticides at different concentrations, PCR amplification was carried out using specific rpfC gene primers which was closely related to resistance, gene and amino acid were sequenced and aligned. The results show that the minimun inhibitory concentration(MIC) of bismerthiazol to all tested strains were 40180mg/L, while the MIC of Benzoic acid1, 3dithiolan2ylidenehydrazide(BADYH, IV) to the strains were 10100mg/L, there was no resistant strains to stretomycin+oxyteracykine. Correlated with the sensitivity to pesticide, the pathogenicity of Xoo strains was improved with the rising resistance to pesticide. The homology alignment results showed that rpfC gene of 10 strains shared 92%-98% identity with the gene of the registration number X97865.1 in the Genebank, while the differences in 11 RpfC protein sequences ranged from 8.3% to 99%. There were six complete functional domains in RpfC protein of strain 200131 with the strongest pathogenicity（Type Ⅵ）and resistance to pesticide. REC reception domain in RpfC protein of strain DHL1 has disappeared , which showed the weakest pathogenicity（Type 0）and resistance to pesticide. Particularly, strains ⑤ (Type Ⅳ), middle pathogenic and resistant,could not form functional domains of RpfC protein.

Key words： japonica rice from Yunnan plateau; Xanthomonas oryzae pv. oryzae; pathotype; resistance to pestcide; analysis of rpfC gene sequence

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