中国主要稻区稻曲病菌的生物学特性及群体遗传多样性
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摘要
【目的】明确采自中国10个水稻主产省份111个稻曲病菌(Ustilaginoidea virens)菌株的生物学特性、群体遗传多样性与地理区域的关系。【方法】采用生物学方法测定111个稻曲病菌菌株的产孢能力、生长速率和致病力,并采用SPSS 20.0软件对稻曲病菌菌株的产孢能力、菌丝生长速率及致病力进行相关性分析。提取111个稻曲病菌基因组DNA,采用3对特异性引物BOX、REP、ERIC对稻曲病菌菌株的全基因组DNA进行PCR扩增,通过聚类分析对其群体遗传多样性进行研究。【结果】111个稻曲病菌菌株的致病力与产孢量的Pearson相关系数为0.765,显著性概率为0.001;致病力与生长速率的Pearson相关系数为0.035,显著性概率为0.685。采用3对特异性引物BOX、ERIC、REP对稻曲病菌菌株的全基因组DNA进行了PCR扩增,群体遗传多样性值分别为0.73、0.62和0.60。111个稻曲病菌菌株的DNA用BOX引物分别扩增出7—13条不等的带谱,DNA指纹相似性在0.70相似水平上,供试菌株被划分为6种基因类群,其中第1类群为优势类型,有57个菌株,占总数的51.4%;来自安徽、四川、广西、湖北和云南的菌株主要划分在类群1和4,第2类群主要是来自江苏的菌株,有18个菌株,占总数的16.2%;第3类群是来自浙江的菌株,有8个菌株,占总数的7.2%;第5类群是来自黑龙江的菌株,有9个菌株,占总数的8.1%,第6类群是来自辽宁的菌株,有12个菌株,占总数的10.8%。稻曲病菌基因类群与地理区域之间有一定的相关性,与致病力、生长速率和产孢量之间没有相关性。【结论】来自中国10个省份的111个稻曲病菌菌株的致病力与产孢量之间具有显著相关性,相关程度为中等;致病力与菌丝的生长速率没有相关性。根据BOX-PCR扩增的稻曲病菌菌株基因组DNA指纹图谱的多态性进行划分的基因类群与地理区域之间显著相关,推测稻曲病菌属于局域性传播;基因类群与生物学特性之间没有相关性。
【Objective】The objective of this study is to reveal the relationship among geographical regions, biological characteristics and genetic diversity of 111 strains of Ustilaginoidea virens from 10 provinces in China.【Method】Sporulation, growth rate and pathogenicity of these strains were evaluated by biological methods, and the relativity was analyzed by using the software SPSS 20.0. DNA was extracted from the strains, the genomic fingerprint profiles of 111 strains were generated by BOX-PCR, REP-PCR and ERIC-PCR, respectively. Their genetic diversities were further investigated by using clustering analysis.【Result】The correlation coefficient of sporulation and pathogenicity was 0.765 and the significance probability was 0.001, the correlation coefficient of growth rate and pathogenicity was 0.035 and the significance probability was 0.685. The genomic fingerprint profiles of 111 strains were generated by BOX-PCR, REP-PCR and ERIC-PCR, respectively. The genetic diversities of the U. virens population were 0.73(in BOX), 0.62(in ERIC) and 0.60(in REP), respectively. The number of polymorphic bands was between 7 and 13 of each examined isolate detected through BOX-PCR analysis. According to cluster analysis of UPGMA, 6 clusters were grouped based on the boundary level of 0.70 average distances. The predominant group was group one which included 57 strains(51.4%). The strains of U. virens from Anhui, Sichuan, Guangxi, Hubei and Yunnan were grouped into group 1 and group 4. The strains of group 2 were mainly from Jiangsu, including 18 strains(16.2%), the strains of group 3 were from Zhejiang, including 8 strains(7.2%), the strains of group 5 were from Heilongjiang, including 9 strains(8.1%), and the strains of group 6 were from Liaoning, including 12 strains(10.8%). There was some correlation between BOX groups and geographic regions, no relationships were observed between BOX groups and their biological characteristics.【Conclusion】The relativity between the sporulation and pathogenicity was assumed to be medium. There was no relativity between the growth rate and pathogenicity. There was a significant relationship between BOX groups and the geographical regions, these results suggested that, naturally, U. virens in China might not be spread over long distances. No significant relationships were observed between BOX groups and their biological characteristics.
【Objective】The objective of this study is to reveal the relationship among geographical regions, biological characteristics and genetic diversity of 111 strains of Ustilaginoidea virens from 10 provinces in China.【Method】Sporulation, growth rate and pathogenicity of these strains were evaluated by biological methods, and the relativity was analyzed by using the software SPSS 20.0. DNA was extracted from the strains, the genomic fingerprint profiles of 111 strains were generated by BOX-PCR, REP-PCR and ERIC-PCR, respectively. Their genetic diversities were further investigated by using clustering analysis.【Result】The correlation coefficient of sporulation and pathogenicity was 0.765 and the significance probability was 0.001, the correlation coefficient of growth rate and pathogenicity was 0.035 and the significance probability was 0.685. The genomic fingerprint profiles of 111 strains were generated by BOX-PCR, REP-PCR and ERIC-PCR, respectively. The genetic diversities of the U. virens population were 0.73(in BOX), 0.62(in ERIC) and 0.60(in REP), respectively. The number of polymorphic bands was between 7 and 13 of each examined isolate detected through BOX-PCR analysis. According to cluster analysis of UPGMA, 6 clusters were grouped based on the boundary level of 0.70 average distances. The predominant group was group one which included 57 strains(51.4%). The strains of U. virens from Anhui, Sichuan, Guangxi, Hubei and Yunnan were grouped into group 1 and group 4. The strains of group 2 were mainly from Jiangsu, including 18 strains(16.2%), the strains of group 3 were from Zhejiang, including 8 strains(7.2%), the strains of group 5 were from Heilongjiang, including 9 strains(8.1%), and the strains of group 6 were from Liaoning, including 12 strains(10.8%). There was some correlation between BOX groups and geographic regions, no relationships were observed between BOX groups and their biological characteristics.【Conclusion】The relativity between the sporulation and pathogenicity was assumed to be medium. There was no relativity between the growth rate and pathogenicity. There was a significant relationship between BOX groups and the geographical regions, these results suggested that, naturally, U. virens in China might not be spread over long distances. No significant relationships were observed between BOX groups and their biological characteristics.
引文
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[2]胡东维,王疏.稻曲病菌侵染机制研究现状与展望.中国农业科学,2012,45(22):4604-4611.Hu D W,Wang S.Progress and perspectives in infection mechanism of Ustilaginoidea virens.Scientia Agricultura Sinica,2012,45(22):4604-4611.(in Chinese)
[3]王大为,王疏,傅俊范.稻曲病研究进展.辽宁农业科学,2004(1):21-24.Wang D W,Wang S,Fu J F.Research advance on false smut of rice.Liaoning Agricultural Sciences,2004(1):21-24.(in Chinese)
[4]黄世文,余柳青.国内稻曲病的研究现状.江西农业学报,2002,14(2):45-51.Huang S W,Yu L Q.Present situation of studies on rice false smut(Ustilaginoidea virens)in China.Acta Agriculturae Jiangxi,2002,14(2):45-51.(in Chinese)
[5]王洪凯,林福呈.稻曲病研究进展.浙江农业学报,2008,20(5):385-390.Wang H K,Lin F C.Research advances in rice false smut,Ustilaginoiden virens.Acta Agriculturae Zhejiangensis,2008,20(5):385-390.(in Chinese)
[6]Koiso Y,Li Y,Iwasaki S,Hanaoka K,Kobayashi T,Sonoda R,Fujita Y,Yaegashi H,Sato Z.Ustiloxins,antimitotic cyclic peptides from false smut balls on rice panicles caused by Ustilaginoidea virens.The Journal of Antibiotics,1994,47(7):765-773.
[7]Li Y,Koiso Y,Kobayashi H,Hashimoto Y,Iwasaki S.Ustiloxins,new antimitotic cyclic peptides:interaction with porcine brain tubulin.Biochemical Pharmacology,1995,49(10):1367-1372.
[8]周永力,樊金娟,曾超珍,刘小舟,王疏,赵开军.稻曲病菌遗传多样性与群体结构的初步分析.植物病理学报,2004,34(5):442-448.Zhou Y L,Fan J J,Zeng C Z,Liu X Z,Wang S,Zhao K J.Preliminary analysis of genetic diversity and population structure of Ustilaginoidea virens.Acta Phytopathologica Sinica,2004,34(5):442-448.(in Chinese)
[9]李燕,刘永锋,张荣胜,俞咪娜,陈志谊.江苏省稻曲病菌的群体遗传结构.江苏农业学报,2011,28(2):296-301.Li Y,Liu Y F,Zhang R S,Yu M N,Chen Z Y.Genetic diversity of Ustilaginoidea virens from rice in Jiangsu Province.Jiangsu Journal of Agricultural Sciences,2011,28(2):296-301.(in Chinese)
[10]张敏,李竞生,刘坚,宋伟,代海霞.四川省籼稻区水稻稻曲病菌遗传多样性分析.植物保护学报,2009,36(2):113-118.Zhang M,Li J S,Liu J,Song W,Dai H X.Preliminary analysis of genetic diversity of Ustilaginoidea virens strains of indica rice from Sichuan Province.Acta Phytophylacica Sinica,2009,36(2):113-118.(in Chinese)
[11]俞咪娜,陈志谊,于俊杰,胡建坤,尹小乐,聂亚锋,刘永锋.来源于同一穗不同稻曲球的稻曲病菌的致病性及遗传多样性.植物病理学报,2013,43(6):561-573.Yu M N,Chen Z Y,Yu J J,Hu J K,Yin X L,Nie Y F,Liu Y F.Genetic diversity and pathogenicity of Ustilaginoidea virens isolated from different rice false smut balls of a diseased spike.Acta Phytopathologica Sinica,2013,43(6):561-573.(in Chinese)
[12]Redondo C,Cubero J,Melgarejo P.Characterization of Penicillium species by ribosomal DNA sequencing and BOX,ERIC and REP-PCR analysis.Mycopathologia,2009,168(1):11-22.
[13]Vicente J G,Roberts S J.Discrimination of Pseudomonas syringae isolates from sweet and wild cherry using rep-PCR.European Journal of Plant Pathology,2007,117(4):383-392.
[14]李燕,尹小乐,刘永锋,于俊杰,陈志谊.稻曲病菌生物学特性与致病力相关性研究.植物病理学报,2012,42(4):353-364.Li Y,Yin X L,Liu Y F,Yu J J,Chen Z Y.Relativity of biological characteristics and pathogenicity of Ustilaginoidea virens.Acta Phytopathologica Sinica,2012,42(4):353-364.(in Chinese)
[15]刘明霞,秦姝,李颖,王疏.稻曲病菌分离技术、培养条件研究.辽宁农业科学,2009(6):20-22.Liu M X,Qin S,Li Y,Wang S.Study on isolation technique and culture condition of Ustilaginoidea virens.Liaoning Agricultural Sciences,2009(6):20-22.(in Chinese)
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[17]李燕,于俊杰,刘永锋,尹小乐,张荣胜,俞咪娜,陈志谊.稻曲病菌产孢能力及致病力测定.中国农业科学,2012,45(20):4166-4177.Li Y,Yu J J,Liu Y F,Yin X L,Zhang R S,Yu M N,Chen Z Y.Determination of sporulation and pathogenicity of Ustilaginoidea virens.Scientia Agricultura Sinica,2012,45(20):4166-4177.(in Chinese)
[18]张君成,陈志谊,张炳欣,刘永锋,陆凡.稻曲病的接种技术研究.植物病理学报,2004,34(5):463-467.Zhang J C,Chen Z Y,Zhang B X,Liu Y F,Lu F.Inoculation techniques used for inducing rice false smut efficiently.Acta Phytopathologica Sinica,2004,34(5):463-467.(in Chinese)
[19]曾列先,陈深,张慧,潘汝谦,杨健源,伍圣远,翟培茹,朱小源.广东水稻白叶枯病菌遗传多样性和小种分化研究.植物病理学报,2009,39(3):231-237.Zeng L X,Chen S,Zhang H,Pan R Q,Yang J Y,Wu S Y,Zhai P R,Zhu X Y.Genetic diversity and variability of Xanthomonas oryzae pv.oryzae in Guangdong.Acta Phytopathologica Sinica,2009,39(3):231-237.(in Chinese)
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