四川省不同地域的核盘菌遗传多样性分析
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摘要
核盘菌(Sclerotinia sclerotiorum)属于世界性分布的植物病原真菌,可以危害油菜等多种经济作物。研究不同地域核盘菌的遗传多样性对了解核盘菌的遗传演化过程和指导病害防控具有重要意义。实验采用序列相关扩增多态性(sequence-related amplified polymorphism,SRAP)标记对四川省17个不同地理来源的66株核盘菌菌株的遗传多样性进行了分析。10对检测引物共获得129个位点,其中123个为多态位点,占95.35%。UPGMA聚类结果显示,在相似性系数为0.7时,66个核盘菌菌株分为5类(Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ),分别包含60、2、2、1和1个菌株。在相似性系数为0.74时,第Ⅰ类又可分为3个亚类(Ⅰ-1、Ⅰ-2、Ⅰ-3),分别包含21、37和2个菌株。聚类及组成分分析结果显示,四川省各地区的核盘菌菌株具有较高的遗传多样性,但其遗传变异与菌株地理来源无明显相关性。
Sclerotinia sclerotiorum is a worldwide pathogenic fungus causing serious diseases on many economically important crops. Studying on the genetic diversity of S. sclerotiorum isolates from different eco-geographical regions is crucially important for understanding the evolution of this fungal pathogen and disease control. In this study,DNA polymorphism of 66 S. sclerotiorum isolates derived from 17 different regions in Sichuan province were detected using sequence-related amplified polymorphism (SRAP) markers. A total of 129 scorable fragments were identified with 10 SRAP primer combinations,among which 123 were polymorphic loci (95.35%). UPMGA (Unweighted Pair Group Method with Arithmetic Mean) indicated that the dendrogram consisted of five groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ) at the genetic similarity coefficient of 0.70,which included 60,2,2,1 and 1 isolates,respectively. Moreover,the groupⅠcontained three sub-groups (I-1,I-2,I-3) at the genetic similarity coefficient of 0.74,which included 21,37 and 2 isolates,respectively. This study showed a rich SRAP polymorphism among the populations of S. sclerotiorum in Sichuan province,but genetic diversity had no significant correlation with geographical location.
Sclerotinia sclerotiorum is a worldwide pathogenic fungus causing serious diseases on many economically important crops. Studying on the genetic diversity of S. sclerotiorum isolates from different eco-geographical regions is crucially important for understanding the evolution of this fungal pathogen and disease control. In this study,DNA polymorphism of 66 S. sclerotiorum isolates derived from 17 different regions in Sichuan province were detected using sequence-related amplified polymorphism (SRAP) markers. A total of 129 scorable fragments were identified with 10 SRAP primer combinations,among which 123 were polymorphic loci (95.35%). UPMGA (Unweighted Pair Group Method with Arithmetic Mean) indicated that the dendrogram consisted of five groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ) at the genetic similarity coefficient of 0.70,which included 60,2,2,1 and 1 isolates,respectively. Moreover,the groupⅠcontained three sub-groups (I-1,I-2,I-3) at the genetic similarity coefficient of 0.74,which included 21,37 and 2 isolates,respectively. This study showed a rich SRAP polymorphism among the populations of S. sclerotiorum in Sichuan province,but genetic diversity had no significant correlation with geographical location.
引文
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[19]Qiu S,Shen B C,Li T T,et al.A method of osmanthus fragrans cultivars identification based on random forest algorithm and SRAP molecular markers(in Chinese)[J].Scientia Silvae Sinicae(林业科学),2018,54(1):32-45.
[20]Matias P,Hedvig K,David L,et al.SRAP technique efficiently generates polymorphisms in Puccinia striiformis isolates[J].Journal of Phytopathology,2010,158(10):708-711.
[21]Li G,Gao M,Yang B,et al.Gene for gene alignment betw een the Brassica and Arabidopsis genomes by direct transcriptome mapping[J].Theoretical and Applied Genetics,2003,107(1):168-180.
[22]Lin Z X,Zhang X L Nie Y C,et al.Construction of a genetic linkage map for cotton based on SRAP[J].Chinese Science Bulletin,2003,48(19):2064-2068.
[23]Chang Y H,Zhang A Y,Liu P,et al.Genetic diversity analysis w ith capillary electrophoresis detection w ith fluorescent ISSR markers on foxtail millet(in Chinese)[J].M olecular Plant Breeding(分子植物育种),2017,15(8):3302-3308.
[2]Hu X J,Qin L,Roberts D P,et al.Characterization of mechanisms underlying degradation of sclerotia of Sclerotinia sclerotiorum by Aspergillus aculeatus Asp-4using a combined qRT-PCR and proteomic approach[J].BMC Genomics,2017,18(1):674-688.
[3]Boland G J,Hall R.Index of host plants of Sclerotinia sclerotiorum[J].Canadian Journal of Plant Pathology,1994,16:93-108.
[4]Liu Y,Huang X Q,Zhang L,et al.Proceedings of the annual meeting of Chinese Society for Plant Pathology(in Chinese)[C].Beijing:China Agricultural Science and Technology Press(北京:中国农业科技出版社),2014.
[5]Ji W Z,Su B.Principles and methodologies of genetic diversity studies[M].Zhejiang:Zhejiang Science and Technology Press(浙江:浙江科学技术出版社),1999.
[6]Li G Q,Wang D B,Jiang D H,et al.RAPD assay of strains of Sclerotinia sclerotiorum and its related species(in Chinese)[J].Acta Phytopathologica Sinica(植物病理学报),2000,30(2):166-170.
[7]Wang X H.Genetic diversity of Phyllosticta species associated w ith Citrus spp.in China(in Chinese)[J].Acta Phytopathologica Sinica(植物病理学报),2018,48(1):46-54.
[8]Xu Y.Identification and genetic diversity of Cenococcum geophilum Fr.strains using PCR-RFLP and RAPD markers(in Chinese)[D].Hohhot:Inner M ongolia Agricultural University(呼和浩特:内蒙古农业大学),2006.
[9]Lu N H,Zheng W M,Wang J F,et al.SSR analysis of population genetic diversity of Puccinia striiformis f.sp.tritici in Longnan region of Gansu,China(in Chinese)[J].Scientia Agricultura Sinica(中国农业科学),2009,42(8):2763-2770.
[10]Chen B Y,Hu Q,Li G Q,et al.Genetic diversity in Sclerotinia sclerotiorum assessed w ith SRAP markers(in Chinese)[J].Biodiversity Science(生物多样性),2010,18(5):509-515.
[11]Karimi E,Safaie N,Shams-Bakhsh M.Assessment of genetic diversity among Sclerotinia sclerotiorum populations in fieled by Rep-PCR[J].Trakia Journal of Sciences,2011,9(2):62-70.
[12]Hemmati R,Javan-Nikkhah M,Linde C.Population genetic structure of Sclerotinia sclerotiorum on canola in Iran[J].European Journal of Plant Pathology,2009,125(4):617-628.
[13]Zhang Q,Chen Y F,Gao X N,et al.SRAP analysis of genetic diversity of Sclerotinia sclerotiorum in Shaanxi province(in Chinese)[J].Acta Agriculturae Boreali-occidentalis Sinica(西北农业学报),2012,21(10):149-155.
[14]Laura A W,Steven T,Berlin D.Genetic variation of Sclerotinia sclerotiorum from multiple crops in the North Central United States[J].PLoS One,2015,10(9):doi:10.1371/journal.pone.0139188.
[15]Sun J M,Irzykowski W,Jedryczka M,et al.Analysis of the genetic structure of Sclerotinia sclerotiorum(Lib.)de Bary populations from different regions and host plants by random amplified polymorphic DNAmarkers[J].Journal of Intergrative Plant Biology,2005,47(4):385-395.
[16]Han G Z,Han F X,Sun J M,et al.Genetic diversity of Sclerotinia sclerotiorum isolates from different regions and host plants by SSR markers(in Chinese)[J].Chinese Journal of Oil Crop Sciences(中国油料作物学报),2014,36(2):244-249.
[17]Li G,Quiros C F.Sequence related amplified polymorphism(SRAP),a new marker system based on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theoretical and Applied Genetics,2001,103(3):455-461.
[18]Li T T,Zhu J R,Qiu S,et al.Fingerprinting construction for superior individuals of Zelkova schneideriana based on CE-SRAP(in Chinese)[J].Journal of Plant Genetic Resources(植物遗传资源学报),2016,17(1):169-176.
[19]Qiu S,Shen B C,Li T T,et al.A method of osmanthus fragrans cultivars identification based on random forest algorithm and SRAP molecular markers(in Chinese)[J].Scientia Silvae Sinicae(林业科学),2018,54(1):32-45.
[20]Matias P,Hedvig K,David L,et al.SRAP technique efficiently generates polymorphisms in Puccinia striiformis isolates[J].Journal of Phytopathology,2010,158(10):708-711.
[21]Li G,Gao M,Yang B,et al.Gene for gene alignment betw een the Brassica and Arabidopsis genomes by direct transcriptome mapping[J].Theoretical and Applied Genetics,2003,107(1):168-180.
[22]Lin Z X,Zhang X L Nie Y C,et al.Construction of a genetic linkage map for cotton based on SRAP[J].Chinese Science Bulletin,2003,48(19):2064-2068.
[23]Chang Y H,Zhang A Y,Liu P,et al.Genetic diversity analysis w ith capillary electrophoresis detection w ith fluorescent ISSR markers on foxtail millet(in Chinese)[J].M olecular Plant Breeding(分子植物育种),2017,15(8):3302-3308.