基于MLST鄱阳湖微囊藻的遗传多样性及其系统发育
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摘要
为了研究鄱阳湖微囊藻的遗传多样性及其系统发育关系,基于7个管家基因(fts Z、gln A、glt X、gyr B、pgi、rec A和tpi),建立了一个多位点序列分型(MLST)方法.对来自鄱阳湖的20株微囊藻分离株进行MLST研究,并构建本地微囊藻MLST基因库.结果表明:这20株藻株具有20种独特的序列型(STs),揭示了微囊藻高水平的遗传多样性(H=0.986).基于7个MLST位点串联序列的系统发育分析显示,微囊藻藻株序列型可分为5个不同的组.与之前日本湖泊研究的237个STs共建的系统发育树比对表明,本研究的STs形成了独立的分枝.该结果说明不同地区的微囊藻是相对稳定的,因此可以用MLST进行明确的表征.
Using the multilocus sequence typing(MLST) based on seven housekeeping loci(fts Z, gln A, glt X, gyr B, pgi, rec A and tpi), twenty Microcystis strain isolated from Lake Poyang, the largest freshwater Lake in China, were examined to study their genetic diversity and phylogenetic relationship. The MLST revealed the 20 unique sequence types(STs) for these 20 Microcystis strains and their higher genetic diversity with H value as 0.986. The phylogenetic analysis based on the seven gene sequences showed that Microcystis strains were divided into five different groups, Phylogenetic tree based on the STs from our research and Japanese lakes revealed that our STs formed the independent branch, indicating that microcysts from different areas is relatively stable. So it can be clearly characterized by MLST.
Using the multilocus sequence typing(MLST) based on seven housekeeping loci(fts Z, gln A, glt X, gyr B, pgi, rec A and tpi), twenty Microcystis strain isolated from Lake Poyang, the largest freshwater Lake in China, were examined to study their genetic diversity and phylogenetic relationship. The MLST revealed the 20 unique sequence types(STs) for these 20 Microcystis strains and their higher genetic diversity with H value as 0.986. The phylogenetic analysis based on the seven gene sequences showed that Microcystis strains were divided into five different groups, Phylogenetic tree based on the STs from our research and Japanese lakes revealed that our STs formed the independent branch, indicating that microcysts from different areas is relatively stable. So it can be clearly characterized by MLST.
引文
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[23]Kaebernick M,Neilan B A,Borner T,et al.Light and the transcriptional response of the microcystin biosynthesis gene cluster[J].Applied and Environmental Microbiology,2000,66(8):3387-3392.
[24]Sevilla E,Martin-Luna B,Vela L,et al.Iron availability affects mcy Dexpression and microcystin-LR synthesis in Microcystis aeruginosa PCC7806[J].Environmental Microbiology,2008,10(10):2476-2483.
[25]Biand E,Escoffier N,Straub C,et al.Spatiotemporal changes in the genetic diversity of a bloom-forming Microcystis aeruginosa(cyanobacteria)population[J].Isme Journal,2009,3(4):419-429.
[26]Tanabe Y,Watanabe M M.Local expansion of a panmictic lineage of water bloom-forming cyanobacterium Microcystis aeruginosa[J].Plos One,2011,6(2):e17085.
[27]Liu Y,Xu Y,Wang Z,et al.Dominance and succession of Microcystis genotypes and morphotypes in Lake Taihu,a large and shallow freshwater lake in China[J].Environmental Pollution,2016,219:399-408.
[2]Mohamed Z A,Deyab M A,Abou-Dobara M I,et al.Occurrence of cyanobacteria and microcystin toxins in raw and treated waters of the Nile River,Egypt:implication for water treatment and human health[J].Environmental Science and Pollution Research,2015,22(15):11716-11727.
[3]Nolen R S.A one-health solution to the toxic algae problem[J].Javma-Journal of the American Veterinary Medical Association,2018,252(8):906-909.
[4]Otsuka S,Suda S,Shibata S,et al.A proposal for the unification of five species of the cyanobacterial genus Microcystis Kutzing ex Lemmermann 1907under the Rules of the Bacteriological Code[J].International Journal of Systematic and Evolutionary Microbiology,2001,51(3):873-879.
[5]Schmidt J R,Wilhelm S W,Boyer G L.The fate of microcystins in the environment and challenges for monitoring[J].Toxins,2014,6(12):3354-3387.
[6]Huo D,Chen Y,Zheng T,et al.Characterization of microcystis(Cyanobacteria)genotypes based on the internal transcribed spacer region of rRNA by next-generation sequencing[J].Frontiers in Microbiology,2018,9:971.
[7]Yang C,Lin F,Li Q,et al.Comparative genomics reveals diversified CRISPR-Cas systems of globally distributed Microcystis aeruginosa,a freshwater bloom-forming cyanobacterium[J].Front Microbiol,2015,6:394.
[8]Tanabe Y,Kasai F,Watanabe M M.Multilocus sequence typing(MLST)reveals high genetic diversity and clonal population structure of the toxic cyanobacterium Microcystis aeruginosa[J].MicrobiologySgm,2007,153(11):3695-3703.
[9]Tanabe Y,Kasai F,Watanabe M M.Fine-scale spatial and temporal genetic differentiation of water bloom-forming cyanobacterium Microcystis aeruginosa:revealed by multilocus sequence typing[J].Environmental Microbiology Reports,2009,1(6):575-582.
[10]Huo D,Cao Q,Wang S Y,et al.The spatial distribution of Microcystis and microcystin and its relationship with environmental factors in Haihe Tianjin City[J].China of Environmental Science,2018,38(10):3897-3903.
[11]Feil E J,Holmes E C,Bessen D E,et al.Recombination within natural populations of pathogenic bacteria:Short-term empirical estimates and long-term phylogenetic consequences[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(1):182-187.
[12]Rozas J,Sanchez-Delbarrio J C,Messeguer X,et al.DnaSP,DNApolymorphism analyses by the coalescent and other methods[J].Bioinformatics,2003,19(18):2496-2507.
[13]Tajima F.Statistical method for testing the neutral mutation hypothesis by DNA polymorphism[J].Genetics Society of America,1989,123(3):585-595.
[14]Zhang N,Zeng L,Shan H,et al.Highly conserved low-copy nuclear genes as effective markers for phylogenetic analyses in angiosperms[J].New Phytologist,2012,195(4):923-937.
[15]Swofford D L.Phylogenetic analysis using parsimony(*and other methods)[Z].2002.
[16]Lam-Tung N,Schmidt H A,Von Haeseler A,et al.IQ-TREE:A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies[J].Molecular Biology and Evolution,2015,32(1):268-274.
[17]Hanage W P,Fraser C,Spratt B G.Fuzzy species among recombinogenic bacteria[J].Bmc.Biology,2005,3:6.
[18]Caugant D A,Levin B R,Selander R K.Genetic Diversity and Temporal Variation in the Escherichia-Coli Population of a Human Host[J].Genetics,1981,98(3):467-490.
[19]Pinchua I V,Bressollier P,Sorokulova I B,et al.Amicoumacin antibiotic production and genetic diversity of Bacillus subtilis strains isolated from different habitats[J].Research in Microbiology,2002,153(5):269-276.
[20]Kurmayer R,Christiansen G,Gumpenberger M,et al.Genetic identification of microcystin ecotypes in toxic cyanobacteria of the genus planktothrix[J].Microbiology-Sgm,2005,151(5):1525-1533.
[21]Christiansen G,Kurmayer R,Liu Q,et al.Transposons inactivate biosynthesis of the nonribosomal peptide microcystin in naturally occurring Planktothrix spp[J].Applied and Environmental Microbiology,2006,72(1):117-123.
[22]Mlouka A,Comte K,Castets A M,et al.The gas vesicle gene cluster from Microcystis aeruginosa and DNA rearrangements that lead to loss of cell buoyancy[J].Journal of Bacteriology,2004,186(8):2355-2365.
[23]Kaebernick M,Neilan B A,Borner T,et al.Light and the transcriptional response of the microcystin biosynthesis gene cluster[J].Applied and Environmental Microbiology,2000,66(8):3387-3392.
[24]Sevilla E,Martin-Luna B,Vela L,et al.Iron availability affects mcy Dexpression and microcystin-LR synthesis in Microcystis aeruginosa PCC7806[J].Environmental Microbiology,2008,10(10):2476-2483.
[25]Biand E,Escoffier N,Straub C,et al.Spatiotemporal changes in the genetic diversity of a bloom-forming Microcystis aeruginosa(cyanobacteria)population[J].Isme Journal,2009,3(4):419-429.
[26]Tanabe Y,Watanabe M M.Local expansion of a panmictic lineage of water bloom-forming cyanobacterium Microcystis aeruginosa[J].Plos One,2011,6(2):e17085.
[27]Liu Y,Xu Y,Wang Z,et al.Dominance and succession of Microcystis genotypes and morphotypes in Lake Taihu,a large and shallow freshwater lake in China[J].Environmental Pollution,2016,219:399-408.