云南高原异龙湖噬藻体psbA基因遗传多样性研究
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摘要
噬藻体作为一类感染蓝藻的特异性病毒,广泛分布于不同水生态系统中。作为噬藻体的一种靶标基因,编码光合作用反应中心D1蛋白的基因psbA不仅可用于揭示噬藻体的遗传多样性,还可用于探索噬藻体与其宿主蓝藻之间的关系。该文以云南高原富营养化湖泊异龙湖为研究对象,以psbA基因为分子标记,从异龙湖浓缩水样中共获得15条噬藻体psbA环境序列。基于噬藻体的psbA序列构建系统进化树,分析异龙湖水样噬藻体光合作用基因psbA的遗传多样性。研究结果表明:云南高原湖泊异龙湖水体中噬藻体psbA基因类群与海洋、其他淡水湖和日本稻田水体中psbA基因类群不同,与中国东北稻田的生态类群进化距离较近,进化相对独立。异龙湖水体中存在新的噬藻体类群,并且不同季节水样中噬藻体psbA基因序列存在差异,秋季其遗传多样性更加丰富。
Cyanophages, as a kind of peculiar viruse that infect specifically cyanobacteria, exists extensively in various aquatic environments. The gene psbA, encoding the D1 protein involved in photosynthesis reaction center, is a target gene of cyanophage, which not only reveals the genetic diversity of cyanophage, but also explores the relationship between the cyanophage and its host cyanobacteria. In this study, the plateau eutrophic Yilong Lake in Yunnan Province was taken as the research object. A total of 15 cyanophage psbA environmental sequences were obtained by using the psbA gene as a molecular marker from the concentrated water samples of Yilong Lake. Based on the cyanophage psbA sequence, phylogenetic tree was constructed, and the genetic diversity of cyanophage photosynthesis psbA gene in the water samples of Yilong Lake were analyzed. The results show that the psbA gene assemblage of the plateau eutrophic Yilong Lake in Yunnan Province are obviously different from those obtained from marine waters, other freshwater lakes and Japanese paddy floodwater.It is closer to evolutionary distance of paddy water in Northeast China, and the evolution is relatively independent. The novel psbA assembly of cyanophage was found in the plateau eutrophic Yilong Lake. For different seasons, the genetic diversity of psbA gene in cyanophage was more abundant in autumn.
Cyanophages, as a kind of peculiar viruse that infect specifically cyanobacteria, exists extensively in various aquatic environments. The gene psbA, encoding the D1 protein involved in photosynthesis reaction center, is a target gene of cyanophage, which not only reveals the genetic diversity of cyanophage, but also explores the relationship between the cyanophage and its host cyanobacteria. In this study, the plateau eutrophic Yilong Lake in Yunnan Province was taken as the research object. A total of 15 cyanophage psbA environmental sequences were obtained by using the psbA gene as a molecular marker from the concentrated water samples of Yilong Lake. Based on the cyanophage psbA sequence, phylogenetic tree was constructed, and the genetic diversity of cyanophage photosynthesis psbA gene in the water samples of Yilong Lake were analyzed. The results show that the psbA gene assemblage of the plateau eutrophic Yilong Lake in Yunnan Province are obviously different from those obtained from marine waters, other freshwater lakes and Japanese paddy floodwater.It is closer to evolutionary distance of paddy water in Northeast China, and the evolution is relatively independent. The novel psbA assembly of cyanophage was found in the plateau eutrophic Yilong Lake. For different seasons, the genetic diversity of psbA gene in cyanophage was more abundant in autumn.
引文
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[18] ROCAP G,DISTEL D L,WATERBURY J B,et al.Resolution of Prochlorococcus and Synechococcus Ecotypes by Using 16S-23S Ribosomal DNA Internal Transcribed Spacer Sequences[J].Applied and Environmental Microbiology,2002,68(3):1180-1191.
[19] 彭玉辅.滇池水域微藻种群变化、食藻原生动物和溶藻细菌分离纯化及初步研究[D].昆明:昆明理工大学,2012.[PENG Yu-fu.Changes of Phyloplankton Population of Lake Dianchi,Isolation and Preliminary Study on Protozoan and Algicidal Bacteria[D].Kunming:Kunming University of Science and Technology,2012.]
[20] SHARON I,TZAHOR S,WILLIAMSON S,et al.Viral Photosynthetic Reaction Center Genes and Transcripts in the Marine Environment[J].Multidisciplinary Journal of Microbial Ecology,2007,1(6):492-501.
[21] WANG X Z,JING R Y,LIU J J,et al.Narrow Distribution of CyanophagepsbA Genes Observed in Two Paddy Waters of Northeast China by an Incubation Experiment[J].Virologica Sinica,2016,31 (2):188-191.
[22] WANG G H,MURASE J,ASAKAWA S,et al.Novel Cyanophage Photosynthetic Gene psbA in the Floodwater of a Japanese Rice Field[J].FEMS Microbiology Ecology,2009,70(1):79-86.
[23] SANDAA R A,CLOKIE M,MANN N H.Photosynthetic Genes in Viral Populations With a Large Genomic Size Range From Norwegian Coastal Waters[J].FEMS Microbiology Ecology,2008,63(1):2-11.
[24] CHENARD C,SUTTLE CA.Phylogenetic Diversity of Sequences of Cyanophage Photosynthetic Gene psbA in Marine and Freshwaters[J].Applied and Environmental Microbiology,2008,74(17):5317-5324.
[25] THERIOT E C,ASHWORTH M P,NAKOV T,et al.Dissecting Signal and Noise in Diatom Chloroplast Protein Encoding Genes With Phylogenetic Information Profiling[J].Molecular Phylogenetics and Evolution,2015,89:28-36.
[26] WILHELM S W,MATTESON A R.Freshwater and Marine Virioplankton:a Brief Overview of Commonalities and Differences[J].Freshwater Biology,2008,53(6):1076-1089.
[27] ZEIDENER G,PRESTON C M,DELONG E F,et al.Molecular Diversity Among Marine Picophytoplankton as Revealed by psbA Analyses[J].Environmental Microbiology,2010,5(3):212-216.
[28] ZHONG X,JACQUET S.Prevalence of Viral Photosynthetic and Capsid Protein Genes From Cyanophages in Two Large and Deep Perialpine Lakes[J].Applied and Environmental Microbiology,2013,79 (23) :7169-7178.
[29] ZHONG Y,CHEN F,WILHELM S W,et al.Phylogenetic Diversity of Marine Cyanophage Isolates and Natural Virus Communities as Revealed by Sequences of Viral Capsid Assembly Protein Gene g20[J].Applied and Environmental Microbiology,2002,68(4) :1576-1584.
[30] 吴阿娜,朱梦杰,汤琳等.淀山湖蓝藻水华高发期叶绿素a动态及相关环境因子分析[J].湖泊科学,2011,23(1):67-72.[WU A-na,ZHU Meng-jie,TANG Lin,et al.Dynamics of Chlorophyll-a and Analysis of Environmental Factors in Lake Dianshan During Summer Autumn[J].Journal of Lake Sciences,2011,23(1):67-72.]
[2] 张奇亚.噬藻体生物多样性的研究动态[J].微生物学通报,2014,41(3):545-559.[ZHANG Qi-ya.Advances in Studies on Biodiversity of Cyanophages[J].Microbiology China,2014,41(3):545-559.]
[3] WEI D,FU C,RAYTCHEVA D,et al.Visualizing Virus Assembly Intermediates Inside Marine Cyanobacteria [J].Nature,2013,502(7473):707-710.
[4] OU T,GAO X C,LI S H,et al.Genome Analysisand Gene nblA Identification of Microcystis aeruginosa Myovirus (MaMV-DC) Reveal the Evidence for HorizontalGene Transfer Events Between Cyanomyovirus and Host[J].Journal of General Virology,2015,96(12):3681.
[5] 荆瑞勇,KIMURA M,王光华.不同自然环境下噬藻体g20基因多样性研究进展[J].微生物学报,2013,53(11):1149-1157.[JING Rui-yong,KIMURA M,WANG Guang-hua.Genetic Diversity of Capsid Assembly Protein Genes (g20) of Cyanophage in Different Natural Environment:A Review [J].Acta Microbiologica Sinica,2013,53(11):1149-1157.]
[6] BUTINA T V,POTAPOV S A,BELYKH O I,et al.Genetic Diversity of Cyanophages of the Myoviridae Family as a Constituent of the Associated Community of the Baikal Sponge Lubomirskia baicalensis[J].Russian Journal of Genetics,2015,51(3):313-317.
[7] XU Y,ZHANG R,WANG N,et al.Novel Phage-host Interactions and Evolution as Revealed by a Cyanomyovirus Isolated From an Estuarine Environment[J].Environmental Microbiology,2018,20 (8):2974 -2989.
[8] CHEN F,WANG K,HUANG S J,et al.Diverse and Dynamic Populations of Cyanobacterial Podoviruses in the Chesapeake Bay Unveiled Through DNA Polymerase Gene Sequences[J].Environmental Microbiology,2009,11(11):2884-2892.
[9] 汪岷,闫群.噬藻体遗传多样性的研究进展[J].中国海洋大学学报,2010,40(8):73-79.[WANG Min,YAN Qun.Advances in Research on Genetic Diversity of Cyanophage[J].Periodical of Ocean University of China,2010,40(8):73-79.]
[10] 李樾,刘婷婷,刘丽,等.基于psbA基因的噬藻体遗传多样性研究进展[J].生态与农村环境学报,2016,32(3):417-423.[LI Yue,LIU Ting-ting,LIU Li,et al.Progress of Research on Genetic Diversity of Cyanophage Based on psbA Gene[J].Journal of Ecology and Rural Environment,2016,32(3):417-423.]
[11] 荆瑞勇,曹焜,刘俊杰,等.东北稻田水体噬藻体psbA基因多样性[J].微生物学报,2017,57(1)131-139.[JING Rui-yong,CAO Kun,LIU Jun-jie,et al.Genetic Diversity of psbA of Cyanophage from Paddy Floodwater in Northeast China [J].Acta Microbiologica Sinica,2017,57(1)131-139.]
[12] SULLIVAN M B,LINDELL D,LEE J A,et al.Prevalence and Evolution of Core Photosystem Ⅱ Genes in Marine Cyanobacterial Viruses and Their Hosts[J].PLoS Biology,2006,4(8):e234.
[13] 闫群.青岛近海海域噬藻体g20基因多样性的研究[D].青岛:中国海洋大学,2010.[YAN Qun.g20 Gene Diversity of Cyanophage Community in the Yellow Sea Coastal Area Around Qingdao [D].Qingdao:Ocean University of China,2010.]
[14] JING R Y,LIU J J,YU Z H,et al.Phylogenetic Distribution of the Capsid Assembly Protein Gene (g20) of Cyanophage in Paddy Floodwaters in Northeast China[J].PLoS One,2014,9(2):e88634.
[15] WANG M N,GE X Y,WU Y Q,et al.Genetic Diversity and Temporal Dynamics of Phytoplankton Viruses in East Lake,China[J].Virologica Sinica,2015,30 (4):290-300.
[16] GARCIA M J,ACINAS S G,ANTON A I,et al.Use of the 16S-23S Ribosomal Genes Spacer Region in Studies of Prokaryotic Diversity[J].Journal of Microbiological Methods,1999,36(1/2):55-64.
[17] FERRIS M J,KUHL M,WIELAND A,et al.Cyanobacterial Ecotypes in Different Optical Microenvironments of a 68Degrees C Hot Spring Mat Community Revealed by 16S-23S rRNA Internal Transcribed Spacer Region Variation[J].Applied and Environmental Microbiology,2003,69(5):2893-2898.
[18] ROCAP G,DISTEL D L,WATERBURY J B,et al.Resolution of Prochlorococcus and Synechococcus Ecotypes by Using 16S-23S Ribosomal DNA Internal Transcribed Spacer Sequences[J].Applied and Environmental Microbiology,2002,68(3):1180-1191.
[19] 彭玉辅.滇池水域微藻种群变化、食藻原生动物和溶藻细菌分离纯化及初步研究[D].昆明:昆明理工大学,2012.[PENG Yu-fu.Changes of Phyloplankton Population of Lake Dianchi,Isolation and Preliminary Study on Protozoan and Algicidal Bacteria[D].Kunming:Kunming University of Science and Technology,2012.]
[20] SHARON I,TZAHOR S,WILLIAMSON S,et al.Viral Photosynthetic Reaction Center Genes and Transcripts in the Marine Environment[J].Multidisciplinary Journal of Microbial Ecology,2007,1(6):492-501.
[21] WANG X Z,JING R Y,LIU J J,et al.Narrow Distribution of CyanophagepsbA Genes Observed in Two Paddy Waters of Northeast China by an Incubation Experiment[J].Virologica Sinica,2016,31 (2):188-191.
[22] WANG G H,MURASE J,ASAKAWA S,et al.Novel Cyanophage Photosynthetic Gene psbA in the Floodwater of a Japanese Rice Field[J].FEMS Microbiology Ecology,2009,70(1):79-86.
[23] SANDAA R A,CLOKIE M,MANN N H.Photosynthetic Genes in Viral Populations With a Large Genomic Size Range From Norwegian Coastal Waters[J].FEMS Microbiology Ecology,2008,63(1):2-11.
[24] CHENARD C,SUTTLE CA.Phylogenetic Diversity of Sequences of Cyanophage Photosynthetic Gene psbA in Marine and Freshwaters[J].Applied and Environmental Microbiology,2008,74(17):5317-5324.
[25] THERIOT E C,ASHWORTH M P,NAKOV T,et al.Dissecting Signal and Noise in Diatom Chloroplast Protein Encoding Genes With Phylogenetic Information Profiling[J].Molecular Phylogenetics and Evolution,2015,89:28-36.
[26] WILHELM S W,MATTESON A R.Freshwater and Marine Virioplankton:a Brief Overview of Commonalities and Differences[J].Freshwater Biology,2008,53(6):1076-1089.
[27] ZEIDENER G,PRESTON C M,DELONG E F,et al.Molecular Diversity Among Marine Picophytoplankton as Revealed by psbA Analyses[J].Environmental Microbiology,2010,5(3):212-216.
[28] ZHONG X,JACQUET S.Prevalence of Viral Photosynthetic and Capsid Protein Genes From Cyanophages in Two Large and Deep Perialpine Lakes[J].Applied and Environmental Microbiology,2013,79 (23) :7169-7178.
[29] ZHONG Y,CHEN F,WILHELM S W,et al.Phylogenetic Diversity of Marine Cyanophage Isolates and Natural Virus Communities as Revealed by Sequences of Viral Capsid Assembly Protein Gene g20[J].Applied and Environmental Microbiology,2002,68(4) :1576-1584.
[30] 吴阿娜,朱梦杰,汤琳等.淀山湖蓝藻水华高发期叶绿素a动态及相关环境因子分析[J].湖泊科学,2011,23(1):67-72.[WU A-na,ZHU Meng-jie,TANG Lin,et al.Dynamics of Chlorophyll-a and Analysis of Environmental Factors in Lake Dianshan During Summer Autumn[J].Journal of Lake Sciences,2011,23(1):67-72.]
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