菌群条件下铜绿微囊藻休眠体复苏期间光合效率变化
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摘要
将3株冲天湖底泥优势菌(Exiguobacterium.sp013、Bacillus.spD06与Bacillus.spD24)按不同配比与铜绿微囊藻休眠体用底泥包埋,在梯度温度下(10、15、20和25℃)进行复苏模拟实验.结果表明,在15、20和25℃条件下,菌组和无菌组铜绿微囊藻休眠体均能从底泥复苏进入上覆水体,但菌组复苏率显著高于无菌组(p<0.05).在10℃条件下,菌组铜绿微囊藻休眠体能复苏,而无菌组不能.在20和25℃时,菌组对铜绿微囊藻休眠体光合系统的光合效率F_v/F_m无显著提升作用(p> 0.05),而在10和15℃时,菌组能显著提升铜绿微囊藻休眠体的光合效率F_v/F_m(p<0.05),且菌组Mix-4的提升能力显著强于其他菌组和无菌组.基因测序和KEGG富集分析发现,15℃时菌组Mix-4差异表达的基因显著富集于Cell cycle通路中,包括氧化磷酸化(ATP生成反应)通路和内质网蛋白合成通路,同时与光合作用相关的蛋白基因psaB、psbA及rbcL的表达量均发生显著上调,这可能是Mix-4菌群在较低温度下(15℃)提升铜绿微囊藻休眠体光合效率F_v/F_m并促进其复苏的分子机理.
To explore the effects of the three dominant benthic freshwater bacteria( Exiguobacterium.sp013,Bacillus.spD06 and Bacillus.spD24) on the recruitment of dormant Microcystis aeruginosa from the sediment,and to find potential factors and mechanism,simulation on the recruitment of M. aeruginosa cells from sediment were performed in the presence or absence of the three bacterial strains. Three strains of the dominant bacteria isolated from Lake Chongtian and dormant M. aeruginosa were embedded in the sediment according to different bacterial ratio,and the recruitment experiments were conducted under different temperatures(10,15,20 and 25 ℃). Results show that the dormant M. aeruginosa cells in both of the bacterial and aseptic groups recruited from the sediment into the overlying water under the temperature of 15,20 and 25 ℃,and the recruitment rate of bacterial groups were significantly higher than that of aseptic groups( p<0.05). At 10 ℃,the recruitment of dormant M. aeruginosa cells takes place in bacterial groups,but not in aseptic groups. At 10 ℃ and 15 ℃,the photosynthetic efficiency( F_v/F_m) of the dormant M. aeruginosa were significantly enhanced by mixed bacterial group( Mix-4),which was significantly stronger than other bacterial and aseptic group. When the temperature was 20 ℃ and 25 ℃,there were no significant effect on the photosynthetic efficiency( F_v/F_m) of dormant M. aeruginosa cells(p>0.05) induced by mixed bacterial group. Then,the differentially expressed genes were found that were significantly enriched in the cell cycle pathway,including the oxidative phosphorylation( ATP generation reaction) pathway and endoplasmic reticulum protein synthesis pathway at 15 ℃ for Mix-4 groups by gene sequencing and KEGG enrichment analysis. At the same time,the expression levels of psa B,psb A and rbc L genes related to photosynthesis significantly increased( UP). These might be the molecular mechanism,which mixed bacteria Mix-4 enhanced photosynthetic efficiency( F_v/F_m) of dormant M.aeruginosa at lower temperature(15 ℃)and promoted its recruitment.
To explore the effects of the three dominant benthic freshwater bacteria( Exiguobacterium.sp013,Bacillus.spD06 and Bacillus.spD24) on the recruitment of dormant Microcystis aeruginosa from the sediment,and to find potential factors and mechanism,simulation on the recruitment of M. aeruginosa cells from sediment were performed in the presence or absence of the three bacterial strains. Three strains of the dominant bacteria isolated from Lake Chongtian and dormant M. aeruginosa were embedded in the sediment according to different bacterial ratio,and the recruitment experiments were conducted under different temperatures(10,15,20 and 25 ℃). Results show that the dormant M. aeruginosa cells in both of the bacterial and aseptic groups recruited from the sediment into the overlying water under the temperature of 15,20 and 25 ℃,and the recruitment rate of bacterial groups were significantly higher than that of aseptic groups( p<0.05). At 10 ℃,the recruitment of dormant M. aeruginosa cells takes place in bacterial groups,but not in aseptic groups. At 10 ℃ and 15 ℃,the photosynthetic efficiency( F_v/F_m) of the dormant M. aeruginosa were significantly enhanced by mixed bacterial group( Mix-4),which was significantly stronger than other bacterial and aseptic group. When the temperature was 20 ℃ and 25 ℃,there were no significant effect on the photosynthetic efficiency( F_v/F_m) of dormant M. aeruginosa cells(p>0.05) induced by mixed bacterial group. Then,the differentially expressed genes were found that were significantly enriched in the cell cycle pathway,including the oxidative phosphorylation( ATP generation reaction) pathway and endoplasmic reticulum protein synthesis pathway at 15 ℃ for Mix-4 groups by gene sequencing and KEGG enrichment analysis. At the same time,the expression levels of psa B,psb A and rbc L genes related to photosynthesis significantly increased( UP). These might be the molecular mechanism,which mixed bacteria Mix-4 enhanced photosynthetic efficiency( F_v/F_m) of dormant M.aeruginosa at lower temperature(15 ℃)and promoted its recruitment.
引文
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Brunberg A K,Blomqvist P.2002.Benthic overwintering of Microcystis colonies under different environmental conditions[J].J Plankton Res,24:1247-1252
Brunberg A K,Blomqvist P.2003.Recruitment of Microcystis(Cyanophycae)from lake sediments:The importance of littoral inocula[J].J Phycol,39(1):58-63
Cáceres O,Reynolds C S.1984.Some effects of artificially enhanced anoxia on the Microcystis aeruginosa Kütz emend Elenkin with special reference to the initiation of its annual growth cycle in lake[J].Archivfur Hydrobiologia,99:379-397
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Cires S,Wormer L,Agha R,et al.2013.Overwintering populations of Anabaena,Aphanizomenon and Microcystis as potential inocula for summer blooms[J].J Plankton Res,35(6):1254-1266
Dorais P,Stoichev T,Basto M C P,et al.2014.Cyanobacterium Microcystis aeruginosa response to pentachlorophenol and comparison with that of the microalga Chlorella vulgaris[J].Water Res,52:63-72
Fontana A,Patrone V,Puglisi E,et al.2016.Effects of geographic area,feedstock,temperature,and operating time on microbial communities of sixfull-scale biogas plants[J].Bioresour Technol,218:980-990
Hodoki Y,Ohbayashi K,Kobayashi Y,et al.2011.Temporal variation in cyanobacteria species composition and photosynthetic activity in experimentally induced blooms[J].J Plankton Res,33:1410-1416
Huang D W,Sherman B T,Lempicki R A.2009.Bioinformatics enrichment tools:paths towards the comprehensive functional analysis of largr gene lists[J].Nucleic Acids Res,37(1):1-13
Huang D W,Sherman B T,Lempicki R A.2009.Systematic and integrative analysis of large gene lists using DAVIDE bioinformatics resources[J].Nature Protocols,4(1):44-57
Ihle T,Jahnichen S,Benndorf J.2005.Wax and wane of Microcystis(Cyanophyceae)and Microcystins in lake sediment:a case syudy in Quitzdorf Reservoir(Germany)[J].J Phycol,41:479-488
Jahnichen S,Long B M,Petzoldt T.2011.Microcystin production by Microcystis aeruginosa:Direct regulation by multiple environmental factors[J].Harmful Algae,12:95-104
Karlson A M L,Nascimento F J A,Sanna S,et al.2012.Benthic fauna affects recruitment from sediments of the harmful cyanobacterium Nodularia spumigena[J].Harmful Algae,20:126-131
Lehman P W,Teh S J,Boyer G L,et al.2010.Initial impacts of Microcystis aeruginosa blooms on the aquatic food web in the San Francisco Estuary[J].Hydrobiologia,637:229-248
Ma J R,Qin B Q,Paerl H W,et al.2016.The persistence of cyanobacterial(Microcystis spp.)blooms throughout winter in Lake Taihu,China[J].Limnol Oceanogr,61:711-722
Misson B,Latour D.2013.Influence of light,sediment mixing,temperature and duration of the benthic life phase on the benthic recruitment of Microcystis[J].Journal of Plankton Research,34(2):113-119
Neilan B A,Pearson L A,Muenchhoff J,et al.2013.Environmental conditions that influence toxin biosynthesis in cyanobacteria[J].Environ Microbiol,15(5):1239-1253
Paerl H W,Otten T G.2013.Blooms bite the hand that feeds them[J].Science,342:433-434
Preston T,Stewart W D P,Reynolds C S.1980.Bloom-forming cyanobacterium Microcystis aeruginosa overwinters on sediment surface[J].Nature,288:365-367
Qian H F,Pan X J,Chen J,et al.2016.Analyses of gene expression and physiological changes in Microcystis aeruginosa reveal the phytotoxicities of three environmental pollutants[J].Ecotoxicology,21:847-859
Reynolds C S,Jaworski G H M,Cmiech H A,et al.1981.On the annual cycle of the blue-green alga Microcystis aeruginosa Kütz.emend.Elenkin[J].Philosophical Transactions of the Royal Society B:Biological Sciencess,293:419-477
Robert Edward Lee.2008.Phycology[M].Cambridge:Cambridge University Press
Rossetti V,Schirrmeister B E,Bernasconi M V.2010.The evolutionary path to terminal differentiation and division of labor in cyanobacteria[J].Journal of Theoretical Biology,262(1):23-34
Schone K,Jahnichen S,Ihle T,et al.2010.Arriving in better shape:Benthic Microcystis as inoculum for pelagic growth[J].Harmful Algae,9(5):494-503
Stahl-Delbanco A,Hansson L A,Gyllstr9m M.2003.Recruitment of resting stages may induce blooms of Microcystis at low N:P ratios[J].Journal of Plankton Research,25(9):1099-1106
Sun Q Q,Zhu W,Li M,et al.2016.Morphological changes of Microcystis aeruginosa colonies in culture[J].Journal of Limnology,75(1):14-23
苏玉萍,李艳芳,钟厚璋,等.2012.山仔水库沉积物蓝藻复苏试验研究[J].环境科学学报,32(2):341-348
Takamura N,Yasuno M,Sugahara K.1984.Overwintering of Microcystis aeruginosa Kütz in a shallow lake[J].Journal of Plankton Research,6(6):1019-1029
Tan X,Kong F X,Zhang M Y,et al.2009.Effects of temperature on phytoplankton recruitment and variations in phytoplankton community composition[J].J Freshwater Ecol,24:339-341
Thomas R H,Walsby A E.1986.The effects of temperatures on recovery of buoyancy by Microcystis[J].Microbiology,132(6):1665-1672
Trapnell C,Williams B A,Pertea G,et al.2010.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nat Biotech,28:511-515
Tsujimura S,Tsukada H,Nakahara H,et al.2000.Seasonal variations of Microcystis populations in sediments of lake Biwa Japan[J].Hydrobiologia,434(1/3):183-192
Tsukada H A.2006.Study on the life history and the factor affecting the dominance of Microcystis in eutrophic lakes[D].Kyoto:Kyoto University
谭啸,孔繁翔,于洋,等.2009.升温过程对藻类复苏和群落演替的影响[J].中国环境科学,29(6):578-582
陶益,孔繁翔,曹焕生,等.2005.太湖底泥水华蓝藻复苏的模拟[J].湖泊科学,17(3):231-236
Verspagen J M H,Snelder E O F,Visser P M,et al.2004.Recruitment of benthic Microcystis(Cyanophyceae)to the water column:internal buoyancy changes or susupension?[J].Journal of Phycology,40:260-270
Wan N,Tang J,Li Q M,et al.2008.The responses of Microcystis to sediment environments and the assessment for its overwintering a simulation study in a novel device[J].Fresenius Environmental Bulletin,17(12b):2146-2151
Yamamoto Y.2009.Effect of temperature on recruitment of cyanobacteria from sediment and bloom formation in a shallow pond[J].Plankton&Benthos Research,4:95-103
Yamamoto Y.2010.Contribution of bioturbation by the red swamp crayfish Procambarus clarkia to the recruitment of bloom-forming cyanobacteria from sediment[J].Journal of Limnology,69(1):102-111
Yang C Y,Yi L,Zhou Y Y,et al.2015.Illumina sequencing-based analysis of free-living bacterial community dynamics during an Akashiwo sanguine bloom in Xiamen sea,China[J].Science Rep,5:8476-8486
邹万生,王智,宋齐生,等.2017.两株浅层底泥菌对铜绿微囊藻休眠体复苏的调控作用[J].环境科学学报,37(11):4160-4169
Boltovskoy D,Correa N,Bordet F,et al.2013.Toxic Microcystis(Cyanobacteria)inhibit recruitment of the bloom-enhancing invasive bivalve Limnoperna fortune[J].Freshwater Biol,58:1968-1981
Brunberg A K,Blomqvist P.2002.Benthic overwintering of Microcystis colonies under different environmental conditions[J].J Plankton Res,24:1247-1252
Brunberg A K,Blomqvist P.2003.Recruitment of Microcystis(Cyanophycae)from lake sediments:The importance of littoral inocula[J].J Phycol,39(1):58-63
Cáceres O,Reynolds C S.1984.Some effects of artificially enhanced anoxia on the Microcystis aeruginosa Kütz emend Elenkin with special reference to the initiation of its annual growth cycle in lake[J].Archivfur Hydrobiologia,99:379-397
Cao H S,Tao Y,Kong F X,et al.2008.Relationship between temperature and cyanobacterial recruitment from sediments in laboratory and field studies[J].J Freshwater Ecol,23(3):405-412
Cires S,Wormer L,Agha R,et al.2013.Overwintering populations of Anabaena,Aphanizomenon and Microcystis as potential inocula for summer blooms[J].J Plankton Res,35(6):1254-1266
Dorais P,Stoichev T,Basto M C P,et al.2014.Cyanobacterium Microcystis aeruginosa response to pentachlorophenol and comparison with that of the microalga Chlorella vulgaris[J].Water Res,52:63-72
Fontana A,Patrone V,Puglisi E,et al.2016.Effects of geographic area,feedstock,temperature,and operating time on microbial communities of sixfull-scale biogas plants[J].Bioresour Technol,218:980-990
Hodoki Y,Ohbayashi K,Kobayashi Y,et al.2011.Temporal variation in cyanobacteria species composition and photosynthetic activity in experimentally induced blooms[J].J Plankton Res,33:1410-1416
Huang D W,Sherman B T,Lempicki R A.2009.Bioinformatics enrichment tools:paths towards the comprehensive functional analysis of largr gene lists[J].Nucleic Acids Res,37(1):1-13
Huang D W,Sherman B T,Lempicki R A.2009.Systematic and integrative analysis of large gene lists using DAVIDE bioinformatics resources[J].Nature Protocols,4(1):44-57
Ihle T,Jahnichen S,Benndorf J.2005.Wax and wane of Microcystis(Cyanophyceae)and Microcystins in lake sediment:a case syudy in Quitzdorf Reservoir(Germany)[J].J Phycol,41:479-488
Jahnichen S,Long B M,Petzoldt T.2011.Microcystin production by Microcystis aeruginosa:Direct regulation by multiple environmental factors[J].Harmful Algae,12:95-104
Karlson A M L,Nascimento F J A,Sanna S,et al.2012.Benthic fauna affects recruitment from sediments of the harmful cyanobacterium Nodularia spumigena[J].Harmful Algae,20:126-131
Lehman P W,Teh S J,Boyer G L,et al.2010.Initial impacts of Microcystis aeruginosa blooms on the aquatic food web in the San Francisco Estuary[J].Hydrobiologia,637:229-248
Ma J R,Qin B Q,Paerl H W,et al.2016.The persistence of cyanobacterial(Microcystis spp.)blooms throughout winter in Lake Taihu,China[J].Limnol Oceanogr,61:711-722
Misson B,Latour D.2013.Influence of light,sediment mixing,temperature and duration of the benthic life phase on the benthic recruitment of Microcystis[J].Journal of Plankton Research,34(2):113-119
Neilan B A,Pearson L A,Muenchhoff J,et al.2013.Environmental conditions that influence toxin biosynthesis in cyanobacteria[J].Environ Microbiol,15(5):1239-1253
Paerl H W,Otten T G.2013.Blooms bite the hand that feeds them[J].Science,342:433-434
Preston T,Stewart W D P,Reynolds C S.1980.Bloom-forming cyanobacterium Microcystis aeruginosa overwinters on sediment surface[J].Nature,288:365-367
Qian H F,Pan X J,Chen J,et al.2016.Analyses of gene expression and physiological changes in Microcystis aeruginosa reveal the phytotoxicities of three environmental pollutants[J].Ecotoxicology,21:847-859
Reynolds C S,Jaworski G H M,Cmiech H A,et al.1981.On the annual cycle of the blue-green alga Microcystis aeruginosa Kütz.emend.Elenkin[J].Philosophical Transactions of the Royal Society B:Biological Sciencess,293:419-477
Robert Edward Lee.2008.Phycology[M].Cambridge:Cambridge University Press
Rossetti V,Schirrmeister B E,Bernasconi M V.2010.The evolutionary path to terminal differentiation and division of labor in cyanobacteria[J].Journal of Theoretical Biology,262(1):23-34
Schone K,Jahnichen S,Ihle T,et al.2010.Arriving in better shape:Benthic Microcystis as inoculum for pelagic growth[J].Harmful Algae,9(5):494-503
Stahl-Delbanco A,Hansson L A,Gyllstr9m M.2003.Recruitment of resting stages may induce blooms of Microcystis at low N:P ratios[J].Journal of Plankton Research,25(9):1099-1106
Sun Q Q,Zhu W,Li M,et al.2016.Morphological changes of Microcystis aeruginosa colonies in culture[J].Journal of Limnology,75(1):14-23
苏玉萍,李艳芳,钟厚璋,等.2012.山仔水库沉积物蓝藻复苏试验研究[J].环境科学学报,32(2):341-348
Takamura N,Yasuno M,Sugahara K.1984.Overwintering of Microcystis aeruginosa Kütz in a shallow lake[J].Journal of Plankton Research,6(6):1019-1029
Tan X,Kong F X,Zhang M Y,et al.2009.Effects of temperature on phytoplankton recruitment and variations in phytoplankton community composition[J].J Freshwater Ecol,24:339-341
Thomas R H,Walsby A E.1986.The effects of temperatures on recovery of buoyancy by Microcystis[J].Microbiology,132(6):1665-1672
Trapnell C,Williams B A,Pertea G,et al.2010.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nat Biotech,28:511-515
Tsujimura S,Tsukada H,Nakahara H,et al.2000.Seasonal variations of Microcystis populations in sediments of lake Biwa Japan[J].Hydrobiologia,434(1/3):183-192
Tsukada H A.2006.Study on the life history and the factor affecting the dominance of Microcystis in eutrophic lakes[D].Kyoto:Kyoto University
谭啸,孔繁翔,于洋,等.2009.升温过程对藻类复苏和群落演替的影响[J].中国环境科学,29(6):578-582
陶益,孔繁翔,曹焕生,等.2005.太湖底泥水华蓝藻复苏的模拟[J].湖泊科学,17(3):231-236
Verspagen J M H,Snelder E O F,Visser P M,et al.2004.Recruitment of benthic Microcystis(Cyanophyceae)to the water column:internal buoyancy changes or susupension?[J].Journal of Phycology,40:260-270
Wan N,Tang J,Li Q M,et al.2008.The responses of Microcystis to sediment environments and the assessment for its overwintering a simulation study in a novel device[J].Fresenius Environmental Bulletin,17(12b):2146-2151
Yamamoto Y.2009.Effect of temperature on recruitment of cyanobacteria from sediment and bloom formation in a shallow pond[J].Plankton&Benthos Research,4:95-103
Yamamoto Y.2010.Contribution of bioturbation by the red swamp crayfish Procambarus clarkia to the recruitment of bloom-forming cyanobacteria from sediment[J].Journal of Limnology,69(1):102-111
Yang C Y,Yi L,Zhou Y Y,et al.2015.Illumina sequencing-based analysis of free-living bacterial community dynamics during an Akashiwo sanguine bloom in Xiamen sea,China[J].Science Rep,5:8476-8486
邹万生,王智,宋齐生,等.2017.两株浅层底泥菌对铜绿微囊藻休眠体复苏的调控作用[J].环境科学学报,37(11):4160-4169