北极海域优势种微单胞藻(Micromonas sp.)与其关联菌群的相互作用
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摘要
以夏季北极海域的优势微藻-微单胞藻(Micromonas sp.,CMPP2099)及其关联菌群为切入点,对这两个重要生态学角色之间的相互关系进行了探索研究。实验选择两种不同生长状态的微单胞藻(指数生长期和稳定生长期);并针对微单胞藻的这两种生长状态,添加外源细菌(单一外源细菌和自然外源细菌)作为实验组。研究发现,无论添加细菌与否,都不会对微单胞藻的生长产生影响,微单胞藻均保持着正常稳定的生长状态,且微单胞藻能够促进细菌的增长。稳定生长期实验组的细菌数量明显高于指数生长期的细菌数量。微单胞藻由于其不具胶体形成的细胞特性,在藻菌共生中并没有发现透明胞外聚合颗粒物(transparent exopolymer particles,TEP)大量存在。添加外源细菌对培养液中的微单胞藻生长、细菌的数量、溶解有机碳(dissolved organic carbon,DOC)、透明胞外聚合颗粒物(TEP)和溶解游离氨基酸(dissovled free amino acid,DFAA)的浓度并无影响,而仅仅由于其所在状态不同存在着差异。该研究初次开展的北极中心海域优势藻种及关联菌群之间关系研究,对极地海洋生态系统中的微食物环有着重要的意义,并为在极地海域中的微食物环-优势浮游植物-微生物种群之间的生态协作提供了实验和理论依据。
The predominant picoplankton of the Arctic is Micromonas sp.,which plays a pivotal role in biogeochemical cycles and is an important link showing that the microbial loop affects organic carbon transfer to higher trophic levels and to deep-sea outputs. As one of the key ecological roles in the Arctic Ocean ecosystem,the interactions between Micromonas sp. and associated bacterioplankton were explored. A series of inocula experiments was undertaken with isolated CCMP2099 Micromonas sp. in an axenic treatment and in one inoculated with marine bacteria.There was no significant effect on the growth of Micromonas sp.,whether treated with bacteria or not. The microalgae grew regularly during the experiments and vice versa promoted bacterial growth. Bacterial abundance of the stationary phase was higher than the exponential phase,although it fluctuated occasionally. Adding exogenous bacteria had no significant effect on algal growth,bacterial numbers,dissolved organic carbon( DOC),transparent exopolymer particles( TEP),and dissolved free amino acids( DFAA). Based on this preliminary study,the interaction between Micromonas sp. and its associated bacteria was discussed with regard to TEP formation,DOC,and DFAA.Algae-bacterial interaction has a significant effect on the microbial loop in the Arctic ecosystem. The results of this study provide an experimental and theoretical basis for microbial loop,phytoplankton,and key microbial community coupling interactions in the Arctic ecosystem.
The predominant picoplankton of the Arctic is Micromonas sp.,which plays a pivotal role in biogeochemical cycles and is an important link showing that the microbial loop affects organic carbon transfer to higher trophic levels and to deep-sea outputs. As one of the key ecological roles in the Arctic Ocean ecosystem,the interactions between Micromonas sp. and associated bacterioplankton were explored. A series of inocula experiments was undertaken with isolated CCMP2099 Micromonas sp. in an axenic treatment and in one inoculated with marine bacteria.There was no significant effect on the growth of Micromonas sp.,whether treated with bacteria or not. The microalgae grew regularly during the experiments and vice versa promoted bacterial growth. Bacterial abundance of the stationary phase was higher than the exponential phase,although it fluctuated occasionally. Adding exogenous bacteria had no significant effect on algal growth,bacterial numbers,dissolved organic carbon( DOC),transparent exopolymer particles( TEP),and dissolved free amino acids( DFAA). Based on this preliminary study,the interaction between Micromonas sp. and its associated bacteria was discussed with regard to TEP formation,DOC,and DFAA.Algae-bacterial interaction has a significant effect on the microbial loop in the Arctic ecosystem. The results of this study provide an experimental and theoretical basis for microbial loop,phytoplankton,and key microbial community coupling interactions in the Arctic ecosystem.
引文
1 Lovejoy C,Vincent W F,Bonilla S,et al.Distribution,phylogeny,and growth of cold-adapted picoprasinophytes in arctic seas[J].Journal of Phycology,2007,43(1):78-89.
2 Bano N,Hollibaugh J T.Phylogenetic composition of bacterioplankton assemblages from the Arctic Ocean[J].Applied and Environmental Microbiology,2002,68(2):505-518.
3 Garneau M,Vincent W F,Alonso-Sáez L,et al.Prokaryotic community structure and heterotrophic production in a river-influenced coastal arctic ecosystem[J].Aquatic Microbial Ecology,2006,42:27-40.
4 Brinkmeyer R,Knittel K,Jürgens J,et al.Diversity and structure of bacterial communities in arctic versus Antarctic pack ice[J].Applied and Environmental Microbiology,2003,69(11):6610-6619.
5 Brown M V,Bowman J P.A molecular phylogenetic survey of sea-ice microbial communities(SIMCO)[J].FEMS Microbiology Ecology,2001,35(3):267-275.
6 Groudieva T,Kambourova M,Yusef H,et al.Diversity and cold-active hydrolytic enzymes of culturable bacteria associated with Arctic sea ice,Spitzbergen[J].Extremophiles,2004,8(6):475-488.
7 Ferrier M,Martin J L,Rooney-Varga J N.Stimulation of Alexandrium fundyense growth by bacterial assemblages from the Bay of Fundy[J].Journal of Applied Microbiology,2002,92(4):706-716.
8 Grossart H P,Kirboe T,Tang K,et al.Bacterial colonization of particles:growth and interactions[J].Applied and Environmental Microbiology,2003,69(6):3500-3509.
9王新,周立红,郑天凌,等.塔玛亚历山大藻藻际细菌溶藻过程[J].生态学报,2007,27(7):2864-2871.
10翟春梅,刘常宏,吕路.铜绿微囊藻与藻际细菌Ma-B1菌株的相互作用[J].环境科学研究,2014,27(7):704-710.
11解冬雪,潘伟斌.溶藻细菌L7(Algicidal Bacteria Strain L7)与中肋骨条藻(Skeletonema costatum)的相互作用[J].环境保护科学,2014,40(4):60-64.
12张化俊,彭云,张夙,等.溶藻细菌BS01产二异丁氧基苯基对塔玛亚历山大藻生长的影响[J].微生物学报,2015,55(7):834-842.
13 Kaczmarska I,Ehrman J M,Bates S S,et al.Diversity and distribution of epibiotic bacteria on Pseudo-nitzschia multiseries(Bacillariophyceae)in culture,and comparison with those on diatoms in native seawater[J].Harmful Algae,2005,4(4):725-741.
14苗祯,杜宗军,李会荣,等.5株北极微藻藻际环境的细菌多样性[J].生态学报,2015,35(5):1587-1600.
15 Bell W,Mitchell R.Chemotactic and growth responses of marine bacteria to algal extracellular products[J].Biological Bulletin,1972,143(2):265-277.
16 Natrah F M I,Bossier P,Sorgeloos P,et al.Significance of microalgal-bacterial interactions for aquaculture[J].Reviews in Aquaculture,2014,6(1):48-61.
17 Krediet C J,Ritchie K B,Paul V J,et al.Coral-associated micro-organisms and their roles in promoting coral health and thwarting diseases[J].Proceedings:Biological Sciences,2013,280(1755):20122328.
18 Takemura A F,Chien D M,Polz M F.Associations and dynamics of Vibrionaceae in the environment,from the genus to the population level[J].Frontiers in Microbiology,2014,5:38.
19 Van Oostende N,Moerdijk-Poortvliet T C W,Boschker H T S,et al.Release of dissolved carbohydrates by Emiliania huxleyi and formation of transparent exopolymer particles depend on algal life cycle and bacterial activity[J].Environmental Microbiology,2013,15(5):1514-1531.
20 Simon M,Grossart H P,Schweitzer B,et al.Microbial ecology of organic aggregates in aquatic ecosystems[J].Aquatic Microbial Ecology,2002,28(2):175-211.
21 Smith D C,Steward G F,Long R A,et al.Bacterial mediation of carbon fluxes during a diatom bloom in a mesocosm[J].Deep Sea Research PartⅡ,1995,42(1):75-97.
22 Grossart H P,Ploug H.Microbial degradation of organic carbon and nitrogen on diatom aggregates[J].Limnology and Oceanography,2001,46(2):267-277.
23 Middelboe M,Borch N H,Kirchman D L.Bacterial utilization of dissolved free amino acids,dissolved combined amino acids and ammonium in the Delaware Bay estuary:effects of carbon and nitrogen limitation[J].Marine Ecology Progress Series,1995,128(1-3):109-120.
24 CCMP2099[Z].Bigelow(National Center for Marine Algae and Microbiota).https://ncma.bigelow.org/ccmp2099[2015-2-10].
25 Cottrell M T,Suttle C A.Production of axenic cultures of Micromonas pusilla(prasinophyceae)using antibiotic[J].Journal of Phycology,1993,29(3):385-387.
26 Porter K G,Feig Y S.The use of DAPI for identifying and counting aquatic microflora[J].Limnology and Oceanography,1980,25(5):943-948.
27侯建军,黄邦钦,戴相辉.赤潮藻细胞计数方法比较研究[J].中国公共卫生,2004,20(8):907-908.
28 Alldredge A L,Passow U,Logan B E.The abundance and significance of a class of large,transparent organic particles in the ocean[J].Deep Sea Research Part I,1993,40(6):1131-1140.
29林伟.几种海洋微藻的无菌化培养[J].海洋科学,2000,24(10):4-6.
30 Passow U.Production of transparent exopolymer particles(TEP)by phyto-and bacterioplankton[J].Marine Ecology Progress Series,2002,236:1-12.
31 Stoderegger K,Herndl G J.Production and release of bacterial capsular material and its subsequent utilization by marine bacterioplankton[J].Limnology and Oceanography,1998,43(5):877-884.
32 Stoderegger K E,Herndl G J.Production of exopolymer particles by marine bacterioplankton under contrasting turbulence conditions[J].Marine E-cology Progress Series,1999,189:9-16.
33林伟,陈騳,刘秀云.海洋微藻除菌及除菌与自然带菌微藻生长特点比较[J].海洋与湖沼,2000,31(6):647-652.
34 Lancelot C,Rousseau V.Ecology of Phaeocystis-dominated ecosystems:the key role of colony forms[M]//Green J C,Leadbeater B S C.The Haptophyte Algae,The Systematics Association.Oxford:Oxford University Press,1994:229-245.
35 Grossart H P,Czub G,Simon M.Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea[J].Environmental Microbiology,2006,8(6):1074-1084.
36 Grossart H P,Levold F,Allgaier M,et al.Marine diatom species harbour distinct bacterial communities[J].Environmental Microbiology,2005,7(6):860-873.
37 Riemann L,Steward G F,Azam F.Dynamics of bacterial community composition and activity during a mesocosm diatom bloom[J].Applied and Environmental Microbiology,2000,66(2):578-587.
38 Kirchman D L,Keel R G,Simon M,et al.Biomass and production of heterotrophic bacterioplankton in the oceanic subarctic Pacific[J].Deep Sea Research Part I,1993,40(5):967-988.
39 Schfer H,Abbas B,Witte H,et al.Genetic diversity of‘satellite’bacteria present in cultures of marine diatoms[J].FEMS Microbiology Ecology,2002,42(1):25-35.
40 Grossart H P,Berman T,Simon M,et al.Occurrence and microbial dynamics of macroscopic organic aggregates(lake snow)in Lake Kinneret,Israel,in fall[J].Aquatic Microbial Ecology,1998,14:59-67.
41 Passow U,Alldredge A L,Logan B E.The role of particulate carbohydrate exudates in the flocculation of diatom blooms[J].Deep Sea Research I,1994,41(2):335-357.
42 Wetz M S,Wheeler P A.Release of dissolved organic matter by coastal diatoms[J].Limnology and Oceanography,2007,52(2):798-807.
43 Sasaki H,Miyamura T,Saitoh S I,et al.Seasonal variation of absorption by particles and colored dissolved organic matter(CDOM)in Funka Bay,southwestern Hokkaido,Japan[J].Estuarine,Coastal and Shelf Science,2005,64(2-3):447-458.
44 Hammer K D,Eberlein K.Parallel experiments with Thalassiosira rotula in outdoor plastic tanks:development of dissolved free amino acids during an algae bloom[J].Marine Chemistry,1981,10(6):533-544.
45 Poulet S A,Martin-Jézéquel V.Relationships between dissolved free amino acids,chemical composition and growth of the marine diatom Chaetoceros debile[J].Marine Biology,1983,77(1):93-100.
46 Grdes A,Iversen M H,Grossart H P,et al.Diatom-associated bacteria are required for aggregation of Thalassiosira weissflogii[J].The ISME Journal,2011,5(3):436-445.
47 Gogou A,Repeta D J.Particulate-dissolved transformations as a sink for semi-labile dissolved organic matter:chemical characterization of high molecular weight dissolved and surface-active organic matter in seawater and in diatom cultures[J].Marine Chemistry,2010,121(1-4):215-223.
48 Engel A,Thoms S,Riebesell U,et al.Polysaccharide aggregation as a potential sink of marine dissolved organic carbon[J].Nature,2004,428(6986):929-932.
49王将克,陈水挟,钟月明,等.氨基酸生物地球化学[M].北京:科学出版社,1991.
50 Mulholland M R,Gobler C J,Lee C.Peptide hydrolysis,amino acid oxidation,and nitrogen uptake in communities seasonally dominated by Aureococcus anophagefferens[J].Limnology and Oceanography,2002,47(4):1094-1108.
2 Bano N,Hollibaugh J T.Phylogenetic composition of bacterioplankton assemblages from the Arctic Ocean[J].Applied and Environmental Microbiology,2002,68(2):505-518.
3 Garneau M,Vincent W F,Alonso-Sáez L,et al.Prokaryotic community structure and heterotrophic production in a river-influenced coastal arctic ecosystem[J].Aquatic Microbial Ecology,2006,42:27-40.
4 Brinkmeyer R,Knittel K,Jürgens J,et al.Diversity and structure of bacterial communities in arctic versus Antarctic pack ice[J].Applied and Environmental Microbiology,2003,69(11):6610-6619.
5 Brown M V,Bowman J P.A molecular phylogenetic survey of sea-ice microbial communities(SIMCO)[J].FEMS Microbiology Ecology,2001,35(3):267-275.
6 Groudieva T,Kambourova M,Yusef H,et al.Diversity and cold-active hydrolytic enzymes of culturable bacteria associated with Arctic sea ice,Spitzbergen[J].Extremophiles,2004,8(6):475-488.
7 Ferrier M,Martin J L,Rooney-Varga J N.Stimulation of Alexandrium fundyense growth by bacterial assemblages from the Bay of Fundy[J].Journal of Applied Microbiology,2002,92(4):706-716.
8 Grossart H P,Kirboe T,Tang K,et al.Bacterial colonization of particles:growth and interactions[J].Applied and Environmental Microbiology,2003,69(6):3500-3509.
9王新,周立红,郑天凌,等.塔玛亚历山大藻藻际细菌溶藻过程[J].生态学报,2007,27(7):2864-2871.
10翟春梅,刘常宏,吕路.铜绿微囊藻与藻际细菌Ma-B1菌株的相互作用[J].环境科学研究,2014,27(7):704-710.
11解冬雪,潘伟斌.溶藻细菌L7(Algicidal Bacteria Strain L7)与中肋骨条藻(Skeletonema costatum)的相互作用[J].环境保护科学,2014,40(4):60-64.
12张化俊,彭云,张夙,等.溶藻细菌BS01产二异丁氧基苯基对塔玛亚历山大藻生长的影响[J].微生物学报,2015,55(7):834-842.
13 Kaczmarska I,Ehrman J M,Bates S S,et al.Diversity and distribution of epibiotic bacteria on Pseudo-nitzschia multiseries(Bacillariophyceae)in culture,and comparison with those on diatoms in native seawater[J].Harmful Algae,2005,4(4):725-741.
14苗祯,杜宗军,李会荣,等.5株北极微藻藻际环境的细菌多样性[J].生态学报,2015,35(5):1587-1600.
15 Bell W,Mitchell R.Chemotactic and growth responses of marine bacteria to algal extracellular products[J].Biological Bulletin,1972,143(2):265-277.
16 Natrah F M I,Bossier P,Sorgeloos P,et al.Significance of microalgal-bacterial interactions for aquaculture[J].Reviews in Aquaculture,2014,6(1):48-61.
17 Krediet C J,Ritchie K B,Paul V J,et al.Coral-associated micro-organisms and their roles in promoting coral health and thwarting diseases[J].Proceedings:Biological Sciences,2013,280(1755):20122328.
18 Takemura A F,Chien D M,Polz M F.Associations and dynamics of Vibrionaceae in the environment,from the genus to the population level[J].Frontiers in Microbiology,2014,5:38.
19 Van Oostende N,Moerdijk-Poortvliet T C W,Boschker H T S,et al.Release of dissolved carbohydrates by Emiliania huxleyi and formation of transparent exopolymer particles depend on algal life cycle and bacterial activity[J].Environmental Microbiology,2013,15(5):1514-1531.
20 Simon M,Grossart H P,Schweitzer B,et al.Microbial ecology of organic aggregates in aquatic ecosystems[J].Aquatic Microbial Ecology,2002,28(2):175-211.
21 Smith D C,Steward G F,Long R A,et al.Bacterial mediation of carbon fluxes during a diatom bloom in a mesocosm[J].Deep Sea Research PartⅡ,1995,42(1):75-97.
22 Grossart H P,Ploug H.Microbial degradation of organic carbon and nitrogen on diatom aggregates[J].Limnology and Oceanography,2001,46(2):267-277.
23 Middelboe M,Borch N H,Kirchman D L.Bacterial utilization of dissolved free amino acids,dissolved combined amino acids and ammonium in the Delaware Bay estuary:effects of carbon and nitrogen limitation[J].Marine Ecology Progress Series,1995,128(1-3):109-120.
24 CCMP2099[Z].Bigelow(National Center for Marine Algae and Microbiota).https://ncma.bigelow.org/ccmp2099[2015-2-10].
25 Cottrell M T,Suttle C A.Production of axenic cultures of Micromonas pusilla(prasinophyceae)using antibiotic[J].Journal of Phycology,1993,29(3):385-387.
26 Porter K G,Feig Y S.The use of DAPI for identifying and counting aquatic microflora[J].Limnology and Oceanography,1980,25(5):943-948.
27侯建军,黄邦钦,戴相辉.赤潮藻细胞计数方法比较研究[J].中国公共卫生,2004,20(8):907-908.
28 Alldredge A L,Passow U,Logan B E.The abundance and significance of a class of large,transparent organic particles in the ocean[J].Deep Sea Research Part I,1993,40(6):1131-1140.
29林伟.几种海洋微藻的无菌化培养[J].海洋科学,2000,24(10):4-6.
30 Passow U.Production of transparent exopolymer particles(TEP)by phyto-and bacterioplankton[J].Marine Ecology Progress Series,2002,236:1-12.
31 Stoderegger K,Herndl G J.Production and release of bacterial capsular material and its subsequent utilization by marine bacterioplankton[J].Limnology and Oceanography,1998,43(5):877-884.
32 Stoderegger K E,Herndl G J.Production of exopolymer particles by marine bacterioplankton under contrasting turbulence conditions[J].Marine E-cology Progress Series,1999,189:9-16.
33林伟,陈騳,刘秀云.海洋微藻除菌及除菌与自然带菌微藻生长特点比较[J].海洋与湖沼,2000,31(6):647-652.
34 Lancelot C,Rousseau V.Ecology of Phaeocystis-dominated ecosystems:the key role of colony forms[M]//Green J C,Leadbeater B S C.The Haptophyte Algae,The Systematics Association.Oxford:Oxford University Press,1994:229-245.
35 Grossart H P,Czub G,Simon M.Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea[J].Environmental Microbiology,2006,8(6):1074-1084.
36 Grossart H P,Levold F,Allgaier M,et al.Marine diatom species harbour distinct bacterial communities[J].Environmental Microbiology,2005,7(6):860-873.
37 Riemann L,Steward G F,Azam F.Dynamics of bacterial community composition and activity during a mesocosm diatom bloom[J].Applied and Environmental Microbiology,2000,66(2):578-587.
38 Kirchman D L,Keel R G,Simon M,et al.Biomass and production of heterotrophic bacterioplankton in the oceanic subarctic Pacific[J].Deep Sea Research Part I,1993,40(5):967-988.
39 Schfer H,Abbas B,Witte H,et al.Genetic diversity of‘satellite’bacteria present in cultures of marine diatoms[J].FEMS Microbiology Ecology,2002,42(1):25-35.
40 Grossart H P,Berman T,Simon M,et al.Occurrence and microbial dynamics of macroscopic organic aggregates(lake snow)in Lake Kinneret,Israel,in fall[J].Aquatic Microbial Ecology,1998,14:59-67.
41 Passow U,Alldredge A L,Logan B E.The role of particulate carbohydrate exudates in the flocculation of diatom blooms[J].Deep Sea Research I,1994,41(2):335-357.
42 Wetz M S,Wheeler P A.Release of dissolved organic matter by coastal diatoms[J].Limnology and Oceanography,2007,52(2):798-807.
43 Sasaki H,Miyamura T,Saitoh S I,et al.Seasonal variation of absorption by particles and colored dissolved organic matter(CDOM)in Funka Bay,southwestern Hokkaido,Japan[J].Estuarine,Coastal and Shelf Science,2005,64(2-3):447-458.
44 Hammer K D,Eberlein K.Parallel experiments with Thalassiosira rotula in outdoor plastic tanks:development of dissolved free amino acids during an algae bloom[J].Marine Chemistry,1981,10(6):533-544.
45 Poulet S A,Martin-Jézéquel V.Relationships between dissolved free amino acids,chemical composition and growth of the marine diatom Chaetoceros debile[J].Marine Biology,1983,77(1):93-100.
46 Grdes A,Iversen M H,Grossart H P,et al.Diatom-associated bacteria are required for aggregation of Thalassiosira weissflogii[J].The ISME Journal,2011,5(3):436-445.
47 Gogou A,Repeta D J.Particulate-dissolved transformations as a sink for semi-labile dissolved organic matter:chemical characterization of high molecular weight dissolved and surface-active organic matter in seawater and in diatom cultures[J].Marine Chemistry,2010,121(1-4):215-223.
48 Engel A,Thoms S,Riebesell U,et al.Polysaccharide aggregation as a potential sink of marine dissolved organic carbon[J].Nature,2004,428(6986):929-932.
49王将克,陈水挟,钟月明,等.氨基酸生物地球化学[M].北京:科学出版社,1991.
50 Mulholland M R,Gobler C J,Lee C.Peptide hydrolysis,amino acid oxidation,and nitrogen uptake in communities seasonally dominated by Aureococcus anophagefferens[J].Limnology and Oceanography,2002,47(4):1094-1108.