Comparison of community composition between Microcystis colony-attached and free-living bacteria, and among bacteria attached with Microcystis colonies of various sizes in culture

  • Qiang Wu
  • Yapeng Zhang
  • Yemei Li
  • Jing Li
  • Xiaohong Zhang
  • Pengfu LiEmail author


A better understanding of the distribution pattern of bacterial community in the Microcystis phycosphere will aid in elucidating the role of bacteria in the formation of cyanobacterial bloom. In the present study, we aimed to compare community composition between Microcystis colony-attached and free-living bacteria, as well as among bacteria attached with Microcystis colonies of various sizes in culture. In the exponentially growing cyanobacterial cultures, Proteobacteria was the most dominant phylum in each colony-attached bacterial community, whereas Bacteroidetes was the most dominant phylum in each free-living bacterial community. The analysis using an indirect PCA model and Bray–Curtis dissimilarity index indicated that the dissimilarity between colony-attached and free-living bacterial communities was greater in the exponentially growing cyanobacterial cultures, and it became smaller in the stationary cultures of Microcystis. In the exponential growth phase of Microcystis, the relative abundance of Proteobacteria in colony-attached bacterial communities tended to decrease with decreasing colony size, whereas the relative abundance of Bacteroidetes in colony-attached bacterial communities tended to increase. In the exponential growth phase of Microcystis, the community composition dissimilarity among bacteria attached with Microcystis colonies of various sizes could be ranked in a descending order as follows: > 100 µm versus < 50 µm; 50–100 µm versus < 50 µm; and > 100 µm versus 50–100 µm. Our data indicated that the community composition of Microcystis colony-attached bacteria was different from that of free-living bacteria, and the colony size of Microcystis played an important role in structuring the community composition of Microcystis-attached bacteria.


Bacterial community Microcystis Colony-attached communities Free-living communities Colonies of various sizes 



This study was financially supported by the National Natural Science Foundation of China (No. 31270447).

Supplementary material

10452_2019_9702_MOESM1_ESM.docx (25 kb)
Supplementary material 1 (DOCX 24 kb)
10452_2019_9702_MOESM2_ESM.tif (1.3 mb)
Fig. S1 Colony surface areas of M. aeruginosa, M. flos-aquae and M. wesenbergii in the exponential and stationary phase. Each point represents mean ± standard deviation (n = 3) (TIFF 1351 kb)
10452_2019_9702_MOESM3_ESM.tif (5.5 mb)
Fig. S2 Epifluorescence microscopy of the bacteria attached to the Microcystis colony stained with DAPI. Scale bar = 10 µm. White arrows indicate some bacterial cells (TIFF 5625 kb)
10452_2019_9702_MOESM4_ESM.tif (1.5 mb)
Fig. S3 Light (a) and epifluorescence (b) microscopy of an inorganic particle from the cyanobacterial culture stained with DAPI. Scale bar = 10 µm. The inorganic particle is indicated by white arrow. No fluorescence dot was observed in the epifluorescence microscopy (TIFF 1531 kb)
10452_2019_9702_MOESM5_ESM.tif (12.6 mb)
Fig. S4 Rarefaction curves based on OTUs (a) and Shannon index (b) for both colony-attached and free-living bacterial communities in the exponential (1) and stationary (2) cyanobacterial cultures. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii; CA, colony-attached bacteria; FL, free-living bacteria (TIFF 12,859 kb)
10452_2019_9702_MOESM6_ESM.tif (10.7 mb)
Fig. S5 Rarefaction curves based on OTUs (a) and Shannon index (b) for bacteria attached with Microcystis colonies of various sizes in the exponential (1) and stationary (2) cyanobacterial cultures. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii (TIFF 10,962 kb)
10452_2019_9702_MOESM7_ESM.tif (5.9 mb)
Fig. S6 Phylogenetic composition of Proteobacteria in colony-attached and free-living bacterial communities. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii; CA, colony-attached bacteria; FL, free-living bacteria (TIFF 6015 kb)
10452_2019_9702_MOESM8_ESM.tif (5.9 mb)
Fig. S7 Phylogenetic composition of Alphaproteobacteria in colony-attached and free-living bacterial communities. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii; CA, colony-attached bacteria; FL, free-living bacteria (TIFF 6079 kb)
10452_2019_9702_MOESM9_ESM.tif (4.4 mb)
Fig. S8 The dissimilarity between colony-attached and free-living bacteria at the OTU level based on Bray–Curtis dissimilarity index (TIFF 4526 kb)
10452_2019_9702_MOESM10_ESM.tif (9.6 mb)
Fig. S9 Relative abundance of the classes found in the bacterial communities attached with Microcystis colonies of various sizes. The classes at relative abundance of < 1% were included in others. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii. Microcystis colonies with size of > 100 µm, 50–100 µm and < 50 µm are represented by b, m and s, respectively (TIFF 9801 kb)
10452_2019_9702_MOESM11_ESM.tif (8.5 mb)
Fig. S10 The dissimilarity among bacterial communities attached with Microcystis colonies of various sizes at the OTU level based on Bray–Curtis dissimilarity index. MA, M. aeruginosa; MF, M. flos-aquae; MW, M. wesenbergii (TIFF 8735 kb)


  1. Bagatini IL, Eiler A, Bertilsson S, Klaveness D, Tessarolli LP, Vieira AAH (2014) Host-specificity and dynamics in bacterial communities associated with bloom-forming freshwater phytoplankton. PLoS ONE 9:e85950Google Scholar
  2. Benyamina S, Baldacci-Cresp F, Couturier J, Chibani K, Hopkins J, Bekki A, Lajudie P, Rouhier N, Jacquot J, Alloing G (2013) Two Sinorhizobium meliloti glutaredoxins regulate iron metabolism and symbiotic bacteroid differentiation. Environ Microbiol 15:795–810Google Scholar
  3. Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120Google Scholar
  4. Briand E, Humbert J-F, Tambosco K, Bormans M, Gerwick WH (2016) Role of bacteria in the production and degradation of Microcystis cyanopeptides. Microbiologyopen 5:469–478Google Scholar
  5. Brunberg AK (1999) Contribution of bacteria in the mucilage of Microcystis spp. (Cyanobacteria) to benthic and pelagic bacterial production in a hypereutrophic lake. FEMS Microbiol Ecol 29:13–22Google Scholar
  6. Cai YF, Shi LM, Li PF, Xing P, Yu Y, Kong FX (2009) Composition of bacterial community related to degrading the exopolysaccharide from the cyanobacterium Microcystis aeruginosa. J Lake Sci 21:369–374Google Scholar
  7. Cai H, Jiang H, Krumholz LR, Yang Z (2014) Bacterial community composition of size-fractioned aggregates within the phycosphere of cyanobacterial blooms in a eutrophic freshwater lake. PLoS ONE 9:e102879Google Scholar
  8. Cao HS, Yang Z (2010) Variation in colony size of Microcystis aeruginosa in a eutrophic lake during recruitment and bloom formation. J Freshw Ecol 25:331–335Google Scholar
  9. Cao XY, Zhou YY, Wang ZC, Song CL (2016) The contribution of attached bacteria to Microcystis bloom: evidence from field investigation and microcosm experiment. Geomicrobiol J 33:607–617Google Scholar
  10. Commault AS, Laczka O, Siboni N, Tamburic B, Crosswell JR, Seymour JR, Ralph PJ (2017) Electricity and biomass production in a bacteria-Chlorella based microbial fuel cell treating wastewater. J Power Sources 356:299–309Google Scholar
  11. Du JJ, Zhao GY, Wang FY, Zhao D, Chen XX, Zhang SR, Jia Y, Tian XJ (2013) Growth stimulation of Microcystis aeruginosa by a bacterium from hyper-eutrophic water (Taihu Lake, China). Aquat Ecol 47:303–313Google Scholar
  12. Dziallas C, Grossart HP (2011) Temperature and biotic factors influence bacterial communities associated with the cyanobacterium Microcystis sp. Environ Microbiol 13:1632–1641Google Scholar
  13. Fan Q, Xiao HJ, Wu Q, Wang SJ, Li PF (2017) Characterization of epiphytic bacteria associated with colonial Microcystis. J Lake Sci 29:617–624Google Scholar
  14. Fulton RS, Paerl HW (1987) Effects of colonial morphology on zooplankton utilization of algal resources during blue-green algal (Microcystis aeruginosa) blooms. Limnol Oceanogr 32:634–644Google Scholar
  15. Gan NQ, Xiao Y, Zhu L, Wu ZX, Liu J, Hu CL, Song LR (2012) The role of microcystins in maintaining colonies of bloom-forming Microcystis spp. Environ Microbiol 14:730–742Google Scholar
  16. García-Cayuela T, Korany AM, Bustos I, de Cadiñanos PG, Requena T, Peláez C, Martínez-Cuesta MC (2014) Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggregation phenotype. Food Res Int 57:44–50Google Scholar
  17. García-Salamanca A, Molina-Henares MA, Dillewijn PV, Solano J, Pizarro-Tobías P, Roca A, Duque E, Ramos JL (2013) Bacterial diversity in the rhizosphere of maize and the surrounding carbonate-rich bulk soil. Microb Biotechnol 6:36–44Google Scholar
  18. Gómez-Acata S, Esquivel-Ríos I, Pérez-Sandoval MV, Navarro-Noya Y, Rojas-Valdez A, Thalasso F, Luna-Guido M, Dendooven L (2017) Bacterial community structure within an activated sludge reactor added with phenolic compounds. Appl Microbiol Biotechnol 101:3405–3414Google Scholar
  19. Grant MA, Kazamia E, Cicuta P, Smith AG (2014) Direct exchange of vitamin B12 is demonstrated by modelling the growth dynamics of algal-bacterial cocultures. ISME J 8:1418–1427Google Scholar
  20. Grossart HP, Levold F, Allgaier M, Simon M, Brinkhoff T (2005) Marine diatom species harbour distinct bacterial communities. Environ Microbiol 7:860–873Google Scholar
  21. Haichar FZ, Marol C, Berge O, Rangelcastro JI, Prosser JI, Balesdent J, Heulin T, Achouak W (2008) Plant host habitat and root exudates shape soil bacterial community structure. ISME J 2:1221–1230Google Scholar
  22. Hayat R, Ali S, Amara U, Khalid R, Ahmed I (2010) Soil beneficial bacteria and their role in plant growth promotion: a review. Ann Microbiol 60:579–598Google Scholar
  23. Jiang LJ, Yang LY, Xiao L, Shi XL, Gao G, Qin BQ (2007) Quantitative studies on phosphorus transference occuring between Microcystis aeruginosa and its attached bacterium (Pseudomonas sp.). Hydrobiologia 581:161–165Google Scholar
  24. Jones KL, Mikulski CM, Barnhorst A, Doucette GJ (2010) Comparative analysis of bacterioplankton assemblages from Karenia brevis bloom and nonbloom water on the west Florida shelf (Gulf of Mexico, USA) using 16S rRNA gene clone libraries. FEMS Microbiol Ecol 73:468–485Google Scholar
  25. Kazamia E, Czesnick H, Nguyen TTV, Croft MT, Sherwood E, Sasso S, Hodson SJ, Warren MJ, Smith AG (2012) Mutualistic interactions between vitamin B12-dependent algae and heterotrophic bacteria exhibit regulation. Environ Microbiol 14:1466–1476Google Scholar
  26. Kodama M, Doucette GJ, Green DH (2006) Relationships between bacteria and harmful algae. In: Granéli E, Turner JT (eds) Ecology of harmful algae. Springer, Berlin, pp 243–255Google Scholar
  27. Leloup M, Nicolau R, Pallier V, Yéprémian C, Feuillade-cathalifaud G (2013) Organic matter produced by algae and cyanobacteria: quantitative and qualitative characterization. J Environ Sci (China) 25:1089–1097Google Scholar
  28. Li YX, Li DH (2012) Physiological variations of bloom-forming Microcystis (Cyanophyceae) related to colony size changes during blooms. Phycologia 51:599–603Google Scholar
  29. Li H, Zhang Q, Wang XL, Ma XY, Lin KF, Liu YD, Gu JD, Lu SG, Shi L, Lu Q, Shen TT (2012a) Biodegradation of benzene homologues in contaminated sediment of the East China Sea. Bioresour Technol 124:129–136Google Scholar
  30. Li L, Gao NY, Deng Y, Yao JJ, Zhang KJ (2012b) Characterization of intracellular and extracellular algae organic matters (AOM) of Microcystic aeruginosa and formation of AOM-associated disinfection byproducts and odor and taste compounds. Water Res 46:1233–1240Google Scholar
  31. Li M, Zhu W, Gao L, Huang JY, Li L (2013a) Seasonal variations of morphospecies composition and colony size of Microcystis in a shallow hypertrophic lake (Lake Taihu, China). Fresenius Environ Bull 22:3474–3483Google Scholar
  32. Li M, Zhu W, Gao L, Lu L (2013b) Changes in extracellular polysaccharide content and morphology of Microcystis aeruginosa at different specific growth rates. J Appl Phycol 25:1023–1030Google Scholar
  33. Li Q, Lin FB, Yang C, Wang JP, Lin Y, Shen MY, Park MS, Li T, Zhao JD (2018) A large-scale comparative metagenomic study reveals the functional interactions in six bloom-forming Microcystis-epibiont communities. Front Microbiol 9:746Google Scholar
  34. Lu Y, Rosencrantz D, Liesack W, Conrad R (2006) Structure and activity of bacterial community inhabiting rice roots and the rhizosphere. Environ Microbiol 8:1351–1360Google Scholar
  35. Maruyama T, Kato K, Yokoyama A, Tanaka T, Hiraishi A, Park HD (2003) Dynamics of microcystin-degrading bacteria in mucilage of Microcystis. Microb Ecol 46:279–288Google Scholar
  36. Mori H, Maruyama F, Kato H, Toyoda A, Dozono A, Ohtsubo Y, Nagata Y, Fujiyama A, Tsuda M, Kurokawa K (2014) Design and experimental application of a novel non-degenerate universal primer set that amplifies prokaryotic 16S rRNA genes with a low possibility to amplify eukaryotic rRNA genes. DNA Res 21:217–227Google Scholar
  37. Nakagawa M, Takamura Y, Yagi O (1987) Isolation and characterization of the slime from a cyanobacterium, Microcystis aeruginosa K-3A. Agric Biol Chem 51:329–337Google Scholar
  38. Otsuka S, Suda S, Li R, Matsumoto S, Watanabe MM (2000) Morphological variability of colonies of Microcystis morphospecies in culture. J Gen Appl Microbiol 46:39–50Google Scholar
  39. Parveen B, Ravet V, Djediat C, Mary I, Quiblier C, Debroas D, Humbert JF (2013) Bacterial communities associated with Microcystis colonies differ from free-living communities living in the same ecosystem. Environ Microbiol Rep 5:716–724Google Scholar
  40. Porter KG, Feig YS (1980) The use of DAPI for identifying and counting aquatic microflora. Limnol Oceanogr 25:943–948Google Scholar
  41. Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:590–596Google Scholar
  42. Ramani A, Rein K, Shetty KG, Jayachandran K (2012) Microbial degradation of microcystin in Florida’s freshwaters. Biodegradation 23:35–45Google Scholar
  43. Rickard AH, Gilbert P, High NJ, Kolenbrander PE, Handley PS (2003) Bacterial coaggregation: an integral process in the development of multi-species biofilms. Trends Microbiol 11:94–100Google Scholar
  44. Rippka R (1988) Isolation and purification of cyanobacteria. Methods Enzymol 167:3–27Google Scholar
  45. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541Google Scholar
  46. Shen H, Niu Y, Xie P, Tao M, Yang X (2011) Morphological and physiological changes in Microcystis aeruginosa as a result of interactions with heterotrophic bacteria. Freshw Biol 56:1065–1080Google Scholar
  47. Shi LM, Cai YF, Yang HL, Xing P, Li PF, Kong LD, Kong FX (2009) Phylogenetic diversity and specificity of bacteria associated with Microcystis aeruginosa and other cyanobacteria. J Environ Sci (China) 21:1581–1590Google Scholar
  48. Shi LM, Cai YF, Wang XY, Li PF, Yu Y, Kong FX (2010) Community structure of bacteria associated with Microcystis colonies from cyanobacterial blooms. J Freshw Ecol 25:193–203Google Scholar
  49. Shi LM, Cai YF, Kong FX, Yu Y (2012) Specific association between bacteria and buoyant Microcystis colonies compared with other bulk bacterial communities in the eutrophic Lake Taihu, China. Environ Microbiol Rep 4:669–678Google Scholar
  50. Shi LM, Huang YX, Zhang M, Yu Y, Lu YP, Kong FX (2017) Bacterial community dynamics and functional variation during the long-term decomposition of cyanobacterial blooms in-vitro. Sci Total Environ 598:77–86Google Scholar
  51. Świątczak P, Cydzikkwiatkowska A, Rusanowska P (2017) Microbiota of anaerobic digesters in a full-scale wastewater treatment plant. Arch Environ Prot 43:53–60Google Scholar
  52. Tang XM, Chao JY, Yi Gong, Wang YP, Wilhelm SW, Gao G (2017) Spatiotemporal dynamics of bacterial community composition in large shallow eutrophic Lake Taihu: high overlap between free-living and particle-attached assemblages. Limnol Oceanogr 62:1366–1382Google Scholar
  53. Teeling H, Fuchs BM, Becher D, Klockow C, Gardebrecht A, Bennke CM, Kassabgy M, Huang S, Mann AJ, Waldmann J, Weber M, Klindworth A, Otto A, Lange J, Bernhardt J, Reinsch C, Hecker M, Peplies J, Bockelmann FD, Callies U, Gerdts G, Wichels A, Wiltshire KH, Glockner FO, Schweder T, Amann R (2012) Substrate-controlled succession of marine bacterioplankton populations induced by a phytoplankton bloom. Science 336:608–611Google Scholar
  54. Tillett D, Neilan BA (2000) Xanthogenate nucleic acid isolation from cultured and environmental cyanobacteria. J Phycol 36:251–258Google Scholar
  55. Timonen S, Sinkko H, Sun H, Sietiö OM, Rinta-kanto JM, Kiheri H, Heinonsalo J (2017) Ericoid roots and mycospheres govern plant-specific bacterial communities in boreal forest humus. Microb Ecol 73:939–953Google Scholar
  56. Vriezen JAC, Bruijn FJD, Nüsslein K (2013) Identification and characterization of a NaCl-responsive genetic locus involved in survival during desiccation in Sinorhizobium meliloti. Appl Environ Microbiol 79:5693–5700Google Scholar
  57. Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267Google Scholar
  58. Wang XY, Xie MJ, Wu W, Shi LM, Luo L, Li PF (2013) Differential sensitivity of colonial and unicellular Microcystis strains to an algicidal bacterium Pseudomonas aeruginosa. J Plankton Res 35:1172–1176Google Scholar
  59. Wang ZC, Li DH, Cao XY, Song CL, Zhou YY (2014) Photosynthetic adaptation mechanism of Microcystis (Cyanophyceae) related to changes of colony size in a eutrophic lake. Phycologia 53:552–560Google Scholar
  60. Wang WJ, Zhang YL, Shen H, Xie P, Yu J (2015) Changes in the bacterial community and extracellular compounds associated with the disaggregation of Microcystis colonies. Biochem Syst Ecol 61:62–66Google Scholar
  61. Wang WJ, Shen H, Shi PL, Chen J, Ni LY, Xie P (2016) Experimental evidence for the role of heterotrophic bacteria in the formation of Microcystis colonies. J Appl Phycol 28:1111–1123Google Scholar
  62. Worm J, Søndergaard M (1998) Dynamics of heterotrophic bacteria attached to Microcystis spp. (Cyanobacteria). Aquat Microb Ecol 14:19–28Google Scholar
  63. Wu XD, Kong FX (2009) Effects of light and wind speed on the vertical distribution of Microcystis aeruginosa colonies of different sizes during a summer bloom. Int Rev Hydrobiol 94:258–266Google Scholar
  64. Xie ML, Ren ML, Yang C, Yi HS, Li Z, Li T, Zhao JD (2016) Metagenomic analysis reveals symbiotic relationship among bacteria in Microcystis-dominated community. Front Microbiol 7:56Google Scholar
  65. Xu N, Tan G, Wang H, Gai X (2016) Effect of biochar additions to soil on nitrogen leaching, microbial biomass and bacterial community structure. Eur J Soil Biol 74:1–8Google Scholar
  66. Yang HL, Cai YF, Xia M, Wang XY, Shi LM, Li PF, Kong FX (2011) Role of cell hydrophobicity on colony formation in Microcystis (Cyanobacteria). Int Rev Hydrobiol 96:141–148Google Scholar
  67. Yang CY, Wang Q, Simon PN, Liu JY, Liu LC, Dai XZ, Zhang XH, Kuang JL, Igarashi Y, Pan XJ, Luo F (2017) Distinct network interactions in particle-associated and free-living bacterial communities during a Microcystis aeruginosa bloom in a plateau lake. Front Microbiol 8:1202Google Scholar
  68. Yu GL, Song LR, Li RH (2007) Taxonomic notes on water bloom forming Microcystis species (Cyanophyta) from China-an example from samples of the Dianchi Lake. Acta Phytotaxon Sin 45:727–741Google Scholar
  69. Yuan L, Zhu W, Xiao L, Yang L (2009) Phosphorus cycling between the colonial cyanobacterium Microcystis aeruginosa and attached bacteria, Pseudomonas. Aquat Ecol 43:859–866Google Scholar
  70. Yuan Y, Wang SY, Liu Y, Li BK, Wang B, Peng YZ (2015) Long-term effect of pH on short-chain fatty acids accumulation and microbial community in sludge fermentation systems. Bioresour Technol 197:56–63Google Scholar
  71. Zhang M, Kong FX, Tan X, Yang Z, Cao HS, Xing P (2007) Biochemical, morphological, and genetic variations in Microcystis aeruginosa due to colony disaggregation. World J Microbiol Biotechnol 23:663–670Google Scholar
  72. Zhang YZ, Wang ET, Li M, Li QQ, Zhang YM, Zhao SJ, Jia XL, Zhang LH, Chen WF, Chen WX (2011) Effects of rhizobial inoculation, cropping systems and growth stages on endophytic bacterial community of soybean roots. Plant Soil 347:147–161Google Scholar
  73. Zhao LF, Lu L, Li M, Xu Z, Zhu W (2011) Effects of Ca and Mg levels on colony formation and EPS content of cultured M. aeruginosa. Procedia Environ Sci 10:1452–1458Google Scholar
  74. Zhu L, Zancarini A, Louati I, De Cesare S, Duval C, Tambosco K, Bernard C, Debroas D, Song LR, Leloup J, Humbert J-F (2016) Bacterial communities associated with four cyanobacterial genera display structural and functional differences: evidence from an experimental approach. Front Microbiol 7:1662Google Scholar
  75. Zuo N, He J, Ma X, Peng Y, Li X (2016) Phosphorus removal performance and population structure of phosphorus-accumulating organisms in HA-A/A-MCO sludge reduction process. Bioengineered 7:327–333Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Pharmaceutical Biotechnology, School of Life SciencesNanjing UniversityNanjingChina

Personalised recommendations