Journal of Applied Phycology

, Volume 26, Issue 3, pp 1483–1492 | Cite as

The interactions between Chlorella vulgaris and algal symbiotic bacteria under photoautotrophic and photoheterotrophic conditions

Article

Abstract

A Chlorella vulgaris ATCC 13482 culture was semi-continuously cultivated for 18 months in a 4-L photobioreactor and formed associated consortia with other symbionts. Three symbiotic bacterial strains were isolated on heterotrophic medium agar plates. Based on 16S rDNA analysis, they were found to show closest similarity to Pseudomonas alcaligenes, Elizabethkingia miricola and Methylobacterium radiotolerans. C. vulgaris was co-cultured with each bacterial strain, and it was found that the symbiotic bacterium Pseudomonas sp. had a growth-promoting effect on C. vulgaris while the other two inhibited algal growth. The interactions between C. vulgaris and Pseudomonas sp. were further investigated under different cultivation conditions. The co-culture resulted in 1.4 times greater algal cell concentration than that of C. vulgaris alone under photoautotrophic condition. In contrast, the algal cell concentration was lower in the co-culture compared with single algal culture when glucose was supplied in the medium (photoheterotrophic). Under both cultivation conditions, the number of Pseudomonas sp. increased at the beginning of experiment, and then decreased. However, the bacterial number decreased to almost zero under photoheterotrophic conditions, while the growth of bacteria went into a stationary phase under photoautotrophic conditions. The chlorophyll content in C. vulgaris cell was higher in co-culture than in single algal culture. Algal cells in photoautotrophic condition showed higher photosynthetic efficiency compared to those in photoheterotrophic condition. Extracellular organic carbon dissolved in the medium continuously increased under photoautotrophic condition. The mutualistic and competing relationships between C. vulgaris and symbiotic bacteria observed in this study could aid our understanding of algae–bacteria interactions in nature as well as broadening its practical applications.

Keywords

Microalgae Chlorella vulgaris Symbiotic bacteria Interactions 

Supplementary material

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ESM 1(DOC 679 kb)

References

  1. Barranguet C, Veuger B, Van Beusekom SAM, Marvan P, Sinke JJ, Admiraal W (2005) Divergent composition of algal–bacterial biofilms developing under various external factors. Eur J Phycol 40:1–8CrossRefGoogle Scholar
  2. Borde X, Guieysse B, Delgado O, Munoz R, Hatti-Kaul R, Nugier-Chauvin C, Patin H, Mattiasson B (2003) Synergistic relationships in algal–bacterial microcosms for the treatment of aromatic pollutants. Bioresour Technol 86:293–300PubMedCrossRefGoogle Scholar
  3. Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306PubMedCrossRefGoogle Scholar
  4. Choi O, Das A, Yu CP, Hu ZQ (2010a) Nitrifying bacterial growth inhibition in the presence of algae and cyanobacteria. Biotechnol Bioeng 107:1004–1011PubMedCrossRefGoogle Scholar
  5. Choi SP, Nguyen MT, Sim SJ (2010b) Enzymatic pretreatment of Chlamydomonas reinhardtii biomass for ethanol production. Bioresour Technol 101:5330–5336PubMedCrossRefGoogle Scholar
  6. Cole JJ (1982) Interactions between bacteria and algae in aquatic ecosystems. Annu Rev Ecol Syst 13:291–314CrossRefGoogle Scholar
  7. Cosgrove J, Borowitzka MA (2011) Chlorophyll fluorescence terminology: an introduction. In: Suggett DJ, Prášil O, Borowitzka MA (eds) Chlorophyll a fluorescence in aquatic sciences. Springer, Dordrecht, pp 1–17Google Scholar
  8. Croft MT, Lawrence AD, Raux-Deery E, Warren MJ, Smith AG (2005) Algae acquire vitamin B12 through a symbiotic relationship with bacteria. Nature 438:90–93PubMedCrossRefGoogle Scholar
  9. Czerpak R, Krotke A, Mical A (1999) Comparison of stimulatory effect of auxins and cytokinins on protein, saccharides and chlorophylls content in Chlorella pyrenoidosa Chick. Pol Arch Hydrobiol 46:71–82Google Scholar
  10. de-Bashan LE, Moreno M, Hernandez JP, Bashan Y (2002) Removal of ammonium and phosphorus ions from synthetic wastewater by the microalgae Chlorella vulgaris coimmobilized in alginate beads with the microalgae growth-promoting bacterium Azospirillum brasilense. Water Res 36:2941–2948PubMedCrossRefGoogle Scholar
  11. DellaGreca M, Zarrelli A, Fergola P, Cerasuolo M, Pollio A, Pinto G (2010) Fatty acids released by Chlorella vulgaris and their role in interference with Pseudokirchneriella subcapitata: experiments and modelling. J Chem Ecol 36:339–349PubMedCrossRefGoogle Scholar
  12. Fukami K, Nishijima T, Ishida Y (1997) Stimulative and inhibitory effects of bacteria on the growth of microalgae. Hydrobiologia 358:185–191CrossRefGoogle Scholar
  13. Gong YM, Hu HH, Gao Y, Xu XD, Gao H (2011) Microalgae as platforms for production of recombinant proteins and valuable compounds: progress and prospects. J Ind Microbiol Biotechnol 38:1879–1890PubMedCrossRefGoogle Scholar
  14. Gonzalez LE, Bashan Y (2000) Increased growth of the microalga Chlorella vulgaris when coimmobilized and cocultured in alginate beads with the plant-growth-promoting bacterium Azospirillum brasilense. Appl Environ Microbiol 66:1527–1531PubMedCentralPubMedCrossRefGoogle Scholar
  15. Guillard RRL (2005) Purification methods for microalgae. In: Andersen RA (ed) Algal culturing techniques. Elsevier Academic Press, Burlington, pp 117–132Google Scholar
  16. Gurung TB, Urabe J, Nakanishi M (1999) Regulation of the relationship between phytoplankton Scenedesmus acutus and heterotrophic bacteria by the balance of light and nutrients. Aquat Microb Ecol 17:27–35CrossRefGoogle Scholar
  17. Hernandez JP, De-Bashan LE, Rodriguez DJ, Rodriguez Y, Bashan Y (2009) Growth promotion of the freshwater microalga Chlorella vulgaris by the nitrogen-fixing, plant growth-promoting bacterium Bacillus pumilus from and zone soils. Eur J Soil Biol 45:88–93CrossRefGoogle Scholar
  18. Hodoki Y, Ohbayashi K, Kobayashi Y, Okuda N, Nakano S (2011) Temporal variation in cyanobacteria species composition and photosynthetic activity in experimentally induced blooms. J Plankton Res 33:1410–1416CrossRefGoogle Scholar
  19. Huo YX, Cho KM, Rivera JGL, Monte E, Shen CR, Yan Y, Liao JC (2011) Conversion of proteins into biofuels by engineering nitrogen flux. Nat Biotechnol 29:346–351PubMedCrossRefGoogle Scholar
  20. Hyenstrand P, Burkert U, Pettersson A, Blomqvist P (2000) Competition between the green alga Scenedesmus and the cyanobacterium Synechococcus under different modes of inorganic nitrogen supply. Hydrobiologia 435:91–98CrossRefGoogle Scholar
  21. Imai I, Ishida Y, Sakaguchi K, Hata Y (1995) Algicidal marine-bacteria isolated from northern Hiroshima bay, Japan. Fisheries Sci 61:628–636Google Scholar
  22. Jung SW, Kim BH, Katano T, Kong DS, Han MS (2008) Pseudomonas fluorescens HYK0210-SK09 offers species-specific biological control of winter algal blooms caused by freshwater diatom Stephanodiscus hantzschii. J Appl Microbiol 105:186–195PubMedCrossRefGoogle Scholar
  23. Lee BK, Katano T, Kitamura SI, Oh MJ, Han MS (2008) Monitoring of algicidal bacterium, Alteromonas sp. strain A14 in its application to natural Cochlodinium polykrikoides blooming seawater using fluorescence in situ hybridization. J Microbiol 46:274–282PubMedCrossRefGoogle Scholar
  24. Lee YK, Shen H (2004) Basic culturing techniques. In: Richmond A (ed) Handbook of microalgal culture: biotechnology and applied phycology. Blackwell Publishing, Oxford, pp 40–56Google Scholar
  25. Liu JQ, Lewitus AJ, Kempton JW, Wilde SB (2008) The association of algicidal bacteria and raphidophyte blooms in South Carolina brackish detention ponds. Harmful Algae 7:184–193CrossRefGoogle Scholar
  26. Mazur H, Konop A, Synak R (2001) Indole-3-acetic acid in the culture medium of two axenic green microalgae. J Appl Phycol 13:35–42CrossRefGoogle Scholar
  27. Mouget JL, Dakhama A, Lavoie MC, de la Noue J (1995) Algal growth enhancement by bacteria — is consumption of photosynthetic oxygen involved. Fems Microbiol Ecol 18:35–43CrossRefGoogle Scholar
  28. Munoz R, Köllner C, Guieysse B, Mattiasson B (2004) Photosynthetically oxygenated salicylate biodegradation in a continuous stirred tank photobioreactor. Biotechnol Bioeng 87:797–803PubMedCrossRefGoogle Scholar
  29. Munoz R, Guieysse B (2006) Algal-bacterial processes for the treatment of hazardous contaminants: a review. Water Res 40:2799–2815PubMedCrossRefGoogle Scholar
  30. Ortiz-Marquez JCF, Nascimento MD, Dublan MD, Curatti L (2012) Association with an ammonium-excreting bacterium allows diazotrophic culture of oil-rich eukaryotic microalgae. Appl Environ Microbiol 78:2345–2352PubMedCentralPubMedCrossRefGoogle Scholar
  31. Paerl HW (1976) Specific associations of blue-green algae Anabaena and Aphanizomenon with bacteria in freshwater blooms. J Phycol 12:431–435Google Scholar
  32. Parmar A, Singh NK, Pandey A, Gnansounou E, Madamwar D (2011) Cyanobacteria and microalgae: a positive prospect for biofuels. Bioresour Technol 102:10163–10172PubMedCrossRefGoogle Scholar
  33. Paul C, Pohnert G (2011) Interactions of the algicidal bacterium Kordia algicida with diatoms: regulated protease excretion for specific algal lysis. Plos One 6:e21032PubMedCentralPubMedCrossRefGoogle Scholar
  34. Pratt R, Daniels TC, Eiler JJ, Gunnison JB, Kumler WD, Oneto JF, Strait LA, Spoehr HA, Hardin GJ, Milner HW, Smith JH, Strain HH (1944) Chlorellin, an antibacterial substance from Chlorella. Science 99:351–352PubMedCrossRefGoogle Scholar
  35. Rier ST, Stevenson RJ (2002) Effects of light, dissolved organic carbon, and inorganic nutrients on the relationship between algae and heterotrophic bacteria in stream periphyton. Hydrobiologia 489:179–184CrossRefGoogle Scholar
  36. Rivas MO, Vargas P, Riquelme CE (2010) Interactions of Botryococcus braunii cultures with bacterial biofilms. Microbial Ecol 60:628–635CrossRefGoogle Scholar
  37. Sapp M, Schwaderer AS, Wiltshire KH, Hoppe HG, Gerdts G, Wichels A (2007) Species-specific bacterial communities in the phycosphere of microalgae? Microbial Ecol 53:683–699CrossRefGoogle Scholar
  38. Sarmento H, Gasol JM (2012) Use of phytoplankton-derived dissolved organic carbon by different types of bacterioplankton. Environ Microbiol 14:2348–2360PubMedCrossRefGoogle Scholar
  39. Seyedsayamdost MR, Carr G, Kolter R, Clardy J (2011) Roseobacticides: small molecule modulators of an algal–bacterial symbiosis. J Am Chem Soc 133:18343–18349PubMedCentralPubMedCrossRefGoogle Scholar
  40. Su YY, Mennerich A, Urban B (2012) Synergistic cooperation between wastewater-born algae and activated sludge for wastewater treatment: influence of algae and sludge inoculation ratios. Bioresour Technol 105:67–73PubMedCrossRefGoogle Scholar
  41. Suminto, Hirayama K (1997) Application of a growth-promoting bacteria for stable mass culture of three marine microalgae. Hydrobiologia 358:223–230Google Scholar
  42. Tarakhovskaya ER, Maslov YI, Shishova MF (2007) Phytohormones in algae. Russ J Plant Physio 54:163–170CrossRefGoogle Scholar
  43. Terauchi AM, Peers G, Kobayashi MC, Niyogi KK, Merchant SS (2010) Trophic status of Chlamydomonas reinhardtii influences the impact of iron deficiency on photosynthesis. Photosynth Res 105:39–49PubMedCentralPubMedCrossRefGoogle Scholar
  44. Tison DL, Lingg AJ (1979) Dissolved organic matter utilization and oxygen uptake in algal–bacterial microcosms. Can J Microbiol 25:1315–1320PubMedCrossRefGoogle Scholar
  45. Vance BD (1987) Phytohormone effects on cell-division in Chlorella-pyrenoidosa chick (TX-7-11-05) (Chlorellaceae). J Plant Growth Regul 5:169–173CrossRefGoogle Scholar
  46. Watanabe K, Takihana N, Aoyagi H et al (2005) Symbiotic association in Chlorella culture. Fems Microbiol Ecol 51:187–196PubMedCrossRefGoogle Scholar
  47. Zhong WH, Li YX, Sun KD, Jin J, Li X, Zhang F, Chen J (2011) Aerobic degradation of methyl tert-butyl ether in a closed symbiotic system containing a mixed culture of Chlorella ellipsoidea and Methylibium petroleiphilum PM1. J Hazard Mater 185:1249–1255PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Department of Chemical and Biomolecular EngineeringNUS Environment Research Institute, National University of SingaporeSingaporeSingapore
  2. 2.Department of Chemical and Biomolecular EngineeringNational University of SingaporeSingaporeSingapore

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