Abstract
Fatty acid profiles were used to characterize nutritional pathways in Chlamydomonas sp. isolated from an acidic mining lake (pH 2.7). Surprisingly, profiles of Chlamydomonas sp. grown in the lab under photoautotrophic, mixotrophic, and heterotrophic conditions at in situ deep strata lake water temperatures (8°C) were very similar, polyunsaturated fatty acids including α-linolenic acid (18:3ω3) and 16:4ω3 along with palmitic acid (16:0) being most abundant. Therefore, heterotrophic growth of Chlamydomonas sp. at low temperatures can result in high concentrations of polyunsaturated fatty acids, as previously only described for some psychrophilic bacteria. By contrast, the cultivation of isolated Chlamydomonas sp. at 20°C, reflecting surface water temperatures, provided fatty acid patterns characteristic of the nutrition strategy applied: the concentration of polyunsaturated fatty acids decreased when the growth pathway changed from photoautotrophic via mixotrophic to heterotrophic. Total fatty acid concentration also diminished in this order. Principal component analysis confirmed the significance of FA profiling to mirror nutritional pathways. Lake-water analysis revealed low concentrations of dissolved organic carbon, mainly consisting of polymeric fulvic acids that are unable to support heterotrophic growth of Chlamydomonas sp. Polymeric fulvic acids present in the deeper strata of the lake turned out to be formed in situ on the basis of organic monomers including reduced sulfur-containing ones, as revealed by thermochemolysis and pyrolysis. Growth of Chlamydomonas sp. in the deep chlorophyll maximum is therefore assumed to mainly result from photosynthesis, despite very low photon densities. Phytol-including metabolites proved to be significant biomarkers to indicate the nutritional pathway of Chlamydomonas sp. α, ω-Dicarboxylic acids—light-induced degradation products of unsaturated fatty acids—appeared to be good indicators of photooxidative alterations to the algal species under study.
Similar content being viewed by others
References
BA Annous MF Kozempel MJ Kurantz (1999) ArticleTitleChanges in membrane fatty acid composition of Pediococcus sp. strain NRRL B-2354 in response to growth conditions and its effect on thermal resistance. Appl Environ Microbiol 65 2857–2862 Occurrence Handle1:CAS:528:DyaK1MXktlemtrg%3D Occurrence Handle10388676
V Bissinger J Jander J Tittel (2000) ArticleTitleA new medium free of organic carbon to cultivate organisms from extremely acidic mining lakes (pH 2.7). Acta Hydrochim Hydrobiol 28 310–312 Occurrence Handle10.1002/1521-401X(200012)28:6<310::AID-AHEH310>3.3.CO;2-8 Occurrence Handle1:CAS:528:DC%2BD3MXis1yrs7k%3D
C Blodau A Peine S Hoffmann S Pfeiffer (2000) ArticleTitleOrganic matter diagenesis in acidic mining lakes. Acta Hydrochim Hydrobiol 28 123–135 Occurrence Handle10.1002/1521-401X(200003)28:3<123::AID-AHEH123>3.0.CO;2-A Occurrence Handle1:CAS:528:DC%2BD3cXkslOmtL4%3D
PE Cassin (1974) ArticleTitleIsolation, growth, and physiology of acidophilic Chlamydomonas. J Phycol 10 439–447 Occurrence Handle1:CAS:528:DyaE2MXhs1OgtLo%3D
MR Droop (1974) Heterotrophy of carbon. WDP Stewart (Eds) Algal Physiology and Biochemistry, vol. 10 Blackwell Scientific Publications Oxford 530–559
P Drouin D Prevost H Antoun (2000) ArticleTitlePhysiological adaptation to low temperatures of strains of Rhizobium leguminosarum bv. viciae associated with Lathyrus spp. FEMS Microbiol Ecol 32 111–120 Occurrence Handle10.1016/S0168-6496(00)00020-9 Occurrence Handle1:CAS:528:DC%2BD3cXjt1ejtrk%3D Occurrence Handle10817864
W Eichenberger (1976) ArticleTitleLipids of Chlamydomonas reinhardtii under different growth conditions. Phytochemistry 15 459–463 Occurrence Handle10.1016/S0031-9422(00)88947-5 Occurrence Handle1:CAS:528:DyaE28XktFOgsbg%3D
NR Filho FM Lancas (1995) ArticleTitleIdentification of FAMEs using ECL values and a three-dimensional Kovats retention index system. J High Resol Chromatogr 18 167–170
R Frömmichen M Koschorreck K Wendt-Potthoff K Friese (2001) Neutralization of acidic mining lakes via in situ stimulation of bacteria. A Leeson B Peyton VS Magar (Eds) Bioremediation of Inorganic Compounds, vol. 9 Batelle OH 43–52
R Frömmichen S Kellner K Friese (2003) ArticleTitleSediment conditioning with organic and/or inorganic carbon sources as a first step in alkalinity generation of acid mine pit lake water (pH 2–3). Environ Sci Technol 37 1414–1421 Occurrence Handle10.1021/es026131c
A Fuggi G Pinto A Pollio R Taddei (1988) ArticleTitleThe role of glycerol in osmoregulation of the acidophilic alga Dunaliella acidophilla (Volvocales, Chlorophyta). Effect of solute stress on photosynthetic respiration and glycerol synthesis. Phycologia 27 439–446
A Gauthier S Derenne C Largeau L Dupont E Guillon J Dumonceau M Aplincourt (2003) ArticleTitleComparative study of ligno-cellulosic material from wheat straw and of pure and mixed standard compounds via solid state 13C NMR spectroscopy, conventional pyrolysis and TMAH thermochemolysis. J Anal Appl Pyrol 67 277–293 Occurrence Handle10.1016/S0165-2370(02)00067-0 Occurrence Handle1:CAS:528:DC%2BD3sXhsF2ns74%3D
W Geller H Klapper W Salomons (1998) Acid Mining Lakes—Acid Mine Drainage, Limnology, and Reclamation SeriesTitleEnviron Sci Series Springer Berlin
D Ghoshal HD Music A Goyal (2002) ArticleTitleDissolved inorganic carbon concentration mechanism in Chlamydomonas moewusii. Plant Physiol Biochem 40 299–305 Occurrence Handle10.1016/S0981-9428(02)01384-0 Occurrence Handle1:CAS:528:DC%2BD38XivValsLY%3D
C Giroud W Eichenberger (1989) ArticleTitleLipids of Chlamydomonas reinhardtii - incorporation of [C-14]acetate, [C-14]palmitate, and [C-14]oleate into different lipids and evidence for lipid-linked desaturation of fatty acids. Plant Cell Physiol 30 121–128 Occurrence Handle1:CAS:528:DyaL1MXhtlWqsLc%3D
C Giroud A Gerber W Eichenberger (1988) ArticleTitleLipids of Chlamydomonas reinhardtii. Analysis of molecular species and intracellular site(s) of biosynthesis. Plant Cell Physiol 29 587–595 Occurrence Handle1:CAS:528:DyaL1cXkslSjs74%3D
T Gorecki J Poerschmann (2001) ArticleTitleIn-column pyrolysis: A new approach to an old problem. Anal Chem 73 2012–2017 Occurrence Handle10.1021/ac000913b Occurrence Handle1:CAS:528:DC%2BD3MXitVyku7g%3D Occurrence Handle11354483
JB Guckert (1996) ArticleTitleToxicity assessment by community analysis. J Microbiol Methods 25 101–112 Occurrence Handle10.1016/0167-7012(95)00096-8 Occurrence Handle1:CAS:528:DyaK28XksFSht7c%3D
DB Hedrick A Peacock JR Stephen SJ Macnaughton J Brüggeann DC White (2000) ArticleTitleMeasuring soil microbial community diversity using polar lipid fatty acid and denaturing gradient gel electrophoresis data. J Microbiol Methods 41 235–248 Occurrence Handle10.1016/S0167-7012(00)00157-3 Occurrence Handle1:CAS:528:DC%2BD3cXlvV2mtL0%3D Occurrence Handle10958969
PB Heifetz B Forster CB Osmond LJ Giles JE Boynton (2000) ArticleTitleEffects of acetate on facultative autotrophy in Chlamydomonas reinhardtii assessed by photosynthetic measurements and stable isotope analyses. Plant Physiol 122 1439–1445 Occurrence Handle10.1104/pp.122.4.1439 Occurrence Handle1:CAS:528:DC%2BD3cXktFWjs78%3D Occurrence Handle10759539
P Herzsprung K Friese G Packroff M Schimmele K Wendt-Potthoff M Winkler (1998) ArticleTitleVertical and annual distribution of ferric and ferrous iron in acidic mining lakes. Acta Hydrochim Hydrobiol 26 253–262 Occurrence Handle10.1002/(SICI)1521-401X(199809)26:5<253::AID-AHEH253>3.3.CO;2-J Occurrence Handle1:CAS:528:DyaK1cXmsVagu74%3D
PW Hochachka GN Somero (2002) Biochemical Adaptation—Mechanism and Process in Physiological Evolution Oxford University Press Oxford
A Hubert K-D Wenzel W Engewald G Schürmann (2001) ArticleTitleAccelerated solvent extraction—more efficient extraction of POPs and PAHs from real contaminated plant and soil samples. Rev Anal Chem 20 101–144 Occurrence Handle1:CAS:528:DC%2BD3MXmslWlt7o%3D
H Keweloh HJ Heipieper (1996) ArticleTitleTrans unsaturated fatty acids in bacteria. Lipids 31 129–137 Occurrence Handle1:CAS:528:DyaK28XhtFOmt7Y%3D Occurrence Handle8835399
G Klyachko-Gurvicha LN Tsoglina J Douchab J Kopetskiib IB Shebalina VE Semenenko (1999) ArticleTitleDesaturation of fatty acids as an adaptive response to shifts in light intensity. Physiol Plant 107 240–247 Occurrence Handle10.1034/j.1399-3054.1999.100212.x
M Koschorreck R Frömmichen P Herzsprung J Tittel K Wendt-Potthoff (2002) ArticleTitleFunctions of straw for in situ remediation of acidic mining lakes. Wat Air Soil Pollut Focus 2 97–109 Occurrence Handle10.1023/A:1019991326498 Occurrence Handle1:CAS:528:DC%2BD3sXhslKhsrw%3D
MA Lag L Meza-Basso J Fernandez R Cristi M Romero (1991) ArticleTitleLipid composition of chloroplasts from cold-acclimated and non-acclimated Nothofagus dombeyi. Phytochemistry 30 763–768 Occurrence Handle10.1016/0031-9422(91)85248-X
U Langer T Günther (2001) ArticleTitleEffects of alkaline dust deposits of phosphate fertilizer production on microbial biomass and enzyme activities in grassland soils. Environ Pollut 112 321–327 Occurrence Handle10.1016/S0269-7491(00)00148-2 Occurrence Handle1:CAS:528:DC%2BD3MXit12lurs%3D Occurrence Handle11291438
D Lessmann A Fyson B Nixdorf (2000) ArticleTitlePhytoplankton of the extremely acidic mining lakes of Lusatia (Germany) with pH <3. Hydrobiologia 433 123–128 Occurrence Handle10.1023/A:1004018722898
M Ma W Zhu Z Wang GJ Witkamp (2003) ArticleTitleAccumulation, assimilation and growth inhibition of copper on freshwater algae (Scenedesmus subspicatus 86.81 SAG) in the presence of EDTA and fulvic acid. Aquatic Toxicol 63 221–228 Occurrence Handle10.1016/S0166-445X(02)00179-0 Occurrence Handle1:CAS:528:DC%2BD3sXjtVKhsLY%3D
AE Maanni G Dubertret M-J Delrieu O Roche A Tremolières (1998) ArticleTitleMutants of Chlamydomonas reinhardtii affected in phosphatidylglycerol metabolism and thylakoid biogenesis. Plant Physiol Biochem 36 609–619 Occurrence Handle10.1016/S0981-9428(98)80009-0
A McSpadden B Gardener A Lilley (1997) Application of common statistical tools. J van Elsas J Trevors E Wellington (Eds) Modern Soil Microbiology Dekker New York
Meier, J (2001) Untersuchungen zum mikrobiellen Schwefelkreislauf in sauren Tagebau-Restseen der Niederlausitz (Brandenburg). UFZ-Bericht 21/2001, Dissertation, ISSN 0948–9452
J Morgan C Winstanley (1997) Microbial biomarkers. JD van Elsas JT Trevors EM Wellington (Eds) Modern Soil Microbiology Dekker New York
Y Murakami M Tsuyama Y Kobayashi H Kodama K Iba (2000) ArticleTitleTrienoic fatty acids and plant tolerance. Science 287 476–479 Occurrence Handle10.1126/science.287.5452.476 Occurrence Handle1:CAS:528:DC%2BD3cXntl2nug%3D%3D Occurrence Handle10642547
DS Nichols TA McMeekin (2002) ArticleTitleBiomarker techniques to screen for bacteria that produce polyunsaturated fatty acids. J Microbiol Methods 48 161–170 Occurrence Handle10.1016/S0167-7012(01)00320-7 Occurrence Handle1:CAS:528:DC%2BD38XkvFWk Occurrence Handle11777566
WH Nichols (1973) Growth media–freshwater. JR Stein (Eds) Handbook of Phycological Methods: Culture Methods and Growth Measurements Cambridge University Press Cambridge, UK 7–24
B Nixdorf A Fyson H Krumbeck (2001) ArticleTitleReview: plant life in extremely acidic waters. Environ Exp Bot 46 203–211 Occurrence Handle10.1016/S0098-8472(01)00104-6 Occurrence Handle1:CAS:528:DC%2BD3MXnslWgu7w%3D
G Packroff S Woelfl (2000) ArticleTitleA review on the occurrence and taxonomy of heterotrophic protists in extreme acidic environments of pH values <3. Hydrobiologia 433 153–156 Occurrence Handle10.1023/A:1004039309694
C Pepe H Sayer P Bacelon (1999) ArticleTitleDetermination of positions of double bonds in conjugated alkadienes by means of gas chromatography/mass spectrometry of dimethyl disulfide derivatives [letter]. Eur Mass Spectrom 5 285–287 Occurrence Handle1:CAS:528:DC%2BD3cXhtValsbY%3D
J Perry A Liebhold M Rosenberg J Dungan M Miriti A Jakomulska S Citron-Pousty (2002) ArticleTitleIllustrations and guidelines for selecting statistical methods for quantifying spatial pattern in ecological data. Ecogr 25 578–600 Occurrence Handle10.1034/j.1600-0587.2002.250507.x
I Pöhler DF Wenderoth K Wendt-Potthoff MG Höfle (2002) ArticleTitleBacterioplankton community structure and dynamics in enclosures during bio-remediation experiments in an acid mining lake. Water Air Soil Pollut Focus 2 111–121 Occurrence Handle10.1023/A:1019951612385
J Pörschmann T Welsch R Herzschuh W Engewald K Müller (1982) ArticleTitleAnalysis of fatty acids by combined application of chemical, chromatographic and spectroscopic methods. J Chromatogr 241 73–87 Occurrence Handle10.1016/S0021-9673(00)82391-2
J Pörschmann F-D Kopinke M Remmler K Mackenzie W Geyer S Mothes (1996) ArticleTitleHyphenated techniques for characterizing coal wastewaters and associated sediments. J Chromatogr A 750 287–301 Occurrence Handle10.1016/0021-9673(96)00403-7
NA Pronina NB Rogova S Furnadzhieva GL Klyachko-Gurvich (1998) ArticleTitleEffect of CO2 concentration on the fatty acid composition of lipids in Chlamydomonas reinhardtii cia-3, a mutant deficient in CO2 -concentrating mechanism. Russian J Plant Physiol 45 447–455 Occurrence Handle1:CAS:528:DyaK1cXltVyksb8%3D
J-F Rontani (1998) ArticleTitlePhotodegradation of unsaturated fatty acids in senescent cells of phytoplankton: photoproduct structural identification and mechanistic aspects. J Photochem Photobiol A: Chem 114 37–44 Occurrence Handle10.1016/S1010-6030(98)00201-9 Occurrence Handle1:CAS:528:DyaK1cXhvFCrurc%3D
J-F Rontani PC Bonin JK Volkman (1999) ArticleTitleBiodegradation of free phytol by bacterial communities isolated from marine sediments under aerobic and denitrifying conditions. Appl Environ Microbiol 65 5484–5492 Occurrence Handle1:CAS:528:DyaK1MXnvVGjtbY%3D Occurrence Handle10584007
T Rosswall E Kvillner (1978) ArticleTitlePrincipal components and factor analysis for the description of microbial populations. Adv Microb Ecol 2 1–48
N Sato K Sonoike A Kawaguchi M Tsuzuki (1996) ArticleTitleContribution of lowered unsaturation levels of chloroplast lipids to high temperature tolerance of photosynthesis in Chlamydomonas reinhardtii. J Photochem Photobiol B 36 333–337 Occurrence Handle10.1016/S1011-1344(96)07389-7 Occurrence Handle1:CAS:528:DyaK2sXksVSntQ%3D%3D
V Spitzer (1997) ArticleTitleStructure analysis of fatty acids by gas chromatography–low resolution electron impact mass spectrometry of their 4,4-dimethyloxazoline derivatives—a review. Prog Lipid Res 35 387–409 Occurrence Handle10.1016/S0163-7827(96)00011-2
K Stransky T Jursik A Vitek (1992) ArticleTitleAn improved method of characterizing fatty acids by equivalent chain length values. J High Resol Chromatogr 15 730–740 Occurrence Handle1:CAS:528:DyaK3sXht1Cjsrk%3D
K Stransky T Jursik A Vitek (1997) ArticleTitleStandard equivalent chain length values of monoenic and polyenic (methylene interrupted) fatty acids. J High Resol Chromatogr 20 143–158 Occurrence Handle1:CAS:528:DyaK2sXivFOit7w%3D
R Tate (2000) Soil Microbiology Wiley & Sons New York
H Tatsuzawa E Takizawa M Wada K Yamamoto (1996) ArticleTitleFatty acid and lipid composition of the acidophilic green alga Chlamydomonas sp. J Phycol 32 598–601 Occurrence Handle1:CAS:528:DyaK28XlvVWisbg%3D
RH Thompson (1996) ArticleTitleSimplifying fatty acid analyses in multicomponent foods with a standard set of isothermal GLC conditions coupled with ECL determinations. J Chromatogr Sci 34 495–504 Occurrence Handle1:CAS:528:DyaK28XmsFOitbg%3D
J Tittel V Bissinger B Zippel U Gaedke E Bell A Lorke N Kamjunke (2003) ArticleTitleMixotrophs combine resource use to out-compete specialists: implications for aquatic food webs. Proc Natl Acad Sci USA 100 1276–1278 Occurrence Handle10.1073/pnas.2130696100
A Trémolières (1998) Glycerolipids: composition, biosynthesis and function in Chlamydomonas. JD Rochaix M Goldschmidt-Clermont S Merchant (Eds) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas, vol. 7 Kluwer Academic Dordrecht 415–431
BE van Dongen S Schouten JSS Damste (2002) ArticleTitleCarbon isotope variability in monosaccharides and lipids of aquatic algae and terrestrial plants. Mar Ecol Progr Ser 232 83–92 Occurrence Handle1:CAS:528:DC%2BD38XlsFOquro%3D
E van Hunnik G Amoroso D Sultemeyer (2002) ArticleTitleUptake of CO2 and bicarbonate by intact cells and chloroplasts of Tetraedron minimum and Chlamydomonas noctigama. Planta 215 763–769 Occurrence Handle10.1007/s00425-002-0817-2 Occurrence Handle1:CAS:528:DC%2BD38XotVGksrs%3D Occurrence Handle12244441
R Vazhappilly F Chen (1998) ArticleTitleHeterotrophic production potential of omega-3 polyunsaturated fatty acids by microalgae and algae-like microorganisms. Bot Mar 41 553–558 Occurrence Handle1:CAS:528:DyaK1cXnvVahtL8%3D
B Vigneault A Percot M Lafleur PGC Campbell (2000) ArticleTitlePermeability changes in model and phytoplankton membranes in the presence of aquatic humic substances. Environ Sci Technol 34 3907–3913 Occurrence Handle10.1021/es001087r Occurrence Handle1:CAS:528:DC%2BD3cXlsVSls7k%3D
JK Volkman SM Barrett SI Blackburn MP Mansour EL Sikes F Gelin (1998) ArticleTitleMicroalgal biomarkers: A review of recent research developments. Org Geochem 29 1163–1179 Occurrence Handle10.1016/S0146-6380(98)00062-X Occurrence Handle1:CAS:528:DyaK1MXpslyrtg%3D%3D
Z-Y Wen F Chen (2000) ArticleTitleHeterotrophic production of eicosapentaenoid acid by the diatom Nitzschia laevis: Effects of silicate and glucose. J Ind Microbiol Biotechnol 25 218–224 Occurrence Handle10.1038/sj.jim.7000056 Occurrence Handle1:CAS:528:DC%2BD3MXlvF2ntg%3D%3D
K Wendt-Potthoff R Frömmichen P Herzsprung M Koschorreck (2002) ArticleTitleMicrobial Fe(III) reduction in acidic mining lake sediments after addition of organic substrate and lime. Wat Air Soil Pollut Focus 2 81–96 Occurrence Handle10.1023/A:1019959814202 Occurrence Handle1:CAS:528:DC%2BD3sXhslKhsr4%3D
K Wendt-Potthoff M Koschorreck (2002) ArticleTitleFunctional groups and activities of bacteria in a highly acidic volcanic mountain stream and lake in Patagonia, Argentina. Microb Ecol 43 92–106 Occurrence Handle10.1007/s00248-001-1030-8 Occurrence Handle1:CAS:528:DC%2BD38XjslGhtro%3D Occurrence Handle11984632
DC White HC Pinkart DB Ringelberg (1997) Biomass measurements: biochemical approaches. CJ Hurst (Eds) Manual of Environmental Microbiology ASM Press Washington 91–101
K Wollmann R Deneke B Nixdorf G Packroff (2000) ArticleTitleDynamics of planktonic food webs in three mining lakes across a pH gradient (pH 2–4). Hydrobiologia 433 3–14 Occurrence Handle10.1023/A:1004060732467 Occurrence Handle1:CAS:528:DC%2BD3cXosFehsrg%3D
K Yamamoto R Murakami Y Takamura (1998) ArticleTitleIsoprenoid quinone, cellular fatty acid composition and diaminopimelic acid isomers of newly classified thermophilic anaerobic Gram-positive bacteria. FEMS Microbiol Lett 161 351–358 Occurrence Handle10.1016/S0378-1097(98)00094-9 Occurrence Handle1:CAS:528:DyaK1cXhvFKjsL0%3D
Acknowledgments
We are most grateful to Jana Jost, Marion Hoyer, Evelyn Becker, and Ursula Bachmann for their skillful technical assistance. Thanks are due to Norbert Kamjunke, Jörg Tittel, and Jutta Meier for their valuable input comprising unpublished data on bacterial populations of Lake 111 and from field observations. This work was kindly funded by a European Union Marie Curie Development Host Fellowship (HPMD-CT-2001-00103) and a scholarship received by E.S. from UFZ, Magdeburg, Department of Inland Water Research.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Poerschmann, J., Spijkerman, E. & Langer, U. Fatty Acid Patterns in Chlamydomonas sp. as a Marker for Nutritional Regimes and Temperature under Extremely Acidic Conditions. Microb Ecol 48, 78–89 (2004). https://doi.org/10.1007/s00248-003-0144-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00248-003-0144-6