Abstract
Phospholipid fatty acid (PLFA) analysis is a method that has been used for over 35 years and can be useful to the microbial ecologist by quantitatively providing: (1) an estimate of viable microbial biomass by measuring the amount of cellular membrane, (2) an outline of community structure by identifying signature PLFA biomarkers indicative of prokaryotic and eukaryotic taxa, and (3) an indicator of microbial physiological status by analyzing for known stress indicators. Additionally, a unique aspect of using PLFA for investigating microbial communities is that microbial lipids can be quantitatively extracted from almost any sample matrix. Once extracted and prepared the PLFAs can then be analyzed via established and mature techniques such as gas chromatography and mass spectroscopy. The quantitative nature of the resulting data allows for the application of statistics that authenticate microbial biomass and composition differences across an environment or between treatments.
David C. White was deceased at the time of publication
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References
Tollefson TS, McKercher RB (1983) The degradation of 14C-labelled phosphatidyl choline in soil. Soil Biol Biochem 15:145–148
Frostegård Å, Tunlid A, Bååth E (2011) Use and misuse of PLFA measurements in soils. Soil Biol Biochem 43:1621–1625
Hedrick DB, Peacock AD, White DC (2007) Biochemical approaches to microbial biomass measurements and community structure analysis. In: Hurst CJ, Crawford RL, Knudsen GR, McInerney MJ, Stetzenbach LD (eds) Manual of environmental microbiology, 3rd edn. ASM Press, Washington
Bailey VL, Peacock AD, Smith JL, Bolton H Jr (2002) Relationships between soil microbial biomass determined by chloroform fumigation-extraction, substrate-induced respiration, and phospholipids fatty acid analysis. Soil Biol Biochem 34:1385–1389
White DC, Bobbie RJ, Herron JS, King JD, Morrison SJ (1979) Biochemical measurements of microbial mass and activity from environmental samples. In: Costerton JW, Colwell RR (eds) Native Aquatic Bacteria: Enumeration, Activity and Ecology, vol 695, ASTM STP. American Society for Testing Materials, Philadelphia, pp 69–81
Tunlid A, White DC (1992) Biochemical analysis of biomass, community structure, nutritional status and metabolic activity of the microbial community in soil. In: Bollag J-M, Stotzky G (eds) Soil biochemistry, vol 7. Marcel Dekker Inc., New York, pp 229–262
Federle TW, Dobbins DC, Thornton-Manning JR, Jones DD (1986) Microbial biomass, activity, and community structure in subsurface soils. Groundwater 24:365–374
Findlay RH, Dobbs FC (1993) Quantitative description of microbial communities using lipid analysis. In: Kemp PF, Sherr BF, Sherr EB, Cole JJ (eds) Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca Raton, pp 271–284
Kieft TL, Ringelberg DB, White DC (1994) Changes in ester-linked phospholipid fatty acid profiles of subsurface bacteria during starvation and desiccation in a porous medium. Appl Environ Microbiol 60:3294–3299
Frostegård A, Tunlid A, Bååth E (1996) Changes in microbial community structure during long-term incubation in two soils experimentally contaminated with metals. Soil Biol Biochem 28:55–63
Stephen JR, Chang Y-J, Gan Y-D, Peacock A, Pfiffner SM, Barcelona MJ, White DC, Macnaughton SJ (1999) Microbial characterization of a JP-4 fuel contaminated site using a combined lipid biomarker/PCR-DGGE based approach. Environ Microbiol 1:231–241
Willers C, Jansen van Rensburg PJ, Claassens S (2015) Phospholipid fatty acid profiling of microbial communities – a review of interpretations and recent application. J Appl Microbiol 119(5):1207–1218
Frostegård A, Tunlid A, Bååth E (2011) Use and misuse of PLFA measurements in soils. Soil Biol Biochem 43(8):1621–1625
Willers C, Jansen van Rensburg PJ, Claassens S (2015) Microbial signature lipid biomarker analysis – an approach that is still preferred, even amid various method modifications. J Appl Microbiol 118:1251–1263
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 31:911–917
Guckert JB, Antworth CP, Nichols PD, White DC (1985) Phospholipid ester-linked fatty acid profiles as reproducible assays for changes in prokaryotic community structure of estuarine sediments. FEMS Microbiol Ecol 31:147–158
Christie WW (1989) Gas chromatography and lipids. The Oily Press, Ayr, Scotland
Hedrick DB, Peacock AD, White DC (2005) Fatty Acid Profiles in Soil Microorganisms. In: Margesin R, Schinner F (eds) R. Manual for Monitoring and Assessing Soil Bioremediation, Springer-Verlag, Heidelberg, pp 251–258
Dunkelblum E, Tan SH, Silk PJ (1985) Double-bond location in monounsaturated fatty acids by dimethyl disulfide derivatization and mass spectrometry: application to analysis of fatty acids in pheromone glands of four lepidoptera. Journal of Chem Ecol 11(3):265–277
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Peacock, A.D., White, D.C. (2016). Microbial Biomass and Community Composition Analysis Using Phospholipid Fatty Acids. In: McGenity, T., Timmis, K., Nogales , B. (eds) Hydrocarbon and Lipid Microbiology Protocols. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2016_213
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DOI: https://doi.org/10.1007/8623_2016_213
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