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Applied and Environmental Microbiology, June 2006, p. 4419-4422, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00191-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Thermophilic Temperature Optimum for Crenarchaeol Synthesis and Its Implication for Archaeal Evolution

Chuanlun L. Zhang,1* Ann Pearson,2 Yi-Liang Li,1,{dagger} Gary Mills,1 and Juergen Wiegel3

Savannah River Ecology Laboratory, University of Georgia, Aiken, South Carolina 29802,1 Department of Earth and Planetary Sciences, Harvard University, Cambridge, Massachusetts 02138,2 Department of Microbiology, University of Georgia, Athens, Georgia 306053

Received 24 January 2006/ Accepted 20 March 2006


   ABSTRACT
Top
 Abstract
Introduction
 Lipid extraction and liquid...
 Distribution of crenarchaeol in...
 References
 
The isoprenoid lipid crenarchaeol is widespread in hot springs of California and Nevada. Terrestrial and marine data together suggest a maximum relative abundance of crenarchaeol at ~40°C. This warm temperature optimum may have facilitated colonization of the ocean by (hyper)thermophilic Archaea and the major marine radiation of Crenarchaeota.


   INTRODUCTION
Top
 Abstract
 Introduction
Lipid extraction and liquid...
 Distribution of crenarchaeol in...
 References
 
Crenarchaeota synthesize membranes in which the major lipids are glycerol dialkyl glycerol tetraethers (GDGTs) (5, 14, 15). In thermophilic archaea (8, 26) and in the ocean (22, 23) the number of cyclopentane rings increases with increasing surface water temperature (20, 22, 23). Crenarchaeol is an unusual GDGT first identified in marine sediments (1, 2, 4, 10, 18, 21, 24, 29, 30). It contains four cyclopentyl rings and, uniquely among GDGTs, it also contains one cyclohexyl ring (1). Results from molecular models show that the cyclohexyl moiety adds molecular volume, thereby enhancing flexibility at lower temperatures (1). However, contrary to physical expectations, we observed that crenarchaeol is not a more abundant fraction of total GDGTs in samples from progressively colder environments (references 1 and 24 and this study).

Recently, crenarchaeol was found in high relative abundance in terrestrial hot springs (19). Therefore, we sought to explore the distribution of crenarchaeol in environmental samples across a broad range of temperatures. The marine and terrestrial data for the relative abundance of crenarchaeol now span psychrophilic to hyperthermophilic communities ranging from <10°C to 87°C, indicating a broad biosynthetic distribution of the compound. The data suggest an evolutionary history of crenarchaeol that may be longer and more complex than its distribution in the modern ocean indicates.


   Lipid extraction and liquid chromatography-mass spectrometry.
Top
 Abstract
 Introduction
 Lipid extraction and liquid...
Distribution of crenarchaeol in...
 References
 
Total lipids were extracted from mat material collected from hot springs by using established procedures (19, 30). The lipids were acid hydrolyzed and screened by high-performance liquid chromatography-mass spectrometry. The column was Zorbax NH2 (custom 2.1 mm by 150 mm; 5-µm particle size; isocratic 1.3% isopropanol in hexane; 30°C). Conditions for atmospheric-pressure chemical ionization-mass spectromety were as stated previously (11, 19).


   Distribution of crenarchaeol in California and Nevada hot springs.
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 Abstract
 Introduction
 Lipid extraction and liquid...
 Distribution of crenarchaeol in...
References
 
Seventeen samples were analyzed from 11 hot-spring locations in California and Nevada and from one spring in Thailand (Table 1). Additional data for marine samples from Santa Monica Basin, Santa Barbara Basin, and Cariaco Basin and from the literature are shown in Table 2.


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  		TABLE 1. Data for hot springs in United States and Thailand, including relative abundance of GDGTs I to VII as determined by liquid chromatography-mass spectrometry

 

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  		TABLE 2. Relative abundances of GDGTs in marine samplesa

 
For the hot-spring samples, correlations were explored between the relative abundances of all GDGTs and each of the environmental parameters, including temperature, pH, and alkalinity. Crenarchaeol (I) (Fig. 1) was the only GDGT that showed a negative slope with temperature (R2 = 0.64) (Fig. 2a). GDGT II exhibited no relationship to temperature (R2 = 0.03), and the correlation lines for all other numbers of cyclopentyl rings (III to VII) had positive slopes (R2 = 0.45 to 0.57) (Fig. 2a).


Figure 1
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  		FIG. 1. Chromatograms of GDGTs from hot-spring and marine sediments. (a) Microbial mat from Nevada hot spring Hard To Find-2 (Table 1); 58.1°C; pH 6.2. (b) A typical marine sediment (Santa Monica Basin; sea surface temperature, 16°C). (c) Sediment from California hot spring Surprise Valley (SV-1); 86.5°C; pH 5.8. Compounds are identified using the roman numeral conventions (23) II, m/z 1,302, no rings; III, m/z 1,300, one cyclopentyl ring; IV, m/z 1,298, two cyclopentyl rings; V, m/z 1,296, three cyclopentyl rings; I, crenarchaeol, m/z 1,292, four cyclopentyl rings, one cyclohexyl ring; VI, m/z 1,292, crenarchaeol regioisomer; VII (new convention), m/z 1,294, four cyclopentyl rings. In panel a, I accounted for about 24% and VII accounted for about 13% of the total GDGTs, respectively; in panel c, traces of I (2%) were detectable, but VII dominated (28 to 30%) (Table 1).

 

Figure 2
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  		FIG. 2. Distribution of GDGTs as a function of temperature. (a) Relative abundances of GDGTs in hot-spring microbial mats between 36°C and 87°C. Linear trend lines are shown (R2 values are reported in the text). Data for compound VI are not shown, as VI is always ~10% of the magnitude of I. (b) Ratio of I to II in samples from hot springs (Table 1) and marine sediments (Table 2) versus temperature. Polynomial curve fit; y = –0.00037x2 + 0.0315x + 0.141; R2 = 0.70.

 
Our discovery of crenarchaeol in a wide range of terrestrial hot springs has significant ecological and evolutionary implications. Crenarchaeol is abundant in marine environments and accounts for up to 46% of total GDGTs in marine sediments having temperatures below 20°C (24) and up to 60% of total GDGTs in surface waters at 26°C (12). There is also approximately 60% crenarchaeol in the GDGTs of the symbiont Cenarchaeum symbiosium (1, 21).

In hot springs, two of our samples contained 100% of the total detectable GDGTs as crenarchaeol (Dixie Valley and Seven Devils) (Table 1), and previous work showed that a third sample (40°C) contained crenarchaeol in 22:1 ratio to GDGT II (19). The abundance of crenarchaeol decreased to the 40 to 60% threshold between 50 and 65°C, causing the GDGT profiles of these hot springs (Fig. 1a) to resemble the GDGT distributions in marine sediments (Fig. 1b). GDGTs from the 87°C spring in Surprise Valley [samples SV (0 to 1 cm) and SV (1 to 2 cm) (Table 1)] are noticeably different (Fig. 1c) but still contain detectable traces of crenarchaeol (Table 1). When the abundance of crenarchaeol (I) is normalized to GDGT II (which has no temperature correlation) and plotted as a function of temperature, the data from terrestrial and marine systems together fit a second-order polynomial with significant correlation (R2 = 0.70) (Fig. 2b). These results suggest that the marine archaea live on the low-temperature biosynthetic end of what may be a normal distribution centered around 40 to 45°C (Fig. 2b). Indeed, sediments underlying cooler marine waters always appear to contain relatively smaller amounts of crenarchaeol than samples obtained from tropical latitudes (24) (Table 2 and Fig. 2b). Most importantly, however, the data suggest that the optimum temperature for organisms that produce crenarchaeol is greater than either the present or past temperature of surface seawater.

It also is possible that pH, in addition to temperature, may affect the abundance of crenarchaeol in a manner similar to pH control of sterols in eukaryotes (9). These variables, however, cannot easily be decoupled for the organisms living in California or Nevada hot springs, as the lower-temperature springs tend to have higher pH values. A combined temperature-pH effect could explain the absence of the cyclohexyl ring in cultured thermophilic archaea, because these species commonly grow at low pH (26) rather than at the higher pH values measured here. Based on our findings and the observation that cyclohexyl rings have not been found in any Euryarchaeota, we hypothesize that crenarchaeol is a primitive phenotypic feature that is specific to the Crenarchaeota. While a temperature effect of the cyclohexyl ring has been reasonably explained (1), the effect of pH has not yet been rigorously examined.

Experimental studies using pure cultures of thermophilic archaea have demonstrated that the number of cyclopentyl rings in GDGTs increases with growth temperature (8, 26). Because the cyclohexyl ring has not been found in cultured thermophiles, its presence in the nonthermophilic Crenarchaeota has been attributed solely to the significant decrease in growth temperature experienced by the Crenarchaeota when they were colonizing marine waters, perhaps around 112 million years ago in the Cretaceous Period (1, 16). Our findings differ significantly from the view that crenarchaeol, or specifically, its unique cyclohexyl ring, is an evolutionary product of this relatively recent event (16, 22). The apparent ~40 to 45°C temperature optimum for crenarchaeol exceeds the warmest sea surface temperatures (27 to 36°C), even in the Cretaceous Period (22). We suggest that crenarchaeol could be an original and ancient biochemical property of the thermophilic Crenarchaeota, which occupy a deep branching point in the tree of life (3, 7).


   ACKNOWLEDGMENTS
 
We thank Christopher Romanek, Rick Socki, and Sue Lutz for help with field work in Nevada and Lily Eurwilaichitr for providing samples from Thailand. We thank Susan Carter and Sunita Shah for laboratory assistance. The owners of hot springs in Surprise Valley, Gerlach, and other locations showed great hospitality and supported our sampling efforts.

This research was supported by National Science Foundation grant MCB-0348180 to C.L.Z. and by NSF grant OCE-0241363 to A.P. The U.S. Department of Energy also supported this research through Financial Assistance Award DE-FC09-96SR18546 to the University of Georgia Research Foundation (C.L.Z. and G.M.).


   FOOTNOTES
 
* Corresponding author. Mailing address: Savannah River Ecology Laboratory, University of Georgia, Aiken, SC 29802. Phone: (803) 725-5299. Fax: (803) 725-3309. E-mail: zhang{at}srel.edu. Back

{dagger} Present address: Center for Biomarker Analysis, University of Tennessee, Knoxville, TN 37932-2575. Back


   REFERENCES
Top
 Abstract
 Introduction
 Lipid extraction and liquid...
 Distribution of crenarchaeol in...
 References
 

  1. Damsté, J. S. S., E. C. Hopmans, S. Schouten, A. C. T. van Duin, and J. A. J. Geenevasen. 2002. Crenarchaeol: the characteristic glycerol dibiphytanyl glycerol tetraether membrane lipid of cosmopolitan pelagic Crenarchaeota. J. Lipid Res. 43:1641-1651.[Abstract/Free Full Text]
  2. Damsté, J. S. S., W. Rijpstra, E. C. Hopmans, F. Prahl, S. Wakeham, and S. Schouten. 2002. Distribution of membrane lipids of planktonic Crenarchaeota in the Arabian Sea. Appl. Environ. Microbiol. 68:2997-3002.[Abstract/Free Full Text]
  3. Dawson, S. C., N. R. Pace, and E. F. DeLong. 2000. Phylogenetic and ecological perspectives on uncultured Crenarchaeota and Korarchaeota. In M. Dworkin, S. Falkow, E. Rosenberg, K.-H. Schleifer, and E. Stackebrandt (ed.), The prokaryotes—an evolving electronic resource for the microbiological community. Springer, New York, N.Y. [online.] http://link.springer-ny.com/link/service/books/10125/.
  4. DeLong, E. F., L. L. King, R. Massana, H. Cittone, A. Murray, C. Schleper, and S. G. Wakeham. 1998. Dibiphytanyl ether lipids in nonthermophilic crenarchaeotes. Appl. Environ. Microbiol. 64:1133-1138.[Abstract/Free Full Text]
  5. De Rosa, M., and A. Gambacorta. 1988. The lipids of archaebacteria. Prog. Lipid Res. 27:153-175.[CrossRef][Medline]
  6. Emeis, K. C., U. Struck, H. M. Schulz, R. Rosenberg, S. Bernasconi, H. Erlenkeuser, T. Sakamoto, and F. Martinez-Ruiz. 2000. Temperature and salinity variations of Mediterranean Sea surface waters over the last 16,000 years from records of planktonic stable oxygen isotopes and alkenone unsaturation ratios. Palaeogeogr. Palaeoclimatol. Palaeoecol. 158:259-280.
  7. Forterre, P. 2002. Evolution of the Archaea. Theor. Pop. Biol. 61:409-422.[CrossRef][Medline]
  8. Gliozzi, A., G. Paoli, M. DeRosa, and A. Gambacorta. 1983. Effect of isoprenoid cyclization on the transition temperature of lipids in thermophilic archaeabacteria. Biochim. Biophys. Acta 735:234-242.[CrossRef]
  9. Haines, T. H. 2001. Do sterols reduce proton and sodium leaks through lipid bilayers? Prog. Lip. Res. 40:299-324.[CrossRef][Medline]
  10. Hoefs, M. J. L., S. Schouten, J. W. deLeeuw, L. L. King, S. G. Wakeham, and J. S. S. Damsté. 1997. Ether lipids of planktonic Archaea in the marine water column. Appl. Environ. Microbiol. 63:3090-3095.[Abstract]
  11. Hopmans, E. C., S. Schouten, R. D. Pancost, M. T. J. van der Meer, and J. S. S. Damsté. 2000. Analysis of intact tetraether lipids in archaeal cell material and sediments by high performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. Rapid Comm. Mass Spectr. 14:585-589.
  12. Ingalls, A. E., S. R. Shah, R. L. Hansman, L. I. Aluwihare, G. M. Santos, E. R. M. Druffel, and A. Pearson. Quantifying archaeal community autotrophy in the mesopelagic ocean using natural radiocarbon. Proc. Natl. Acad. Sci. USA, in press.
  13. Kara, A. B., A. J. Wallcraft, and H. E. Hurlburt. 2005. Sea surface temperature sensitivity to water turbidity from simulations of the turbid Black Sea using HYCOM. J. Phys. Ocean 35:33-54.[CrossRef]
  14. Koga, Y., and H. Morii. 2005. Recent advances in structural research on ether lipids from archaea including comparative and physiological aspects. Biosci. Biotechnol. Biochem. 69:2019-2034.[CrossRef][Medline]
  15. Koga, Y., M. Nishihara, H. Morii, and M. Akagawa-Matsushita. 1993. Ether polar lipids of methanogenic bacteria: structures, comparative aspects, and biosynthesis. Microbiol. Rev. 57:164-182.[Abstract/Free Full Text]
  16. Kuypers, M. M. M., P. Blokker, J. Erbacher, H. Kinkel, R. D. Pancost, S. Schouten, and J. S. S. Damsté. 2001. Massive expansion of marine archaea during a mid-Cretaceous oceanic anoxic event. Science 293:92-94.[Abstract/Free Full Text]
  17. Lea, D. W., D. K. Pak, L. C. Peterson, and K. A. Hughen. 2003. Synchroneity of tropical and high-latitude Atlantic temperatures over the last glacial termination. Science 301:1361-1364.[Abstract/Free Full Text]
  18. Pearson, A., A. P. McNichol, B.-C. Benitez-Nelson, J. M. Hayes, and T. I. Eglington. 2001. Origins of lipid biomarkers in Santa Monica Basin surface sediment: a case study using compound-specific 14C analysis. Geochem. Cosmochim. Acta 65:3123-3137.
  19. Pearson, A., Z. Huang, A. E. Ingalls, C. S. Romanek, J. Wiegel, K. H. Freeman, R. H. Smittenberg, and C. L. Zhang. 2004. Non-marine crenarchaeol in Nevada hot springs. Appl. Environ. Microbiol. 70:5229-5237.[Abstract/Free Full Text]
  20. Powers, L. A., J. P. Werne, T. C. Johnson, E. C. Hopmans, J. S. S. Damsté, and S. Schouten. 2004. Crenarchaeotal membrane lipids in lake sediments: a new paleotemperature proxy for continental paleoclimate reconstruction? Geology 32:613-616.[Abstract/Free Full Text]
  21. Preston, C. M., K. Y. Wu, T. F. Molinski, and E. F. DeLong. 1996. A psychrophilic crenarchaeon inhabits a marine sponge: Cenarchaeum symbiosum gen. nov., sp. nov. Proc. Natl. Acad. Sci. USA 93:6241-6246.[Abstract/Free Full Text]
  22. Schouten, S., E. C. Hopmans, A. Forster, Y. van Breugel, M. M. M. Kuypers, and J. S. S. Damsté. 2003. Extremely high sea-surface temperatures at low latitudes during the middle Cretaceous as revealed by archaeal membrane lipids Geology 31:1069-1072.[Abstract/Free Full Text]
  23. Schouten, S., E. C. Hopmans, E. Schefuß, and J. S. S. Damsté. 2002. Distributional variations in marine crenarchaeotal membrane lipids: a new tool for reconstructing ancient sea water temperatures? Earth Planet. Sci. Lett. 204:265-274.
  24. Schouten, S., E. C. Hopmans, R. D. Pancost, and J. S. S. Damsté. 2000. Widespread occurrence of structurally diverse tetraether membrane lipids: evidence for the ubiquitous presence of low-temperature relatives of hyperthermophiles. Proc. Natl. Acad. Sci. USA 97:14421-14426.[Abstract/Free Full Text]
  25. Smittenberg, R. H., R. D. Pancost, E. C. Hopmans, M. Paetzel, and J. S. S. Damsté. 2004. A 400-year record of environmental change in an euxinic fjord as revealed by the sedimentary biomarker record. Palaeogeogr. Palaeoclimatol. Palaeoecol. 202:331-351.[CrossRef]
  26. Uda, I., A. Sugai, Y. H. Itoh, and T. Itoh. 2001. Variation on molecular species of polar lipids from Thermoplasma acidophilum depends on growth temperature. Lipids 36:103-105.[Medline]
  27. Wakeham, S. G., and J. A. Beier. 1991. Fatty acid and sterol biomarkers as indicators of particulate matter source and alteration processes in the Black Sea. Deep Sea Res. 38:S943-S968.[CrossRef]
  28. Wakeham, S. G., C. Lee, J. W. Farrington, and R. B. Gagosian. 1984. Biogeochemistry of particulate organic matter in the oceans: results from sediment trap experiments. Deep Sea Res. 31:509-528.[CrossRef]
  29. Wakeham, S. G., E. C. Hopmans, S. Schouten, and J. S. S. Damsté. 2004. Archaeal lipids and anaerobic oxidation of methane in euxinic water columns: a comparative study of the Black Sea and Cariaco Basin. Chem. Geol. 205:427-442.[CrossRef]
  30. Zhang, C. L., R. D. Pancost, Y. Qian, R. Sassen, and S. A. Macko. 2003. Archaeal lipid biomarkers and isotopic evidence of anaerobic methane oxidation associated with gas hydrates in the Gulf of Mexico. Org. Geochem. 34:827-834.

Applied and Environmental Microbiology, June 2006, p. 4419-4422, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.00191-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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