Developing a genetic approach to target cyanobacterial producers of heterocyte glycolipids in the environment

Heterocytous cyanobacteria are important players in the carbon and nitrogen cycle. They can fix dinitrogen by using heterocytes, specialized cells containing the oxygen-sensitive nitrogenase enzyme surrounded by a thick polysaccharide and glycolipid layer which prevents oxygen diffusion and nitrogenase inactivation. Heterocyte glycolipids can be used to detect the presence of heterocytous cyanobacteria in present-day and past environments, providing insight into the functioning of the studied ecosystems. However, due to their good preservation throughout time, heterocyte glycolipids are not ideal to detect and study living communities, instead methods based on DNA are preferred. Currently cyanobacteria can be detected using untargeted genomic approaches such as metagenomics, or they can be specifically targeted by, for example, the use of primers that preferentially amplify their 16S rRNA gene or their nifH gene in the case of nitrogen fixing cyanobacteria. However, since not all cyanobacterial nitrogen fixers are heterocytous, there is currently no fast gene-based method to specifically detect and distinguish heterocytous cyanobacteria. Here, we developed a PCR-based method to specifically detect heterocytous cyanobacteria by designing primers targeting the gene (hglT) encoding the enzyme responsible for the last step in the biosynthesis of heterocyte glycolipid (i.e., a glycosyltransferase). We designed several primer sets using the publicly available sequences of 23 heterocytous cyanobacteria, after testing them on DNA extracts of 21 heterocyte-forming and 7 non-heterocyte forming freshwater cyanobacteria. The best primer set was chosen and successfully used to confirm the presence of heterocytous cyanobacteria in a marine environmental sample

Holocene subsurface temperature variability in the eastern Antarctic continental margin

We reconstructed subsurface (∼45-200m water depth) temperature variability in the eastern Antarctic continental margin during the late Holocene, using an archaeal lipid-based temperature proxy (TEX 86 L). Our results reveal that subsurface temperature changes were probably positively coupled to the variability of warmer, nutrient-rich Modified Circumpolar Deep Water (MCDW, deep water of the Antarctic circumpolar current) intrusion onto the continental shelf. The TEX 86 L record, in combination with previously published climatic records, indicates that this coupling was probably related to the thermohaline circulation, seasonal variability in sea ice extent, sea temperature, and wind associated with high frequency climate dynamics at low-latitudes such as internal El Nio Southern Oscillation (ENSO). This in turn suggests a linkage between centennial ENSO-like variability at low-latitudes and intrusion variability of MCDW into the eastern Antarctic continental shelf, which might have further impact on ice sheet evolution. Copyright 2012 by the American Geophysical Union

Rapid sulfurisation of highly branched isoprenoid (HBI) alkenes in sulfidic Holocene sediments from Ellis Fjord, Antarctica

Author Posting. © Elsevier B.V., 2007. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Organic Geochemistry 38 (2007): 128-139, doi:10.1016/j.orggeochem.2006.08.003.Samples of particulate organic matter from the water column and anoxic Holocene sediment layers from the Small Meromictic Basin (SMB) in Ellis Fjord (eastern Antarctica) were analyzed to study the early incorporation of reduced inorganic sulfur species into highly branched isoprenoid (HBI) alkenes. HBIs were not detected in the water column samples from austral winter, whereas compounds containing the C25 HBI skeleton were abundant in all analyzed Holocene sediment layers. The structure of the C25:2 HBI alkene together with its enriched stable carbon isotopic composition suggest that the HBI alkene is produced by a diatom or diatoms probably belonging to the Navicula genus present in the sea-ice which covers the area most of the year. Within just 500 years of deposition, all of the HBI alkene was sulfurised. A mixture of products was formed, including components tentatively identified as a C25 HBI thiane and three S-containing dimers composed of two C25:1 HBI skeletons linked together by a sulfide bond. Most of the HBI alkene, however, was converted to polar S-containing compounds. The observed reaction rate for sulfurisation the C25:2 HBI alkene is the highest observed so far in natural systems. Sterols and other lipids known to be prone to sulfurisation were only minimally sulfurised under these depositional conditions. The reason for this is presently unclear.Funding for the collection of the sediment and water samples (by MJLC and CW) was provided by ASAC grant 1166 to JKV. This work was further supported by a grant from the Netherlands Organization for Scientific Research (NWO; Netherlands Antarctic Research Proposals 851.20.006 to JSSD)

Rapid Climate Changes in the Westernmost Mediterranean (Alboran Sea) Over the Last 35 kyr: New Insights From Four Lipid Paleothermometers (UK'37, TEXH86, RI-OH', and LDI)

The westernmost Mediterranean is one of the most sensitive areas to global climate change and high sedimentation rates allow recording high frequency variability. We present a high-resolution paleotemperature reconstruction over the last 35 kyr using, for the first time, four independent organic sea surface temperature (SST) proxies (U-37(K'), TEX86H, RI-OH' and LDI) based on alkenones, (hydroxy) isoprenoid GDGTs, and long-chain diols. We also generated a delta O-18 of planktonic foraminifera G. bulloides record together with records of bulk parameters (total organic carbon content, delta C-13(org)) and the accumulation rates of different biomarkers to provide insights into terrestrial input and primary producers. All derived-SST records showed similar trends over the last 35 kyr, revealing abrupt temperature variations during the last seven Dansgaard-Oeschger (D/O) cycles, the three Heinrich (H) events, the Last Glacial Maximum, and the Younger Dryas. H3 is recognized as the coldest event, while H1 was recorded by all SST proxies as the most abrupt one. In general, TEX86H-, RI-OH'- and LDI-SST estimates were lower than those obtained from U-37(K'). The LDI paleothermometer recorded the largest range of absolute SSTs over the whole period (ca. 20 degrees C) followed by RI-OH' (ca. 16 degrees C). TEX86H, RI-OH' and LDI proxies reflected sudden SST changes during the D/O 6 and 5 particularly well. Low BIT values and the abundance of C-32 1,15-diol in range with typical marine values indicated only minor input of continental organic matter. Accumulation rates of different lipid biomarkers were generally modulated by D/O cycles, suggesting enhanced productivity during D/O interstadials and the Bolling-Allerod period

Organohalide-respiring Desulfoluna species isolated from marine environments

The online version of this article (https://doi.org/10.1038/s41396-019-0573-y) contains supplementary material, which is available to authorized usersThe genus Desulfoluna comprises two anaerobic sulfate-reducing strains, D. spongiiphila AA1T and D. butyratoxydans MSL71T, of which only the former was shown to perform organohalide respiration (OHR). Here we isolated a third strain, designated D. spongiiphila strain DBB, from marine intertidal sediment using 1,4-dibromobenzene and sulfate as the electron acceptors and lactate as the electron donor. Each strain harbors three reductive dehalogenase gene clusters (rdhABC) and corrinoid biosynthesis genes in their genomes, and dehalogenated brominated but not chlorinated organohalogens. The Desulfoluna strains maintained OHR in the presence of 20?mM sulfate or 20?mM sulfide, which often negatively affect other organohalide-respiring bacteria. Strain DBB sustained OHR with 2\% oxygen in the gas phase, in line with its genetic potential for reactive oxygen species detoxification. Reverse transcription-quantitative PCR revealed differential induction of rdhA genes in strain DBB in response to 1,4-dibromobenzene or 2,6-dibromophenol. Proteomic analysis confirmed expression of rdhA1 with 1,4-dibromobenzene, and revealed a partially shared electron transport chain from lactate to 1,4-dibromobenzene and sulfate, which may explain accelerated OHR during concurrent sulfate reduction. Versatility in using electron donors, de novo corrinoid biosynthesis, resistance to sulfate, sulfide and oxygen, and concurrent sulfate reduction and OHR may confer an advantage to marine Desulfoluna strains.We thank Johanna Gutleben and Maryam Chaib de Mares for sediment sampling, W. Irene C. Rijpstra for fatty acid analysis, and Andreas Marquardt (Proteomics Centre of the University of Konstanz) for proteomic analyses. We acknowledge the China Scholarship Council (CSC) for the support to PP and YL. The authors thank BE-BASIC funds (grants F07.001.05 and F08.004.01) from the Dutch Ministry of Economic Affairs, ERC grant (project 323009), the Gravitation grant (project 024.002.002) and the UNLOCK project (NRGWI.obrug.2018.005) of the Netherlands Ministry of Education, Culture and Science and the Netherlands Science Foundation (NWO), and National Natural Science Foundation of China (project No.51709100) for funding.info:eu-repo/semantics/publishedVersio

The spread of marine anoxia on the northern Tethys margin during the Paleocene-Eocene Thermal Maximum

Records of the paleoenvironmental changes that occurred during the Paleocene-Eocene Thermal Maximum (PETM) are preserved in sedimentary rocks along the margins of the former Tethys Ocean and Peri-Tethys. This paper presents new geochemical data that constrain paleoproductivity, sediment delivery, and seawater redox conditions, from three sites that were located in the Peri-Tethys region. Trace and major element, iron speciation, and biomarker data indicate that water column anoxia was established during episodes when inputs of land-derived higher plant organic carbon and highly weathered detrital clays and silts became relatively higher. Anoxic conditions are likely to have been initially caused by two primary processes: (i) oxygen consumption by high rates of marine productivity, initially stimulated by the rapid delivery of terrestrially derived organic matter and nutrients, and (ii) phosphorus regeneration from seafloor sediments. The role of the latter process requires further investigation before its influence on the spread of deoxygenated seawater during the PETM can be properly discerned. Other oxygen-forcing processes, such as temperature/salinity-driven water column stratification and/or methane oxidation, are considered to have been relatively less important in the study region. Organic carbon enrichments occur only during the initial stages of the PETM as defined by the negative carbon isotope excursions at each site. The lack of observed terminal stage organic carbon enrichment does not support a link between PETM climate recovery and the sequestration of excess atmospheric CO2 as organic carbon in this region; such a feedback may, however, have been important in the early stages of the PETM

An interlaboratory study of TEX86 and BIT analysis of sediments, extracts and standard mixtures.

Two commonly used proxies based on the distribution of glycerol dialkyl glycerol tetraethers (GDGTs) are the TEX86 (TetraEther indeX of 86 carbon atoms) paleothermometer for sea surface temperature reconstructions and the BIT (Branched Isoprenoid Tetraether) index for reconstructing soil organic matter input to the ocean. An initial round-robin study of two sediment extracts, in which 15 laboratories participated, showed relatively consistent TEX86 values (reproducibility ±3-4°C when translated to temperature) but a large spread in BIT measurements (reproducibility ±0.41 on a scale of 0-1). Here we report results of a second round-robin study with 35 laboratories in which three sediments, one sediment extract, and two mixtures of pure, isolated GDGTs were analyzed. The results for TEX86 and BIT index showed improvement compared to the previous round-robin study. The reproducibility, indicating interlaboratory variation, of TEX86 values ranged from 1.3 to 3.0°C when translated to temperature. These results are similar to those of other temperature proxies used in paleoceanography. Comparison of the results obtained from one of the three sediments showed that TEX86 and BIT indices are not significantly affected by interlaboratory differences in sediment extraction techniques. BIT values of the sediments and extracts were at the extremes of the index with values close to 0 or 1, and showed good reproducibility (ranging from 0.013 to 0.042). However, the measured BIT values for the two GDGT mixtures, with known molar ratios of crenarchaeol and branched GDGTs, had intermediate BIT values and showed poor reproducibility and a large overestimation of the "true" (i.e., molar-based) BIT index. The latter is likely due to, among other factors, the higher mass spectrometric response of branched GDGTs compared to crenarchaeol, which also varies among mass spectrometers. Correction for this different mass spectrometric response showed a considerable improvement in the reproducibility of BIT index measurements among laboratories, as well as a substantially improved estimation of molar-based BIT values. This suggests that standard mixtures should be used in order to obtain consistent, and molar-based, BIT values

Advances in methods for detection of anaerobic ammonium oxidizing (anammox) bacteria

Anaerobic ammonium oxidation (anammox), the biochemical process oxidizing ammonium into dinitrogen gas using nitrite as an electron acceptor, has only been recognized for its significant role in the global nitrogen cycle not long ago, and its ubiquitous distribution in a wide range of environments has changed our knowledge about the contributors to the global nitrogen cycle. Currently, several groups of methods are used in detection of anammox bacteria based on their physiological and biochemical characteristics, cellular chemical composition, and both 16S rRNA gene and selective functional genes as biomarkers, including hydrazine oxidoreductase and nitrite reductase encoding genes hzo and nirS, respectively. Results from these methods coupling with advances in quantitative PCR, reverse transcription of mRNA genes and stable isotope labeling have improved our understanding on the distribution, diversity, and activity of anammox bacteria in different environments both natural and engineered ones. In this review, we summarize these methods used in detection of anammox bacteria from various environments, highlight the strengths and weakness of these methods, and also discuss the new development potentials on the existing and new techniques in the future

Fossilization and degradation of archaeal intact polar tetraether lipids in deeply buried marine sediments (Peru Margin)

Glycerol dibiphytanyl glycerol tetraether (GDGT) lipids are part of the cellular membranes of Thaumarchaeota, an archaeal phylum composed of aerobic ammonia oxidizers, and are used in the paleotemperature proxy TEX86. GDGTs in live cells possess polar head groups and are called intact polar lipids (IPL-GDGTs). Their transformation to core lipids (CL) by cleavage of the head group was assumed to proceed rapidly after cell death, but it has been suggested that some of these IPL-GDGTs can, just like the CL-GDGTs, be preserved over geological timescales. Here, we examined IPL-GDGTs in deeply buried (0.2–186 mbsf, ~2.5 Myr) sediments from the Peru Margin. Direct measurements of the most abundant IPL-GDGT, IPL-crenarchaeol, specific for Thaumarchaeota, revealed depth profiles, which differed per head group. Shallow sediments (1 m) marine sediments, which only reported glycosidic and no phosphate-containing IPL-GDGTs. TEX86 values of CL-GDGTs did not markedly change with depth, and the TEX86 of IPL-derived GDGTs decreased only when the proportions of monohexose- to dihexose-GDGTs changed, likely due to the enhanced preservation of the monohexose GDGTs. Our results support the hypothesis that in situ GDGT production and differential IPL degradation in sediments is not substantially affecting TEX86 paleotemperature estimations based on CL–GDGTs and indicates that likely only a small amount of IPL-GDGTs present in deeply buried sediments is part of cell membranes of active archaea. The amount of archaeal biomass in the deep biosphere based on these IPLs may have been substantially overestimated

Anoxic in situ production of bacterial GMGTs in the water column and surficial bottom sediments of a meromictic tropical crater lake: Implications for lake paleothermometry

Branched glycerol monoalkyl glycerol tetraethers (brGMGTs) are membrane-spanning lipids which were initially identified in marine settings and have more recently been found in peats and lake sediments. In the latter settings, their abundance relative to that of the branched glycerol dialkyl glycerol tetraethers (brGDGTs) appears to increase under warmer climate conditions. Furthermore, their relative distribution in bottom sediments from an altitudinal transect of East African lakes has been linked to local mean annual air temperature (MAAT). To shed light on their sources and identify potential environmental factors driving their production in lakes, we investigated their occurrence and distribution in the water column, surficial lake-bottom sediments, and catchment soils of Lake Chala, a permanently stratified (meromictic) tropical crater lake in East Africa. Data from the water column comprise both suspended particulate matter (SPM) collected monthly at eight depth intervals between 0 and 80 m for 17 consecutive months, and settling particles collected monthly at a depth of 35 m over a period of 53 months. In catchment soils, brGMGTs are either relatively scarce or absent. In the water column, their occurrence is dominated by four of the seven known brGMGTs and mostly limited to the anoxic zone. Whereas the spatial distribution and concentration of brGMGTs in SPM were strongly influenced by the seasonal cycle of strong water-column stratification alternating with deep mixing, this trend is not consistent on the multi-year timescale of the settling-particles record. Moreover, their relative distribution in both SPM and settling particles did not respond to temperature variation during the studied interval. In contrast to the water column, surficial lake-bottom sediments contain the full suite of known brGMGTs in a distinct and spatially uniform fractional abundance, strongly suggesting that the majority of brGMGT production takes place there. BrGMGT-inferred MAAT for the Chala area reasonably estimates measured MAAT and conforms to the East African lake brGMGT calibration. Our results have important implications for the use of brGMGTs as proxy for MAAT, in that outside of the equatorial zone the presumably greater offset between bottom-water temperature and MAAT may create a bias towards cold-season air temperature. Regardless, the more singular source of brGMGTs in lake sediments is an advantage for paleotemperature reconstruction compared to brGDGTs, which several studies now indicate to have a mixed aquatic and terrestrial source of which the relative contribution cannot easily be disentangled or assumed to have been constant through time. (C) 2021 The Author(s). Published by Elsevier Ltd