Philippa Kaur

Overmann, J., Abt, B. & Sikorski, J. Present and future of culturing bacteria. Annual Review of Microbiology 71, 711–730 (2017).CAS 

Google Scholar 
O’Leary, N. A. et al. Reference sequence (RefSeq) database at NCBI: current status, taxonomic expansion, and functional annotation. Nucleic Acids Research 44, D733 (2016).
Google Scholar 
Bobay, L. M. & Ochman, H. Biological species are universal across life’s domains. Genome Biology and Evolution 9, 491–501 (2017).
Google Scholar 
Magnabosco, C., Moore, K., Wolfe, J. & Fournier, G. Dating phototrophic microbial lineages with reticulate gene histories. Geobiology 16, 179–189 (2018).CAS 

Google Scholar 
Louca, S. et al. Function and functional redundancy in microbial systems. Nature Ecology & Evolution 2, 936–943 (2018).ADS 

Google Scholar 
Jain, C., Rodriguez-R, L. M., Phillippy, A. M., Konstantinidis, K. T. & Aluru, S. High throughput ANI analysis of 90 K prokaryotic genomes reveals clear species boundaries. Nature Communications 9, 5114 (2018).ADS 

Google Scholar 
Zhu, Q. et al. Phylogenomics of 10,575 genomes reveals evolutionary proximity between domains bacteria and archaea. Nature Communications 10, 5477 (2019).ADS 
CAS 

Google Scholar 
Royalty, T.M. & Steen, A.D. Quantitatively partitioning microbial genomic traits among taxonomic ranks across the microbial tree of life. mSphere 4 (2019).Murray, C. S., Gao, Y. & Wu, M. Re-evaluating the evidence for a universal genetic boundary among microbial species. Nature Communications 12, 4059 (2021).ADS 
CAS 

Google Scholar 
Powell, S. et al. eggNOG v4.0: nested orthology inference across 3686 organisms. Nucleic Acids Research 42, D231–D239 (2014).CAS 

Google Scholar 
Stoddard, S. F., Smith, B. J., Hein, R., Roller, B. R. & Schmidt, T. M. rrnDB: improved tools for interpreting rRNA gene abundance in bacteria and archaea and a new foundation for future development. Nucleic Acids Research 43, D593–D598 (2014).
Google Scholar 
Douglas, G. M. et al. Picrust2 for prediction of metagenome functions. Nature Biotechnology 38, 685–688 (2020).CAS 

Google Scholar 
Wemheuer, F. et al. Tax4Fun2: prediction of habitat-specific functional profiles and functional redundancy based on 16S rRNA gene sequences. Environmental Microbiome 15, 1–12 (2020).
Google Scholar 
Louca, S., Parfrey, L. W. & Doebeli, M. Decoupling function and taxonomy in the global ocean microbiome. Science 353, 1272–1277 (2016).ADS 
CAS 

Google Scholar 
Wu, D. et al. A phylogeny-driven genomic encyclopaedia of bacteria and archaea. Nature 462, 1056–1060 (2009).ADS 
CAS 

Google Scholar 
Louca, S. & Pennell, M. W. A general and efficient algorithm for the likelihood of diversification and discrete-trait evolutionary models. Systematic Biology 69, 545–556 (2020).
Google Scholar 
Tyson, G. W. et al. Community structure and metabolism through reconstruction of microbial genomes from the environment. Nature 428, 37–43 (2004).ADS 
CAS 

Google Scholar 
Sharon, I. & Banfield, J. F. Genomes from metagenomics. Science 342, 1057–1058 (2013).ADS 
CAS 

Google Scholar 
Parks, D. H. et al. Recovery of nearly 8,000 metagenome-assembled genomes substantially expands the tree of life. Nature Microbiology 2, 1533–1542 (2017).CAS 

Google Scholar 
Chen, L. X., Anantharaman, K., Shaiber, A., Eren, A. M. & Banfield, J. F. Accurate and complete genomes from metagenomes. Genome Research 30, 315–333 (2020).CAS 

Google Scholar 
Konstantinidis, K. T. & Tiedje, J. M. Genomic insights that advance the species definition for prokaryotes. Proceedings of the National Academy of Sciences 102, 2567–2572 (2005).ADS 
CAS 

Google Scholar 
Kim, M., Oh, H. S., Park, S. C. & Chun, J. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Journal of Systematic and Evolutionary Microbiology 64, 346–351 (2014).CAS 

Google Scholar 
Shapiro, B.J. What microbial population genomics has taught us about speciation. In Polz, M.F. & Rajora, O.P. (eds.) Population Genomics: Microorganisms, 31–47 (Springer International Publishing, Cham, Switzerland, 2019).Olm, M. R. et al. Consistent metagenome-derived metrics verify and delineate bacterial species boundaries. mSystems 5, e00731–19 (2020).CAS 

Google Scholar 
Lagkouvardos, I., Overmann, J. & Clavel, T. Cultured microbes represent a substantial fraction of the human and mouse gut microbiota. Gut Microbes 8, 493–503 (2017).
Google Scholar 
Zhang, Z., Wang, J., Wang, J., Wang, J. & Li, Y. Estimate of the sequenced proportion of the global prokaryotic genome. Microbiome 8, 1–9 (2020).
Google Scholar 
Aramaki, T. et al. KofamKOALA: KEGG Ortholog assignment based on profile HMM and adaptive score threshold. Bioinformatics 36, 2251–2252 (2019).
Google Scholar 
Mira, A., Ochman, H. & Moran, N. A. Deletional bias and the evolution of bacterial genomes. Trends in Genetics 17, 589–596 (2001).CAS 

Google Scholar 
Morris, J. J., Lenski, R. E. & Zinser, E. R. The Black Queen Hypothesis: evolution of dependencies through adaptive gene loss. MBio 3, e00036–12 (2012).
Google Scholar 
Giovannoni, S. J., Cameron Thrash, J. & Temperton, B. Implications of streamlining theory for microbial ecology. ISME Journal 8, 1553–1565 (2014).
Google Scholar 
Nayfach, S., Shi, Z. J., Seshadri, R., Pollard, K. S. & Kyrpides, N. C. New insights from uncultivated genomes of the global human gut microbiome. Nature 568, 505–510 (2019).ADS 
CAS 

Google Scholar 
Gary, P.R. Adjusting for nonresponse in surveys. In Smart, J.C. (ed.) Higher Education: Handbook of Theory and Research, chap. 8, 411–449 (Springer, Dordrecht, Netherlands, 2007).Maguire, F. et al. Metagenome-assembled genome binning methods with short reads disproportionately fail for plasmids and genomic islands. Microbial Genomics 6, mgen000436 (2020).
Google Scholar 
Huerta-Cepas, J. et al. eggnog 5.0: a hierarchical, functionally and phylogenetically annotated orthology resource based on 5090 organisms and 2502 viruses. Nucleic Acids Research 47, D309–D314 (2019).CAS 

Google Scholar 
Abdel-Hamid, A.M., Solbiati, J.O., Cann, I.K.O., Sariaslani, S. & Gadd, G.M. Insights into lignin degradation and its potential industrial applications, vol. 82, chap. 1, 1–28 (Academic Press, 2013).El-Bondkly, A.M. Sequence analysis of industrially important genes from trichoderma. In Biotechnology and biology of Trichoderma, chap. 28, 377–392 (Elsevier, 2014).Dawood, A. & Ma, K. Applications of microbial β-mannanases. Frontiers in Bioengineering and Biotechnology 8 (2020).Khelaifia, S., Raoult, D. & Drancourt, M. A versatile medium for cultivating methanogenic archaea. PLOS ONE 8, e61563 (2013).ADS 
CAS 

Google Scholar 
Khelaifia, S. et al. Aerobic culture of methanogenic archaea without an external source of hydrogen. European Journal of Clinical Microbiology & Infectious Diseases 35, 985–991 (2016).CAS 

Google Scholar 
Michał, B. et al. Phymet2: a database and toolkit for phylogenetic and metabolic analyses of methanogens. Environmental Microbiology Reports 10, 378–382 (2018).
Google Scholar 
Albright, S. & Louca, S. Trait biases in microbial reference genomes, figshare., https://doi.org/10.6084/m9.figshare.c.6055004.v1 (2022).Castelle, C. J. & Banfield, J. F. Major new microbial groups expand diversity and alter our understanding of the tree of life. Cell 172, 1181–1197 (2018).CAS 

Google Scholar 
Murray, A. E. et al. Roadmap for naming uncultivated archaea and bacteria. Nature Microbiology 5, 987–994 (2020).CAS 

Google Scholar 
Palleroni, N. J. Prokaryotic diversity and the importance of culturing. Antonie van Leeuwenhoek 72, 3–19 (1997).CAS 

Google Scholar 
Langille, M. G. et al. Predictive functional profiling of microbial communities using 16S rRNA marker gene sequences. Nature Biotechnology 31, 814–821 (2013).CAS 

Google Scholar 
Tran, P. Q. et al. Depth-discrete metagenomics reveals the roles of microbes in biogeochemical cycling in the tropical freshwater Lake Tanganyika. The ISME Journal 15, 1971–1986 (2021).CAS 

Google Scholar 
Kroeger, M. E. et al. New biological insights into how deforestation in amazonia affects soil microbial communities using metagenomics and metagenome-assembled genomes. Frontiers in Microbiology 9, 1635 (2018).
Google Scholar 
Nathani, N. M. et al. 309 metagenome assembled microbial genomes from deep sediment samples in the Gulfs of Kathiawar Peninsula. Scientific Data 8, 194 (2021).
Google Scholar 
Irazoqui, J. M., Eberhardt, M. F., Adjad, M. M., Amadio, A. F. & Collado, M. C. Identification of key microorganisms in facultative stabilization ponds from dairy industries, using metagenomics. PeerJ 10, e12772 (2022).
Google Scholar 
Hwang, Y. et al. Leave no stone unturned: individually adapted xerotolerant Thaumarchaeota sheltered below the boulders of the Atacama Desert hyperarid core. Microbiome 9, 234 (2021).CAS 

Google Scholar 
Tully, B., Wheat, C. G., Glazer, B. T. & Huber, J. A dynamic microbial community with high functional redundancy inhabits the cold, oxic subseafloor aquifer. ISME Journal 12, 1–16 (2018).CAS 

Google Scholar 
Vanwonterghem, I., Jensen, P. D., Rabaey, K. & Tyson, G. W. Genome-centric resolution of microbial diversity, metabolism and interactions in anaerobic digestion. Environmental Microbiology 18, 3144–3158 (2016).CAS 

Google Scholar 
Glasl, B. et al. Comparative genome-centric analysis reveals seasonal variation in the function of coral reef microbiomes. The ISME Journal 14, 1435–1450 (2020).
Google Scholar 
Robbins, S. J. et al. A genomic view of the reef-building coral Porites lutea and its microbial symbionts. Nature Microbiology 4, 2090–2100 (2019).
Google Scholar 
Engelberts, J. P. et al. Characterization of a sponge microbiome using an integrative genome-centric approach. The ISME Journal 14, 1100–1110 (2020).CAS 

Google Scholar 
Bowerman, K. L. et al. Disease-associated gut microbiome and metabolome changes in patients with chronic obstructive pulmonary disease. Nature Communications 11, 5886 (2020).ADS 
CAS 

Google Scholar 
Chen, Y. J. et al. Hydrodynamic disturbance controls microbial community assembly and biogeochemical processes in coastal sediments. The ISME Journal 16, 750–763 (2022).CAS 

Google Scholar 
Hugerth, L. W. et al. Metagenome-assembled genomes uncover a global brackish microbiome. Genome Biology 16, 279 (2015).
Google Scholar 
Alneberg, J. et al. Ecosystem-wide metagenomic binning enables prediction of ecological niches from genomes. Communications Biology 3, 119 (2020).
Google Scholar 
Di Cesare, A. et al. Genomic comparison and spatial distribution of different Synechococcus phylotypes in the Black Sea. Frontiers in Microbiology 11, 1979 (2020).
Google Scholar 
van Vliet, D. M. et al. The bacterial sulfur cycle in expanding dysoxic and euxinic marine waters. Environmental Microbiology 23, 2834–2857 (2021).
Google Scholar 
Dalcin Martins, P. et al. Enrichment of novel Verrucomicrobia, Bacteroidetes, and Krumholzibacteria in an oxygen-limited methane- and iron-fed bioreactor inoculated with Bothnian Sea sediments. MicrobiologyOpen 10, e1175 (2021).CAS 

Google Scholar 
Stewart, R. D. et al. Compendium of 4,941 rumen metagenome-assembled genomes for rumen microbiome biology and enzyme discovery. Nature Biotechnology 37, 953–961 (2019).CAS 

Google Scholar 
Segura-Wang, M., Grabner, N., Koestelbauer, A., Klose, V. & Ghanbari, M. Genome-resolved metagenomics of the chicken gut microbiome. Frontiers in Microbiology 12, 726923 (2021).
Google Scholar 
Ruuskanen, M. O. et al. Microbial genomes retrieved from High Arctic lake sediments encode for adaptation to cold and oligotrophic environments. Limnology and Oceanography 65, S233–S247 (2020).CAS 

Google Scholar 
Haas, S., Desai, D. K., LaRoche, J., Pawlowicz, R. & Wallace, D. W. R. Geomicrobiology of the carbon, nitrogen and sulphur cycles in Powell Lake: a permanently stratified water column containing ancient seawater. Environmental Microbiology 21, 3927–3952 (2019).CAS 

Google Scholar 
Spasov, E. et al. High functional diversity among Nitrospira populations that dominate rotating biological contactor microbial communities in a municipal wastewater treatment plant. The ISME Journal 14, 1857–1872 (2020).CAS 

Google Scholar 
Vigneron, A. et al. Genomic evidence for sulfur intermediates as new biogeochemical hubs in a model aquatic microbial ecosystem. Microbiome 9, 46 (2021).CAS 

Google Scholar 
Galambos, D., Anderson, R. E., Reveillaud, J. & Huber, J. A. Genome-resolved metagenomics and metatranscriptomics reveal niche differentiation in functionally redundant microbial communities at deep-sea hydrothermal vents. Environmental Microbiology 21, 4395–4410 (2019).CAS 

Google Scholar 
Stewart, R. D. et al. Assembly of 913 microbial genomes from metagenomic sequencing of the cow rumen. Nature Communications 9, 870 (2018).ADS 

Google Scholar 
Xing, P. et al. Stratification of microbiomes during the holomictic period of Lake Fuxian, an alpine monomictic lake. Limnology and Oceanography 65, S134–S148 (2020).
Google Scholar 
Zhang, S., Hu, Z. & Wang, H. Metagenomic analysis exhibited the co-metabolism of polycyclic aromatic hydrocarbons by bacterial community from estuarine sediment. Environment International 129, 308–319 (2019).CAS 

Google Scholar 
Lin, Y., Wang, L., Xu, K., Li, K. & Ren, H. Revealing taxon-specific heavy metal-resistance mechanisms in denitrifying phosphorus removal sludge using genome-centric metaproteomics. Microbiome 9, 67 (2021).CAS 

Google Scholar 
Liu, L. et al. High-quality bacterial genomes of a partial-nitritation/anammox system by an iterative hybrid assembly method. Microbiome 8, 155 (2020).CAS 

Google Scholar 
Kantor, R. S. et al. Bioreactor microbial ecosystems for thiocyanate and cyanide degradation unravelled with genome-resolved metagenomics. Environmental Microbiology 17, 4929–4941 (2015).CAS 

Google Scholar 
Zhou, Z. et al. Gammaproteobacteria mediating utilization of methyl-, sulfur- and petroleum organic compounds in deep ocean hydrothermal plumes. The ISME Journal 14, 3136–3148 (2020).CAS 

Google Scholar 
Reysenbach, A. L. et al. Complex subsurface hydrothermal fluid mixing at a submarine arc volcano supports distinct and highly diverse microbial communities. Proceedings of the National Academy of Sciences 117, 32627–32638 (2020).ADS 
CAS 

Google Scholar 
Hou, J. et al. Microbial succession during the transition from active to inactive stages of deep-sea hydrothermal vent sulfide chimneys. Microbiome 8, 102 (2020).CAS 

Google Scholar 
Campanaro, S. et al. Metagenomic analysis and functional characterization of the biogas microbiome using high throughput shotgun sequencing and a novel binning strategy. Biotechnology for Biofuels 9, 26 (2016).
Google Scholar 
Singleton, C. M. et al. Connecting structure to function with the recovery of over 1000 high-quality metagenome-assembled genomes from activated sludge using long-read sequencing. Nature Communications 12, 2009 (2021).CAS 

Google Scholar 
Diamond, S. et al. Mediterranean grassland soil C–N compound turnover is dependent on rainfall and depth, and is mediated by genomically divergent microorganisms. Nature Microbiology 4, 1356–1367 (2019).CAS 

Google Scholar 
Rasigraf, O. et al. Microbial community composition and functional potential in Bothnian Sea sediments is linked to Fe and S dynamics and the quality of organic matter. Limnology and Oceanography 65, S113–S133 (2020).CAS 

Google Scholar 
Rissanen, A. J. et al. Vertical stratification patterns of methanotrophs and their genetic controllers in water columns of oxygen-stratified boreal lakes. FEMS Microbiology Ecology 97, fiaa252 (2021).CAS 

Google Scholar 
Campanaro, S. et al. New insights from the biogas microbiome by comprehensive genome-resolved metagenomics of nearly 1600 species originating from multiple anaerobic digesters. Biotechnology for Biofuels 13, 25 (2020).CAS 

Google Scholar 
Almeida, A. et al. A unified catalog of 204,938 reference genomes from the human gut microbiome. Nature Biotechnology 39, 105–114 (2021).CAS 

Google Scholar 
Zhou, Z. et al. Genome- and community-level interaction insights into carbon utilization and element cycling functions of hydrothermarchaeota in hydrothermal sediment. mSystems 5 (2020).Pachiadaki, M. G. et al. Charting the complexity of the marine microbiome through single-cell genomics. Cell 179, 1623–1635.e11 (2019).CAS 

Google Scholar 
Martijn, J., Vosseberg, J., Guy, L., Offre, P. & Ettema, T. J. G. Deep mitochondrial origin outside the sampled alphaproteobacteria. Nature 557, 101–105 (2018).ADS 
CAS 

Google Scholar 
Greenlon, A. et al. Global-level population genomics reveals differential effects of geography and phylogeny on horizontal gene transfer in soil bacteria. Proceedings of the National Academy of Sciences 116, 15200–15209 (2019).ADS 
CAS 

Google Scholar 
Hervé, V. et al. Phylogenomic analysis of 589 metagenome-assembled genomes encompassing all major prokaryotic lineages from the gut of higher termites. PeerJ 8, e8614 (2020).
Google Scholar 
von Appen, W.J. The expedition PS114 of the research vessel POLARSTERN to the Fram Strait in 2018. Tech. Rep., Alfred Wegener Institute for Polar and Marine Research (2018).Dombrowski, N., Seitz, K. W., Teske, A. P. & Baker, B. J. Genomic insights into potential interdependencies in microbial hydrocarbon and nutrient cycling in hydrothermal sediments. Microbiome 5, 106 (2017).
Google Scholar 
Yu, J. et al. Dna-stable isotope probing shotgun metagenomics reveals the resilience of active microbial communities to biochar amendment in oxisol soil. Frontiers in Microbiology 11, 587972 (2020).
Google Scholar 
Forster, S. C. et al. A human gut bacterial genome and culture collection for improved metagenomic analyses. Nature Biotechnology 37, 186–192 (2019).CAS 

Google Scholar 
Gharechahi, J. et al. Metagenomic analysis reveals a dynamic microbiome with diversified adaptive functions to utilize high lignocellulosic forages in the cattle rumen. The ISME Journal 15, 1108–1120 (2021).CAS 

Google Scholar 
Meier, D. V., Imminger, S., Gillor, O. & Woebken, D. Distribution of mixotrophy and desiccation survival mechanisms across microbial genomes in an arid biological soil crust community. mSystems 6, e00786–20 (2021).CAS 

Google Scholar 
Haro-Moreno, J. M. et al. Dysbiosis in marine aquaculture revealed through microbiome analysis: reverse ecology for environmental sustainability. FEMS Microbiology Ecology 96, fiaa218 (2020).CAS 

Google Scholar 
Haro-Moreno, J. M. et al. Fine metagenomic profile of the Mediterranean stratified and mixed water columns revealed by assembly and recruitment. Microbiome 6, 128 (2018).
Google Scholar 
Dong, X. et al. Metabolic potential of uncultured bacteria and archaea associated with petroleum seepage in deep-sea sediments. Nature Communications 10, 1816 (2019).ADS 

Google Scholar 
Poghosyan, L. et al. Metagenomic profiling of ammonia- and methane-oxidizing microorganisms in two sequential rapid sand filters. Water Research 185, 116288 (2020).CAS 

Google Scholar 
Paula, D. M., Jeroen, F., Hugh, M. & Meng, M. L. & J., W.M. Wetland sediments host diverse microbial taxa capable of cycling alcohols. Applied and Environmental Microbiology 85, 00189–19 (2019).
Google Scholar 
Aromokeye, D. A. et al. Crystalline iron oxides stimulate methanogenic benzoate degradation in marine sediment-derived enrichment cultures. The ISME Journal 15, 965–980 (2021).CAS 

Google Scholar 
Borchert, E. et al. Deciphering a marine bone-degrading microbiome reveals a complex community effort. mSystems 6, e01218–20 (2021).CAS 

Google Scholar 
Osvatic, J. T. et al. Global biogeography of chemosynthetic symbionts reveals both localized and globally distributed symbiont groups. Proceedings of the National Academy of Sciences 118, e2104378118 (2021).CAS 

Google Scholar 
Boeuf, D. et al. Biological composition and microbial dynamics of sinking particulate organic matter at abyssal depths in the oligotrophic open ocean. Proceedings of the National Academy of Sciences 116, 11824–11832 (2019).ADS 
CAS 

Google Scholar 
Woodcroft, B. J. et al. Genome-centric view of carbon processing in thawing permafrost. Nature 560, 49–54 (2018).ADS 
CAS 

Google Scholar 
Alqahtani, M. F. et al. Enrichment of Marinobacter sp. and halophilic homoacetogens at the biocathode of microbial electrosynthesis system inoculated with Red Sea brine pool. Frontiers in Microbiology 10, 2563 (2019).
Google Scholar 
Haroon, M. F., Thompson, L. R., Parks, D. H., Hugenholtz, P. & Stingl, U. A catalogue of 136 microbial draft genomes from Red Sea metagenomes. Scientific Data 3, 160050 (2016).CAS 

Google Scholar 
Vavourakis, C. D. et al. A metagenomics roadmap to the uncultured genome diversity in hypersaline soda lake sediments. Microbiome 6, 1–18 (2018).
Google Scholar 
Cabello-Yeves, P. J. et al. Microbiome of the deep Lake Baikal, a unique oxic bathypelagic habitat. Limnology and Oceanography 65, 1471–1488 (2020).ADS 
CAS 

Google Scholar 
Vavourakis, C. D. et al. Metagenomes and metatranscriptomes shed new light on the microbial-mediated sulfur cycle in a siberian soda lake. BMC Biology 17, 69 (2019).
Google Scholar 
Waterworth, S. C., Isemonger, E. W., Rees, E. R., Dorrington, R. A. & Kwan, J. C. Conserved bacterial genomes from two geographically isolated peritidal stromatolite formations shed light on potential functional guilds. Environmental Microbiology Reports 13, 126–137 (2021).CAS 

Google Scholar 
Huddy, R. J. et al. Thiocyanate and organic carbon inputs drive convergent selection for specific autotrophic Afipia and Thiobacillus strains within complex microbiomes. Frontiers in Microbiology 12, 643368 (2021).
Google Scholar 
Emerson, J. B. et al. Diverse sediment microbiota shape methane emission temperature sensitivity in Arctic lakes. Nature Communications 12, 5815 (2021).ADS 
CAS 

Google Scholar 
Chiri, E. et al. Termite gas emissions select for hydrogenotrophic microbial communities in termite mounds. Proceedings of the National Academy of Sciences 118, e2102625118 (2021).CAS 

Google Scholar 
Gong, G., Zhou, S., Luo, R., Gesang, Z. & Suolang, S. Metagenomic insights into the diversity of carbohydrate-degrading enzymes in the yak fecal microbial community. BMC Microbiology 20, 302 (2020).
Google Scholar 
Zhou, S. et al. Characterization of metagenome-assembled genomes and carbohydrate-degrading genes in the gut microbiota of Tibetan pig. Frontiers in Microbiology 11, 595066 (2020).
Google Scholar 
Tully, B. J., Graham, E. D. & Heidelberg, J. F. The reconstruction of 2,631 draft metagenome-assembled genomes from the global oceans. Scientific Data 5, 170203 (2018).CAS 

Google Scholar 
Lavrinienko, A. et al. Two hundred and fifty-four metagenome-assembled bacterial genomes from the bank vole gut microbiota. Scientific Data 7, 312 (2020).CAS 

Google Scholar 
Peng, X. et al. Genomic and functional analyses of fungal and bacterial consortia that enable lignocellulose breakdown in goat gut microbiomes. Nature Microbiology 6, 499–511 (2021).CAS 

Google Scholar 
Dudek, N. K. et al. Novel microbial diversity and functional potential in the marine mammal oral microbiome. Current Biology 27, 3752–3762.e6 (2017).CAS 

Google Scholar 
Pinto, A. J. et al. Metagenomic evidence for the presence of comammox nitrospira-like bacteria in a drinking water system. mSphere 1, e00054–15 (2015).
Google Scholar 
Zaremba-Niedzwiedzka, K. et al. Asgard archaea illuminate the origin of eukaryotic cellular complexity. Nature 541, 353–358 (2017).ADS 
CAS 

Google Scholar 
Nobu, M. K. et al. Catabolism and interactions of uncultured organisms shaped by eco-thermodynamics in methanogenic bioprocesses. Microbiome 8, 111 (2020).CAS 

Google Scholar 
Butterfield, C. N. et al. Proteogenomic analyses indicate bacterial methylotrophy and archaeal heterotrophy are prevalent below the grass root zone. PeerJ 4, e2687 (2016).
Google Scholar 
Castelle, C. J. et al. Protein family content uncovers lineage relationships and bacterial pathway maintenance mechanisms in DPANN Archaea. Frontiers in Microbiology 12, 660052 (2021).
Google Scholar 
Alteio, L. V. et al. Complementary metagenomic approaches improve reconstruction of microbial diversity in a forest soil. mSystems 5, e00768–19 (2020).
Google Scholar 
Shaiber, A. et al. Functional and genetic markers of niche partitioning among enigmatic members of the human oral microbiome. Genome Biology 21, 292 (2020).
Google Scholar 
Jungbluth, S. P., Amend, J. P. & Rappé, M. S. Metagenome sequencing and 98 microbial genomes from Juan de Fuca Ridge flank subsurface fluids. Scientific Data 4, 170037 (2017).CAS 

Google Scholar 
Sheik, C. S. et al. Dolichospermum blooms in Lake Superior: DNA-based approach provides insight to the past, present and future of blooms. Journal of Great Lakes Research 48, 1191–1205 (2022).CAS 

Google Scholar 
Barnum, T. P. et al. Genome-resolved metagenomics identifies genetic mobility, metabolic interactions, and unexpected diversity in perchlorate-reducing communities. The ISME Journal 12, 1568–1581 (2018).CAS 

Google Scholar 
Julian, D. et al. Coastal ocean metagenomes and curated metagenome-assembled genomes from Marsh Landing, Sapelo Island (Georgia, USA). Microbiology Resource Announcements 8, e00934–19 (2019).
Google Scholar 
Breister, A. M. et al. Soil microbiomes mediate degradation of vinyl ester-based polymer composites. Communications Materials 1, 101 (2020).ADS 

Google Scholar 
Fu, H., Uchimiya, M., Gore, J. & Moran, M. A. Ecological drivers of bacterial community assembly in synthetic phycospheres. Proceedings of the National Academy of Sciences 117, 3656–3662 (2020).ADS 
CAS 

Google Scholar 
Nobu, M. K. et al. Thermodynamically diverse syntrophic aromatic compound catabolism. Environmental Microbiology 19, 4576–4586 (2017).CAS 

Google Scholar 
Pasolli, E. et al. Extensive unexplored human microbiome diversity revealed by over 150,000 genomes from metagenomes spanning age, geography, and lifestyle. Cell 176, 649–662 (2019).CAS 

Google Scholar 
Nayfach, S. et al. A genomic catalog of Earth’s microbiomes. Nature Biotechnology 39, 499–509 (2021).CAS 

Google Scholar 
Li, Z. et al. Deep sea sediments associated with cold seeps are a subsurface reservoir of viral diversity. The ISME Journal 15, 2366–2378 (2021).CAS 

Google Scholar 
Bay, S. K. et al. Trace gas oxidizers are widespread and active members of soil microbial communities. Nature Microbiology 6, 246–256 (2021).CAS 

Google Scholar 
Seyler, L. M., Trembath-Reichert, E., Tully, B. J. & Huber, J. A. Time-series transcriptomics from cold, oxic subseafloor crustal fluids reveals a motile, mixotrophic microbial community. The ISME Journal 15, 1192–1206 (2021).CAS 

Google Scholar 
Herold, M. et al. Integration of time-series meta-omics data reveals how microbial ecosystems respond to disturbance. Nature Communications 11, 5281 (2020).ADS 
CAS 

Google Scholar 
Dong, X. et al. Thermogenic hydrocarbon biodegradation by diverse depth-stratified microbial populations at a Scotian Basin cold seep. Nature Communications 11, 5825 (2020).ADS 
CAS 

Google Scholar 
Thompson, L. R. et al. Metagenomic covariation along densely sampled environmental gradients in the Red Sea. The ISME Journal 11, 138–151 (2017).CAS 

Google Scholar 
Dominik, S., Daniela, Z., Anja, P., Katharina, R. & Rolf, D. Metagenome-assembled genome sequences from different wastewater treatment stages in Germany. Microbiology Resource Announcements 10, e00504–21 (2021).
Google Scholar 
Langwig, M. V. et al. Large-scale protein level comparison of Deltaproteobacteria reveals cohesive metabolic groups. The ISME Journal 16, 307–320 (2022).CAS 

Google Scholar 
Rezaei Somee, M. et al. Distinct microbial community along the chronic oil pollution continuum of the Persian Gulf converge with oil spill accidents. Scientific Reports 11, 11316 (2021).ADS 
CAS 

Google Scholar 
Gilroy, R. et al. Metagenomic investigation of the equine faecal microbiome reveals extensive taxonomic diversity. PeerJ 10, e13084 (2022).
Google Scholar 
Bhattarai, B., Bhattacharjee, A. S., Coutinho, F. H. & Goel, R. K. Viruses and their interactions with bacteria and archaea of hypersaline Great Salt Lake. Frontiers in Microbiology 12, 701414 (2021).
Google Scholar 
Liu, L. et al. Microbial diversity and adaptive strategies in the Mars-like Qaidam Basin, North Tibetan Plateau, China. Environmental Microbiology Reports (2022).Lin, H. et al. Mercury methylation by metabolically versatile and cosmopolitan marine bacteria. The ISME Journal 15, 1810–1825 (2021).CAS 

Google Scholar 
Martnez-Pérez, C. et al. Lifting the lid: nitrifying archaea sustain diverse microbial communities below the Ross Ice Shelf. SSRN (2020).Zhang, L. et al. Metagenomic insights into the effect of thermal hydrolysis pre-treatment on microbial community of an anaerobic digestion system. Science of The Total Environment 791, 148096 (2021).ADS 
CAS 

Google Scholar 
Starr, E. P. et al. Stable-isotope-informed, genome-resolved metagenomics uncovers potential cross-kingdom interactions in rhizosphere soil. mSphere 6, e00085–21 (2021).CAS 

Google Scholar 
Matthew, C. et al. Archaeal and bacterial metagenome-assembled genome sequences derived from pig feces. Microbiology Resource Announcements 11, 01142–21 (2022).
Google Scholar 
Wang, Y., Zhao, R., Liu, L., Li, B. & Zhang, T. Selective enrichment of comammox from activated sludge using antibiotics. Water Research 197, 117087 (2021).CAS 

Google Scholar 
Gilroy, R. et al. Extensive microbial diversity within the chicken gut microbiome revealed by metagenomics and culture. PeerJ 9, e10941 (2021).
Google Scholar 
Chen, Y. H. et al. Salvaging high-quality genomes of microbial species from a meromictic lake using a hybrid sequencing approach. Communications Biology 4, 996 (2021).CAS 

Google Scholar 
Beach, N. K., Myers, K. S., Donohue, T. J. & Noguera, D. R. Metagenomes from 25 low-abundance microbes in a partial nitritation anammox microbiome. Microbiology Resource Announcements 11, 00212–22 (2022).CAS 

Google Scholar 
Solanki, V. et al. Glycoside hydrolase from the GH76 family indicates that marine Salegentibacter sp. Hel_I_6 consumes alpha-mannan from fungi. The ISME Journal 16, 1818–1830 (2022).CAS 

Google Scholar 
Hiraoka, S. et al. Diverse DNA modification in marine prokaryotic and viral communities. Nucleic Acids Research 50, 1531–1550 (2022).CAS 

Google Scholar 
Haryono, M.A.S. et al. Recovery of high quality metagenome-assembled genomes from full-scale activated sludge microbial communities in a tropical climate using longitudinal metagenome sampling. Frontiers in Microbiology 13 (2022).Rodrguez-Ramos, J.A. et al. Microbial genome-resolved metaproteomic analyses frame intertwined carbon and nitrogen cycles in river hyporheic sediments. Research Square (2021).Kim, M., Cho, H. & Lee, W. Y. Distinct gut microbiotas between southern elephant seals and Weddell seals of Antarctica. Journal of Microbiology 58, 1018–1026 (2020).CAS 

Google Scholar 
Voorhies, A. A. et al. Cyanobacterial life at low O2: community genomics and function reveal metabolic versatility and extremely low diversity in a Great Lakes sinkhole mat. Geobiology 10, 250–267 (2012).CAS 

Google Scholar 
McDaniel, E. A. et al. Tbasco: trait-based comparative ‘omics identifies ecosystem-level and niche-differentiating adaptations of an engineered microbiome. ISME Communications 2, 111 (2022).
Google Scholar 
Wang, W. et al. Contrasting bacterial and archaeal distributions reflecting different geochemical processes in a sediment core from the Pearl River Estuary. AMB Express 10, 16 (2020).
Google Scholar 
Mandakovic, D. et al. Genome-scale metabolic models of Microbacterium species isolated from a high altitude desert environment. Scientific Reports 10, 5560 (2020).ADS 
CAS 

Google Scholar 
Wang, Y. et al. Seasonal prevalence of ammonia-oxidizing archaea in a full-scale municipal wastewater treatment plant treating saline wastewater revealed by a 6-year time-series analysis. Environmental Science & Technology 55, 2662–2673 (2021).ADS 
CAS 

Google Scholar 
Bulzu, P. A. et al. Casting light on Asgardarchaeota metabolism in a sunlit microoxic niche. Nature Microbiology 4, 1129–1137 (2019).CAS 

Google Scholar 
Karen, J. et al. Hydrogen-oxidizing bacteria are abundant in desert soils and strongly stimulated by hydration. mSystems 5, e01131–20 (2020).
Google Scholar 
Rust, M. et al. A multiproducer microbiome generates chemical diversity in the marine sponge Mycale hentscheli. Proceedings of the National Academy of Sciences 117, 9508–9518 (2020).ADS 
CAS 

Google Scholar 
Podowski, J. C., Paver, S. F., Newton, R. J. & Coleman, M. L. Genome streamlining, proteorhodopsin, and organic nitrogen metabolism in freshwater nitrifiers. mBio 13, e02379–21 (2022).
Google Scholar 
Coutinho, F. H. et al. New viral biogeochemical roles revealed through metagenomic analysis of Lake Baikal. Microbiome 8, 163 (2020).CAS 

Google Scholar 
Philippi, M. et al. Purple sulfur bacteria fix N2 via molybdenum-nitrogenase in a low molybdenum Proterozoic ocean analogue. Nature Communications 12, 4774 (2021).ADS 
CAS 

Google Scholar 
Katie, S. et al. Eight metagenome-assembled genomes provide evidence for microbial adaptation in 20,000- to 1,000,000-year-old Siberian permafrost. Applied and Environmental Microbiology 87, e00972–21 (2021).
Google Scholar 
Mert, K. et al. Unexpected abundance and diversity of phototrophs in mats from morphologically variable microbialites in Great Salt Lake, Utah. Applied and Environmental Microbiology 86, e00165–20 (2020).
Google Scholar 
Patin, N. V. et al. Gulf of Mexico blue hole harbors high levels of novel microbial lineages. The ISME Journal 15, 2206–2232 (2021).CAS 

Google Scholar 
Wang, J., Tang, X., Mo, Z. & Mao, Y. Metagenome-assembled genomes from Pyropia haitanensis microbiome provide insights into the potential metabolic functions to the seaweed. Frontiers in Microbiology 13, 857901 (2022).
Google Scholar 
Burgsdorf, I. et al. Lineage-specific energy and carbon metabolism of sponge symbionts and contributions to the host carbon pool. The ISME Journal 16, 1163–1175 (2022).CAS 

Google Scholar 
Suarez, C. et al. Disturbance-based management of ecosystem services and disservices in partial nitritation-anammox biofilms. npj Biofilms and Microbiomes 8, 47 (2022).CAS 

Google Scholar 
Kumar, D. et al. Textile industry wastewaters from Jetpur, Gujarat, India, are dominated by Shewanellaceae, Bacteroidaceae, and Pseudomonadaceae harboring genes encoding catalytic enzymes for textile dye degradation. Frontiers in Environmental Science 9, 720707 (2021).ADS 

Google Scholar 
Seitz, V. A. et al. Variation in root exudate composition influences soil microbiome membership and function. Applied and Environmental Microbiology 88, e00226–22 (2022).
Google Scholar 
Lindner, B. G. et al. Toward shotgun metagenomic approaches for microbial source tracking sewage spills based on laboratory mesocosms. Water Research 210, 117993 (2022).CAS 

Google Scholar 
Yancey, C. E. et al. Metagenomic and metatranscriptomic insights into population diversity of microcystis blooms: Spatial and temporal dynamics of mcy genotypes, including a partial operon that can be abundant and expressed. Applied and Environmental Microbiology 88, e02464–21 (2022).
Google Scholar 
Liu, L. et al. Charting the complexity of the activated sludge microbiome through a hybrid sequencing strategy. Microbiome 9, 205 (2021).CAS 

Google Scholar 
Speth, D. R. et al. Microbial communities of Auka hydrothermal sediments shed light on vent biogeography and the evolutionary history of thermophily. The ISME Journal 16, 1750–1764 (2022).CAS 

Google Scholar 
Blyton, M. D. J., Soo, R. M., Hugenholtz, P. & Moore, B. D. Maternal inheritance of the koala gut microbiome and its compositional and functional maturation during juvenile development. Environmental Microbiology 24, 475–493 (2022).CAS 

Google Scholar 
Nuccio, E. E. et al. Niche differentiation is spatially and temporally regulated in the rhizosphere. The ISME Journal 14, 999–1014 (2020).CAS 

Google Scholar 
Jaffe, A. L. et al. Long-term incubation of lake water enables genomic sampling of consortia involving planctomycetes and candidate phyla radiation bacteria. mSystems 7, e00223–22 (2022).
Google Scholar 
Cabral, L. et al. Gut microbiome of the largest living rodent harbors unprecedented enzymatic systems to degrade plant polysaccharides. Nature Communications 13, 629 (2022).ADS 
CAS 

Google Scholar 
Blyton, M. D. J., Soo, R. M., Hugenholtz, P. & Moore, B. D. Characterization of the juvenile koala gut microbiome across wild populations. Environmental Microbiology 24, 4209–4219 (2022).CAS 

Google Scholar 
Xu, B. et al. A holistic genome dataset of bacteria, archaea and viruses of the Pearl River estuary. Scientific Data 9, 49 (2022).MathSciNet 
CAS 

Google Scholar 
Royo-Llonch, M. et al. Compendium of 530 metagenome-assembled bacterial and archaeal genomes from the polar Arctic Ocean. Nature Microbiology 6, 1561–1574 (2021).CAS 

Google Scholar 
Sun, J., Prabhu, A., Aroney, S. T. N. & Rinke, C. Insights into plastic biodegradation: community composition and functional capabilities of the superworm (Zophobas morio) microbiome in styrofoam feeding trials. Microbial Genomics 8, 000842 (2022).CAS 

Google Scholar 
Kim, M. et al. Higher pathogen load in children from Mozambique vs. USA revealed by comparative fecal microbiome profiling. ISME Communications 2, 74 (2022).ADS 

Google Scholar 
Kelly, J. B., Carlson, D. E., Low, J. S. & Thacker, R. W. Novel trends of genome evolution in highly complex tropical sponge microbiomes. Microbiome 10, 164 (2022).CAS 

Google Scholar 
Bray, M. S. et al. Phylogenetic and structural diversity of aromatically dense pili from environmental metagenomes. Environmental Microbiology Reports 12, 49–57 (2020).CAS 

Google Scholar 
Cabello-Yeves, P. J. et al. α-cyanobacteria possessing form IA RuBisCO globally dominate aquatic habitats. The ISME Journal 16, 2421–2432 (2022).CAS 

Google Scholar 
Berben, T. et al. The Polar Fox Lagoon in Siberia harbours a community of Bathyarchaeota possessing the potential for peptide fermentation and acetogenesis. Antonie van Leeuwenhoek 115, 1229–1244 (2022).CAS 

Google Scholar 
Tamburini, F. B. et al. Short- and long-read metagenomics of urban and rural South African gut microbiomes reveal a transitional composition and undescribed taxa. Nature Communications 13, 926 (2022).ADS 
CAS 

Google Scholar 
Kantor, R. S., Miller, S. E. & Nelson, K. L. The water microbiome through a pilot scale advanced treatment facility for direct potable reuse. Frontiers in Microbiology 10, 993 (2019).
Google Scholar 
Muratore, D. et al. Complex marine microbial communities partition metabolism of scarce resources over the diel cycle. Nature Ecology & Evolution 6, 218–229 (2022).
Google Scholar 
Zhou, Y. L., Mara, P., Cui, G. J., Edgcomb, V. P. & Wang, Y. Microbiomes in the challenger deep slope and bottom-axis sediments. Nature Communications 13, 1515 (2022).ADS 
CAS 

Google Scholar 
Zhang, H. et al. Metagenome sequencing and 768 microbial genomes from cold seep in South China Sea. Scientific Data 9, 480 (2022).CAS 

Google Scholar 
Zhuang, J. L., Zhou, Y. Y., Liu, Y. D. & Li, W. Flocs are the main source of nitrous oxide in a high-rate anammox granular sludge reactor: insights from metagenomics and fed-batch experiments. Water Research 186, e116321 (2020).
Google Scholar 
Shiffman, M. E. et al. Gene and genome-centric analyses of koala and wombat fecal microbiomes point to metabolic specialization for eucalyptus digestion. PeerJ 5, 4075 (2017).
Google Scholar 
Murphy, S. M. C., Bautista, M. A., Cramm, M. A. & Hubert, C. R. J. Diesel and crude oil biodegradation by cold-adapted microbial communities in the Labrador Sea. Applied and Environmental Microbiology 87, e00800–21 (2021).ADS 
CAS 

Google Scholar 
Suarez, C. et al. Metagenomic evidence of a novel family of anammox bacteria in a subsea environment. Environmental Microbiology 24, 2348–2360 (2022).CAS 

Google Scholar 
Dharamshi, J.E. et al. Genomic diversity and biosynthetic capabilities of sponge-associated chlamydiae. The ISME Journal (2022).Florian, P. O., Hugo, R. & Mathieu, A. Recovery of metagenome-assembled genomes from a human fecal sample with pacific biosciences high-fidelity sequencing. Microbiology Resource Announcements 11, e00250–22 (2022).
Google Scholar 
Bloom, S. M. et al. Cysteine dependence of Lactobacillus iners is a potential therapeutic target for vaginal microbiota modulation. Nature Microbiology 7, 434–450 (2022).CAS 

Google Scholar 
Aylward, F. O. et al. Diel cycling and long-term persistence of viruses in the ocean’s euphotic zone. Proceedings of the National Academy of Sciences 114, 11446–11451 (2017).ADS 
CAS 

Google Scholar 
Parks, D. H., Imelfort, M., Skennerton, C. T., Hugenholtz, P. & Tyson, G. W. CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Research 25, 1043–1055 (2015).CAS 

Google Scholar 
Bowers, R. M. et al. Minimum information about a single amplified genome (MISAG) and a metagenome-assembled genome (MIMAG) of bacteria and archaea. Nature biotechnology 35, 725 (2017).CAS 

Google Scholar 
Chaumeil, P. A., Mussig, A. J., Hugenholtz, P. & Parks, D. H. GTDB-Tk: a toolkit to classify genomes with the Genome Taxonomy Database. Bioinformatics 36, 1925–1927 (2020).CAS 

Google Scholar 
Louca, S. The rates of global bacterial and archaeal dispersal. ISME Journal 16, 159–167 (2021).ADS 

Google Scholar 
Ondov, B. D. et al. Mash: fast genome and metagenome distance estimation using minhash. Genome Biology 17, 132 (2016).
Google Scholar 
Müllner, D. fastcluster: Fast hierarchical, agglomerative clustering routines for R and Python. Journal of Statistical Software 53, 1–18 (2013).
Google Scholar 
Kinene, T., Wainaina, J., Maina, S., Boykin, L.M. & Kliman, R.M. Methods for rooting trees, vol. 3, 489–493 (Academic Press, Oxford, 2016).Louca, S. & Doebeli, M. Efficient comparative phylogenetics on large trees. Bioinformatics 34, 1053–1055 (2018).CAS 

Google Scholar 
Rees, J. A. & Cranston, K. Automated assembly of a reference taxonomy for phylogenetic data synthesis. Biodiversity Data Journal 5, e12581 (2017).
Google Scholar 
Heck, K. et al. Evaluating methods for purifying cyanobacterial cultures by qPCR and high-throughput Illumina sequencing. Journal of Microbiological Methods 129, 55–60 (2016).CAS 

Google Scholar 
Cornet, L. et al. Consensus assessment of the contamination level of publicly available cyanobacterial genomes. PLOS ONE 13, e0200323 (2018).
Google Scholar 
Alneberg, J. et al. Genomes from uncultivated prokaryotes: a comparison of metagenome-assembled and single-amplified genomes. Microbiome 6, 173 (2018).
Google Scholar 
Eddy, S. R. Accelerated profile HMM searches. PLoS Computational Biology 7, e1002195 (2011).ADS 
MathSciNet 
CAS 

Google Scholar 
Buchfink, B., Xie, C. & Huson, D. H. Fast and sensitive protein alignment using DIAMOND. Nature Methods 12, 59–60 (2014).
Google Scholar 
Pedregosa, F. et al. Scikit-learn: Machine Learning in Python. Journal of Machine Learning Research 12, 2825–2830 (2011).MathSciNet 
MATH 

Google Scholar  More

Costanza, R. et al. Twenty years of ecosystem services: How far have we come and how far do we still need to go?. Ecosyst. Serv. 28, 1–16. https://doi.org/10.1016/j.ecoser.2017.09.008 (2017).Article 

Google Scholar 
Harwell, M. A. et al. Conceptual framework for assessing ecosystem health. Integr. Environ. Assess. Manag. 15, 544–564. https://doi.org/10.1002/ieam.4152 (2019).Article 

Google Scholar 
Halpern, B. S. et al. A global map of human impact on marine ecosystems. Science 319, 948–952. https://doi.org/10.1126/science.1149345 (2008).Article 
ADS 
CAS 

Google Scholar 
Roca, G. et al. Response of seagrass indicators to shifts in environmental stressors: A global review and management synthesis. Ecol. Ind. 63, 310–323. https://doi.org/10.1016/j.ecolind.2015.12.007 (2016).Article 

Google Scholar 
Westgate, M. J., Likens, G. E. & Lindenmayer, D. B. Adaptive management of biological systems: A review. Biol. Cons. 158, 128–139. https://doi.org/10.1016/j.biocon.2012.08.016 (2013).Article 

Google Scholar 
Logan, M. et al. Ecosystem health report cards: An overview of frameworks and analytical methodologies. Ecol. Indic. 113, 105834. https://doi.org/10.1016/j.ecolind.2019.105834 (2020).Article 

Google Scholar 
Dennison, W. C., Lookingbill, T. R., Carruthers, T. J., Hawkey, J. M. & Carter, S. L. An eye-opening approach to developing and communicating integrated environmental assessments. Front. Ecol. Environ. 5, 307–314. https://doi.org/10.1890/1540-9295(2007)5[307:AEATDA]2.0.CO;2 (2007).Article 

Google Scholar 
Harwell, M. A. et al. A framework for an ecosystem integrity report card: examples from south Florida show how an ecosystem report card links societal values and scientific information. Bioscience 49, 543–556. https://doi.org/10.2307/1313475 (1999).Article 

Google Scholar 
Collier, C. J. et al. An evidence-based approach for setting desired state in a complex Great Barrier Reef seagrass ecosystem: A case study from Cleveland Bay. Environ. Sustain. Indic. 7, 100042. https://doi.org/10.1016/j.indic.2020.100042 (2020).Article 

Google Scholar 
Coles, R. G. et al. Seagrass: Ecology, Uses and Threats (Nova Science Publishers, Inc., 2011).
Google Scholar 
Grech, A. et al. A comparison of threats, vulnerabilities and management approaches in global seagrass bioregions. Environ. Res. Lett. 7, 024006. https://doi.org/10.1088/1748-9326/7/2/024006 (2012).Article 
ADS 

Google Scholar 
Lambert, V. M. et al. Connecting targets for catchment sediment loads to ecological outcomes for seagrass using multiple lines of evidence. Mar. Pollut. Bull. https://doi.org/10.1016/j.marpolbul.2021.112494 (2021).Article 

Google Scholar 
Adams, M. P. et al. Predicting seagrass decline due to cumulative stressors. Environ. Model. Softw. 130, 104717. https://doi.org/10.1016/j.envsoft.2020.104717 (2020).Article 

Google Scholar 
Chartrand, K. M., Szabó, M., Sinutok, S., Rasheed, M. A. & Ralph, P. J. Living at the margins: The response of deep-water seagrasses to light and temperature renders them susceptible to acute impacts. Mar. Environ. Res. 136, 126–138. https://doi.org/10.1016/j.marenvres.2018.02.006 (2018).Article 
CAS 

Google Scholar 
Chartrand, K., Bryant, C., Carter, A., Ralph, P. & Rasheed, M. Light thresholds to prevent dredging impacts on the Great Barrier Reef seagrass, Zostera muelleri spp. capricorni. Front. Mar. Sci. 3, 17. https://doi.org/10.3389/fmars.2016.00106 (2016).Article 

Google Scholar 
Abal, E. & Dennison, W. Seagrass depth range and water quality in southern Moreton Bay, Queensland, Australia. Mar. Freshwater Res. 47, 763–771. https://doi.org/10.1071/MF9960763 (1996).Article 
CAS 

Google Scholar 
Dennison, W. et al. Assessing water quality with submersed aquatic vegetation: Habitat requirements as barometers of Chesapeake Bay health. Bioscience 43, 86–94. https://doi.org/10.2307/1311969 (1993).Article 

Google Scholar 
Carter, A. B., Collier, C., Coles, R., Lawrence, E. & Rasheed, M. A. Community-specific, “desired” states for seagrasses through cycles of loss and recovery. J. Environ. Manag. 314, 115059. https://doi.org/10.1016/j.jenvman.2022.115059 (2022).Article 

Google Scholar 
Kaldy, J. E., Brown, C. A. & Pacella, S. R. Carbon limitation in response to nutrient loading in an eelgrass mesocosm: Influence of water residence time. Mar. Ecol. Prog. Ser. 689, 1–17. https://doi.org/10.3354/meps14061 (2022).Article 
CAS 

Google Scholar 
Carter, A. B. et al. A spatial analysis of seagrass habitat and community diversity in the Great Barrier Reef World Heritage Area. Sci. Rep. https://doi.org/10.1038/s41598-021-01471-4 (2021).Article 

Google Scholar 
Kenworthy, W. J., Wyllie-Echeverria, S., Coles, R. G., Pergent, G. & Pergent-Martini, C. Seagrasses: Biology, Ecology and Conservation 595–623 (Springer, 2006).
Google Scholar 
Hayes, M. A. et al. The differential importance of deep and shallow seagrass to nekton assemblages of the great barrier reef. Diversity 12, 292. https://doi.org/10.3390/d12080292 (2020).Article 

Google Scholar 
Marsh, H., O’Shea, T. J. & Reynolds, J. E. III. Ecology and Conservation of the Sirenia: Dugongs and Manatees Vol. 18 (Cambridge University Press, 2011).Book 

Google Scholar 
Scott, A. L. et al. The role of herbivory in structuring tropical seagrass ecosystem service delivery. Front. Plant Sci. 9, 1–10. https://doi.org/10.3389/fpls.2018.00127 (2018).Article 

Google Scholar 
York, P. H., Macreadie, P. I. & Rasheed, M. A. Blue carbon stocks of Great Barrier Reef deep-water seagrasses. Biol. Lett. 14, 20180529. https://doi.org/10.1098/rsbl.2018.0529 (2018).Article 
CAS 

Google Scholar 
Unsworth, R. K., Collier, C. J., Waycott, M., Mckenzie, L. J. & Cullen-Unsworth, L. C. A framework for the resilience of seagrass ecosystems. Mar. Pollut. Bull. 100, 34–46. https://doi.org/10.1016/j.marpolbul.2015.08.016 (2015).Article 
CAS 

Google Scholar 
Madden, C. J., Rudnick, D. T., McDonald, A. A., Cunniff, K. M. & Fourqurean, J. W. Ecological indicators for assessing and communicating seagrass status and trends in Florida Bay. Ecol. Ind. 9, S68–S82. https://doi.org/10.1016/j.ecolind.2009.02.004 (2009).Article 
CAS 

Google Scholar 
York, P. et al. Dynamics of a deep-water seagrass population on the Great Barrier Reef: Annual occurrence and response to a major dredging program. Sci. Rep. 5, 13167. https://doi.org/10.1038/srep13167 (2015).Article 
ADS 
CAS 

Google Scholar 
Rasheed, M. A., McKenna, S. A., Carter, A. B. & Coles, R. G. Contrasting recovery of shallow and deep water seagrass communities following climate associated losses in tropical north Queensland, Australia. Mar. Pollut. Bull. 83, 491–499. https://doi.org/10.1016/j.marpolbul.2014.02.013 (2014).Article 
CAS 

Google Scholar 
Smith, T., Chartrand, K., Wells, J., Carter, A. & Rasheed, M. Seagrasses in Port Curtis and Rodds Bay 2019 Annual long-term monitoring and whole port survey. 71, https://www.tropwater.com/wp-content/uploads/2022/10/20-64-Annual-Seagrass-monitoring-in-Port-Curtis-and-Rodds-Bay-2019.pdf (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 20/64, James Cook University, Cairns, 2020).Ruaro, R., Gubiani, E. A., Hughes, R. M. & Mormul, R. P. Global trends and challenges in multimetric indices of biological condition. Ecol. Indic. 110, 105862. https://doi.org/10.1016/j.ecolind.2019.105862 (2020).Article 

Google Scholar 
Kilminster, K. et al. Unravelling complexity in seagrass systems for management: Australia as a microcosm. Sci. Total Environ. 534, 97–109. https://doi.org/10.1016/j.scitotenv.2015.04.061 (2015).Article 
ADS 
CAS 

Google Scholar 
Collier, C. J., Chartrand, K., Honchin, C., Fletcher, A. & Rasheed, M. Light thresholds for seagrasses of the GBR: a synthesis and guiding document. Including knowledge gaps and future priorities. 41, http://nesptropical.edu.au/wp-content/uploads/2016/05/NESP-TWQ-3.3-FINAL-REPORTa.pdf (Report to the National Environmental Science Programme, Cairns, 2016).Bryant, C., Jarvis, J. C., York, P. & Rasheed, M. Gladstone Healthy Harbour Partnership Pilot Report Card; ISP011: Seagrass., 74, https://researchonline.jcu.edu.au/44549/ (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 14/53, James Cook University, Cairns, 2014).McIntosh, E. J. et al. Designing report cards for aquatic health with a whole-of-system approach: Gladstone Harbour in the Great Barrier Reef. Ecol. Ind. 102, 623–632. https://doi.org/10.1016/j.ecolind.2019.03.012 (2019).Article 

Google Scholar 
Birch, W. & Birch, M. Succession and pattern of tropical intertidal seagrasses in Cockle Bay, Queensland, Australia: A decade of observations. Aquat. Bot. 19, 343–367. https://doi.org/10.1016/0304-3770(84)90048-2 (1984).Article 

Google Scholar 
Rasheed, M. A. Recovery and succession in a multi-species tropical seagrass meadow following experimental disturbance: The role of sexual and asexual reproduction. J. Exp. Mar. Biol. Ecol. 310, 13–45. https://doi.org/10.1016/j.jembe.2004.03.022 (2004).Article 

Google Scholar 
Christiaen, B., Lehrter, J., Goff, J. & Cebrian, J. Functional implications of changes in seagrass species composition in two shallow coastal lagoons. Mar. Ecol. Prog. Ser. 557, 11. https://doi.org/10.3354/meps11847 (2016).Article 

Google Scholar 
Hyndes, G. A., Kendrick, A. J., MacArthur, L. D. & Stewart, E. Differences in the species- and size-composition of fish assemblages in three distinct seagrass habitats with differing plant and meadow structure. Mar. Biol. 142, 1195–1206. https://doi.org/10.1007/s00227-003-1010-2 (2003).Article 

Google Scholar 
Ray, B. R., Johnson, M. W., Cammarata, K. & Smee, D. L. Changes in seagrass species composition in Northwestern Gulf of Mexico Estuaries: Effects on associated seagrass Fauna. PLoS ONE 9, e107751. https://doi.org/10.1371/journal.pone.0107751 (2014).Article 
ADS 
CAS 

Google Scholar 
Ondiviela, B. et al. The role of seagrasses in coastal protection in a changing climate. Coast. Eng. 87, 11. https://doi.org/10.1016/j.coastaleng.2013.11.005 (2014).Article 

Google Scholar 
Lavery, P. S., Mateo, M. -Á., Serrano, O. & Rozaimi, M. Variability in the carbon storage of seagrass habitats and its implications for global estimates of blue carbon ecosystem service. PLoS ONE 8, e73748. https://doi.org/10.1371/journal.pone.0073748 (2013).Article 
ADS 
CAS 

Google Scholar 
Coles, R. G. et al. The Great Barrier Reef World Heritage Area seagrasses: Managing this iconic Australian ecosystem resource for the future. Estuar. Coast. Shelf Sci. 153, A1–A12. https://doi.org/10.1016/j.ecss.2014.07.020 (2015).Article 
ADS 

Google Scholar 
Smith, T. M., Reason, C., McKenna, S. & Rasheed, M. A. Seagrasses in Port Curtis and Rodds Bay 2020. Annual long-term monitoring. 54, https://www.dropbox.com/s/f5yb6bjjpbvc1f2/21%2016%20Smith%20et%20al%202021%20Annual%20Seagrass%20monitoring%20in%20Port%20Curtis%20and%20Rodds%20Bay%202020_Final%20version.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/16, James Cook University, Cairns, 2021).Windle, J., Rolfe, J. & Pascoe, S. Assessing recreational benefits as an economic indicator for an industrial harbour report card. Ecol. Ind. 80, 224–231. https://doi.org/10.1016/j.ecolind.2017.05.036 (2017).Article 

Google Scholar 
Scott, A. & Rasheed, M. A. Port of Karumba long-term annual seagrass monitoring 2020. 28, https://www.dropbox.com/s/fwtys67ljssbp9t/21%2005%20Scott%20%26%20Rasheed%202021%20FINAL%202020%20Karumba%20Long-term%20seagrass%20monitoring%20report%20low%20res.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/05, James Cook University, Cairns, 2021).
Google Scholar 
Smith, T., Reason, C., McKenna, S. & Rasheed, M. Port of Weipa long‐term seagrass monitoring program, 2000 ‐ 2020. 49, https://www.dropbox.com/s/ghqy3bmn9p8jbsi/20%2058%20Smith%20et%20al%202020%20Port%20of%20Weipa%20Annual%20Long%20Term%20Seagrass%20Monitoring%20Report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 20/58, James Cook University, Cairns, 2020).Reason, C. L., Smith, T. M. & Rasheed, M. A. Seagrass habitat of Cairns Harbour and Trinity Inlet: Cairns Shipping Development Program and Annual Monitoring Report 2020. 54, https://www.dropbox.com/s/m7xtrytjjip3a42/21%2009%20Final_Cairns%20Harbour%20Seagrass%20Monitoring%20Report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/09, James Cook University, Cairns, 2021).Reason, C. L., York, P. H. & Rasheed, M. A. Seagrass habitat of Mourilyan Harbour: Annual monitoring report – 2020. 36, https://www.dropbox.com/s/kg3toxmlifh62tg/21%2010%20Mourilyan%20Harbour%20seagrass%20monitoring%20report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/10, James Cook University, Cairns, 2021).McKenna, S., Wilkinson, J., Chartrand, K. & Van De Wetering, C. Port of Townsville Seagrass Monitoring Program: 2020. 62, https://www.dropbox.com/s/n8nsx8ts93fgr36/21%2014%20Final%20POTL%20Annual%20Seagrass%20Report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/14, James Cook University, Cairns, 2021).McKenna, S. A., van de Wetering, C., Wilkinson, J. & Rasheed, M. A. Port of Abbot Point long-term seagrass monitoring program: 2020. 35, https://www.dropbox.com/s/l5a5l7pkikcjrfb/21%2025%20McKenna%20et%20al%20Port%20of%20Abbot%20Point%20Long-term%20seagrass%20Monitoring%20report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/25, James Cook University, Cairns, 2021).York, P. H. & Rasheed, M. A. Annual Seagrass Monitoring in the Mackay-Hay Point Region – 2020. 42, https://www.dropbox.com/s/u45yezm3984lw1a/21%2020%20Hay%20Point%20and%20Mackay%20Seagrass%20Final%20Report%202020.pdf?dl=0 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/20, James Cook University, Cairns, 2021).van de Wetering, C., Carter, A. B. & Rasheed, M. A. Mackay-Whitsunday-Isaac Seagrass Monitoring 2017–2020: Marine Inshore South Zone. 30, https://researchonline.jcu.edu.au/70923/ (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/06, James Cook University, Cairns, 2021).Carter, A. B. et al. Torres Strait Seagrass 2021 Report Card. 76, https://researchonline.jcu.edu.au/70797/ (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 21/13, James Cook University, Cairns, 2021).Gladstone Ports Corporation. Port of Gladstone. https://www.gpcl.com.au/port-of-gladstone (2022).Sawynok, B., Venables, B. & Pinto, U. Incorporating a fish recruitment indicator into a health report card: A case study from Gladstone Harbour, Australia. Ecol. Indic. 115, 106329. https://doi.org/10.1016/j.ecolind.2020.106329 (2020).Article 

Google Scholar 
Pascoe, S. et al. Developing a social, cultural and economic report card for a regional industrial harbour. PLoS ONE 11, e0148271. https://doi.org/10.1371/journal.pone.0148271 (2016).Article 
CAS 

Google Scholar 
Chartrand, K. M., Bryant, C. V., Sozou, A., Ralph, P. J. & Rasheed, M. A. Final Report: Deep‐water seagrass dynamics ‐ Light requirements, seasonal change and mechanisms of recruitment. 67, https://www.dropbox.com/sh/mo8dcq1322qv5c3/AAAgu3lEnJsLgxdawXaOltu-a/2017?dl=0&preview=17+16+Final+Report+Deep-water+seagrass+dynamics.pdf&subfolder_nav_tracking=1 (Centre for Tropical Water & Aquatic Ecosystem Research (TropWATER) Publication 17/16, James Cook University, Cairns, 2017).Kirkman, H. Decline of seagrass in northern areas of Moreton Bay, Queensland. Aquat. Bot. 5, 63–76. https://doi.org/10.1016/0304-3770(78)90047-5 (1978).Article 

Google Scholar 
Mellors, J. E. An evaluation of a rapid visual technique for estimating seagrass biomass. Aquat. Bot. 42, 67–73. https://doi.org/10.1016/0304-3770(91)90106-F (1991).Article 

Google Scholar 
Emmer, I. et al. Methodology for tidal wetland and seagrass restoration VM0033, version 2.0. https://verra.org/wp-content/uploads/2018/03/VM0033-Methodology-for-Tidal-Wetland-and-Seagrass-Restoration-v2.0-30Sep21-1.pdf (2021). More

Teuling, A. J., Seneviratne, S. I., Williams, C. & Troch, P. A. Observed timescales of evapotranspiration response to soil moisture. Geophys. Res. Lett. 33, L23403 (2006).Gao, H. et al. Climate controls how ecosystems size the root zone storage capacity at catchment scale. Geophys. Res. Lett. 41, 7916–7923 (2014).Article 

Google Scholar 
Milly, P. C. D. Climate, soil water storage, and the average annual water balance. Water Resour. Res. 30, 2143–2156 (1994).Article 

Google Scholar 
Hahm, W. J. et al. Low subsurface water storage capacity relative to annual rainfall decouples Mediterranean plant productivity and water use from rainfall variability. Geophys. Res. Lett. 46, 6544–6553 (2019).Article 

Google Scholar 
Seneviratne, S. I. et al. Investigating soil moisture–climate interactions in a changing climate: a review. Earth Sci. Rev. 99, 125–161 (2010).Article 

Google Scholar 
Thompson, S. E. et al. Comparative hydrology across AmeriFlux sites: the variable roles of climate, vegetation, and groundwater. Water Resour. Res. 47, W00J07 (2011).Fan, Y., Miguez-Macho, G., Jobbágy, E. G., Jackson, R. B. & Otero-Casal, C. Hydrologic regulation of plant rooting depth. Proc. Natl Acad. Sci. USA 114, 10572–10577 (2017).Article 

Google Scholar 
Hain, C. R., Crow, W. T., Anderson, M. C. & Tugrul Yilmaz, M. Diagnosing neglected soil moisture source–sink processes via a thermal infrared-based two-source energy balance model. J. Hydrometeorol. 16, 1070–1086 (2015).Article 

Google Scholar 
Rempe, D. M. & Dietrich, W. E. Direct observations of rock moisture, a hidden component of the hydrologic cycle. Proc. Natl Acad. Sci. USA 115, 2664–2669 (2018).Article 

Google Scholar 
Dawson, T. E., Jesse Hahm, W. & Crutchfield-Peters, K. Digging deeper: what the critical zone perspective adds to the study of plant ecophysiology. N. Phytol. 226, 666–671 (2020).Article 

Google Scholar 
McCormick, E. L. et al. Widespread woody plant use of water stored in bedrock. Nature 597, 225–229 (2021).Article 

Google Scholar 
Maxwell, R. M. & Condon, L. E. Connections between groundwater flow and transpiration partitioning. Science 353, 377–380 (2016).Article 

Google Scholar 
Schlemmer, L., Schär, C., Lüthi, D. & Strebel, L. A groundwater and runoff formulation for weather and climate models. J. Adv. Model. Earth Syst. 10, 1809–1832 (2018).Article 

Google Scholar 
Teuling, A. J. et al. Contrasting response of European forest and grassland energy exchange to heatwaves. Nat. Geosci. 3, 722–727 (2010).Article 

Google Scholar 
Koirala, S. et al. Global distribution of groundwater–vegetation spatial covariation. Geophys. Res. Lett. 44, 4134–4142 (2017).Article 

Google Scholar 
Esteban, E. J. L., Castilho, C. V., Melgaço, K. L. & Costa, F. R. C. The other side of droughts: wet extremes and topography as buffers of negative drought effects in an Amazonian forest. N. Phytol. 229, 1995–2006 (2021).Article 

Google Scholar 
Liu, Y., Konings, A. G., Kennedy, D. & Gentine, P. Global coordination in plant physiological and rooting strategies in response to water stress. Glob. Biogeochem. Cycles 35, e2020GB006758 (2021).Article 

Google Scholar 
Schenk, H. J. & Jackson, R. B. The global biogeography of roots. Ecol. Monogr. 72, 311–328 (2002).Article 

Google Scholar 
Canadell, J. et al. Maximum rooting depth of vegetation types at the global scale. Oecologia 108, 583–595 (1996).Article 

Google Scholar 
Weaver, J. E. & Darland, R. W. Soil–root relationships of certain native grasses in various soil types. Ecol. Monogr. 19, 303–338 (1949).Article 

Google Scholar 
Chitra-Tarak, R. et al. Hydraulically-vulnerable trees survive on deep-water access during droughts in a tropical forest. N. Phytol. 231, 1798–1813 (2021).Article 

Google Scholar 
Schenk, H. J. & Jackson, R. B. Mapping the global distribution of deep roots in relation to climate and soil characteristics. Geoderma 126, 129–140 (2005).Article 

Google Scholar 
Franklin, O. et al. Organizing principles for vegetation dynamics. Nat. Plants 6, 444–453 (2020).Article 

Google Scholar 
Kleidon, A. & Heimann, M. A method of determining rooting depth from a terrestrial biosphere model and its impacts on the global water and carbon cycle. Glob. Change Biol. 4, 275–286 (1998).Article 

Google Scholar 
Schymanski, S. J., Sivapalan, M., Roderick, M. L., Hutley, L. B. & Beringer, J. An optimality-based model of the dynamic feedbacks between natural vegetation and the water balance. Water Resour. Res. 45, W01412 (2009).Wang-Erlandsson, L. et al. Global root zone storage capacity from satellite-based evaporation. Hydrol. Earth Syst. Sci. 20, 1459–1481 (2016).Article 

Google Scholar 
Knapp, A. K. & Smith, M. D. Variation among biomes in temporal dynamics of aboveground primary production. Science 291, 481–484 (2001).Article 

Google Scholar 
Anderson, M. A two-source time-integrated model for estimating surface fluxes using thermal infrared remote sensing. Remote Sens. Environ. 60, 195–216 (1997).Article 

Google Scholar 
Hain, C. R. & Anderson, M. C. Estimating morning change in land surface temperature from MODIS day/night observations: applications for surface energy balance modeling. Geophys. Res. Lett. 44, 9723–9733 (2017).Article 

Google Scholar 
Tumber-Dávila, S. J., Schenk, H. J., Du, E. & Jackson, R. B. Plant sizes and shapes above- and belowground and their interactions with climate. New Phytol. https://nph.onlinelibrary.wiley.com/doi/abs/10.1111/nph.18031 (2022).Harmonized World Soil Database Version 1.0 (FAO, 2008).Wieder, W. Regridded Harmonized World Soil Database Version 1.2 (ORNL DAAC, 2014); https://doi.org/10.3334/ORNLDAAC/1247Balland, V., Pollacco, J. A. P. & Arp, P. A. Modeling soil hydraulic properties for a wide range of soil conditions. Ecol. Model. 219, 300–316 (2008).Article 

Google Scholar 
Agee, E. et al. Root lateral interactions drive water uptake patterns under water limitation. Adv. Water Resour. 151, 103896 (2021).Article 

Google Scholar 
Krakauer, N. Y., Li, H. & Fan, Y. Groundwater flow across spatial scales: importance for climate modeling. Environ. Res. Lett. 9, 034003 (2014).Article 

Google Scholar 
Stoy, P. C. et al. Reviews and syntheses: turning the challenges of partitioning ecosystem evaporation and transpiration into opportunities. Biogeosciences 16, 3747–3775 (2019).Article 

Google Scholar 
Jackson, R. B., Moore, L. A., Hoffmann, W. A., Pockman, W. T. & Linder, C. R. Ecosystem rooting depth determined with caves and DNA. Proc. Natl Acad. Sci. USA 96, 11387–11392 (1999).Article 

Google Scholar 
Pelletier, J. D. et al. A gridded global data set of soil, intact regolith, and sedimentary deposit thicknesses for regional and global land surface modeling. J. Adv. Model. Earth Syst. 8, 41–65 (2016).Article 

Google Scholar 
Parmesan, C. & Hanley, M. E. Plants and climate change: complexities and surprises. Ann. Bot. 116, 849–864 (2015).Article 

Google Scholar 
Pendergrass, A. G., Knutti, R., Lehner, F., Deser, C. & Sanderson, B. M. Precipitation variability increases in a warmer climate. Sci. Rep. 7, 17966 (2017).Siebert, S. et al. Development and validation of the global map of irrigation areas. Hydrol. Earth Syst. Sci. 9, 535–547 (2005).Article 

Google Scholar 
Friedl, M. A. et al. MODIS Collection 5 global land cover: Algorithm refinements and characterization of new datasets. Remote Sens. Environ. 114, 168–182 (2010).Article 

Google Scholar 
Olson, D. M. et al. Terrestrial ecoregions of the world: a new map of life on Earth. BioScience 51, 933–938 (2001).Article 

Google Scholar 
Mu, Q., Heinsch, F. A., Zhao, M. & Running, S. W. Development of a global evapotranspiration algorithm based on MODIS and global meteorology data. Remote Sens. Environ. 111, 519–536 (2007).Article 

Google Scholar 
Fisher, J. B. et al. ECOSTRESS: NASA’s next generation mission to measure evapotranspiration from the international space station. Water Resour. Res. 56, e2019WR026058 (2020).Article 

Google Scholar 
Davis, T. W. et al. Simple process-led algorithms for simulating habitats (SPLASH v.1.0): robust indices of radiation, evapotranspiration and plant-available moisture. Geosci. Model Dev. 10, 689–708 (2017).Article 

Google Scholar 
Weedon, G. P. et al. The WFDEI meteorological forcing data set: WATCH forcing data methodology applied to ERA-Interim reanalysis data. Water Resour. Res. 50, 7505–7514 (2014).Article 

Google Scholar 
Orth, R., Koster, R. D. & Seneviratne, S. I. Inferring soil moisture memory from streamflow observations using a simple water balance model. J. Hydrometeorol. 14, 1773–1790 (2013).Article 

Google Scholar 
Stocker, B. cwd v.1.0: R package for cumulative water deficit calculation. Zenodo https://doi.org/10.5281/zenodo.5359053 (2021).Zhang, Y. et al. Model-based analysis of the relationship between sun-induced chlorophyll fluorescence and gross primary production for remote sensing applications. Remote Sens. Environ. 187, 145–155 (2016).Article 

Google Scholar 
Duveiller, G. et al. A spatially downscaled sun-induced fluorescence global product for enhanced monitoring of vegetation productivity. Earth Syst. Sci. Data 12, 1101–1116 (2020).Article 

Google Scholar 
Joiner, J. et al. Global monitoring of terrestrial chlorophyll fluorescence from moderate-spectral-resolution near-infrared satellite measurements: methodology, simulations, and application to GOME-2. Atmos. Meas. Tech. 6, 2803–2823 (2013).Article 

Google Scholar 
Köhler, P., Guanter, L. & Joiner, J. A linear method for the retrieval of sun-induced chlorophyll fluorescence from GOME-2 and SCIAMACHY data. Atmos. Meas. Tech. 8, 2589–2608 (2015).Article 

Google Scholar 
Jiang, B. et al. Validation of the surface daytime net radiation product from version 4.0 GLASS product suite. IEEE Geosci. Remote Sens. Lett. 16, 509–513 (2019).Article 

Google Scholar 
Muggeo, V. M. R. Estimating regression models with unknown break-points. Stat. Med. 22, 3055–3071 (2003).Article 

Google Scholar 
Gilleland, E. & Katz, R. W. extRemes 2.0: an extreme value analysis package in R. J. Stat. Softw. 72, 1–39 (2016).Marthews, T. R., Dadson, S. J., Lehner, B., Abele, S. & Gedney, N. High-resolution global topographic index values for use in large-scale hydrological modelling. Hydrol. Earth Syst. Sci. 19, 91–104 (2015).Article 

Google Scholar 
Etopo1, Global 1 Arc-Minute Ocean Depth and Land Elevation from the US National Geophysical Data Center (NGDC) (National Geophysical Data Center, NESDIS, NOAA and US Department of Commerce, 2011); https://doi.org/10.5065/D69Z92Z5Beven, K. J. & Kirkby, M. J. A physically based, variable contributing area model of basin hydrology. Hydrol. Sci. J. 24, 43–69 (1979).Article 

Google Scholar 
Hansen, M. C., Townshend, J. R. G., DeFries, R. S. & Carroll, M. Estimation of tree cover using MODIS data at global, continental and regional/local scales. Int. J. Remote Sens. 26, 4359–4380 (2005).Article 

Google Scholar 
Stocker, B. D. Global rooting zone water storage capacity and rooting depth estimates. Zenodo https://doi.org/10.5281/zenodo.5515246 (2021).Stocker, B. stineb/mct: v3.0: re-submission to Nature Geoscience. Zenodo https://doi.org/10.5281/zenodo.6239187 (2022). More

Waters, C. N. et al. The Anthropocene is functionally and stratigraphically distinct from the Holocene. Science 351, 137. https://doi.org/10.1126/science.aad2622 (2016).Article 
CAS 

Google Scholar 
Thomas, J. A. & Morris, M. G. Patterns, mechanisms and rates of extinction among invertebrates in the United Kingdom. Phil. Trans. R. Soc. Lond. B 344, 47–54 (1994).Article 
ADS 

Google Scholar 
Thomas, J. A. et al. Comparative losses of british butterflies, birds, and plants and the global extinction crisis. Science 303, 1879–1881. https://doi.org/10.1126/science.1095046 (2004).Article 
ADS 
CAS 

Google Scholar 
van Klink, R. et al. Meta-analysis reveals declines in terrestrial but increases in freshwater insect abundances. Science 368, 417–420. https://doi.org/10.1126/science.aax9931 (2020).Article 
ADS 
CAS 

Google Scholar 
Hallmann, C. A. et al. More than 75 percent decline over 27 years in total flying insect biomass in protected areas. PLoS ONE 12, 21. https://doi.org/10.1371/journal.pone.0185809 (2017).Article 
CAS 

Google Scholar 
Barmentlo, S. H. et al. Experimental evidence for neonicotinoid driven decline in aquatic emerging insects. Proc. Natl. Acad. Sci. USA 118, 8. https://doi.org/10.1073/pnas.2105692118j1of8 (2021).Article 

Google Scholar 
Ehlers, B. K., Bataillon, T. & Damgaard, C. F. Ongoing decline in insect-pollinated plants across Danish grasslands. Biol. Lett. 17, 20210493. https://doi.org/10.1098/rsbl.2021.0493 (2021).Article 

Google Scholar 
Seibold, S. et al. Arthropod decline in grasslands and forests is associated with landscape-level drivers. Nature 574, 671–674. https://doi.org/10.1038/s41586-019-1684-3 (2019).Article 
ADS 
CAS 

Google Scholar 
Cardoso, P. et al. Scientists’ warning to humanity on insect extinctions. Biol. Conserv. 242, 108426. https://doi.org/10.1016/j.biocon.2020.108426 (2020).Article 

Google Scholar 
Montgomery, G. A. et al. Is the insect apocalypse upon us? How to find out. Biol. Conserv. 241, 6. https://doi.org/10.1016/j.biocon.2019.108327 (2020).Article 

Google Scholar 
Jactel, H. et al. Insect decline: immediate action is needed. C. R. Biol. 343, 267–293. https://doi.org/10.5802/crbiol.37 (2020).Article 

Google Scholar 
Owens, A. C. S. et al. Light pollution is a driver of insect declines. Biol. Conserv. 241, 9. https://doi.org/10.1016/j.biocon.2019.108259 (2020).Article 

Google Scholar 
Sanchez-Bayo, F. & Wyckhuys, K. A. G. Worldwide decline of the entomofauna: A review of its drivers. Biol. Conserv. 232, 8–27. https://doi.org/10.1016/j.biocon.2019.01.020 (2019).Article 

Google Scholar 
Michalko, R., Pekar, S. & Entling, M. H. An updated perspective on spiders as generalist predators in biological control. Oecologia https://doi.org/10.1007/s00442-018-4313-1 (2018).Article 

Google Scholar 
Nyffeler, M., Sterling, W. & Dean, D. How spiders make a living. Environ. Entomol. 23, 1357–1367 (1994).Article 

Google Scholar 
Branco, V. V. & Cardoso, P. An expert-based assessment of global threats and conservation measures for spiders. Glob. Ecol. Conserv. 24, 15. https://doi.org/10.1016/j.gecco.2020.e01290 (2020).Article 

Google Scholar 
Gobbi, M., Fontaneto, D. & De Bernardi, F. Influence of climate changes on animal communities in space and time: The case of spider assemblages along an alpine glacier foreland. Glob. Change Biol. 12, 1985–1992. https://doi.org/10.1111/j.1365-2486.2006.01236.x (2006).Article 
ADS 

Google Scholar 
Mammola, S., Goodacre, S. L. & Isaia, M. Climate change may drive cave spiders to extinction. Ecography 41, 233–243. https://doi.org/10.1111/ecog.02902 (2018).Article 

Google Scholar 
Potapov, A. M. et al. Functional losses in ground spider communities due to habitat structure degradation under tropical land-use change. Ecology 101, e02957. https://doi.org/10.1002/ecy.2957 (2020).Article 

Google Scholar 
Kormann, U. et al. Local and landscape management drive trait-mediated biodiversity of nine taxa on small grassland fragments. Divers. Distrib. 21, 1204–1217. https://doi.org/10.1111/ddi.12324 (2015).Article 

Google Scholar 
Hogg, B. N. & Daane, K. M. Ecosystem services in the face of invasion: the persistence of native and nonnative spiders in an agricultural landscape. Ecol. Appl. 21, 565–576. https://doi.org/10.1890/10-0496.1 (2011).Article 

Google Scholar 
Galle, R., Happe, A. K., Baillod, A. B., Tscharntke, T. & Batary, P. Landscape configuration, organic management, and within-field position drive functional diversity of spiders and carabids. J. Appl. Ecol. 56, 63–72. https://doi.org/10.1111/1365-2664.13257 (2019).Article 

Google Scholar 
Pekár, S. Spiders (Araneae) in the pesticide world: An ecotoxicological review. Pest. Manage. Sci. 68, 1438–1446. https://doi.org/10.1002/ps.3397 (2012).Article 
CAS 

Google Scholar 
Bommarco, R., Miranda, F., Bylund, H. & Bjorkman, C. Insecticides suppress natural enemies and increase pest damage in cabbage. J. Econ. Entomol. 104, 782–791. https://doi.org/10.1603/ec10444 (2011).Article 
CAS 

Google Scholar 
Outhwaite, C. L., Gregory, R. D., Chandler, R. E., Collen, B. & Isaac, N. J. B. Complex long-term biodiversity change among invertebrates, bryophytes and lichens. Nature Ecol. Evol. 4, 384–392. https://doi.org/10.1038/s41559-020-1111-z (2020).Article 

Google Scholar 
Rix, M. G. et al. Where have all the spiders gone? The decline of a poorly known invertebrate fauna in the agricultural and arid zones of southern Australia. Austral Entomol. 56, 14–22. https://doi.org/10.1111/aen.12258 (2017).Article 

Google Scholar 
Nyffeler, M. & Bonte, D. Where have all the spiders gone? Observations of a dramatic population density decline in the once very abundant garden spider, Araneus diadematus (Araneae: Araneidae), in the Swiss Midland. Insects 11, 12. https://doi.org/10.3390/insects11040248 (2020).Article 

Google Scholar 
Bowden, J. J., Hansen, O. L. P., Olsen, K., Schmidt, N. M. & Høye, T. T. Drivers of inter-annual variation and long-term change in High-Arctic spider species abundances. Polar Biol. 41, 1635–1649. https://doi.org/10.1007/s00300-018-2351-0 (2018).Article 

Google Scholar 
Samu, F., Németh, J. & Kiss, B. Assessment of the efficiency of a hand-held suction device for sampling spiders: Improved density estimation or oversampling?. Ann. Appl. Biol. 130, 371–378. https://doi.org/10.1111/j.1744-7348.1997.tb06840.x (1997).Article 

Google Scholar 
Nentwig, W. et al. Spiders of Europe. Version 07.2022. https://www.araneae.nmbe.ch (2022).Heimer, S. & Nentwig, W. Spinnen Mitteleuropas (Paul Parey, 1991).
Google Scholar 
Samu, F. & Szinetár, C. On the nature of agrobiont spiders. J. Arachnol. 30, 389–402. https://doi.org/10.1636/0161-8202(2002)030[0389:Otnoas]2.0.Co;2 (2002).Article 

Google Scholar 
Buchar, J. & Růžička, V. Catalogue of Spiders of the Czech Republic (Peres, 2002).
Google Scholar 
Samu, F. A general data model for databases in experimental animal ecology. Acta Zool. Acad. Sci. Hung. 45, 273–290 (1999).
Google Scholar 
Laliberté, E., Legendre, P. & Shipley, B. FD: Measuring Functional Diversity from Multiple Traits, and Other Tools for Functional Ecology. R package version 1.0–12. (2014).Bates, D., Mächler, M., Bolker, B. & Walker, S. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48. https://doi.org/10.18637/jss.v067.i01 (2015).Article 

Google Scholar 
Zuur, A., Ieno, E., Walker, N., Saveliev, A. & Smith, G. Mixed Effects Models and Extensions in Ecology with R (Springer, 2009).Book 
MATH 

Google Scholar 
Vegan. Community Ecology Package. R package Version 2.5–6. The Comprehensive R Archive Network (2019).ter Braak, C. J. F. & Smilauer, P. Canoco Reference Manual and User’s Guide: Software for Ordination, Version 5.1x. (Microcomputer Power, 2018).McRae, L., Deinet, S. & Freeman, R. The diversity-weighted living planet index: Controlling for taxonomic bias in a global biodiversity indicator. PLoS ONE 12, e0169156. https://doi.org/10.1371/journal.pone.0169156 (2017).Article 
CAS 

Google Scholar 
Toju, H. & Baba, Y. G. DNA metabarcoding of spiders, insects, and springtails for exploring potential linkage between above- and below-ground food webs. Zool. Lett. 4, 12. https://doi.org/10.1186/s40851-018-0088-9 (2018).Article 

Google Scholar 
Dirzo, R. et al. Defaunation in the anthropocene. Science 345, 401–406. https://doi.org/10.1126/science.1251817 (2014).Article 
ADS 
CAS 

Google Scholar 
Lister, B. C. & Garcia, A. Climate-driven declines in arthropod abundance restructure a rainforest food web. Proc. Natl. Acad. Sci. USA 115, E10397–E10406. https://doi.org/10.1073/pnas.1722477115 (2018).Article 
ADS 
CAS 

Google Scholar 
Harwood, J. D., Sunderland, K. D. & Symondson, W. O. C. Monoclonal antibodies reveal the potential of the tetragnathid spider Pachygnatha degeeri (Araneae: Tetragnathidae) as an aphid predator. Bull. Entomol. Res. 95, 161–167. https://doi.org/10.1079/BER2004346 (2005).Article 
CAS 

Google Scholar 
Samu, F., Beleznai, O. & Tholt, G. A potential spider natural enemy against virus vector leafhoppers in agricultural mosaic landscapes: Corroborating ecological and behavioral evidence. Biol. Control. 67, 390–396. https://doi.org/10.1016/j.biocontrol.2013.08.016 (2013).Article 

Google Scholar 
Biteniekyté, M. & Relys, V. Epigeic spider communities of a peat bog and adjacent habitats. Rev. Iber. Aracnol. 15, 81–87 (2008).
Google Scholar 
Michalko, R., Kosulic, O., Hula, V. & Surovcova, K. Niche differentiation of two sibling wolf spider species, Pardosa lugubris and Pardosa alacris, along a canopy openness gradient. J. Arachnol. 44, 46–51 (2016).Article 

Google Scholar 
Nyffeler, M. & Birkhofer, K. An estimated 400–800 million tons of prey are annually killed by the global spider community. Naturwissenschaften 104, 30. https://doi.org/10.1007/s00114-017-1440-1 (2017).Article 
CAS 

Google Scholar 
Sohlström, E. H. et al. Future climate and land-use intensification modify arthropod community structure. Agric. Ecosyst. Environ. 327, 107830. https://doi.org/10.1016/j.agee.2021.107830 (2022).Article 
CAS 

Google Scholar 
Sallé, A. et al. Climate change alters temperate forest canopies and indirectly reshapes arthropod communities. Front. For. Glob. Change 4, 710854 (2021).Article 

Google Scholar 
Høye, T. T. et al. Nonlinear trends in abundance and diversity and complex responses to climate change in Arctic arthropods. Proc. Natl. Acas. Sci. USA 118, e2002557117 (2021).Article 

Google Scholar 
Tscharntke, T., Klein, A. M., Kruess, A., Steffan-Dewenter, I. & Thies, C. Landscape perspectives on agricultural intensification and biodiversity: Ecosystem service management. Ecol. Lett. 8, 857–874. https://doi.org/10.1111/j.1461-0248.2005.00782.x (2005).Article 

Google Scholar 
Kleijn, D., Rundlöf, M., Scheper, J., Smith, H. G. & Tscharntke, T. Does conservation on farmland contribute to halting the biodiversity decline?. Trends Ecol. Evol. 26, 474–481. https://doi.org/10.1016/j.tree.2011.05.009 (2011).Article 

Google Scholar 
Swinbank, A. The European Union’s Common Agricultural Policy (CAP) The New Palgrave Dictionary of Economics 1–9 (Palgrave Macmillan, 2016).
Google Scholar 
Wissinger, S. Cyclic colonization in predictably ephemeral habitats: A template for biological control in annual crop systems. Biol. Control 10, 4–15 (1997).Article 

Google Scholar 
Samu, F., Szita, É. & Botos, E. Short- and longer-term colonization of alfalfa by spiders: A case study into the succession of perennial fields. In European Arachnology 2008 (eds Nentwig, W. et al.) 153–163 (Natural History Museum, 2010).
Google Scholar 
Samu, F., Horváth, A., Neidert, D., Botos, E. & Szita, É. Metacommunities of spiders in grassland habitat fragments of an agricultural landscape. Basic Appl. Ecol. 31, 92–103. https://doi.org/10.1016/j.baae.2018.07.009 (2018).Article 

Google Scholar  More

Our approach uses hypothetical 30 cm fixed depth samples taken at three successive time points (t0, t1, and t2) with prescribed changes in SOC (1.4% to 1.6%) and BD (1.5–1.1 g cm−3) over these time points (Table 1). The 30 cm soil depth is the common international standard for sampling and analysis required for SOC stock assessment and adhered to by carbon accounting and market organizations6,18. The changes we adopted (a 27% decrease in BD and a 14% increase in SOC) while relatively large, are consistent with those reported in the literature. For example, Reganold and Palmer reported a 25% decrease in BD (1.2–0.9 g cm−3) in neighboring farms with differing management practices23, and Syswerda et al. observed a 17% increase in SOC concentration (10.4–12.2 g C kg soil−1) when converting from a conventionally to organically managed row crop rotation21.Table 1 Hypothetical changes in bulk density (BD) and soil organic carbon (SOC) concentration in 30 cm fixed depth samples at time points t0, t1 and t2 along with calculated values of SOC stock and total soil mass and mineral soil mass.Full size tableIn Table 1, the total soil mass, mineral soil mass, and the SOC stock of the fixed depth samples were calculated by equations as described in the introduction from our prescribed changes in BD and SOC values.ScenariosWe compared hypothetical ESM correction scenarios with our 30 cm fixed depth sample at each time point (Table 2, Figs. 2, 3).Table 2 Hypothetical ESM scenarios showing variation with depth for bulk density (BD) and soil organic carbon (SOC) at each sampling time point, along with the sample depth intervals investigated.Full size tableFigure 2Flow chart of the definition, sampling, and SOC stock correction for a theoretical data set at time points t0, t1, and t2 for scenarios s1 with linear distributions of BD and SOC and s2 with a linear increase in BD and exponential decrease in SOC with depth. Scenario s2 is sampled at (a) 10 cm, (b) 15 cm, and (c) 30 cm intervals.Full size imageFigure 3Scenarios (S1 and S2), showing (a) bulk density variation (BD, g cm−3), and (b) soil organic carbon (SOC, %) variation by depth (0–30 cm) at each time point (t0, t1, and t2). For scenario 2, the single 30 cm depth interval was used (2c). See Table 1 and 2 for details.Full size imageScenario 1We carried out the ESM correction on a 30 cm sample and assumed that the sample was homogenous throughout the profile, with constant SOC and BD values at each time point.To correct for the error in SOC stock estimation when using fixed depth soil sampling, we used  Eqs.2a, 2b and 2c that consider changes in BD28,35. The adjusted soil depth resulting from the change in BD is calculated as:$${mathrm{M}}_{mathrm{n}}= {mathrm{M}}_{mathrm{i}}$$
(2a)
$${mathrm{D}}_{mathrm{a}}*{mathrm{BD}}_{mathrm{n}}*left(1-mathrm{k}*{mathrm{SOC}}_{mathrm{n}}right)={mathrm{D}}_{mathrm{i}}*{mathrm{BD}}_{mathrm{i}}*left(1-mathrm{k}*{mathrm{SOC}}_{mathrm{i}}right)$$
(2b)
$${mathrm{D}}_{mathrm{a}}={mathrm{D}}_{mathrm{i}}*frac{{mathrm{BD}}_{mathrm{i}}}{{mathrm{BD}}_{mathrm{n}}}*frac{1-mathrm{k}*{mathrm{SOC}}_{mathrm{i}}}{1-mathrm{k}*{mathrm{SOC}}_{mathrm{n}}}$$
(2c)
where Mi = Initial mineral soil mass per area (left[frac{M}{{L}^{2}}right]) , Mn = New mineral soil mass per area (left[frac{M}{{L}^{2}}right]) , Da = Adjusted soil surface depth (left[Lright]) , BDi = Initial bulk density (left[frac{M}{{L}^{3}}right]) , BDn = New bulk density (left[frac{M}{{L}^{3}}right]) , SOCi = Initial SOC as a decimal percent (left[frac{M}{M}right]) , SOCn = New SOC as a decimal percent (left[frac{M}{M}right]) , Di = Initial depth (left[Lright]).To conform with Eq. (2a), an increase in SOC over time results in a displacement of some soil mineral mass from the sample, whereas a decrease in SOC over time requires some soil mineral mass to be replaced34. Multiplying the BD by the mineral fraction of the soil (left(1-mathrm{k}*{mathrm{SOC}}right)) for each time point allowed us to compare equivalent mineral mass28. The effect of a change in SOC on mineral mass is small, with a 1% change in SOC equating to approximately a 2% change in apparent depth. This adjustment relates SOC per unit of mineral mass of the fine fraction ( 2 mm)20. The corrected apparent depth can then be used to calculate the corrected SOC stock of a single layer, fixed depth sample (Eq. 3).$$SO{C}_{stock}={D}_{a}*BD*SOC$$
(3)
Scenario 2In ESM correction scenarios 2a, 2b, and 2c, we imposed variable, dynamic BD and SOC values with depth over time (Table 2, Figs. 2, 3). To investigate these profiles, we determined the SOC and BD values throughout the soil depth by separating the soil into one (1) cm depth increments (i.e., 0–1 cm, 1–2 cm, etc.). We refer to this calculated incremental profile as the scenario 2 baseline. We assumed that our prescribed SOC concentration varied with depth following an exponential decay. To represent this decay, we simulated the global average distribution of SOC concentration with depth on crop land36, following the distribution from Hobley and Wilson37 (Eq. 4),$$SOCleft(dright)=SO{C}_{infty }+left(SO{C}_{o}-SO{C}_{infty }right)times {e}^{-dk}$$
(4)
where SOC (d) is the SOC concentration at depth (d), ({SOC}_{infty }) is the infinity SOC concentration, SOC0 is the SOC concentration at the soil surface, and k is the decay rate. We solved for the decay rate, initial SOC0, and infinity ({SOC}_{infty }) to fit the global average distribution for the 30 cm profile36 and then scaled the SOC concentration to our 30 cm fixed depth sample’s average SOC (1.4%) at t0 (Fig. 3).In scenarios 2a, 2b, and 2c, the BD increased linearly with depth38,39. At the initial time point (t0), we varied the BD values by ± 10% of the BD average over the 30 cm depth, such that for example, BD at t0 (profile average of 1.5 g cm−3) was 1.35 g cm−3 and 1.65 g cm−3 for the upper (0–1 cm) and lower (29–30 cm) depth increment, respectively. For each sequential time point, as the average BD decreased, the soil expanded. To determine the expansion, the depth of the initial sample (e.g., at t0) that filled the 30 cm depth in the subsequent sample (e.g., at t1) was calculated as the initial depth multiplied by the ratio of the average initial BD over the average new BD (e.g., 1.5/1.3 = 1.15 for t0/t1).The linear increase in BD with depth of each following time point maintained the average BD of scenario 1. We then varied the new BD by ± the percent change in the average BD between the time periods (see annotated scripts “main.R” and “functions.R” in Supplementary Material 1 for the development of the theoretical dataset). We then divided each initial BD increment (using soil mass for every 1 cm depth increment) by the new BD in the expanded increment (using soil mass for every  > 1 cm depth increment) to determine the expanded depth of each increment. The SOC value at the initial time represented the same, now expanded, ( > 1 cm) increments, as SOC is a ratio of mass. We used a linear decay rate that was twice that of the percent change in BD between time points to maintain an average BD that was consistent with scenario 1. To model the subsequent fixed depth sample, the BD and SOC concentration values of this expanded soil profile were then interpolated back to the 30 × 1 cm increments of the scenario 2 baseline depth. This calculation preserved the prescribed average BD of the new time point by only expanding the initial SOC concentration.We adjusted the SOC concentration of the next time point to maintain the average SOC concentrations of the 30 cm fixed depth sample, (see annotated scripts “main.R” and “functions.R” in Supplementary Material 1). Because the BD changed between time points and because the SOC stock in the 30 cm fixed depth sample was known, we determined the change in SOC stock between time points by subtracting the average SOC stock in the prior sample from the new sample. We then weighted this change across the 30 cm profile using the distribution of the global soil SOC in the top 30 cm to simulate SOC stratification with reduced tillage or agricultural intensification40. We then multiplied this change by the BD to convert back to SOC concentration and added the delta ((Delta )) SOC value to the prior sample. A worked example is shown in Supplementary Material 2 “Correction Example”.At each time point we split the soil profile at 10 cm and 15 cm depth intervals to create samples for scenarios 2a (3 soil intervals) and 2b (2 soil intervals), respectively. Note that scenario 2c is mathematically equivalent to scenario 1—with only one sample depth interval (30 cm) the sample contains no data on varying SOC or BD. The samples for 2a and 2b were generated by summing the total mass per area and SOC stock values from the scenario 2 baseline to produce single sample values of total soil mass per area and SOC concentration values per depth interval (as would be determined in a laboratory) and calculating BD and mineral mass.In scenario 2, any required additional mineral mass and the associated SOC values were ‘placed’ at the base of the sample to represent a soil profile that had expanded below the fixed 30 cm depth. To account for this, we calculated the increase in adjusted sample depth and accumulated additional soil mineral mass with the lowest sample depth interval of each split sample (Eqs. 5 and 6).$$mathrm{Delta D}={D}_{a}-{D}_{i}$$
(5)
$$SO{C}_{stock}={(D}_{1}*B{D}_{1}*SO{C}_{1}+dots + {(D}_{j}+Delta D)*B{D}_{j}*SO{C}_{j}))*10^2 (mathrm{g}/mathrm{cm}^{2})/(mathrm{Mg}/mathrm{ha})$$
(6)
where (mathrm{ Delta D}) is the apparent change in depth needed to generate the same mineral mass of the initial sample and the subscript j is the number of sample depth intervals from 1 to j.Varying BD linearly with depth introduces additional complexity in the calculation of the apparent depth. Each sample depth interval may expand (or contract in cases not explored here) at differing rates. Here, the over or under sampling of soil mineral mass is no longer constant with depth and the correction for apparent depth (Da) is estimated with linear interpolation using the BD of each sampling depth interval (i.e., 10 cm, 15 cm, or 30 cm). To do so, we calculated the mineral mass in each depth interval, determined their difference between the initial sample time point and new sample time point, and converted the change in mineral mass to a depth, where:$${mathrm{D}}_{mathrm{a}}={mathrm{D}}_{mathrm{i}}+frac{left(mathrm{sum}left({mathrm{D}}_{mathrm{ij}}*{mathrm{BD}}_{mathrm{ij}}*(1-mathrm{k}*{mathrm{soc}}_{mathrm{ij}}right))- mathrm{sum}left({mathrm{D}}_{mathrm{nj}}*{mathrm{BD}}_{mathrm{nj}}*(1-mathrm{k}*{mathrm{soc}}_{nmathrm{j}})right)right)}{{mathrm{BD}}_{{mathrm{nj}}_{mathrm{bottom}}}*1-mathrm{k}*{mathrm{soc}}_{n{mathrm{j}}_{mathrm{bottom}}}}$$
(7)
where jbottom is the lowest sample depth interval, and other terms are as previous. Using Eqs. (5), (6), and (7), with variable BD and SOC values, SOC stock can be corrected using samples split into the 10 cm and 15 cm sampling depth intervals. More

Gartland, L. A., Firth, J. A., Laskowski, K. L., Jeanson, R. & Ioannou, C. C. Sociability as a personality trait in animals: Methods, causes and consequences. Biol. Rev. https://doi.org/10.1111/brv.12823 (2021).Article 

Google Scholar 
Bergmüller, R. & Taborsky, M. Adaptive behavioural syndromes due to strategic niche specialization. BMC Ecol. 7, 12. https://doi.org/10.1186/1472-6785-7-12 (2007).Article 

Google Scholar 
Massen, J. J. M., Sterck, E. H. M. & de Vos, H. Close social associations in animals and humans: Functions and mechanisms of friendship. Behaviour 147, 1379–1412. https://doi.org/10.1163/000579510X528224 (2010).Article 

Google Scholar 
Haller, J., Harold, G., Sandi, C. & Neumann, I. D. Effects of adverse early-life events on aggression and anti-social behaviours in animals and humans. J. Neuroendocrinol. 26, 724–738. https://doi.org/10.1111/jne.12182 (2014).Article 
CAS 

Google Scholar 
Carlson Bruce, A. Early life experiences have complex and long-lasting effects on behavior. Proc. Natl. Acad. Sci. 114, 11571–11573. https://doi.org/10.1073/pnas.1716037114 (2017).Article 
ADS 
CAS 

Google Scholar 
Zablocki-Thomas, P. B. et al. Personality and performance are affected by age and early life parameters in a small primate. Ecol. Evol. 8, 4598–4605. https://doi.org/10.1002/ece3.3833 (2018).Article 

Google Scholar 
Langenhof, M. R. & Komdeur, J. Why and how the early-life environment affects development of coping behaviours. Behav. Ecol. Sociobiol. 72, 34–34. https://doi.org/10.1007/s00265-018-2452-3 (2018).Article 

Google Scholar 
Daros, R. R., Costa, J. H. C., von Keyserlingk, M. A. G., Hötzel, M. J. & Weary, D. M. Separation from the dam causes negative judgement bias in dairy calves. PLoS One 9, e98429. https://doi.org/10.1371/journal.pone.0098429 (2014).Article 
ADS 
CAS 

Google Scholar 
Grześkowiak, ŁM. et al. Impact of early-life events on the susceptibility to Clostridium difficile colonisation and infection in the offspring of the pig. Gut Microbes 10, 251–259. https://doi.org/10.1080/19490976.2018.1518554 (2019).Article 

Google Scholar 
Schmauss, C., Lee-McDermott, Z. & Medina, L. R. Trans-generational effects of early life stress: The role of maternal behavior. Sci. Rep. 4, 4873. https://doi.org/10.1038/srep04873 (2014).Article 
ADS 
CAS 

Google Scholar 
Brask, J. B., Ellis, S. & Croft, D. P. Animal social networks: An introduction for complex systems scientists. J. Complex Netw. 9, cnab001. https://doi.org/10.1093/comnet/cnab001 (2021).Article 

Google Scholar 
Almeling, L., Hammerschmidt, K., Sennhenn-Reulen, H., Freund, A. M. & Fischer, J. Motivational shifts in aging monkeys and the origins of social selectivity. Curr. Biol. 26, 1744–1749. https://doi.org/10.1016/j.cub.2016.04.066 (2016).Article 
CAS 

Google Scholar 
Borgeaud, C., Sosa, S., Sueur, C. & Bshary, R. The influence of demographic variation on social network stability in wild vervet monkeys. Anim. Behav. 134, 155–165. https://doi.org/10.1016/j.anbehav.2017.09.028 (2017).Article 

Google Scholar 
Cantor, M. et al. The importance of individual-to-society feedbacks in animal ecology and evolution. J. Anim. Ecol. 90, 27–44. https://doi.org/10.1111/1365-2656.13336 (2021).Article 

Google Scholar 
Sosa, S., Sueur, C. & Puga-Gonzalez, I. Network measures in animal social network analysis: Their strengths, limits, interpretations and uses. Methods Ecol. Evol. 12, 10–21. https://doi.org/10.1111/2041-210X.13366 (2021).Article 

Google Scholar 
Farine, D. R. & Whitehead, H. Constructing, conducting and interpreting animal social network analysis. J. Anim. Ecol. 84, 1144–1163. https://doi.org/10.1111/1365-2656.12418 (2015).Article 

Google Scholar 
Neethirajan, S. & Kemp, B. Social network analysis in farm animals: Sensor-based approaches. Animals 11, 434. https://doi.org/10.3390/ani11020434 (2021).Article 

Google Scholar 
Whitehead, H. Analyzing Animal Societies. University of Chicago Press, Chicago, IL, USA (2008).
Smith, J. E. & Pinter-Wollman, N. Observing the unwatchable: Integrating automated sensing, naturalistic observations and animal social network analysis in the age of big data. J. Anim. Ecol. 90, 62–75. https://doi.org/10.1111/1365-2656.13362 (2021).Article 

Google Scholar 
Chen, S., Ilany, A., White, B. J., Sanderson, M. W. & Lanzas, C. Spatial-temporal dynamics of high-resolution animal networks: What can we learn from domestic animals?. PLoS One 10, e0129253. https://doi.org/10.1371/journal.pone.0129253 (2015).Article 
CAS 

Google Scholar 
Atton, N., Galef, B. J., Hoppitt, W., Webster, M. M. & Laland, K. N. Familiarity affects social network structure and discovery of prey patch locations in foraging stickleback shoals. Proc. R. Soc. B Biol. Sci. 281, 20140579. https://doi.org/10.1098/rspb.2014.0579 (2014).Article 
CAS 

Google Scholar 
Ilany, A. & Akçay, E. Personality and social networks: A generative model approach. Integr. Comp. Biol. 56, 1197–1205. https://doi.org/10.1093/icb/icw068 (2016).Article 

Google Scholar 
Romano, V. et al. Modeling infection transmission in primate networks to predict centrality-based risk. Am. J. Primatol. 78, 767–779. https://doi.org/10.1002/ajp.22542 (2016).Article 

Google Scholar 
Ren, K., Bernes, G., Hetta, M. & Karlsson, J. Tracking and analysing social interactions in dairy cattle with real-time locating system and machine learning. J. Syst. Archit. 116, 102139. https://doi.org/10.1016/j.sysarc.2021.102139 (2021).Article 

Google Scholar 
Boyland, N. K., Mlynski, D. T., James, R., Brent, L. J. N. & Croft, D. P. The social network structure of a dynamic group of dairy cows: From individual to group level patterns. Appl. Anim. Behav. Sci. 174, 1–10. https://doi.org/10.1016/j.applanim.2015.11.016 (2016).Article 

Google Scholar 
Chopra, K. et al. Proximity interactions in a permanently housed dairy herd: Network structure, consistency, and individual differences. Front. Vet. Sci. 7, 583715 (2020).Article 

Google Scholar 
Šárová, R. et al. Pay respect to the elders: Age, more than body mass, determines dominance in female beef cattle. Anim. Behav. 86, 1315–1323. https://doi.org/10.1016/j.anbehav.2013.10.002 (2013).Article 

Google Scholar 
Foris, B., Haas, H. G., Langbein, J. & Melzer, N. Familiarity influences social networks in dairy cows after regrouping. J. Dairy Sci. 104, 3485–3494. https://doi.org/10.3168/jds.2020-18896 (2021).Article 
CAS 

Google Scholar 
Foris, B., Zebunke, M., Langbein, J. & Melzer, N. Comprehensive analysis of affiliative and agonistic social networks in lactating dairy cattle groups. Appl. Anim. Behav. Sci. 210, 60–67. https://doi.org/10.1016/j.applanim.2018.10.016 (2019).Article 

Google Scholar 
de Freslon, I., Martínez-López, B., Belkhiria, J., Strappini, A. & Monti, G. Use of social network analysis to improve the understanding of social behaviour in dairy cattle and its impact on disease transmission. Appl. Anim. Behav. Sci. 213, 47–54. https://doi.org/10.1016/j.applanim.2019.01.006 (2019).Article 

Google Scholar 
Bolt, S. L., Boyland, N. K., Mlynski, D. T., James, R. & Croft, D. P. Pair housing of dairy calves and age at pairing: Effects on weaning stress, health production and social networks. PLoS One 12, e0166926. https://doi.org/10.1371/journal.pone.0166926 (2017).Article 
CAS 

Google Scholar 
Koene, P. & Ipema, B. Social networks and welfare in future animal management. Animals (Basel) 4, 93–118. https://doi.org/10.3390/ani4010093 (2014).Article 

Google Scholar 
Raussi, S. et al. The formation of preferential relationships at early age in cattle. Behav. Proc. 84, 726–731. https://doi.org/10.1016/j.beproc.2010.05.005 (2010).Article 

Google Scholar 
Weary, D. M., Jasper, J. & Hötzel, M. J. Understanding weaning distress. Appl. Anim. Behav. Sci. 110, 24–41. https://doi.org/10.1016/j.applanim.2007.03.025 (2008).Article 

Google Scholar 
Lopes, P. C., Block, P. & König, B. Infection-induced behavioural changes reduce connectivity and the potential for disease spread in wild mice contact networks. Sci. Rep. 6, 31790. https://doi.org/10.1038/srep31790 (2016).Article 
ADS 
CAS 

Google Scholar 
Ripperger, S. P., Stockmaier, S. & Carter, G. G. Tracking sickness effects on social encounters via continuous proximity sensing in wild vampire bats. Behav. Ecol. 31, 1296–1302. https://doi.org/10.1093/beheco/araa111 (2020).Article 

Google Scholar 
McGuirk, S. M. & Peek, S. F. Timely diagnosis of dairy calf respiratory disease using a standardized scoring system. Anim. Health Res. Rev. 15, 145–147. https://doi.org/10.1017/s1466252314000267 (2014).Article 

Google Scholar 
Callan, R. J. & Garry, F. B. Biosecurity and bovine respiratory disease. Vet. Clin. N. Am. Food Anim. Pract. 18, 57–77. https://doi.org/10.1016/S0749-0720(02)00004-X (2002).Article 

Google Scholar 
Sewio. Tag Leonardo iMU/Personal. https://docs.sewio.net/docs/tag-leonardo-imu-personal-30146967.html (2022).Barker, Z. E. et al. Use of novel sensors combining local positioning and acceleration to measure feeding behavior differences associated with lameness in dairy cattle. J. Dairy Sci. 101, 6310–6321. https://doi.org/10.3168/jds.2016-12172 (2018).Article 
CAS 

Google Scholar 
Team, R. C. R: A Language and Environment for Statistical Computing.Hadfield, J. D. MCMC methods for multi-response generalized linear mixed models: The MCMCglmm R package. J. Stat. Softw. 33, 1–22. https://doi.org/10.18637/jss.v033.i02 (2010).Article 

Google Scholar 
Franks, D. W., Weiss, M. N., Silk, M. J., Perryman, R. J. Y. & Croft, D. P. Calculating effect sizes in animal social network analysis. Methods Ecol. Evol. 12, 33–41. https://doi.org/10.1111/2041-210X.13429 (2021).Article 

Google Scholar 
Bates, D., Mächler, M., Bolker, B. & Walker, S. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48. https://doi.org/10.18637/jss.v067.i01 (2015).Article 

Google Scholar 
Bell, D. C., Atkinson, J. S. & Carlson, J. W. Centrality measures for disease transmission networks. Soc. Netw. 21, 1–21. https://doi.org/10.1016/S0378-8733(98)00010-0 (1999).Article 

Google Scholar 
Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: A practical and powerful approach to multiple testing. J. R. Stat. Soc. Ser. B (Methodol.) 57, 289–300. https://doi.org/10.1111/j.2517-6161.1995.tb02031.x (1995).Article 
MathSciNet 
MATH 

Google Scholar 
PercieduSert, N. et al. Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biol. 18, e3000411. https://doi.org/10.1371/journal.pbio.3000411 (2020).Article 
CAS 

Google Scholar 
Hulbert, L. E. & Moisá, S. J. Stress, immunity, and the management of calves. J. Dairy Sci. 99, 3199–3216. https://doi.org/10.3168/jds.2015-10198 (2016).Article 
CAS 

Google Scholar 
Sweeney, B. C., Rushen, J., Weary, D. M. & de Passillé, A. M. Duration of weaning, starter intake, and weight gain of dairy calves fed large amounts of milk. J. Dairy Sci. 93, 148–152. https://doi.org/10.3168/jds.2009-2427 (2010).Article 
CAS 

Google Scholar 
Rault, J.-L. Friends with benefits: Social support and its relevance for farm animal welfare. Appl. Anim. Behav. Sci. 136, 1–14. https://doi.org/10.1016/j.applanim.2011.10.002 (2012).Article 

Google Scholar 
Ishiwata, T., Kilgour, R. J., Uetake, K., Eguchi, Y. & Tanaka, T. Choice of attractive conditions by beef cattle in a Y-maze just after release from restraint. J. Anim. Sci. 85, 1080–1085. https://doi.org/10.2527/jas.2006-405 (2007).Article 
CAS 

Google Scholar 
Ede, T., von Keyserlingk, M. A. G. & Weary, D. M. Social approach and place aversion in relation to conspecific pain in dairy calves. PLoS One 15, e0232897. https://doi.org/10.1371/journal.pone.0232897 (2020).Article 
CAS 

Google Scholar 
Cantor, M. C., Renaud, D. L., Neave, H. W. & Costa, J. H. C. Feeding behavior and activity levels are associated with recovery status in dairy calves treated with antimicrobials for Bovine Respiratory Disease. Sci. Rep. 12, 4854. https://doi.org/10.1038/s41598-022-08131-1 (2022).Article 
ADS 
CAS 

Google Scholar 
Kappeler, P. M., Cremer, S. & Nunn, C. L. Sociality and health: Impacts of sociality on disease susceptibility and transmission in animal and human societies. Philos. Trans. R. Soc. Lond. B Biol. Sci. 370, 20140116. https://doi.org/10.1098/rstb.2014.0116 (2015).Article 

Google Scholar 
Ezenwa, V. O. et al. Host behaviour–parasite feedback: An essential link between animal behaviour and disease ecology. Proc. R. Soc. B Biol. Sci. 283, 20153078. https://doi.org/10.1098/rspb.2015.3078 (2016).Article 
CAS 

Google Scholar 
Klein, S. L. Parasite manipulation of the proximate mechanisms that mediate social behavior in vertebrates. Physiol. Behav. 79, 441–449. https://doi.org/10.1016/S0031-9384(03)00163-X (2003).Article 
ADS 
CAS 

Google Scholar 
LavistaFerres, J. M. et al. Social connectedness and movements among communities of giraffes vary by sex and age class. Anim. Behav. 180, 315–328. https://doi.org/10.1016/j.anbehav.2021.08.008 (2021).Article 

Google Scholar 
VanderWaal, K. L., Wang, H., McCowan, B., Fushing, H. & Isbell, L. A. Multilevel social organization and space use in reticulated giraffe (Giraffa camelopardalis). Behav. Ecol. 25, 17–26. https://doi.org/10.1093/beheco/art061 (2014).Article 

Google Scholar 
Sato, S. & Wood-Gush, D. G. Observations on creche behaviour in suckler calves. Behav. Process. 15, 333–343. https://doi.org/10.1016/0376-6357(87)90017-9 (1987).Article 
CAS 

Google Scholar 
Lecorps, B., Kappel, S., Weary, D. M. & von Keyserlingk, M. A. G. Social proximity in dairy calves is affected by differences in pessimism. PLoS One 14, e0223746. https://doi.org/10.1371/journal.pone.0223746 (2019).Article 
CAS 

Google Scholar 
Carslake, C., Occhiuto, F., Vázquez-Diosdado, J. A. & Kaler, J. Repeatability and predictability of calf feeding behaviors—Quantifying between- and within-individual variation for precision livestock farming. Front. Vet. Sci. 9, 827124 (2022).Article 

Google Scholar 
Occhiuto, F., Vázquez-Diosdado, J. A., Carslake, C. & Kaler, J. Personality and predictability in farmed calves using movement and space-use behaviours quantified by ultra-wideband sensors. R. Soc. Open Sci. 9, 212019. https://doi.org/10.1098/rsos.212019 (2022).Article 
ADS 

Google Scholar 
Carslake, C., Occhiuto, F., Vázquez-Diosdado, J. A. & Kaler, J. Indication of a personality trait in dairy calves and its link to weight gain through automatically collected feeding behaviours. Sci. Rep. 12, 19425. https://doi.org/10.1038/s41598-022-24076-x (2022).Article 
ADS 
CAS 

Google Scholar 
Planas-Sitjà, I., Deneubourg, J.-L. & Cronin, A. L. Variation in personality can substitute for social feedback in coordinated animal movements. Commun. Biol. 4, 469. https://doi.org/10.1038/s42003-021-01991-9 (2021).Article 

Google Scholar 
Stockmaier, S., Bolnick, D. I., Page, R. A. & Carter, G. G. Sickness effects on social interactions depend on the type of behaviour and relationship. J. Anim. Ecol. 89, 1387–1394. https://doi.org/10.1111/1365-2656.13193 (2020).Article 

Google Scholar 
Smith, L. A., Swain, D. L., Innocent, G. T., Nevison, I. & Hutchings, M. R. Considering appropriate replication in the design of animal social network studies. Sci. Rep. 9, 7208. https://doi.org/10.1038/s41598-019-43764-9 (2019).Article 
ADS 
CAS 

Google Scholar 
Shin, D. H., Kang, H. M. & Seo, S. Social relationships enhance the time spent eating and intake of a novel diet in pregnant Hanwoo (Bos taurus coreanae) heifers. PeerJ 5, e3329. https://doi.org/10.7717/peerj.3329 (2017).Article 
CAS 

Google Scholar  More

The ecological baseline in TirezThe geology and the climate of the Tirez region favored the generation and maintenance of a type of hypersaline habitat characterized by extreme seasonality: the sulfate-chlorine waters, with sodium and magnesium cations, showed significant seasonal variations15. The alkaline pH, the low oxidant value for the redox potential of the water column and the highly reduced sediments imposed extreme conditions (see Table 1 and Supplementary Information for details). This extreme seasonality requires to define a valid representative ecological baseline to compare the ecology of the lagoon between 2002 and 2021 and, in this way, set the basis to proposing our model of ecological succession with increasing dryness as a “time-analog” for early Mars. Taxonomic data from 2002 is a snapshot of the community during one season, so we include in our discussion the results presented by Montoya et al. (2013) from a sample campaign carried out in 2005, because they16 analyzed both water and sediment and during both the wet and dry seasons.We consider here only the results obtained by Montoya et al.16 by gene cloning, since those obtained by isolation and sequencing are not comparable. At the level of large groups, no major seasonal differences were observed: Pseudomonadota, followed by Bacteroidetes, were the dominant phyla, in both water and sediments, and both in the dry and the rainy seasons; although Alphaproteobacteria was the dominant class in water, while Gammaproteobacteria was dominant in sediments (in both dry and rainy seasons). With respect to the archaeal domain, all the identified sequences were affiliated to Halobacteriales order, mainly Halorubrum (water) and Halobacterium (sediment), both within Halobacteriaceae family. We can consider these results presented in Montoya et al.16 as the “ecological baseline” for Tirez, however taken with a grain of salt, because only 43 bacterial and 35 archaeal sequences, including rainy and dry seasons and water and sediments, were considered for analysis.Prokaryotic diversity in 2002As can be expected for an extreme environment, the bacterial diversity detected in 2002 was low, although we cannot exclude the possibility that this may reflect the limitation of DNA sequencing techniques at the time. 59% of the obtained clones in the then-wet sediments corresponded to the Malaciobacter genus. Malaciobacter (previously named17 Arcobacter) is an aerotolerant Epsilonproteobacteria. Species within this genus are moderately halophilic, e.g., M. halophilus, capable to grow in up to 4% NaCl. Even though the role that Malaciobacter can play in the environment is not known, it seems to thrive in aquatic systems, like sewage, with a high organic matter content17: e.g., M. canalis, M. cloacae, or M. defluvii.After Malaciobacter-like clones, the next most numerous group belongs to the phylum Bacillota (27% of the sequenced clones; Table 2). Under stressful environmental conditions, members of the genus Virgibacillus produce endospores, a very useful property in an extreme and variable environment (ionic strength, temperature, light intensity), easy to compare with early Mars. Endospores facilitate species survival, allowing them to overcome drastic negative changes, like dry periods, and to germinate when the conditions are favorable again. The closest identified species was the halotolerant V. halodenitrificans, but with low homology, not far from other halotolerant (e.g., V. dokdonensis) or halophilic (e.g., V. marismortui) species within the same genus. The other Gram-positive clones belong to the order Clostridiales. These clones cluster in two taxonomic units related with the strictly anaerobic genus Tissierella.Despite the abundance of Pseudomonadota, their biodiversity was very low, reduced to only two genera within the Epsilon- and Delta-proteobacteria. Six sequences affiliated to Deltaproteobacteria, and clustered in one OTU (salB38, similarity 96.6% with Desulfotignum), were retrieved. Its presence in anaerobic media rich in sulfates (Table 1) seems reasonable. In fact, sulfate-reducing activity was detected using a specific enrichment assay.Finally, one taxon belonging to the phylum Spirochaetota (previously named Spirochaetes) was identified. The presence of Spirochaetota in this system is not strange because members of the genus Spirochaeta are very often found in mud and anaerobic marine environments rich in sulfates18. Moreover, the closest species to SalB63, although with a low similarity of 87%, was Spirochaeta bajacaliforniensis, a spirochete isolated19 from a microbial mat in Laguna Figueroa (Baja California), an extensive hypersaline lagoon with high gypsum content, very similar, although much bigger, than Tirez lagoon.The diversity within the domain Archaea was very low in 2002. The phylogenetic analysis of 96 clones indicate that they correspond to one specie belonging to the obligate halophile genus Methanohalophilus. Their high similarity (99.3%) with several species of Methanohalophilus, such as M. portucalensis (isolated from sediments of a solar saltern in Portugal), M. mahii (isolated from sediments of the Great Salt Lake), or M. halophilus (isolated20 from a cyanobacterial mat at Hamelin Pool, Australia), makes impossible its adscription to any particular species level. Methanohalophilus is strictly methylotrophic, which is consistent with this environment, given that the methylotrophic methanogenesis pathway, non-competitive at low-salt conditions, is predominant at high saline concentrations21. We further confirmed methanogenic activity in Tirez by the measurement of methane by gas chromatography in enrichment cultures.Prokaryotic diversity in context of other studies between 2002 and 2021It was challenging to establish a timeline for the succession of the populations involved, because the scarcity of data harvested and published so far from Tirez. However, combining our results with the few data available in Montoya et al.16 and Preston et al.22 on samplings carried out on 2005 and 2017, respectively, we can see a clear predominance of the phylum Pseudomonadota: Epsilonproteobacteria, i.e. Arcobacter-like, and Deltaproteobacteria, mainly sulfate-reducing bacteria (this work, sampling 2002), and Gammaproteobacteria16 when Tirez maintained a water film, to eventually a final predominance of Gammaproteobacteria, e.g. Chromatiales and Pseudomonadales, in the dry Tirez (this work, 2020 sampling). The Bacillales order has remained widely represented both in the wet and dry Tirez.Regarding the archaeal domain, the few references available (Refs.16,22; this work) confirm that the members of the Halobacteriaceae family are well adapted to both the humid and dry ecosystems of Tirez, being predominant in both conditions. Preston et al.22 found that the second most abundant group of archaea in the dry sediments of Tirez was the Methermicoccaceae family, within the Methanosarcinales order, Methanomicrobia class. Taking into account the results obtained in the dry Tirez (Preston et al.22; and this work, sample 2020), the methanogenic archaea have decreased drastically through time, probably due to salt stress and the competition with sulfate-reducing bacteria.Prokaryotic diversity in 2021From a metabolic point of view, most of the bacteria present today in the sediment are chemoorganotrophs, anaerobes, and halophilic or halotolerant. Scarce information is available about the predominant OTU, Candidate Division OP1. The OP1 division was one of the main bacterial phyla in a sulfur-rich sample in the deepest analyzed samples from the Red Sea sediments under brine pools23. In addition, the phylogenetically related Candidate division KB1 has been observed in deep-sea hypersaline anoxic basins at Orca Basin (Gulf of Mexico), and other hypersaline environments24. Eight of the nine genera identified show coverage greater than 1% of the sequences: i.e., Rubinisphaera, Halothiobacillus, Thiohalophilus, Anaerobacillus/Halolactibacillus, Halomonas, Halothermothrix, and Aliifodinibius are halophilic or halotolerant genera13,25.Regarding archaea, our analyses reveal archaeal groups that seem to thrive in sediments from extreme environments, e.g., marine brine pools/deep water anoxic basins or hypersaline lakes. The most abundant OTU, Thermoplasmata KTK4A, was found prominent and active in the sediment of Lake Strawbridge, a hypersaline lake in Western Australia26, and in soda-saline lakes in China27. The creation of a Candidatus Haloplasmatales, a novel order to include KTK4A-related Thermoplasmata, has been proposed27. On the other hand, both in the aforementioned soda-saline lakes in China27 and in a sulfur-rich section of the sediments from below the Red Sea brine pools23, retrieved sequences were assigned to Marine Benthic Groups B, D, and E. Finally, in the section of nitrogen-rich sediments from the aforementioned Red Sea brine pools, the unclassified lineage ST-12K10A represented the most abundant archaeal group. In the Tirez Lagoon sediment after desiccation, all Methanomicrobia readings belonged to this group.The significance and implications of an ecosystem characterized in 2021 by high diversity, high inequality, and lack of isolated representatives, resides in that Tirez is today an ecosystem in which many (most) of the species/OTUs present are dormant, and they do not play any metabolic role. Hence the high percentage of raretons, greater than 80% for both bacteria and archaea, which are actually present in the lagoon but with only one or two copies each. Only those species adapted to the conditions imposed by the extreme environment are able to actually thrive, and consequently only a few species carry out all the metabolic activity. We conclude that the microbiota in Tirez today represents an ecosystem with a high resilience capacity in the face of environmental changes that may occur.We want to clearly highlight that the technique available in 2002 to study the microbiota of the Tirez lagoon only allowed to obtain a low-resolution image, but that was the state-of-the-art procedure at the time, and the Tirez lagoon cannot be sampled again with the conditions back in 2002, which no longer exist and are not expected to return. Although we have kept in storage several samples of water and sediment from the 2002 Tirez lagoon, it is reasonable to assume that those laboratory microcosms would have chemically and microbiologically changed during the last 20 years, and as such no longer represent reliable replicas of the original lagoon, so we cannot use them for the purposes of this work. Therefore, we are aware that any comparisons of the 2002 laboratory results with the much more robust results obtained by Illumina in 2021 need to be taken with a grain of salt. With all the precautions required, in a high-level, first-order comparison, the most noticeable difference between 2002 and 2021 is a drastic change in the microbial Tirez population. Only some OTUs within Bacillales (Virgibacillus/Anaerobacillus), sulfate-reducing Deltaproteobacteria (Desulfotignum/Desulfobacteraceae-Desulfovibrio), and Spirochaetes are shared among the 2002 and 2021 samples. This comparison is enough for the purposes of this work, as we are interested in the evolution of the lagoon system as a whole to establish a “time-analog” with the wet-to-dry transition on early Mars, and not in the particular outcome of each and every OTU in Tirez. With the results at hand, we conclude that, since 2002, the lacustrine microbiota has shifted to one more adapted to the extreme conditions in the dry sediments, derived from the gradual and persistent desiccation concluding ca. 7 years ago (i.e., completely desiccated in 2015), such as lack of light, absence of oxygen, and lack of water availability. This shift has likely been triggered because organisms that were originally in the lagoon but at low abundance in 2002 became dominant as they were better adapted to desiccation, and because the incoming of new microorganisms transported by birds or wind28.Lipid biomarkers analysis of the desiccated lake sedimentsThe analysis of cell membrane-derived lipid compounds on the dry lake sediments at present allow to provide another perspective of the microbial communities inhabiting the Tirez lagoon, by contributing additional information about the ecosystem and depositional environment. It is important to note that, analyzing only the 2021 lake sediments, we cannot differentiate between lipidic biomarkers of the microorganisms inhabiting Tirez in 2002 and before from those left behind by the microorganisms living in the dried sediments today. Instead, the analyses of lipid biomarkers provide clues about the different microorganisms that have populated Tirez through time, including both older communities inhabiting the former aqueous system and also younger communities better adapted to the present dry conditions. Thus, the lipid biomarkers analysis can be considered as a time-integrative record of the microbial community inhabiting Tirez during the last decades.Based on the molecular distribution of lipid biomarkers, the presence of gram-positive bacteria was inferred from the relative abundance of the monounsaturated alkanoic acid C18:1[ω9], or iso/anteiso pairs of alkanoic acids from 12 to 17 carbons29 with dominance of i/a-C15:0 and i/a-C17:0 (Fig. 3B). In contrast, generally ubiquitous alkanoic acids such as C16:1[ω7], C18:1[ω7], or C18:2[ω6] suggested a provenance rather related to gram-negative bacteria30. The combined detection of the i/a-C15:0 and i/a-C17:0 acids, with dominance of the iso over the anteiso congeners, together with other biomarkers such as the mid-chain branched 10Me16:0, the monounsaturated C17:1, or the cyclopropyl Cy17:0 and Cy19:0 acids, may be associated with a community of SRB31 in today´s dry sediments of Tirez. Specifically, most of those alkanoic acids have been found in a variety of Deltaproteobacteria and/or Bacteroidota (previously named Bacteroidetes). The presence of archaea was deduced from the detection of prominent peaks of archaeol in the polar fraction32 (Fig. 3C), as well as squalene and relatives (dihydrosqualene and tetrahydrosqualene) in the apolar fraction33 (Fig. 3A). Squalene and a variety of unsaturated derivatives are present in the neutral lipid fractions of many archaea with high abundances in saline lakes34. The relative abundance of autotrophs over heterotrophs35 can be estimated by the ratio of the autotrophically-related pristane and phytane over the both autotrophically- and heterotrophically-produced n-C17 and n-C18 alkanes ([Pr + Ph]/[n-C17 + n-C18]). A ratio of 0.56 in the Tirez sediments suggest the presence of a relevant proportion of heterotrophs in the ancient lacustrine system.Furthermore, the lipid biomarkers analysis was able to detect compounds specific of additional microbial sources, such as cyanobacteria36 (n-C17, C17:1, or 7Me-C15 and 7Me-C17), microalgae and/or diatoms (phytosterols37; or C20:5, and C22:6 alkanoic acids30), and other photoautotrophs (phytol and potentially degradative compounds such as pristane and phytane31). A relatively higher preservation of the cell-membrane remnants (i.e., lipids) compared to the DNA-composing nucleic acids may contribute to explain the lack of detection of cyanobacteria, diatoms and microalgae, and other phototrophs by DNA analysis (a deficit in our results shared with Montoya et al.16, and Preston et al.22). Although abundant in higher plants38, sterols such as those detected here (i.e., the sterols campesterol, stigmasterol, and β-sitosterol, as well as ergosterol) are also major sterols in some microalgal classes37 (such as Bacillariophyceae, Chrysophyceae, Euglenophyceae, Eustigmatophyceae, Raphidophyceae, Xanthophyceae, and Chlorophyceae), cyanobacteria (β-sitosterol), and fungi (ergosterol39).The carbon isotopic composition of lipid biomarkers provides a rapid screening of the carbon metabolism in a system, by recognizing the principal carbon fixation pathways used by autotrophs. The range of δ13C values measured in the Tirez sediments (from − 33.9 to − 16.1‰) denotes a mixed use of different carbon assimilation pathways, involving mostly the reductive pentose phosphate (a.k.a. Calvin–Benson–Bassham or just Calvin) cycle (from − 19 to − 30‰), and in lesser extent the reductive acetyl-CoA (a.k.a. Wood–Ljungdahl) pathway (from − 28 to − 44‰), and/or the reverse tricarboxylic acid (rTCA) cycle (from − 12 to − 21‰).The lipids synthesized by microorganisms using the Calvin or reductive acetyl-CoA pathway are typically depleted relative to the bulk biomass, particularly those produced via de latter pathway. In the dry Tirez sediments, the majority of the lipid compounds are more depleted in 13C than the bulk biomass (Fig. 4). In particular, the branched alkane DiMeC18 (Fig. 4A) and the SRB-indicative 10Me16:0 acid (Fig. 4B) showed the most depleted δ13C values and suggested the use of the reductive acetyl-CoA pathway. The rest of lipid compounds showed isotopic signatures (from − 16.1 to − 31.4‰) compatible with the prevalence of the Calvin pathway. These values may directly reflect the autotrophic activity of microorganisms fixing carbon via the Calvin cycle or heterotrophic activity of microorganisms growing on their remnants. Thus, the saturated and linear alkyl chains of lipids (i.e., n-alkanes, n-alkanoic acids, and n-alkanols) showing the most negative δ13C values (e.g., alkanes n-C17 and C17:1; or acid C18:1[ω7]) reflect prokaryotic sources of Calvin-users autotrophs (e.g., cyanobacteria or purple sulfur bacteria), while the rest of compounds with slightly less negative δ13C values instead stem from the autotrophic activity of eukaryotes also users of the Calvin cycle (unsaturated fatty acids and sterols) or from the metabolism of heterotrophs such as SRB (iso/anteiso-, other branched, and cyclopropyl fatty acids) and haloarchaea (isoprenoids, phytanol, and archaeol). All in all, the compound-specific isotope composition of the dry sediments in the today´s Tirez lagoon may indirectly reflect the prevailing autotrophic mechanisms in the present lacustrine system of Tirez, by showing isotopic signatures of secondary lipids similar to their carbon source40.In addition, the use of a number of lipid molecular ratios or proxies allow further characterization of the lacustrine ecosystem and depositional environment. For example, the average chain length of the n-alkanes (24.1) suggests a relevant presence of eukaryotic biomass in the lacustrine sediments, since long-chained alkanes ( > C20) are known to originate from epicuticular leaf waxes in higher plants41. Highlighting the relevance of eukaryotes and their ecological roles is one of the major contributions of this work, because previous studies on the microbial ecology of hypersaline environments have been focused primarily on prokaryotes42.The proportion of odd n-alkanes of high molecular chain (i.e., n-C27, n-C29, and n-C31) over even n-alkanes of low molecular chain (i.e., n-C15, n-C17, and n-C19) provides an estimate of the relative abundance of terrigenous over aqueous biomass43, which in Tirez is TAR = 1.8. The Paq index may also be used to differentiate the proportion of terrigenous versus aquatic (emergent and submerged) plant biomass44. A Paq of 0.3 in the Tirez sediments from 2021 supported the relative abundance of land plants. Finally, the depositional environment in the lacustrine system of Tirez may be also characterized analyzing the ratio of pristane over phytane (Pr/Ph), which is higher than 1 when phytol degrades to pristane under oxic conditions45. Assuming that both isoprenoids in the Tirez sediments derived from phytol31, according to their similarly depleted δ13C (Fig. 4A), we can conclude that the sediments in the Tirez lagoon were deposited under predominantly oxic conditions (i.e., Pr/Ph ratio of 1.1).In summary, the lipid biomarkers study revealed useful information about the depositional environment and lacustrine ecosystem, including the presence of active or past autotrophic metabolisms involving prokaryotes (e.g., cyanobacteria and purple sulfur bacteria) and eukaryotes (plants, diatoms and other microalgae), as well as heterotrophic metabolisms of likely SRB and haloarchaea growing on Calvin-users exudates. These results are quite in agreement with the microbial community previously reported16 in sediments from the wet and dry seasons: abundant Gammaproteobacteria and Alphaproteobacteria, together with Algae and Cyanobacteria, dinoflagellates and filamentous fungi, Bacillota, Actinomycetota (previously named Actinomycetes), and a halophilic sulfate-reducing Deltaproteobacteria.Tirez as the first astrobiological “time-analog” for early MarsEarly Mars most likely had a diversity of environments in terms of pH, redox conditions, geochemistry, temperature, and so on. Field research in terrestrial analog environments contribute to understand the habitability of this diversity of environments on Mars in the past, because terrestrial analogues are places on Earth characterized by environmental, mineralogical, geomorphological, or geochemical conditions similar to those observed on present or past Mars9. Therefore, so far analogs have been referred to terrestrial locations closely similar to any of the geochemical environments that have been inferred on Mars, i.e., they are “site-analogs” that represent snapshots in time: one specific environmental condition at a very specific place and a very specific time. Because of this, each individual field analog site cannot be considered an adequate representation of the changing martian environmental conditions through time. Here we introduce the concept of astrobiological “time-analog”, referred to terrestrial analogs that may help understand environmental transitions and the related possible ecological successions on early Mars. In this sense, they should be “time-resolved analogs”: dynamic analog environments where we can analyze changes over time. To the best of our knowledge, this is the first study that looks at the environmental microbiology of a Mars astrobiological analog site over a significant and long period of change, and try to understand the ecological successions to put them in the context of martian environmental evolution.As Mars lost most of its surface water at the end of the Hesperian5,9,12, this wet-to-dry global transition can be considered the major environmental perturbation in the geological history of Mars, and therefore merits to be the first one to be assigned a “time-analog” for its better understanding and characterization. The drying of Mars was probably a stepwise process, characterized by multiple transitions between drier and wetter environments12,47, and therefore the seasonal fluctuations and eventual full desiccation of Tirez represent a suitable analog to better understand possible ecological transitions during the global desiccation of most of the Mars’s surface before the Amazonian (beginning 3.2 Ga).To introduce Tirez as the first Mars astrobiological “time-analog” of the wet-to-dry transition on early Mars, the objective of this study was threefold: first, we wanted to identify the dominant prokaryotic microorganisms in the active Tirez lagoon 20 years ago, a unique hypersaline ecosystem with an ionic composition different from that of marine environments, and therefore potentially analogous to ancient saline lacustrine environments on Mars during the Noachian and into the Hesperian46,47. Our results provide a preliminary basis to hypothesize how the microbial communities on the Noachian Mars could have developed in salty environments with dramatically fluctuating water availability. The requirement to deal with important variations in ionic strength and water availability, involving at times the complete evaporation of the water, could have represented additional constraints48 for microorganisms on early Mars.The second objective of this investigation was the identification of the microbial community inhabiting the desiccated Tirez sediments today, after all the water was lost, as a potential analog to desiccated basins on Mars at the end of the Hesperian1,3,4,47. Our results suggest that hypothetical early microbial communities on early Mars, living with relative abundance of liquid water during the Noachian, would have been forced to adapt to increasingly desiccating surface environments, characterized by extreme conditions derived from the persistent dryness and lack of water availability. Our investigation in Tirez suggest that hypothetical microorganisms at the end of the Hesperian would have needed to evolve strategies similar to those of microorganisms on Earth adapted to living at very low water activity49, to thrive in the progressively desiccating sediments.And the third objective of this investigation was the identification of the lipidic biomarkers left behind by the microbial communities in Tirez, as a guide to searching and identifying the potential leftovers of a hypothetical ancient biosphere on Mars. Lipids (i.e., fatty acids and other biosynthesized hydrocarbons) are structural components of cell membranes bearing recognized higher resistance to degradation relative to other biomolecules, thus with potential to reconstruct paleobiology in a broader temporal scale than more labile molecules50. Our results reinforce the notion that lipidic biomarkers should be preferred targets in the search for extinct and/or extant life on Mars precisely because they are so recalcitrant. More

Biodiversity on Earth is threatened by land-use changes, overexploitation of resources, pollution, biological invasions, and current and projected climate change. Understanding how species will respond to these stressors is difficult, in part because stressors don’t occur in isolation, and because responses can trickle through ecological networks due to interactions among species. More