{"paper_id":"4bad3fab-e62e-4556-98d6-5e40873f8cb3","body_text":"Microbial differences in the habitats of lithophytic bryophytes and their relationship with soil nutrients | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Microbial differences in the habitats of lithophytic bryophytes and their relationship with soil nutrients Wenping Meng, Ran Jingcheng, Xu Zhang, Deming Kong, Fang Liu, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4417220/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Lithophytic Bryophyte is a pioneer plant on the exposed rock surface in karst area,and they can alter the microorganisms in the rock habitat. Results: At the species level, the number of species of various microorganisms in the habitat after the rocky moss was planted on the rock surface was Fungi 235, Bacteria 20535, Eukaryota 816, Archaea 503, and Viruses 155, respectively. Compared with the original control soil, the growth of various microorganisms was Fungi 52%, Bacteria 11%, Eukaryota 78%, Archaea 27%, and Viruses 146%. The number of microbial species related to carbon fixation was 2779, nitrogen fixation was 1502, phosphorus metabolism was 1750.Compared with the original control soil, the bryophytes increased by 37%, 49% and 53% respectively after planting the rock surface. Compared with the original soil, the exposed rock surface increased by 20%, nitrogen fixation by 28% and phosphorus metabolism by 31%.Microbial species with significant differences between groups,Acidimimicrobia_bacterium,Acidimimicrobiaceae_bacterium,Acidimimicrobiales_bacterium, Iamiaceae_bacterium_SCSIO_58843 is significantly positively correlated with potassium content in soil,Microcoleus_Sp._PCC_7113 is a significant negative correlated with potassium content in soil.Alphaprotoobjective_bacterium, Solirubrobacteriales_bacterium, Betaproteobjective_bacterium is a significant positive correlated with succinic acid content in soil.Chloroflexi_bacterium is a significant positive correlated with oxalic acid content insoil.Acidobacteria_bacterium,Solirubrobacterales_bacterium,Acidimicrobiaceae_bacterium is a significant negative correlated with malic acid in soil.Gemmatimonadetes_bacterium is a significant negative correlated with oxalic acid. Smaragdicoccus_niigatensis,Gemmatimonadetes_bacterium,Nocardiaceae_bacterium_YC2-7 is significantly negatively correlated with succinic acid in soil. Solirubrobacterales_bacterium,Archangium_gephyra is a significant negative correlated with acetic acid in soil. Conclusions: The lithophytic bryophytes changed the microbial composition structure in the rock surface habitat, significantly increased the number of functional microorganisms, and then increased the accumulation of potassium, phosphorus, organic carbon and malic acid in the habitat, and promoted the positive development of the rock surface ecosystem. lithophytic bryophytes microorganism karst Rock surface habitat Soil nutrients organic acid Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Background Bryophyte are composed of moss, Hepaticae, and Anthrocerotae, with over 20000 species worldwide, distributed in various ecosystems in tropical, freshwater, marine, and Arctic tundra regions[ 1 ].Some bryophytes grow in water, some on the rock surface, some on the soil surface, and some on leaves or wood.[ 2 ].Lithophytic bryophytes are often born on the surface of rocks in a cluster or cushion community. They have the functions of drought resistance, high temperature resistance, water and soil conservation[ 3 ]. They are pioneer plants on the surface of bare rocks. During their growth, they promote rock dissolution and release nutrients through biochemical and physical mechanical effects [ 4 ]. At the same time, the growth of bryophytes is often accompanied by microorganisms. These microbial communities are responsible for the first stage of soil development, forming a stable soil and fixing carbon and nitrogen biological soil layer[ 5 – 7 ]. Although the microbial community can promote the establishment of mosses, mosses affect the lower soil microbial community through leachate, similar to how different litter types affect the soil microbial community[ 8 – 9 ]. Generally, the moss associated bacterial communities are dominated bypresumptively acidophilic bacteria often associated with ombrotrophic or other oligotrophic environments and arecomparable to the bacterial communities of other moss species[ 10 ]. At present,moss soil crusts have become a research hotspot.People have studied the effects of moss crusts in different ecosystems on the composition characteristics of microbial communities in habitats. It was proved that moss crusts can change the microbial community structure of crusts and increase the relative abundance of functional bacterial groups (nitrogen fixation, photosynthesis,metabolic C1 compounds, plant symbiosis, saprophytic,thereby improving the surface soil microenvironment [ 11 ].The species number of soil bacteria in the moss crust system was between 1529–2381, and the species number of soil fungi was between 323–758. The species number of bacterial community was significantly higher than that of fungal community [ 12 ].Cyanobacteria and Actinobacteria are dominant in the moss crust,which is conducive to the stability of crust structure[ 13 ].For the lithophytic moss crusts, the diversity of bacteria and fungi was similar to that of the soil moss crusts, showing that bacteria accounted for more than 80% of the microbial community composition, and the diversity of bacteria was much higher than that of fungi. The dominant bacteria were Proteobateria, Bateroidetes, Actinobacteria and Acidobacteria.The dominant fungi were Ascomycota and Mortierellomycota[ 14 ].In fact, the microbial diversity and community composition in the soil under the moss crust are closely related to soil factors. The composition of soil bacterial community was significantly affected by SOC, SWC, AK and pH, and the diversity was significantly affected by soil pH, SWC, TP, AP, AN and NO 3 − N[ 12 ].Mg 2+ and SWC in soil are the main factors affecting the functional traits of moss, as they can affect the stress of moss photosynthesis, thereby affecting the progress of other physiological activities and the expression of functional traits[ 15 ].In addition, root exudates can also affect soil microbial richness index and mcintosh index. The higher the concentration of root exudates, the higher the microbial diversity index. However, the microbial dominance index was not affected by the concentration of root exudates[ 16 – 17 ]. Although efforts have been made to study the microbial community composition and ecological functions of moss crust habitats in different ecosystems, moss soil crust (referred to as moss crust) is a composite shell layer formed by the interaction between moss plants, pseudoroots, hyphae, secretions, and soil surface particles such as cyanobacteria, fungi, and bacteria, It contains certain soil and soil microorganisms [ 18 – 19 ]. Therefore, studying the effects of moss crusts on soil microorganisms and other characteristics in habitats cannot accurately reflect the functional traits of moss. This makes studying moss related microorganisms and their impact on environmental microorganisms particularly interesting and meaningful. We hypothesized that:(1) The lithophytic bryophytes in the karst area can improve the diversity of the soil microbial community and enrich the community composition structure.(2)Lithophytic bryophytes can increase the richness of functional microbial communities in rock surface soil.(3)The microorganisms related to soil carbon, nitrogen, phosphorus, potassium and organic acids increased due to the existence of lithophytic bryophytes,which led to the increase of carbon, nitrogen, phosphorus, potassium and organic acids in soil. Results Changes of soil microbial community composition on rock surface after the colonization of lithophytic bryophytes The average number of microorganisms in the rock surface of the bryophytes was 22340 ( Hyophila involute 22063, Eurohypnum leptothallum 22779, Didymodon constrictus 22180 ), 22329 in the exposed rock surface and 19925 in the original control soil.These microorganisms mainly include Fungi, Bacteria, Eukaryota, Archaea, Viruses. At the phylum level, bacteria had the largest number of species, followed by Eukaryota, Archaea, Fungi and Viruses ( Fig.1-A ).The top 10 phylum in the abundance of Bacteria were Actinobacteria、Proteobacteria、Acidobacteria、Cyanobacteria、Chloroflexi、Candidatus_Rokubacteria、Gemmatimonadetes_d_Bacteria、unclassified_d__Bacteria、Bacteroidetes、Planctomycetes（Fig.1-B）.The top 10 phylum in the abundance of Archae were Euryarchaeota、Thaumarchaeota、unclassified_d__Archaea、Candidatus_Bathyarchaeota、Crenarchaeota、Candidatus_Woesearchaeota、Candidatus_Thorarchaeota、Candidatus_Micrarchaeota、Candidatus_Korarchaeota、Candidatus_Altiarchaeota（Fig.1-C）.The top 10 phylum in the abundance of Eukaryot were Streptophyta、Chordata、Ascomycota、Arthropoda、Mucoromycota、Bacillariophyta、unclassified_d_Eukaryota、Nematoda、Basidiomycota、Chytridiomycota（Fig.1-D）。The top 10 phylum in the abundance of Viruses were Artverviricota、unclassified_d__Viruses、Nucleocytoviricota、Phixviricota、Uroviricota（Fig.1-E）。The top 10 phylum in the abundance of Fungi were Mucoromycota、Basidiomycota、Chytridiomycota、Zoopagomycota、Microsporidia、Cryptomycota、Blastocladiomycota、Ascomycota（Fig.1-F）。 At the species level, Fungi 155, Bacteria 18460, Eukaryota 459, Archaea 395, Viruses 63 in the original control soil, and Fungi 181, Bacteria 20597, Eukaryota 631, Archaea 455, Viruses 94 in the exposed rock surface（Fig.2-A）. The average species of various microorganisms in the rock surface habitat after the colonization of lithophytic bryophytes were Fungi 235, Bacteria 20535, Eukaryota 816, Archaea 503, Viruses155, which were more than those in the bare rock surface habitat. Compared with the original control soil, the growth of various microorganisms was Fungi 52 %, Bacteria 11 %, Eukaryota 78 %, Archaea 27 %, Viruses 146 %. Among them, Viruses had the largest growth of 146 %, followed by Eukaryota, Fungi, Archaea, and Bacteria had the smallest growth. We can see that the number of various microorganisms in the rock surface increased significantly after the colonization of bryophytes, and the impact on Viruses, Eukaryota and Fungi was greater than that of Bacteria and Archaea. The top 15 species of Bacteria in each treatment group were Actinobacteria_bacterium、Acidobacteria_bacterium、Chloroflexi_bacterium、Acidimicrobiia_bacterium、Deltaproteobacteria_bacterium、Candidatus_Rokubacteria_bacterium、Smaragdicoccus_niigatensis、Alphaproteobacteria_bacterium、Solirubrobacterales_bacterium、Archangium_gephyra、Acidimicrobiaceae_bacterium、Gemmatimonadetes_bacterium、Betaproteobacteria_bacterium、Nocardiaceae_bacterium_YC2-7、Microcoleus_sp._PCC_7113（Fig.2-B）. The top 15 species of Archaea in each treatment group were Euryarchaeota_archaeon、Thaumarchaeota_archaeon、Candidatus_Bathyarchaeota_archaeon、uncultured_archaeon、Nitrososphaeraceae_archaeon、Nitrosopumilales_archaeon、archaeon_HR01、ANME-2_cluster_archaeon、Thermoplasmata_archaeon、Candidatus_Methanoperedens_nitroreducens、Methanosarcinales_archaeon、Candidatus_Woesearchaeota_archaeon、Hadesarchaea_archaeon、Candidatus_Poseidoniales_archaeon、archaeon（Fig.2-C）. The top 15 species of Eukaryota in each treatment group were Physcomitrium_patens、Marchantia_polymorpha、Pseudocrossidium_replicatum、Solanum_lycopersicum、Homo_sapiens、Lupinus_albus、Eucalyptus_grandis、Syntrichia_ruralis、Candida_albicans、Mus_musculus、Pohlia_nutans、Fusarium_oxysporum、Syntrichia_filaris、Beta_vulgaris、Phaeodactylum_tricornutum（Fig.2-D）. The top 15 species of Fungi in each treatment group were Candida_albicans、Fusarium_oxysporum、Powellomyces_hirtus、Fusarium_odoratissimum、Rhinocladiella_mackenziei、Parasitella_parasitica、Oidiodendron_maius、Diversispora_epigaea、Lipomyces_starkeyi、Smittium_culicis、Absidia_glauca、Rhizopus_delemar、Gigaspora_rosea、Spizellomyces_sp._'palustris'、Rhizophagus_clarus（Fig.2-E）。The top 15 species of Viruses in each treatment group were Murine_leukemia_virus、Rhodococcus_phage_REQ1、Circular_genetic_element_sp.、Gordonia_phage_Phendrix、Gordonia_phage_Mollymur、Caudovirales_GX15bay、Streptomyces_phage_Henoccus、Gordonia_virus_Suzy、Mycobacterium_phage_Kumao、Arthrobacter_phage_Shoya、Microbacterium_phage_Rasovi、Streptomyces_phage_Yara、Microbacterium_phage_FuzzBuster、Gordonia_virus_Ghobes、Arthrobacter_virus_Joann（Fig.2-F）。 4.2 The main functions of soil microorganisms in the lithophytic bryophytes rock surface By conducting functional clustering analysis on the overall gene set and selecting the top 50 ranked functions, we found that the main functions of these microorganisms are amino acid metabolism, organic acid metabolism, Metabolism of carbon, nitrogen, phosphorus, sulfur, and methane, Photosynthesis related functions, Secondary metabolite related function, glycometabolism, Enzyme related functions(Pantothenate and CoA biosynthesis) （Fig.3-A）. There are differences in microbial functions between different treatment groups. The difference in microbial functions within Hyophila involute and Eurohypnum leptothallum is small, but they differ greatly from Didymodon constrictus and the control group. However, the difference between Didymodon constrictus and the control group is small (Fig.3-B). It indicated that bryophytes had a significant role in promoting the functional development of rock surface soil, but there were interspecific differences in this contribution. Effects of lithophytic bryophytes on carbon fixation, nitrogen fixation and phosphorus metabolism functional microorganisms in rock surface After planting on the rock surface, the microorganisms related to carbon fixation in the soil were 2779, nitrogen fixation 1502, and phosphorus metabolism 1750.In the original control soil, carbon-fixing microorganisms 2031, nitrogen-fixing 1008, and phosphorus metabolism 1141. Carbon-fixing microorganisms 2444, nitrogen-fixing 1288, and phosphorus metabolism 1490 in the bare rock surface habitat. Compared with the original control soil, the growth rate of carbon-fixing microorganisms, nitrogen fixation and phosphorus metabolism in the soil after bryophyte planting rock surface increased by 37 %, 49 % and 53 %, respectively. Compared with the original soil, the growth rate of carbon-fixing microorganisms in the exposed rock surface was 20 %, nitrogen fixation was 28 %, and phosphorus metabolism was 31 % ( Fig.4-A ). We can see that the number of microorganisms related to carbon fixation, nitrogen fixation and phosphorus metabolism in the habitat was significantly increased after the colonization of the rock surface. Among the top 15 species of microbial abundance related to carbon fixation, nitrogen fixation and phosphorus metabolism, 9 kinds of microorganisms appeared in the three functional groups and accounted for a relatively high proportion, which were respectively:Actinobacteria_bacterium、Acidobacteria_bacterium、Acidimicrobiia_bacterium、Chloroflexi_bacterium、Smaragdicoccus_niigatensis、Solirubrobacterales_bacterium、Deltaproteobacteria_bacterium、Alphaproteobacteria_bacterium、Gemmatimonadetes_bacterium. The unique microbial species in the carbon fixation functional group are Iamiaceae_bacterium_SCSIO_58843、Candidatus_Rokubacteria_bacterium、Betaproteobacteria_bacterium、Thermoleophilia_bacterium（Fig.4-B）. The unique microorganisms in the functional groups related to phosphorus metabolism are Thermoleophilia_bacterium、Gaiella_occulta、Archangium_gephyra（Fig.4-C）. The unique microorganisms in the nitrogen-fixing functional group are Candidatus_Rokubacteria_bacterium、Gaiella_occulta、Nocardiaceae_bacterium_YC2-7、Betaproteobacteria_bacterium、Microcoleus_sp._PCC_7113、Rhizobiales_bacterium（Fig.4-D）. Functional analysis of microbial carbon fixation, nitrogen fixation and phosphorus metabolism in the habitat of lithophytic bryophytes after planting rock surface There are differences in the metabolic pathways related to carbon fixation between the moss colonized rock surface and the exposed rock surface compared to the original soil (Fig.5).There are also interspecific differences among lithophytic bryophytes. The carbon fixation metabolic pathways between Hyophila involute and Eurohypnum leptothallum , Didymodon constrictus are significantly different, while the carbon fixation metabolic pathways between Didymodon constrictus and exposed rock surface habitats are similar. The top 15 metabolic pathways related to carbon fixation were significantly different among the treatment groups. The abundance of metabolic pathways K01681, K01903, K01961, K00031, K01007, K01595, and K01847 in the habitat after the colonization of bryophytes on the rock surface was significantly higher than that in the original control soil. However, the abundance of K01848, K00174, K00297, K00175 and K02446 decreased significantly ( Fig.5-B ). The metabolic pathways related to nitrogen fixing in the habitat of lithophytic bryophytes after planting on the rock surface are different from those in the original control soil and exposed rock surface, while the nitrogen fixing metabolic pathways in the original control soil and exposed rock surface are more similar (Fig.6).There are also differences among different species of moss, and the nitrogen fixation metabolic pathways of Hyophila involute are more similar to those of Eurohypnum leptothallum .The metabolic pathways of K00265, K00362, K00266, K02575, and K00370 in the habitat after the planting of lithophytic bryophytes on the rock surface were significantly higher than those in the original control soil, while K0195 and K00261 were significantly lower than those in the original control soil. K15577, K15578, and K15576 are significantly higher in exposed rock habitats than in rocky moss colonized rock surfaces and the original control soil. The phosphorus metabolism pathway in the habitat of lithophytic bryophytes planted on the rock surface was different from that in the original control soil and exposed rock surface. Moreover, there were differences between the lithophytic mosses, Hyophila involute and Didymodon constrictus having more similar soil phosphorus metabolism functions, the original control soil and the exposed rock surface are similar.K00937, K00951, K00873, K03306, and K01507 were significantly higher in the habitat after lithophytic moss was planted on the rock surface than in the original soil control group, while K01524, K01113, K02039, and K00117 were significantly reduced (Fig.7). The different species of carbon fixation, nitrogen fixation and phosphorus metabolism in rock surface colonized by lithophytic bryophytes Fig.8 Changes of functional microbial communities in the habitat of lithophytic bryophytes after planting rock surface, A : significantly different species of carbon-fixing microorganisms, B : significantly different species of nitrogen-fixing microorganisms, C : significantly different species of phosphorus metabolism There were significant differences in the top 15 species of microorganisms related to carbon fixation among the groups.Smaragdicoccus_niigatensis、Gemmatimonadetes_bacterium、Acidimicrobiaceae_bacterium、Thermoleophilia_ bacterium were significantly higher than those of the original control soil, and the richness of Smaragdicoccus_niigatensis was significantly higher than other groups.However, Actinobacteria_bacterium, Acidobacteria _bacterium, Acidimicrobiia _bacterium, Candidatus_Rokubacteria_bacterium, Iamiaceae_bacterium_SCSIO_ 58843 decreased significantly after the bryophytes colonized rock surface ( Fig.8-A ). The top 15 species of nitrogen-fixing microorganisms were significantly different among the groups. Smaragdicoccus_niigatensis, Gemmatimonadetes_bacterium, Nocardiaceae_bacterium_YC2-7, Microcoleus_sp._PCC_7113 were significantly higher than the original control soil.Among them, the abundance of Smaragdicoccus_niigatensis and Nocardiaceae _bacterium_YC2-7 in Hyophila involute and Eurohypnum leptothallum was higher than that of other treatment groups.However, Actinobacteria_bacterium、Candidatus_Rokubacteria_bacterium、Acidimicrobiia_bacterium、Chloroflexi_bacterium、Deltaproteobacteria_bacterium、 Rubrobacter_taiwanensis、Streptomyces_thermoautotrophicus、Rhizobiales_bacterium 、Streptomyces_lunaelactis decreased significantly compared with the original control soil ( Fig.8-B ). The top 15 species of microorganisms related to phosphorus metabolism were significantly different among groups.Chloroflexi_bacterium、Smaragdicoccus_niigatensis、unclassified_d__unclassified、Gemmatimonadetes_bacterium、Archangium_gephyra increased significantly after the rock surface was colonized by Lithophytes.The abundance of Smaragdicoccus_niigatensis was higher in the habitats of Eurohypnum leptothallum and Hyophila involute than that in the habitats of Didymodon constrictus . However, Actinobacteria_bacterium、Acidobacteria_bacterium、Acidimicrobiia_bacterium、Acidimicrobiales_bacterium、Deltaproteobacteria_bacterium、Acidimicrobiaceae_bacterium、Candidatus_Rokubacteria_bacterium decreased significantly after the colonization of lithophytic bryophytes (Fig.8-C). The relationship between different species and soil physical and chemical properties The correlation between the microorganisms related to carbon fixation, nitrogen fixation,phosphorus metabolism and the contents of nitrogen, phosphorus, potassium, organic carbon, oxalic acid, acetic acid, malic acid and succinic acid in rock surface soil was analyzed. We found that Acidimicrobiia_bacterium, Acidimicrobiaceae_ bacterium and Acidimicrobiales_bacterium in the top 15 microorganisms related to phosphorus were significantly positively correlated with potassium content in soil, and Deltaproteobacteria_bacterium was positively correlated with phosphorus content in soil ( Fig.9-A ).Alphaproteobacteria_bacterium and Solirubrobacterales_bacterium were significantly positively correlated with succinic acid content in soil, and significantly negatively correlated with soil organic carbon. Acidobacteria_bacterium, Solirubrobacterales_bacterium and Acidimicrobiaceae_bacterium were significantly negatively correlated with malic acid in soil. Solirubrobacterales_bacterium and Archangium_gephyra were significantly negatively correlated with acetic acid in soil. Among the top 15 microorganisms related to carbon fixation, Iamiaceae_ bacterium_SCSIO_58843 was significantly positively correlated with potassium content in soil. Gemmatimonadetes_bacterium was positively correlated with soil organic carbon. Chloroflexi_bacterium was significantly positively correlated with oxalic acid content in soil. Acidimicrobiaceae _ bacterium was significantly negatively correlated with malic acid in soil. Betaproteobacteria_bacterium and Solirubrobacterales_bacterium were significantly positively correlated with succinic acid in soil (Figure 9-B). Among the top 15 microorganisms related to nitrogen fixation, Candidatus_ Rokubacteria_bacterium_3112 was positively correlated with potassium content in soil.Microcoleus_sp._PCC_7113 was significantly negatively correlated with potassium content in soil. Candidatus _ Rokubacteria _bacterium_3112, Microcoleus _ sp._PCC_7113, Nocardiaceae_bacterium_YC2-7 were positively correlated with soil nitrogen content but not significant.Gemmatimonadetes_bacterium was significantly negatively correlated with oxalic acid.Smaragdicoccus_niigatensis, Gemmatimonadetes_bacterium, Nocardiaceae_bacterium_YC2-7 were significantly negatively correlated with succinic acid in soil. Solirubrobacterales_bacterium was significant positive correlation with succinic acid ( Fig.9-C). Discussion The average number of microorganisms in the epigenetic soil of the rock surface is 22340, the exposed rock surface is 22329, and the original control soil is 19925. The total number of microorganisms in the lithophytic bryophytes increased by 12% compared with the original control soil, and the number growth rates of various microorganisms were Fungi 52%, Bacteria 11%, Eukaryota 78%, Archaea 27%, Viruses 146%. We can see that the total amount of microorganisms in the rock surface increased after the colonization of bryophytes. Among them, the number of microbial species in Viruses, Eukaryota and Fungi increased more, while Bacteria and Archaea increased, but the increase was not significant. Moreover,the microbial abundance of Cyanobacteria,Gemmatimonadetes_d_Bacteria,Bacteroidetes,Planctomycetes in Bacteria, Candidatus_Woesearchaeota in Archaea, Streptophyta, Arthropoda, Bacillariophyta in Eukaryota, Mucoromycota, Chytridiomycota and Zoopagomycota in Fungi was higher in the rock surface of bryophytes than in the exposed rock surface and the original control soil.We can see that the influence of lithophytic bryophytes on the total amount of various microorganisms in the rock surface is not significant, but it has a significant effect on the diversity of microorganisms in Viruses,Eukaryota and Fungi, and then changes the composition of microbial communities. Although there is no lithophytic bryophytes on the exposed rock surface, its development environment is exactly the same as that of the lithophytic bryophytes, and there is little difference in the total amount of soil microorganisms between the two. The reason for this result may be related to the important influence of soil moisture on the development of soil microorganisms[ 20 ]. In addition, the microbial number of Fungi, Eukaryota, Archaea and Viruses in Hyophila involute was higher than that in Eurohypnum leptothallum and Didymodon constrictus . While the number of Bacteria in Eurohypnum leptothallum was the highest.It shows that there are interspecific differences in microbial diversity and community composition in lithophytic bryophytes.The composition of moss-associated bacterial communities may be driven by host identity[ 21 ], the characteristics of bryophyte gametophytes and secondary metabolism[ 22 – 23 ]. These changes may affect the composition of the moss microbiome[ 24 ]. Hyophila involute like to grow on bare rock surfaces or crevices in karst rocky desertification areas[ 25 ]. The characteristics of this rocky habitat are strong sunlight on sunny days, high temperature on the rock surface, low air humidity, rapid water loss on the rock surface during rainfall, and poor water retention. Hyophila involute can grow in this harsh habitat, indicating that it has special skills of sun resistance, heat resistance, and drought resistance[ 26 ]. This special skill may lead to secretions conducive to the development of microorganisms in Fungi, Eukaryota, Archaea, and Viruses. The effect of vascular plant root exudates on soil microbial community structure has been confirmed[ 27 – 28 ],but how bryophyte secretions affect soil microorganisms has not been clearly reported, and research in this area needs to be strengthened in the future. Changes of carbon-fixing microbial communities and their metabolic pathways in the habitat of lithophytic bryophytes planted on the rock surface After the rock surface was colonized by lithophytic bryophytes, the number of microorganisms related to carbon fixation, nitrogen fixation and phosphorus metabolism was not only increased, but also the abundance of metabolic pathways of them was significantly increased.For example, the abundance of K01681, K01903, K01961, K00031, K01007, K01595 and K01847 metabolic pathways in the carbon-fixing functional community was significantly higher than that in the original control soil after lithophytic bryophytes planted on rock surface.Among them, K01681 is aconitate hydratase, which is a coenzyme that catalyzes the conversion between citric acid and isocitric acid[ 29 ]. K01903 (succinyl-CoA synthetase beta subunit ) is an enzyme in the tricarboxylic acid cycle, under the action of succinate sulfur kinase, high-energy thioester bond hydrolysis, energy transfer to GDP to generate GTP and succinic acid[ 30 ].K01961 (acetyl-CoA carboxylase, biotin carboxylase subunit) is a biotin enzyme that catalyzes the reaction of‘acetyl-CoA + ATP + HCO3 − → malonyl-CoA + ADP + Pi’.The malonyl-CoA is the base of important metabolic reactions such as fatty acid synthesis and acyl chain extension system[ 31 ].K00031(isocitrate dehydrogenase) is a key enzyme linking C-N metabolism[ 32 ]. The assimilation of inorganic nitrogen in plants begins with carbon compounds produced by C metabolism, and isocitrate dehydrogenase can catalyze the reversible reaction of oxidative decarboxylation of isocitrate to form α-ketoglutarate, which is the most important and most regulated rate-limiting enzyme in the tricarboxylic acid cycle[ 33 ].K01595(phosphoenolpyruvate carboxylase)is a cytoplasmic enzyme that catalyzes the β-carboxylation of phosphoenolpyruvate in the presence of HCO 3 − , and uses Mg 2+ as a cofactor to produce oxaloacetic acid and inorganic phosphate[ 34 ].The colonization of lithophytic bryophytes not only increased the number of microorganisms related to carbon fixation in the habitat, but also increased the abundance of enzymes related to citric acid cycle (TCA cycle), pyruvate metabolism, oxalic acid metabolism, and prokaryotic carbon fixation, thereby promoting the carbon fixation pathway in the rock surface habitat. The changes of phosphorus metabolic microbial community and its metabolic pathway in the habitat of lithophytic bryophytes planted on the rock surface The number of microorganisms related to phosphorus metabolism, metabolic pathways K00937, K00951, K00873, K03306 and K01507 in the habitat after the colonization of lithophytic bryophytes on the rock surface were significantly higher than those in the original soil control group.Among them,K00937 (polyphosphate kinase) can synthesize polyphosphate polyP, which contains several to hundreds of orthophosphate residues connected by high-energy phosphate glycosidic bonds to form straight chains, belonging to the polymer form of phosphate[ 35 ].K00873 (pyruvate kinase) is an essential irreversible enzyme in the glycolysis process, mainly catalyzing the conversion of phosphoenolpyruvate to pyruvate, accompanied by the production of two molecules of ATP. It requires the participation of divalent cations and is an exothermic reaction[ 36 ].Under anaerobic conditions, pyruvate undergoes fermentation to produce lactic acid or ethanol. Under aerobic conditions, pyruvate is converted to acetyl CoA and enters mitochondria for the tricarboxylic acid cycle[ 37 ].K03306 (inorganic phosphate transporter, PiT family) can transport phosphorus into the cytoplasm[ 38 – 39 ] or various physiological and biochemical processes in cells[ 40 ].These enzymes are key enzymes involved in phosphate synthesis, phosphate transport, and glycolysis processes. Increasing their abundance promotes the synthesis of phosphorus in soil and the transport of phosphorus within plants. Changes in nitrogen fixing microbial communities and their metabolic pathways in the habitat of lithophytic moss planted on rock surfaces The number of nitrogen fixing related microorganisms and metabolic pathways of K00265, K00362, K00266, K02575, and K00370 in the habitat of lithophytic moss after planting on the rock surface were significantly higher than those in the original control soil.Among them, K00265 glutamate synthase (NADPH) large chain is a key enzyme in bacterial ammonia assimilation process[ 41 ], and the ammonia generated by biological reduction of N 2 is also assimilated through the glutamate synthase cycle[ 42 ]. K00362 nitrate reduction (NADH) large subunit is the process of degrading nitrite into ammonium[ 43 ].K02575 (MFS transporter, NNP family, nitrate/nitrate transporter) plays an important role in maintaining nitrogen balance in bacteria[ 44 ].Nitrite oxidoreductases are a class of enzymes that catalyze nitrite reduction and can degrade NIT to NO or NH 3 .They are key enzymes in the natural nitrogen cycle process[ 43 ].These enzymes are key enzymes in the nitrogen metabolism process, indicating an increase in nitrogen metabolism, alanine, aspartic acid, glutamic acid metabolism, and amino acid biosynthesis activity in the habitat of lithophytic mosses after colonization on rock surfaces. The relationship between microorganisms and the contents of carbon, nitrogen, phosphorus and potassium in the rock surface soil of lithophytic moss planted. Through correlation analysis between different species and nitrogen, phosphorus, potassium and organic carbon in rock surface soil, we found that acidimicrobia_ Bacterium, acidimicrobiaceae_Bacterium, acidimicrobiales_Bacterium, iamiaceae_ Bacterium_SCSIO_58843 was significantly positively correlated with the content of potassium in soil. Deltaproteobacteria_There was a significant positive correlation between bacteria and phosphorus content in soil_Bacteria was positively correlated with soil organic carbon. It is suggested that the increase of these microorganisms will promote the increase of potassium, phosphorus and organic carbon in rock surface soil. However, there was no significant correlation between these species and soil nitrogen content. However, studies have shown that bryophyte cyanobacterial symbionts (BCS) can fix nitrogen[ 45 ], and this symbiont inputs 0.5–10kg n/ha of nitrogen into the northern hemisphere ecosystem every year[ 46 – 47 ].Who plays the main role in moss cyanobacteria symbiosis?Cyanobacteria are a class of autotrophic prokaryotes[ 48 ], and their biological nitrogen fixation is mainly carried out by nitrogenase in heterocysts [ 49 ](Adams&Duggan,2008).Other results suggest that moss N 2 -fixationrates are determined by bacterial community structure, rather than moss traitsor time since deglaciation[ 50 ]。Therefore,the symbionts of bryophytes and cyanobacteria mainly rely on cyanobacteria for nitrogen fixation, while bryophytes contribute less to nitrogen fixation.Bryophytes and cyanobacteria belong to mutualism. Bryophytes can provide nutrients such as carbohydrates and relatively stable microenvironment conditions such as temperature, moisture and humidity for cyanobacteria. The transfer of nitrogen fixed by cyanobacteria to bryophytes can increase the biomass of bryophytes[ 51 ].The number of cyanobacterial heterocysts and nitrogen fixation capacity in moss cyanobacteria symbionts were significantly higher than those in independent autotrophic cyanobacteria. For example, the number of heterocysts in cyanobacteria in symbionts can reach 6–10 times that of independent survival, and the nitrogen fixation rate can increase to 7 times[ 49 ].Therefore,bryophytes can not play the role of nitrogen fixation alone, but need to be combined with cyanobacteria, but it also needs a period of adaptation after combining with cyanobacteria to play the role of nitrogen fixation. In addition, N 2 fixation rate is variable and may be affected by moss species[ 52 – 53 ], nitrogen availability[ 54 ], water[ 55 ],temperature[ 56 ], composition of diazotrophs[ 57 ], abundance of diazotrophs[ 54 ] and activity of diazotrophs[ 58 ] . The relationship between microorganisms and organic acid content in rock surface habitat of lithophytic bryophytes The differential species Alphaproteobacteria_bacterium, Solirubrobacterales_ bacterium and Betaproteobacteria_bacterium among bare rock surface, original control soil and the rock surface planted lithophytic bryophytes were significantly positively correlated with succinic acid in soil.Smaragdicoccus_niigatensis and Gemmatimonadetes_bacterium.Chloroflexi_bacterium was significantly positively correlated with oxalic acid content in soil. However,Nocardiaceae_bacterium_YC2-7 was significantly negatively correlated with succinic acid in soil, and Gemmatimonadetes_bacterium was negatively correlated with oxalic acid in soil.Therefore, the content of succinic acid and oxalic acid in the rock surface habitat was affected by the stone moss. However, Acidobacteria_bacterium, Solirubrobacterales_bacterium and Acidimicrobiaceae _bacterium were significantly negatively correlated with malic acid in soil. The difference species Solirubrobacterales_bacterium and Archangium_ gephyra were significantly negatively correlated with acetic acid in soil, and their abundance increased on the rocky moss planting rock surface, indicating that the content of acetic acid in soil decreased, which was consistent with the detection results of acetic acid content in soil. Conclusion Lithophytic bryophytes can increase the number of microorganisms in rocky habitats, especially significantly affecting the diversity of microorganisms in Viruses, Eukaryota, and Fungi, altering the composition and structure of microbial communities, and enriching the types and quantities of functional microorganisms such as carbon fixation, nitrogen fixation, and phosphorus metabolism in soil.Lithophytic bryophytes increased the abundance expression levels of key enzymes in the processes of citric acid cycle (TCA cycle), pyruvate metabolism, oxalate metabolism, prokaryotic carbon fixation, phosphate synthesis, phosphorus transport, glycolysis, nitrogen metabolism, alanine, aspartic acid, glutamic acid metabolism, and amino acid biosynthesis in rocky habitats, promoting an increase in potassium, phosphorus, organic carbon, and malic acid content in soil. However, stone moss has an impact on soil nitrogen, acetic acid The influence of oxalic acid and succinic acid is not significant. Methods Materials In this study, the dominant species of lithophytic bryophytes in the karst area, Didymodon constrictus , Eurohypnum leptothallum , and Hyophila involuta , were selected as the research objects. The materials required for the experiment were collected from the Puding Karst Rocky Desertification Ecosystem Observation and Research Station of the Chinese Academy of Sciences.The plant specimen was identified by Wenping Meng of Guizhou Botanical Gardon.The specimens are now deposited in the herbarium of Guizhou Botanical Garden. Experimental design The dominant species of lithophytic bryophytes in karst area were collected, such as Eurohypnum leptothallum , Didymodon constrictus and Hyophila involute , cleaned the soil, litter and other impurities in the bryophyte.Select carbonate rock flakes, clean, the surface of a thin layer of soil just cover the rock surface is appropriate.Three kinds of cleaned bryophytes were planted on the surface of rock surface respectively. Hyophila involute (S group), Eurohypnum leptothallum (M group) and Didymodon constrictus (D group), and each moss was set up with 5 replicates.The stone flakes with only soil and no bryophytes on the surface were set as L group.Before the experiment,a certain amount of rock surface soil was taken as the control soil and frozen in the refrigerator. At the end of the experiment, the microorganisms were detected.All the experimental groups were placed in the incubator,set the day 12 hours, light intensity 8000Lux, temperature 25°C,night 12 hours,light intensity 0Lux, temperature 15°C.Water was sprayed once a week. After 12 months, the bryophytes on the rock surface were removed, the soil on the rock surface was collected and stored at − 20°C for further analysis. (2) Detection of soil physical and chemical indicators We used potassium dichromate external heating method to detect soil organic carbon (SOC) content, used perchloric acid-sulfuric acid digestion, Kjeldahl method to determine total nitrogen content,used perchloric acid-sulfuric acid digestion-molybdenum antimony colorimetric method to determine total phosphorus content, and used hydrofluoric acid-perchloric acid digestion method to determine total potassium content. (3)Soil microbial detection methods DNA extraction Total genomic DNA was extracted from soil samples using the Mag-Bind® Soil DNA Kit (Omega Bio-tek,Norcross,GA,U.S.) according to manufacturer’s instructions. Concentration and purity of extracted DNA was determined with TBS-380 and NanoDrop2000, respectively. DNA extract quality was checked on 1% agarose gel. Library construction, and metagenomic sequencing DNA extract was fragmented to an average size of about 400 bp using Covaris M220 (Gene Company Limited,China) for paired-end library construction. Paired-end library was constructed using NEXTFLEX Rapid DNA-Seq (Bioo Scientific, Austin, TX,USA). Adapters containing the full complement of sequencing primer hybridization sites were ligated to the blunt-end of fragments. Paired-end sequencing was performed on Illumina NovaSeq(Illumina Inc., San Diego, CA, USA) at Majorbio Bio-Pharm Technology Co., Ltd. (Shanghai, China) using NovaSeq 6000 S4 Reagent Kit v1.5 (300 cycles) according to the manufacturer’s instructions ( www.illumina.com ). (4)Statistical analysis Sequence quality control and genome assembly The data were analyzed on the free online platform of Majorbio Cloud Platform ( www.majorbio.com ). Briefly, the paired-end Illumina reads were trimmed of adaptors, and low-quality reads (length < 50 bp or with a quality value < 20 or having N bases) were removed by fastp.Contigs with with a length ≥ 300 bp were selected as the final assembling result, and then the contigs were used for further gene prediction and annotation. Gene prediction, taxonomy, and functional annotation Open reading frames (ORFs) from each assembled contig were predicted using Prodigal/MetaGene.The predicted ORFs with a length ≥ 100 bp were retrieved and translated into amino acid sequences using the NCBI translation table. A non-redundant gene catalog was constructed using CD-HIT with 90% sequence identity and 90% coverage. High-quality reads were aligned to the non-redundant gene catalogs to calculate gene abundance with 95% identity using SOAPaligner. Species and functional annotations Representative sequences of non-redundant gene catalog were aligned to NR database with an e-value cutoff of 1e − 5 using Diamond for taxonomic annotations. Cluster of orthologous groups of proteins (COG) annotation for the representative sequences was performed using Diamond against egg NOG database with an e-value cutoff of 1e − 5 .The KEGG annotation was conducted using Diamond against the Kyoto Encyclopedia of Genes and Genomes database with an e-value cutoff of 1e − 5 . Analysis The Meiji biological cloud database was used to analyze the data, and the dominant species and functional composition of the community were studied by column graph visualization method. The community Heatmap analysis tool was used to calculate the species/gene/functional abundance of each sample. Hierarchical clustering of the distance matrix can clearly see the distance of the sample branches.Based on the obtained species and functional abundance data of different groups, one-way analysis of variance is used to conduct hypothesis tests on the species and functions among their microbial communities, evaluate the significance level of species, functions, or gene abundance differences, and obtain the species and functions with significant differences between groups. he correlation between different species and soil physical and chemical indexes was analyzed. Abbreviations C carbon N nitrogen SOC soil organic carbon SWC soil water content AK available potassium content pH acidity and alkalinity TP total phosphorus AP available phosphorus AN available nitrogen NO 3 - N nitrate nitrogen Mg 2+ magnesium ion KEGG the most commonly used international bioinformatics database GDP guanosine diphosphate GTP guanosine triphosphate ATP adenosine triphosphate HCO3 - bicarbonate ion ADP adenosine diphosphate Pi phosphate group NADPH nicotinamide adenine dinucleotide phosphate NO nitric oxide NH 3 ammonia BCS bryophyte cyanobacterial symbionts Declarations Ethics approval and consent to participate The manuscript does not involve the use of any animal or human data or tissues. Statement of collect plant samples The plants involved in the manuscript are not rare and endangered plants and are not included in the IUCN protection list.The plants were collected at the Puding karst rocky desertification ecosystem research and observation station of the Chinese Academy of Sciences. Statement of plant materials and experimental methods The collection of plant materials in the manuscript was in accordance with relevant institutional, national and international guidelines and legislation. The methods involving plant materials are performed in accordance with the institutional, national, and international guidelines and legislation. Consent for publication Not applicable. Availability of data and materials The high-throughput sequencing metagenomic data of microorganisms in the soil underlying three types of moss plants is stored in the China National Center for Bioinformation:https://ngdc.cncb.ac.cn/ PRJCA026083 Competing interest The authors declare no competing interests. Funding This work was supported by Science and Technology Fund of Guizhou (ZK[2023]-236,[2020]1Y074,ZK[2023]-237),Guizhou Forestry Research Project([2023]07),Doctoral Fund of Guizhou Academy of Sciences([2023]01),National Natural Science Foundation of China (32160086),National Nature Science Foundation of China and the Karst Science Research Center of Guizhou Province (U1812401). Authors’ contributions Meng Wenping is responsible for the detection, data analysis and manuscript writing of Bryophyte physiological indicators.Xu Zhang,Kong Deming and Zheng Ting are responsible for the field survey.Qi Tong,Chen Wang,Fang Liu are responsible for the collection and cultivation of Bryophyte. Ran Jingcheng is responsible for revising the manuscript. The author has read and approved the final manuscript. Acknowledge Thank you to Daqing Liang from Meiji Biotechnology for his assistance in data processing and mapping. References Klarenberg IJ,Keuschnig C,Salazar A,Benning LG,Vilhelmsson O.Moss and underlying soil bacterial community structures are linked to moss functional traits. Ecosphere,14(3)(2023). Faisal HMK,Kazuhide K,Akio T.Bacterial-biota dynamics of eight bryophyte species from different ecosystems.Saudi Journal of Biological Sciences,22(2)(2015). 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Warshan D,Bay G,Nahar N,Wardle DA,Nilsson MC,Rasmussen U.Seasonal Variation in nifH Abundance and Expression of Cyanobacterial Communities Associated with Boreal Feather Mosses.The ISME Journal 10(9): 2198(2016). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4417220\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":305696579,\"identity\":\"6847369d-cee4-417a-b67a-803443106c24\",\"order_by\":0,\"name\":\"Wenping Meng\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Botanical Garden\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Wenping\",\"middleName\":\"\",\"lastName\":\"Meng\",\"suffix\":\"\"},{\"id\":305696581,\"identity\":\"a9515029-f053-4075-81e2-b9307ac55a4d\",\"order_by\":1,\"name\":\"Ran Jingcheng\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAuUlEQVRIiWNgGAWjYFCCA4wPPjAwyLGxtx8gWguz4QwGBmM+njMJRFvDJszDwJA4T8LBgDj15o2HnzHOqLmT3ibBkMDwo2IbYS0yB46ZPfhw7Flum3TjAcaeM7cJa5FgOMNuOLPhcG6bzIEEZsY24rSwSfM2HE5nk0gwIE1LAilajhkbzjh22LANGMgHifOLxOGHDz7UHJaXb28/+OBHBRFaGCQOINgHcClCBfwNxKkbBaNgFIyCEQwADnE/FGrLtDIAAAAASUVORK5CYII=\",\"orcid\":\"\",\"institution\":\"Guizhou Academy of Forestry Sciences\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Ran\",\"middleName\":\"\",\"lastName\":\"Jingcheng\",\"suffix\":\"\"},{\"id\":305696582,\"identity\":\"ae7d6087-b0ff-4255-ab7f-8a47414e86be\",\"order_by\":2,\"name\":\"Xu Zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Academy of Forestry Sciences\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xu\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":305696583,\"identity\":\"3d724b86-6f60-4527-b1bb-09282cfde778\",\"order_by\":3,\"name\":\"Deming Kong\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Bijie Forestry Bureau\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Deming\",\"middleName\":\"\",\"lastName\":\"Kong\",\"suffix\":\"\"},{\"id\":305696585,\"identity\":\"3b166052-d5a0-4a4d-8a97-5efc5671e26d\",\"order_by\":4,\"name\":\"Fang Liu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Botanical Garden\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Fang\",\"middleName\":\"\",\"lastName\":\"Liu\",\"suffix\":\"\"},{\"id\":305696586,\"identity\":\"de633927-60b0-4fc5-9489-4c2f7579d70c\",\"order_by\":5,\"name\":\"Qi Tong\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Botanical Garden\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Qi\",\"middleName\":\"\",\"lastName\":\"Tong\",\"suffix\":\"\"},{\"id\":305696587,\"identity\":\"17fe1e89-f4dd-4ed7-9d22-209a2e42360b\",\"order_by\":6,\"name\":\"Chen Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Botanical Garden\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Chen\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":305696588,\"identity\":\"0bf8a57b-5925-4d21-8b85-e0f58d895300\",\"order_by\":7,\"name\":\"Ting Zheng\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guizhou Botanical Garden\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Ting\",\"middleName\":\"\",\"lastName\":\"Zheng\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-05-14 07:26:58\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4417220/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4417220/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":57085529,\"identity\":\"7d6af0eb-e354-48e9-958c-42033045c21c\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":182503,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDiversity of microorganisms in rock surface under different treatment conditions at the phylum level\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/f9c94e9ca3869a35be5fd866.png\"},{\"id\":57085530,\"identity\":\"d2ef451f-fe44-42bd-a40a-dff4f6f77f20\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":559784,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDiversity of microorganisms in rock surface under different treatment conditions at the species level\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/8df2fa6a12420bd362f92717.png\"},{\"id\":57085534,\"identity\":\"7d154726-dc78-426b-abbc-448aa7d71c92\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":589250,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eCluster analysis of microbial functions within the rock surface of bryophytes\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/d3818e88a4f3b26e7ce2c187.png\"},{\"id\":57085533,\"identity\":\"467cfc11-ecaa-44fc-9ee4-07ed0b83ce21\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":255210,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChanges of microorganisms related to carbon fixation, nitrogen fixation and phosphorus metabolism in the habitat of lithophytic bryophytesafter planting on rock surface\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/d20ec001ccd30ecc6ead9eaf.png\"},{\"id\":57085531,\"identity\":\"a7690559-7c1f-4fa9-adcd-2acac67b716b\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":196361,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChanges of carbon fixation KEGG in the habitat after the rock surface of the lithophytic bryophytes, A : KEGG with the top 15 abundances, B : KEGG with significant differences.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/5bf7a7dce9e91e03b9b7ca18.png\"},{\"id\":57085532,\"identity\":\"2a1633c4-ef14-4712-b330-3a02fb01a805\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":132675,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChanges in nitrogen fixing KEGG in the habitat after moss colonization, A: KEGG in the top 15 abundance rankings, B: KEGG with significant differences\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/ae776495cefb000d014d7a7e.png\"},{\"id\":57085535,\"identity\":\"387d1d2f-17b1-4e73-8c8a-4194e1a1dd97\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:25\",\"extension\":\"png\",\"order_by\":7,\"title\":\"Figure 7\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":136878,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChanges in phosphorus metabolism KEGG in the habitat of lithophytic bryophytes after planting on rock surfaces. A: KEGG in the top 15 abundances, B: KEGG with significant differences\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"7.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/75cb9d56242a754f7dc583cb.png\"},{\"id\":57085537,\"identity\":\"b0a661a3-5eae-4d78-8a34-340a7e9ac54c\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:26\",\"extension\":\"png\",\"order_by\":8,\"title\":\"Figure 8\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":476957,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChanges of functional microbial communities in the habitat of lithophytic bryophytes after planting rock surface, A : significantly different species of carbon-fixing microorganisms, B : significantly different species of nitrogen-fixing microorganisms, C : significantly different species of phosphorus metabolism\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"8.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/a6b57e5c227859f2131ac46b.png\"},{\"id\":57085538,\"identity\":\"981ca978-720a-422e-9725-6c1c21403d0f\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:39:26\",\"extension\":\"png\",\"order_by\":9,\"title\":\"Figure 9\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":541536,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eThe relationship between significantly different microorganisms and soil physicochemical properties\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"9.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/3f3cbe55270361dbee0466c6.png\"},{\"id\":57086281,\"identity\":\"a7d1dee9-0544-4ecc-99df-2f61f5808057\",\"added_by\":\"auto\",\"created_at\":\"2024-05-24 11:55:29\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":4168212,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4417220/v1/80638501-8a6c-4ab4-a1e0-d2ca2199fd83.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Microbial differences in the habitats of lithophytic bryophytes and their relationship with soil nutrients\",\"fulltext\":[{\"header\":\"Background\",\"content\":\"\\u003cp\\u003eBryophyte are composed of moss, Hepaticae, and Anthrocerotae, with over 20000 species worldwide, distributed in various ecosystems in tropical, freshwater, marine, and Arctic tundra regions[\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e].Some bryophytes grow in water, some on the rock surface, some on the soil surface, and some on leaves or wood.[\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e].Lithophytic bryophytes are often born on the surface of rocks in a cluster or cushion community. They have the functions of drought resistance, high temperature resistance, water and soil conservation[\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e]. They are pioneer plants on the surface of bare rocks. During their growth, they promote rock dissolution and release nutrients through biochemical and physical mechanical effects [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. At the same time, the growth of bryophytes is often accompanied by microorganisms. These microbial communities are responsible for the first stage of soil development, forming a stable soil and fixing carbon and nitrogen biological soil layer[\\u003cspan additionalcitationids=\\\"CR6\\\" citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e]. Although the microbial community can promote the establishment of mosses, mosses affect the lower soil microbial community through leachate, similar to how different litter types affect the soil microbial community[\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. Generally, the moss associated bacterial communities are dominated bypresumptively acidophilic bacteria often associated with ombrotrophic or other oligotrophic environments and arecomparable to the bacterial communities of other moss species[\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eAt present,moss soil crusts have become a research hotspot.People have studied the effects of moss crusts in different ecosystems on the composition characteristics of microbial communities in habitats. It was proved that moss crusts can change the microbial community structure of crusts and increase the relative abundance of functional bacterial groups (nitrogen fixation, photosynthesis,metabolic C1 compounds, plant symbiosis, saprophytic,thereby improving the surface soil microenvironment [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e].The species number of soil bacteria in the moss crust system was between 1529\\u0026ndash;2381, and the species number of soil fungi was between 323\\u0026ndash;758. The species number of bacterial community was significantly higher than that of fungal community [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e].Cyanobacteria and Actinobacteria are dominant in the moss crust,which is conducive to the stability of crust structure[\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e].For the lithophytic moss crusts, the diversity of bacteria and fungi was similar to that of the soil moss crusts, showing that bacteria accounted for more than 80% of the microbial community composition, and the diversity of bacteria was much higher than that of fungi. The dominant bacteria were Proteobateria, Bateroidetes, Actinobacteria and Acidobacteria.The dominant fungi were Ascomycota and Mortierellomycota[\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e].In fact, the microbial diversity and community composition in the soil under the moss crust are closely related to soil factors. The composition of soil bacterial community was significantly affected by SOC, SWC, AK and pH, and the diversity was significantly affected by soil pH, SWC, TP, AP, AN and NO\\u003csub\\u003e3\\u003c/sub\\u003e\\u003csup\\u003e\\u0026minus;\\u003c/sup\\u003eN[\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e].Mg\\u003csup\\u003e2+\\u003c/sup\\u003eand SWC in soil are the main factors affecting the functional traits of moss, as they can affect the stress of moss photosynthesis, thereby affecting the progress of other physiological activities and the expression of functional traits[\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e].In addition, root exudates can also affect soil microbial richness index and mcintosh index. The higher the concentration of root exudates, the higher the microbial diversity index. However, the microbial dominance index was not affected by the concentration of root exudates[\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eAlthough efforts have been made to study the microbial community composition and ecological functions of moss crust habitats in different ecosystems, moss soil crust (referred to as moss crust) is a composite shell layer formed by the interaction between moss plants, pseudoroots, hyphae, secretions, and soil surface particles such as cyanobacteria, fungi, and bacteria, It contains certain soil and soil microorganisms [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. Therefore, studying the effects of moss crusts on soil microorganisms and other characteristics in habitats cannot accurately reflect the functional traits of moss. This makes studying moss related microorganisms and their impact on environmental microorganisms particularly interesting and meaningful.\\u003c/p\\u003e \\u003cp\\u003eWe hypothesized that:(1) The lithophytic bryophytes in the karst area can improve the diversity of the soil microbial community and enrich the community composition structure.(2)Lithophytic bryophytes can increase the richness of functional microbial communities in rock surface soil.(3)The microorganisms related to soil carbon, nitrogen, phosphorus, potassium and organic acids increased due to the existence of lithophytic bryophytes,which led to the increase of carbon, nitrogen, phosphorus, potassium and organic acids in soil.\\u003c/p\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eChanges of soil microbial community composition on rock surface after the colonization of lithophytic bryophytes\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe average number of microorganisms in the rock surface of the bryophytes was 22340 (\\u003cem\\u003eHyophila involute\\u003c/em\\u003e 22063, \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e 22779, \\u003cem\\u003eDidymodon constrictus\\u0026nbsp;\\u003c/em\\u003e22180 ), 22329 in the exposed rock surface and 19925 in the original control soil.These microorganisms mainly include Fungi, Bacteria, Eukaryota, Archaea, Viruses. At the phylum level, bacteria had the largest number of species, followed by Eukaryota, Archaea, Fungi and Viruses ( Fig.1-A ).The top 10 phylum in the abundance of Bacteria were \\u0026nbsp;Actinobacteria、Proteobacteria、Acidobacteria、Cyanobacteria、Chloroflexi、Candidatus_Rokubacteria、Gemmatimonadetes_d_Bacteria、unclassified_d__Bacteria、Bacteroidetes、Planctomycetes（Fig.1-B）.The top 10 phylum in the abundance of Archae were Euryarchaeota、Thaumarchaeota、unclassified_d__Archaea、Candidatus_Bathyarchaeota、Crenarchaeota、Candidatus_Woesearchaeota、Candidatus_Thorarchaeota、Candidatus_Micrarchaeota、Candidatus_Korarchaeota、Candidatus_Altiarchaeota（Fig.1-C）.The top 10 phylum in the abundance of Eukaryot were Streptophyta、Chordata、Ascomycota、Arthropoda、Mucoromycota、Bacillariophyta、unclassified_d_Eukaryota、Nematoda、Basidiomycota、Chytridiomycota（Fig.1-D）。The top 10 phylum in the abundance of Viruses were Artverviricota、unclassified_d__Viruses、Nucleocytoviricota、Phixviricota、Uroviricota（Fig.1-E）。The top 10 phylum in the abundance of Fungi were Mucoromycota、Basidiomycota、Chytridiomycota、Zoopagomycota、Microsporidia、Cryptomycota、Blastocladiomycota、Ascomycota（Fig.1-F）。\\u003c/p\\u003e\\n\\u003cp\\u003eAt the species level, Fungi 155, Bacteria 18460, Eukaryota 459, Archaea 395, Viruses 63 in the original control soil, and Fungi 181, Bacteria 20597, Eukaryota 631, Archaea 455, Viruses 94 in the exposed rock surface（Fig.2-A）. The average species of various microorganisms in the rock surface habitat after the colonization of lithophytic bryophytes were Fungi 235, Bacteria 20535, Eukaryota 816, Archaea 503, Viruses155, which were more than those in the bare rock surface habitat. Compared with the original control soil, the growth of various microorganisms was Fungi 52 %, Bacteria 11 %, Eukaryota 78 %, Archaea 27 %, Viruses 146 %. Among them, Viruses had the largest growth of 146 %, followed by Eukaryota, Fungi, Archaea, and Bacteria had the smallest growth. We can see that the number of various microorganisms in the rock surface increased significantly after the colonization of bryophytes, and the impact on Viruses, Eukaryota and Fungi was greater than that of Bacteria and Archaea.\\u003c/p\\u003e\\n\\u003cp\\u003eThe top 15 species of Bacteria in each treatment group were Actinobacteria_bacterium、Acidobacteria_bacterium、Chloroflexi_bacterium、Acidimicrobiia_bacterium、Deltaproteobacteria_bacterium、Candidatus_Rokubacteria_bacterium、Smaragdicoccus_niigatensis、Alphaproteobacteria_bacterium、Solirubrobacterales_bacterium、Archangium_gephyra、Acidimicrobiaceae_bacterium、Gemmatimonadetes_bacterium、Betaproteobacteria_bacterium、Nocardiaceae_bacterium_YC2-7、Microcoleus_sp._PCC_7113（Fig.2-B）. The top 15 species of Archaea in each treatment group were Euryarchaeota_archaeon、Thaumarchaeota_archaeon、Candidatus_Bathyarchaeota_archaeon、uncultured_archaeon、Nitrososphaeraceae_archaeon、Nitrosopumilales_archaeon、archaeon_HR01、ANME-2_cluster_archaeon、Thermoplasmata_archaeon、Candidatus_Methanoperedens_nitroreducens、Methanosarcinales_archaeon、Candidatus_Woesearchaeota_archaeon、Hadesarchaea_archaeon、Candidatus_Poseidoniales_archaeon、archaeon（Fig.2-C）. The top 15 species of Eukaryota in each treatment group were Physcomitrium_patens、Marchantia_polymorpha、Pseudocrossidium_replicatum、Solanum_lycopersicum、Homo_sapiens、Lupinus_albus、Eucalyptus_grandis、Syntrichia_ruralis、Candida_albicans、Mus_musculus、Pohlia_nutans、Fusarium_oxysporum、Syntrichia_filaris、Beta_vulgaris、Phaeodactylum_tricornutum（Fig.2-D）. The top 15 species of Fungi in each treatment group were Candida_albicans、Fusarium_oxysporum、Powellomyces_hirtus、Fusarium_odoratissimum、Rhinocladiella_mackenziei、Parasitella_parasitica、Oidiodendron_maius、Diversispora_epigaea、Lipomyces_starkeyi、Smittium_culicis、Absidia_glauca、Rhizopus_delemar、Gigaspora_rosea、Spizellomyces_sp._\\u0026apos;palustris\\u0026apos;、Rhizophagus_clarus（Fig.2-E）。The top 15 species of Viruses in each treatment group were Murine_leukemia_virus、Rhodococcus_phage_REQ1、Circular_genetic_element_sp.、Gordonia_phage_Phendrix、Gordonia_phage_Mollymur、Caudovirales_GX15bay、Streptomyces_phage_Henoccus、Gordonia_virus_Suzy、Mycobacterium_phage_Kumao、Arthrobacter_phage_Shoya、Microbacterium_phage_Rasovi、Streptomyces_phage_Yara、Microbacterium_phage_FuzzBuster、Gordonia_virus_Ghobes、Arthrobacter_virus_Joann（Fig.2-F）。\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e4.2 The main functions of soil microorganisms in the lithophytic bryophytes rock surface\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eBy conducting functional clustering analysis on the overall gene set and selecting the top 50 ranked functions, we found that the main functions of these microorganisms are amino acid metabolism, organic acid metabolism, Metabolism of carbon, nitrogen, phosphorus, sulfur, and methane, Photosynthesis related functions, Secondary metabolite related function, glycometabolism, Enzyme related functions(Pantothenate and CoA biosynthesis) （Fig.3-A）.\\u003c/p\\u003e\\n\\u003cp\\u003eThere are differences in microbial functions between different treatment groups. The difference in microbial functions within \\u003cem\\u003eHyophila involute\\u003c/em\\u003e and \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e is small, but they differ greatly from \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e and the control group. However, the difference between \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e and the control group is small (Fig.3-B). It indicated that bryophytes had a significant role in promoting the functional development of rock surface soil, but there were interspecific differences in this contribution.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEffects of lithophytic bryophytes on carbon fixation, nitrogen fixation and phosphorus metabolism functional microorganisms in rock surface\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAfter planting on the rock surface, the microorganisms related to carbon fixation in the soil were 2779, nitrogen fixation 1502, and phosphorus metabolism 1750.In the original control soil, carbon-fixing microorganisms 2031, nitrogen-fixing 1008, and phosphorus metabolism 1141. Carbon-fixing microorganisms 2444, nitrogen-fixing 1288, and phosphorus metabolism 1490 in the bare rock surface habitat. Compared with the original control soil, the growth rate of carbon-fixing microorganisms, nitrogen fixation and phosphorus metabolism in the soil after bryophyte planting rock surface increased by 37 %, 49 % and 53 %, respectively. Compared with the original soil, the growth rate of carbon-fixing microorganisms in the exposed rock surface was 20 %, nitrogen fixation was 28 %, and phosphorus metabolism was 31 % ( Fig.4-A ). We can see that the number of microorganisms related to carbon fixation, nitrogen fixation and phosphorus metabolism in the habitat was significantly increased after the colonization of the rock surface.\\u003c/p\\u003e\\n\\u003cp\\u003eAmong the top 15 species of microbial abundance related to carbon fixation, nitrogen fixation and phosphorus metabolism, 9 kinds of microorganisms appeared in the three functional groups and accounted for a relatively high proportion, which were respectively:Actinobacteria_bacterium、Acidobacteria_bacterium、Acidimicrobiia_bacterium、Chloroflexi_bacterium、Smaragdicoccus_niigatensis、Solirubrobacterales_bacterium、Deltaproteobacteria_bacterium、Alphaproteobacteria_bacterium、Gemmatimonadetes_bacterium. The unique microbial species in the carbon fixation functional group are Iamiaceae_bacterium_SCSIO_58843、Candidatus_Rokubacteria_bacterium、Betaproteobacteria_bacterium、Thermoleophilia_bacterium（Fig.4-B）. The unique microorganisms in the functional groups related to phosphorus metabolism are Thermoleophilia_bacterium、Gaiella_occulta、Archangium_gephyra（Fig.4-C）. The unique microorganisms in the nitrogen-fixing functional group are Candidatus_Rokubacteria_bacterium、Gaiella_occulta、Nocardiaceae_bacterium_YC2-7、Betaproteobacteria_bacterium、Microcoleus_sp._PCC_7113、Rhizobiales_bacterium（Fig.4-D）.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunctional analysis of microbial carbon fixation, nitrogen fixation and phosphorus metabolism in the habitat of lithophytic bryophytes after planting rock surface\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThere are differences in the metabolic pathways related to carbon fixation between the moss colonized rock surface and the exposed rock surface compared to the original soil (Fig.5).There are also interspecific differences among lithophytic bryophytes. The carbon fixation metabolic pathways between \\u003cem\\u003eHyophila involute\\u003c/em\\u003e and\\u003cem\\u003e\\u0026nbsp;Eurohypnum leptothallum\\u0026nbsp;\\u003c/em\\u003e,\\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e are significantly different, while the carbon fixation metabolic pathways between\\u003cem\\u003e\\u0026nbsp;Didymodon constrictus\\u003c/em\\u003e and exposed rock surface habitats are similar. The top 15 metabolic pathways related to carbon fixation were significantly different among the treatment groups. The abundance of metabolic pathways K01681, K01903, K01961, K00031, K01007, K01595, and K01847 in the habitat after the colonization of bryophytes on the rock surface was significantly higher than that in the original control soil. However, the abundance of K01848, K00174, K00297, K00175 and K02446 decreased significantly ( Fig.5-B ).\\u003c/p\\u003e\\n\\u003cp\\u003eThe metabolic pathways related to nitrogen fixing in the habitat of lithophytic bryophytes after planting on the rock surface are different from those in the original control soil and exposed rock surface, while the nitrogen fixing metabolic pathways in the original control soil and exposed rock surface are more similar (Fig.6).There are also differences among different species of moss, and the nitrogen fixation metabolic pathways of \\u003cem\\u003eHyophila involute\\u003c/em\\u003e are more similar to those of \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e.The metabolic pathways of K00265, K00362, K00266, K02575, and K00370 in the habitat after the planting of lithophytic bryophytes on the rock surface were significantly higher than those in the original control soil, while K0195 and K00261 were significantly lower than those in the original control soil. K15577, K15578, and K15576 are significantly higher in exposed rock habitats than in rocky moss colonized rock surfaces and the original control soil.\\u003c/p\\u003e\\n\\u003cp\\u003eThe phosphorus metabolism pathway in the habitat of lithophytic bryophytes planted on the rock surface was different from that in the original control soil and exposed rock surface. Moreover, there were differences between the lithophytic mosses, \\u003cem\\u003eHyophila involute\\u003c/em\\u003e and \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e having more similar soil phosphorus metabolism functions, the original control soil and the exposed rock surface are similar.K00937, K00951, K00873, K03306, and K01507 were significantly higher in the habitat after lithophytic moss was planted on the rock surface than in the original soil control group, while K01524, K01113, K02039, and K00117 were significantly reduced (Fig.7).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eThe different species of carbon fixation, nitrogen fixation and phosphorus metabolism in rock surface colonized by lithophytic bryophytes\\u003c/strong\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eFig.8 Changes of functional microbial communities in the habitat of lithophytic bryophytes after planting rock surface,\\u0026nbsp;A : significantly different species of carbon-fixing microorganisms, B : significantly different species of nitrogen-fixing microorganisms, C : significantly different species of phosphorus metabolism\\u003c/p\\u003e\\n\\u003cp\\u003eThere were significant differences in the top 15 species of microorganisms related to carbon fixation among the groups.Smaragdicoccus_niigatensis、Gemmatimonadetes_bacterium、Acidimicrobiaceae_bacterium、Thermoleophilia_ bacterium were significantly higher than those of the original control soil, and the richness of Smaragdicoccus_niigatensis was significantly higher than other groups.However, Actinobacteria_bacterium, Acidobacteria _bacterium, Acidimicrobiia _bacterium, Candidatus_Rokubacteria_bacterium, Iamiaceae_bacterium_SCSIO_ 58843 decreased significantly after the bryophytes colonized rock surface ( Fig.8-A ).\\u003c/p\\u003e\\n\\u003cp\\u003eThe top 15 species of nitrogen-fixing microorganisms were significantly different among the groups. Smaragdicoccus_niigatensis, Gemmatimonadetes_bacterium, Nocardiaceae_bacterium_YC2-7, Microcoleus_sp._PCC_7113 were significantly higher than the original control soil.Among them, the abundance of Smaragdicoccus_niigatensis and Nocardiaceae _bacterium_YC2-7 in \\u003cem\\u003eHyophila involute\\u003c/em\\u003e and \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e was higher than that of other treatment groups.However,\\u0026nbsp;Actinobacteria_bacterium、Candidatus_Rokubacteria_bacterium、Acidimicrobiia_bacterium、Chloroflexi_bacterium、Deltaproteobacteria_bacterium、\\u0026nbsp;Rubrobacter_taiwanensis、Streptomyces_thermoautotrophicus、Rhizobiales_bacterium\\u0026nbsp;、Streptomyces_lunaelactis decreased significantly compared with the original control soil ( Fig.8-B ).\\u003c/p\\u003e\\n\\u003cp\\u003eThe top 15 species of microorganisms related to phosphorus metabolism were significantly different among groups.Chloroflexi_bacterium、Smaragdicoccus_niigatensis、unclassified_d__unclassified、Gemmatimonadetes_bacterium、Archangium_gephyra increased significantly after the rock surface was colonized by Lithophytes.The abundance of Smaragdicoccus_niigatensis was higher in the habitats of \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e and \\u003cem\\u003eHyophila involute\\u003c/em\\u003e than that in the habitats of \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e.\\u0026nbsp;However, Actinobacteria_bacterium、Acidobacteria_bacterium、Acidimicrobiia_bacterium、Acidimicrobiales_bacterium、Deltaproteobacteria_bacterium、Acidimicrobiaceae_bacterium、Candidatus_Rokubacteria_bacterium\\u0026nbsp;decreased significantly after the colonization of\\u0026nbsp;lithophytic bryophytes\\u0026nbsp;(Fig.8-C).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eThe relationship between different species and soil physical and chemical properties\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe correlation between the microorganisms related to carbon fixation, nitrogen fixation,phosphorus metabolism and the contents of nitrogen, phosphorus, potassium, organic carbon, oxalic acid, acetic acid, malic acid and succinic acid in rock surface soil was analyzed. We found that Acidimicrobiia_bacterium, Acidimicrobiaceae_ bacterium and Acidimicrobiales_bacterium in the top 15 microorganisms related to phosphorus were significantly positively correlated with potassium content in soil, and Deltaproteobacteria_bacterium was positively correlated with phosphorus content in soil ( Fig.9-A ).Alphaproteobacteria_bacterium and Solirubrobacterales_bacterium were significantly positively correlated with succinic acid content in soil, and significantly negatively correlated with soil organic carbon. Acidobacteria_bacterium, Solirubrobacterales_bacterium and Acidimicrobiaceae_bacterium were significantly negatively correlated with malic acid in soil. Solirubrobacterales_bacterium and Archangium_gephyra were significantly negatively correlated with acetic acid in soil.\\u003c/p\\u003e\\n\\u003cp\\u003eAmong the top 15 microorganisms related to carbon fixation, Iamiaceae_ bacterium_SCSIO_58843 was significantly positively correlated with potassium content in soil. Gemmatimonadetes_bacterium was positively correlated with soil organic carbon. Chloroflexi_bacterium was significantly positively correlated with oxalic acid content in soil. Acidimicrobiaceae _ bacterium was significantly negatively correlated with malic acid in soil. Betaproteobacteria_bacterium and Solirubrobacterales_bacterium were significantly positively correlated with succinic acid in soil (Figure 9-B).\\u003c/p\\u003e\\n\\u003cp\\u003eAmong the top 15 microorganisms related to nitrogen fixation, Candidatus_ Rokubacteria_bacterium_3112 was positively correlated with potassium content in soil.Microcoleus_sp._PCC_7113 was significantly negatively correlated with potassium content in soil. Candidatus _ Rokubacteria _bacterium_3112, Microcoleus _ sp._PCC_7113, Nocardiaceae_bacterium_YC2-7 were positively correlated with soil nitrogen content but not significant.Gemmatimonadetes_bacterium was significantly negatively correlated with oxalic acid.Smaragdicoccus_niigatensis, Gemmatimonadetes_bacterium, Nocardiaceae_bacterium_YC2-7 were significantly negatively correlated with succinic acid in soil. Solirubrobacterales_bacterium was significant positive correlation with succinic acid ( Fig.9-C).\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eThe average number of microorganisms in the epigenetic soil of the rock surface is 22340, the exposed rock surface is 22329, and the original control soil is 19925. The total number of microorganisms in the lithophytic bryophytes increased by 12% compared with the original control soil, and the number growth rates of various microorganisms were Fungi 52%, Bacteria 11%, Eukaryota 78%, Archaea 27%, Viruses 146%. We can see that the total amount of microorganisms in the rock surface increased after the colonization of bryophytes. Among them, the number of microbial species in Viruses, Eukaryota and Fungi increased more, while Bacteria and Archaea increased, but the increase was not significant.\\u003c/p\\u003e \\u003cp\\u003eMoreover,the microbial abundance of Cyanobacteria,Gemmatimonadetes_d_Bacteria,Bacteroidetes,Planctomycetes in Bacteria, Candidatus_Woesearchaeota in Archaea, Streptophyta, Arthropoda, Bacillariophyta in Eukaryota, Mucoromycota, Chytridiomycota and Zoopagomycota in Fungi was higher in the rock surface of bryophytes than in the exposed rock surface and the original control soil.We can see that the influence of lithophytic bryophytes on the total amount of various microorganisms in the rock surface is not significant, but it has a significant effect on the diversity of microorganisms in Viruses,Eukaryota and Fungi, and then changes the composition of microbial communities. Although there is no lithophytic bryophytes on the exposed rock surface, its development environment is exactly the same as that of the lithophytic bryophytes, and there is little difference in the total amount of soil microorganisms between the two. The reason for this result may be related to the important influence of soil moisture on the development of soil microorganisms[\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eIn addition, the microbial number of Fungi, Eukaryota, Archaea and Viruses in \\u003cem\\u003eHyophila involute\\u003c/em\\u003e was higher than that in \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e and \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e. While the number of Bacteria in \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e was the highest.It shows that there are interspecific differences in microbial diversity and community composition in lithophytic bryophytes.The composition of moss-associated bacterial communities may be driven by host identity[\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e], the characteristics of bryophyte gametophytes and secondary metabolism[\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. These changes may affect the composition of the moss microbiome[\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e].\\u003cem\\u003eHyophila involute\\u003c/em\\u003e like to grow on bare rock surfaces or crevices in karst rocky desertification areas[\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. The characteristics of this rocky habitat are strong sunlight on sunny days, high temperature on the rock surface, low air humidity, rapid water loss on the rock surface during rainfall, and poor water retention. \\u003cem\\u003eHyophila involute\\u003c/em\\u003e can grow in this harsh habitat, indicating that it has special skills of sun resistance, heat resistance, and drought resistance[\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e]. This special skill may lead to secretions conducive to the development of microorganisms in Fungi, Eukaryota, Archaea, and Viruses. The effect of vascular plant root exudates on soil microbial community structure has been confirmed[\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e],but how bryophyte secretions affect soil microorganisms has not been clearly reported, and research in this area needs to be strengthened in the future.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eChanges of carbon-fixing microbial communities and their metabolic pathways in the habitat of lithophytic bryophytes planted on the rock surface\\u003c/b\\u003e \\u003c/p\\u003e \\u003cp\\u003eAfter the rock surface was colonized by lithophytic bryophytes, the number of microorganisms related to carbon fixation, nitrogen fixation and phosphorus metabolism was not only increased, but also the abundance of metabolic pathways of them was significantly increased.For example, the abundance of K01681, K01903, K01961, K00031, K01007, K01595 and K01847 metabolic pathways in the carbon-fixing functional community was significantly higher than that in the original control soil after lithophytic bryophytes planted on rock surface.Among them, K01681 is aconitate hydratase, which is a coenzyme that catalyzes the conversion between citric acid and isocitric acid[\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e]. K01903 (succinyl-CoA synthetase beta subunit ) is an enzyme in the tricarboxylic acid cycle, under the action of succinate sulfur kinase, high-energy thioester bond hydrolysis, energy transfer to GDP to generate GTP and succinic acid[\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e].K01961 (acetyl-CoA carboxylase, biotin carboxylase subunit) is a biotin enzyme that catalyzes the reaction of\\u0026lsquo;acetyl-CoA\\u003csup\\u003e+\\u003c/sup\\u003e ATP\\u0026thinsp;+\\u0026thinsp;HCO3\\u003csup\\u003e\\u0026minus;\\u003c/sup\\u003e\\u0026rarr; malonyl-CoA\\u0026thinsp;+\\u0026thinsp;ADP\\u0026thinsp;+\\u0026thinsp;Pi\\u0026rsquo;.The malonyl-CoA is the base of important metabolic reactions such as fatty acid synthesis and acyl chain extension system[\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e].K00031(isocitrate dehydrogenase) is a key enzyme linking C-N metabolism[\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e]. The assimilation of inorganic nitrogen in plants begins with carbon compounds produced by C metabolism, and isocitrate dehydrogenase can catalyze the reversible reaction of oxidative decarboxylation of isocitrate to form α-ketoglutarate, which is the most important and most regulated rate-limiting enzyme in the tricarboxylic acid cycle[\\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e].K01595(phosphoenolpyruvate carboxylase)is a cytoplasmic enzyme that catalyzes the β-carboxylation of phosphoenolpyruvate in the presence of HCO\\u003csub\\u003e3\\u003c/sub\\u003e\\u003csup\\u003e\\u0026minus;\\u003c/sup\\u003e, and uses Mg\\u003csup\\u003e2+\\u003c/sup\\u003e as a cofactor to produce oxaloacetic acid and inorganic phosphate[\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e].The colonization of lithophytic bryophytes not only increased the number of microorganisms related to carbon fixation in the habitat, but also increased the abundance of enzymes related to citric acid cycle (TCA cycle), pyruvate metabolism, oxalic acid metabolism, and prokaryotic carbon fixation, thereby promoting the carbon fixation pathway in the rock surface habitat.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eThe changes of phosphorus metabolic microbial community and its metabolic pathway in the habitat of lithophytic bryophytes planted on the rock surface\\u003c/b\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe number of microorganisms related to phosphorus metabolism, metabolic pathways K00937, K00951, K00873, K03306 and K01507 in the habitat after the colonization of lithophytic bryophytes on the rock surface were significantly higher than those in the original soil control group.Among them,K00937 (polyphosphate kinase) can synthesize polyphosphate polyP, which contains several to hundreds of orthophosphate residues connected by high-energy phosphate glycosidic bonds to form straight chains, belonging to the polymer form of phosphate[\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e].K00873 (pyruvate kinase) is an essential irreversible enzyme in the glycolysis process, mainly catalyzing the conversion of phosphoenolpyruvate to pyruvate, accompanied by the production of two molecules of ATP. It requires the participation of divalent cations and is an exothermic reaction[\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e].Under anaerobic conditions, pyruvate undergoes fermentation to produce lactic acid or ethanol. Under aerobic conditions, pyruvate is converted to acetyl CoA and enters mitochondria for the tricarboxylic acid cycle[\\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e].K03306 (inorganic phosphate transporter, PiT family) can transport phosphorus into the cytoplasm[\\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e39\\u003c/span\\u003e] or various physiological and biochemical processes in cells[\\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e40\\u003c/span\\u003e].These enzymes are key enzymes involved in phosphate synthesis, phosphate transport, and glycolysis processes. Increasing their abundance promotes the synthesis of phosphorus in soil and the transport of phosphorus within plants.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eChanges in nitrogen fixing microbial communities and their metabolic pathways in the habitat of lithophytic moss planted on rock surfaces\\u003c/b\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe number of nitrogen fixing related microorganisms and metabolic pathways of K00265, K00362, K00266, K02575, and K00370 in the habitat of lithophytic moss after planting on the rock surface were significantly higher than those in the original control soil.Among them, K00265 glutamate synthase (NADPH) large chain is a key enzyme in bacterial ammonia assimilation process[\\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e41\\u003c/span\\u003e], and the ammonia generated by biological reduction of N\\u003csub\\u003e2\\u003c/sub\\u003e is also assimilated through the glutamate synthase cycle[\\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e]. K00362 nitrate reduction (NADH) large subunit is the process of degrading nitrite into ammonium[\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e].K02575 (MFS transporter, NNP family, nitrate/nitrate transporter) plays an important role in maintaining nitrogen balance in bacteria[\\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e44\\u003c/span\\u003e].Nitrite oxidoreductases are a class of enzymes that catalyze nitrite reduction and can degrade NIT to NO or NH\\u003csub\\u003e3\\u003c/sub\\u003e.They are key enzymes in the natural nitrogen cycle process[\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e].These enzymes are key enzymes in the nitrogen metabolism process, indicating an increase in nitrogen metabolism, alanine, aspartic acid, glutamic acid metabolism, and amino acid biosynthesis activity in the habitat of lithophytic mosses after colonization on rock surfaces.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eThe relationship between microorganisms and the contents of carbon, nitrogen, phosphorus and potassium in the rock surface soil of lithophytic moss planted.\\u003c/b\\u003e \\u003c/p\\u003e \\u003cp\\u003eThrough correlation analysis between different species and nitrogen, phosphorus, potassium and organic carbon in rock surface soil, we found that acidimicrobia_ Bacterium, acidimicrobiaceae_Bacterium, acidimicrobiales_Bacterium, iamiaceae_ Bacterium_SCSIO_58843 was significantly positively correlated with the content of potassium in soil. Deltaproteobacteria_There was a significant positive correlation between bacteria and phosphorus content in soil_Bacteria was positively correlated with soil organic carbon. It is suggested that the increase of these microorganisms will promote the increase of potassium, phosphorus and organic carbon in rock surface soil.\\u003c/p\\u003e \\u003cp\\u003eHowever, there was no significant correlation between these species and soil nitrogen content. However, studies have shown that bryophyte cyanobacterial symbionts (BCS) can fix nitrogen[\\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e45\\u003c/span\\u003e], and this symbiont inputs 0.5\\u0026ndash;10kg n/ha of nitrogen into the northern hemisphere ecosystem every year[\\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e46\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e47\\u003c/span\\u003e].Who plays the main role in moss cyanobacteria symbiosis?Cyanobacteria are a class of autotrophic prokaryotes[\\u003cspan citationid=\\\"CR48\\\" class=\\\"CitationRef\\\"\\u003e48\\u003c/span\\u003e], and their biological nitrogen fixation is mainly carried out by nitrogenase in heterocysts [\\u003cspan citationid=\\\"CR49\\\" class=\\\"CitationRef\\\"\\u003e49\\u003c/span\\u003e](Adams\\u0026amp;Duggan,2008).Other results suggest that moss N\\u003csub\\u003e2\\u003c/sub\\u003e-fixationrates are determined by bacterial community structure, rather than moss traitsor time since deglaciation[\\u003cspan citationid=\\\"CR50\\\" class=\\\"CitationRef\\\"\\u003e50\\u003c/span\\u003e]。Therefore,the symbionts of bryophytes and cyanobacteria mainly rely on cyanobacteria for nitrogen fixation, while bryophytes contribute less to nitrogen fixation.Bryophytes and cyanobacteria belong to mutualism. Bryophytes can provide nutrients such as carbohydrates and relatively stable microenvironment conditions such as temperature, moisture and humidity for cyanobacteria. The transfer of nitrogen fixed by cyanobacteria to bryophytes can increase the biomass of bryophytes[\\u003cspan citationid=\\\"CR51\\\" class=\\\"CitationRef\\\"\\u003e51\\u003c/span\\u003e].The number of cyanobacterial heterocysts and nitrogen fixation capacity in moss cyanobacteria symbionts were significantly higher than those in independent autotrophic cyanobacteria. For example, the number of heterocysts in cyanobacteria in symbionts can reach 6\\u0026ndash;10 times that of independent survival, and the nitrogen fixation rate can increase to 7 times[\\u003cspan citationid=\\\"CR49\\\" class=\\\"CitationRef\\\"\\u003e49\\u003c/span\\u003e].Therefore,bryophytes can not play the role of nitrogen fixation alone, but need to be combined with cyanobacteria, but it also needs a period of adaptation after combining with cyanobacteria to play the role of nitrogen fixation.\\u003c/p\\u003e \\u003cp\\u003eIn addition, N\\u003csub\\u003e2\\u003c/sub\\u003e fixation rate is variable and may be affected by moss species[\\u003cspan citationid=\\\"CR52\\\" class=\\\"CitationRef\\\"\\u003e52\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e53\\u003c/span\\u003e], nitrogen availability[\\u003cspan citationid=\\\"CR54\\\" class=\\\"CitationRef\\\"\\u003e54\\u003c/span\\u003e], water[\\u003cspan citationid=\\\"CR55\\\" class=\\\"CitationRef\\\"\\u003e55\\u003c/span\\u003e],temperature[\\u003cspan citationid=\\\"CR56\\\" class=\\\"CitationRef\\\"\\u003e56\\u003c/span\\u003e], composition of diazotrophs[\\u003cspan citationid=\\\"CR57\\\" class=\\\"CitationRef\\\"\\u003e57\\u003c/span\\u003e], abundance of diazotrophs[\\u003cspan citationid=\\\"CR54\\\" class=\\\"CitationRef\\\"\\u003e54\\u003c/span\\u003e] and activity of diazotrophs[\\u003cspan citationid=\\\"CR58\\\" class=\\\"CitationRef\\\"\\u003e58\\u003c/span\\u003e] .\\u003c/p\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eThe relationship between microorganisms and organic acid content in rock surface habitat of lithophytic bryophytes\\u003c/h2\\u003e \\u003cp\\u003eThe differential species Alphaproteobacteria_bacterium, Solirubrobacterales_ bacterium and Betaproteobacteria_bacterium among bare rock surface, original control soil and the rock surface planted lithophytic bryophytes were significantly positively correlated with succinic acid in soil.Smaragdicoccus_niigatensis and Gemmatimonadetes_bacterium.Chloroflexi_bacterium was significantly positively correlated with oxalic acid content in soil. However,Nocardiaceae_bacterium_YC2-7 was significantly negatively correlated with succinic acid in soil, and Gemmatimonadetes_bacterium was negatively correlated with oxalic acid in soil.Therefore, the content of succinic acid and oxalic acid in the rock surface habitat was affected by the stone moss.\\u003c/p\\u003e \\u003cp\\u003eHowever, Acidobacteria_bacterium, Solirubrobacterales_bacterium and Acidimicrobiaceae _bacterium were significantly negatively correlated with malic acid in soil. The difference species Solirubrobacterales_bacterium and Archangium_ gephyra were significantly negatively correlated with acetic acid in soil, and their abundance increased on the rocky moss planting rock surface, indicating that the content of acetic acid in soil decreased, which was consistent with the detection results of acetic acid content in soil.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eLithophytic bryophytes can increase the number of microorganisms in rocky habitats, especially significantly affecting the diversity of microorganisms in Viruses, Eukaryota, and Fungi, altering the composition and structure of microbial communities, and enriching the types and quantities of functional microorganisms such as carbon fixation, nitrogen fixation, and phosphorus metabolism in soil.Lithophytic bryophytes increased the abundance expression levels of key enzymes in the processes of citric acid cycle (TCA cycle), pyruvate metabolism, oxalate metabolism, prokaryotic carbon fixation, phosphate synthesis, phosphorus transport, glycolysis, nitrogen metabolism, alanine, aspartic acid, glutamic acid metabolism, and amino acid biosynthesis in rocky habitats, promoting an increase in potassium, phosphorus, organic carbon, and malic acid content in soil. However, stone moss has an impact on soil nitrogen, acetic acid The influence of oxalic acid and succinic acid is not significant.\\u003c/p\\u003e\"},{\"header\":\"Methods\",\"content\":\"\\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMaterials\\u003c/h2\\u003e \\u003cp\\u003eIn this study, the dominant species of lithophytic bryophytes in the karst area, \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e, \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e, and \\u003cem\\u003eHyophila involuta\\u003c/em\\u003e, were selected as the research objects. The materials required for the experiment were collected from the Puding Karst Rocky Desertification Ecosystem Observation and Research Station of the Chinese Academy of Sciences.The plant specimen was identified by Wenping Meng of Guizhou Botanical Gardon.The specimens are now deposited in the herbarium of Guizhou Botanical Garden.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eExperimental design\\u003c/h2\\u003e \\u003cp\\u003eThe dominant species of lithophytic bryophytes in karst area were collected, such as \\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e, \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e and \\u003cem\\u003eHyophila involute\\u003c/em\\u003e, cleaned the soil, litter and other impurities in the bryophyte.Select carbonate rock flakes, clean, the surface of a thin layer of soil just cover the rock surface is appropriate.Three kinds of cleaned bryophytes were planted on the surface of rock surface respectively. \\u003cem\\u003eHyophila involute\\u003c/em\\u003e (S group),\\u003cem\\u003eEurohypnum leptothallum\\u003c/em\\u003e (M group) and \\u003cem\\u003eDidymodon constrictus\\u003c/em\\u003e (D group), and each moss was set up with 5 replicates.The stone flakes with only soil and no bryophytes on the surface were set as L group.Before the experiment,a certain amount of rock surface soil was taken as the control soil and frozen in the refrigerator. At the end of the experiment, the microorganisms were detected.All the experimental groups were placed in the incubator,set the day 12 hours, light intensity 8000Lux, temperature 25\\u0026deg;C,night 12 hours,light intensity 0Lux, temperature 15\\u0026deg;C.Water was sprayed once a week. After 12 months, the bryophytes on the rock surface were removed, the soil on the rock surface was collected and stored at \\u0026minus;\\u0026thinsp;20\\u0026deg;C for further analysis.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e(2) Detection of soil physical and chemical indicators\\u003c/h2\\u003e \\u003cp\\u003eWe used potassium dichromate external heating method to detect soil organic carbon (SOC) content, used perchloric acid-sulfuric acid digestion, Kjeldahl method to determine total nitrogen content,used perchloric acid-sulfuric acid digestion-molybdenum antimony colorimetric method to determine total phosphorus content, and used hydrofluoric acid-perchloric acid digestion method to determine total potassium content.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e(3)Soil microbial detection methods\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eDNA extraction\\u003c/h2\\u003e \\u003cp\\u003eTotal genomic DNA was extracted from soil samples using the Mag-Bind\\u0026reg; Soil DNA Kit (Omega Bio-tek,Norcross,GA,U.S.) according to manufacturer\\u0026rsquo;s instructions. Concentration and purity of extracted DNA was determined with TBS-380 and NanoDrop2000, respectively. DNA extract quality was checked on 1% agarose gel.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eLibrary construction, and metagenomic sequencing\\u003c/h2\\u003e \\u003cp\\u003eDNA extract was fragmented to an average size of about 400 bp using Covaris M220 (Gene Company Limited,China) for paired-end library construction. Paired-end library was constructed using NEXTFLEX Rapid DNA-Seq (Bioo Scientific, Austin, TX,USA). Adapters containing the full complement of sequencing primer hybridization sites were ligated to the blunt-end of fragments. Paired-end sequencing was performed on Illumina NovaSeq(Illumina Inc., San Diego, CA, USA) at Majorbio Bio-Pharm Technology Co., Ltd. (Shanghai, China) using NovaSeq 6000 S4 Reagent Kit v1.5 (300 cycles) according to the manufacturer\\u0026rsquo;s instructions (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ewww.illumina.com\\u003c/span\\u003e\\u003cspan address=\\\"http://www.illumina.com\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e(4)Statistical analysis\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec17\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eSequence quality control and genome assembly\\u003c/h2\\u003e \\u003cp\\u003eThe data were analyzed on the free online platform of Majorbio Cloud Platform (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ewww.majorbio.com\\u003c/span\\u003e\\u003cspan address=\\\"http://www.majorbio.com\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e). Briefly, the paired-end Illumina reads were trimmed of adaptors, and low-quality reads (length\\u0026thinsp;\\u0026lt;\\u0026thinsp;50 bp or with a quality value\\u0026thinsp;\\u0026lt;\\u0026thinsp;20 or having N bases) were removed by fastp.Contigs with with a length\\u0026thinsp;\\u0026ge;\\u0026thinsp;300 bp were selected as the final assembling result, and then the contigs were used for further gene prediction and annotation.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec18\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eGene prediction, taxonomy, and functional annotation\\u003c/h2\\u003e \\u003cp\\u003eOpen reading frames (ORFs) from each assembled contig were predicted using Prodigal/MetaGene.The predicted ORFs with a length\\u0026thinsp;\\u0026ge;\\u0026thinsp;100 bp were retrieved and translated into amino acid sequences using the NCBI translation table.\\u003c/p\\u003e \\u003cp\\u003eA non-redundant gene catalog was constructed using CD-HIT with 90% sequence identity and 90% coverage. High-quality reads were aligned to the non-redundant gene catalogs to calculate gene abundance with 95% identity using SOAPaligner.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eSpecies and functional annotations\\u003c/h2\\u003e \\u003cp\\u003eRepresentative sequences of non-redundant gene catalog were aligned to NR database with an e-value cutoff of 1e\\u003csup\\u003e\\u0026minus;\\u0026thinsp;5\\u003c/sup\\u003e using Diamond for taxonomic annotations. Cluster of orthologous groups of proteins (COG) annotation for the representative sequences was performed using Diamond against egg NOG database with an e-value cutoff of 1e\\u003csup\\u003e\\u0026minus;\\u0026thinsp;5\\u003c/sup\\u003e.The KEGG annotation was conducted using Diamond against the Kyoto Encyclopedia of Genes and Genomes database with an e-value cutoff of 1e\\u003csup\\u003e\\u0026minus;\\u0026thinsp;5\\u003c/sup\\u003e.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec20\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAnalysis\\u003c/h2\\u003e \\u003cp\\u003eThe Meiji biological cloud database was used to analyze the data, and the dominant species and functional composition of the community were studied by column graph visualization method. The community Heatmap analysis tool was used to calculate the species/gene/functional abundance of each sample. Hierarchical clustering of the distance matrix can clearly see the distance of the sample branches.Based on the obtained species and functional abundance data of different groups, one-way analysis of variance is used to conduct hypothesis tests on the species and functions among their microbial communities, evaluate the significance level of species, functions, or gene abundance differences, and obtain the species and functions with significant differences between groups. he correlation between different species and soil physical and chemical indexes was analyzed.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"Abbreviations\",\"content\":\"\\u003cp\\u003eC \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;carbon\\u003c/p\\u003e\\n\\u003cp\\u003eN \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;nitrogen\\u003c/p\\u003e\\n\\u003cp\\u003eSOC \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;soil organic carbon\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eSWC \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;soil water content\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eAK \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; available potassium content\\u003c/p\\u003e\\n\\u003cp\\u003epH \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; acidity and alkalinity\\u003c/p\\u003e\\n\\u003cp\\u003eTP \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; total phosphorus\\u003c/p\\u003e\\n\\u003cp\\u003eAP \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; available phosphorus\\u003c/p\\u003e\\n\\u003cp\\u003eAN \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; available nitrogen\\u003c/p\\u003e\\n\\u003cp\\u003eNO\\u003csub\\u003e3\\u003c/sub\\u003e\\u003csup\\u003e-\\u003c/sup\\u003eN \\u0026nbsp; \\u0026nbsp;nitrate nitrogen\\u003c/p\\u003e\\n\\u003cp\\u003eMg\\u003csup\\u003e2+ \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;\\u0026nbsp;\\u003c/sup\\u003emagnesium ion\\u003c/p\\u003e\\n\\u003cp\\u003eKEGG \\u0026nbsp; the most commonly used international bioinformatics database\\u003c/p\\u003e\\n\\u003cp\\u003eGDP \\u0026nbsp; \\u0026nbsp; guanosine diphosphate\\u003c/p\\u003e\\n\\u003cp\\u003eGTP \\u0026nbsp; \\u0026nbsp; guanosine triphosphate\\u003c/p\\u003e\\n\\u003cp\\u003eATP \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;adenosine triphosphate\\u003c/p\\u003e\\n\\u003cp\\u003eHCO3\\u003csup\\u003e- \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;\\u003c/sup\\u003ebicarbonate ion\\u003c/p\\u003e\\n\\u003cp\\u003eADP \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;adenosine diphosphate\\u003c/p\\u003e\\n\\u003cp\\u003ePi \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;phosphate group\\u003c/p\\u003e\\n\\u003cp\\u003eNADPH \\u0026nbsp; \\u0026nbsp;nicotinamide adenine dinucleotide phosphate\\u003c/p\\u003e\\n\\u003cp\\u003eNO \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;nitric oxide\\u003c/p\\u003e\\n\\u003cp\\u003eNH\\u003csub\\u003e3 \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;\\u003c/sub\\u003eammonia\\u003c/p\\u003e\\n\\u003cp\\u003eBCS \\u0026nbsp; \\u0026nbsp; bryophyte cyanobacterial symbionts\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eEthics approval and consent to participate\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe manuscript does not involve the use of any animal or human data or tissues.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eStatement of collect plant samples\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe plants involved in the manuscript are not rare and endangered plants and are not included in the IUCN protection list.The plants were collected at the Puding karst rocky desertification ecosystem research and observation station of the Chinese Academy of Sciences.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eStatement of plant materials and experimental methods\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe collection of plant materials in the manuscript was in accordance with relevant institutional, national and international guidelines and legislation.\\u003c/p\\u003e\\n\\u003cp\\u003eThe methods involving plant materials are performed in accordance with the institutional, national, and international guidelines and legislation.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and materials\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe high-throughput sequencing metagenomic data of microorganisms in the soil underlying three types of moss plants is stored in the China National Center for Bioinformation:https://ngdc.cncb.ac.cn/\\u003ca href=\\\"https://ngdc.cncb.ac.cn/gsub/submit/bioproject/PRJCA026083/overview\\\"\\u003ePRJCA026083\\u003c/a\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompeting interest\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare no competing interests.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was supported by Science and Technology Fund of Guizhou (ZK[2023]-236,[2020]1Y074,ZK[2023]-237),Guizhou Forestry Research Project([2023]07),Doctoral Fund of Guizhou Academy of Sciences([2023]01),National Natural Science Foundation of China (32160086),National Nature Science Foundation of China and the Karst Science Research Center of Guizhou Province (U1812401).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthors\\u0026rsquo; contributions\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMeng Wenping is responsible for the detection, data analysis and manuscript writing of Bryophyte physiological indicators.Xu Zhang,Kong Deming and Zheng Ting are responsible for the field survey.Qi Tong,Chen Wang,Fang Liu are responsible for the collection and cultivation of Bryophyte. Ran Jingcheng is responsible for revising the manuscript. The author has read and approved the final manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledge\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThank you to Daqing Liang from Meiji Biotechnology for his assistance in data processing and mapping.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eKlarenberg IJ,Keuschnig C,Salazar A,Benning LG,Vilhelmsson O.Moss and underlying soil bacterial community structures are linked to moss functional traits. 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Compared with the original control soil, the growth of various microorganisms was Fungi 52%, Bacteria 11%, Eukaryota 78%, Archaea 27%, and Viruses 146%. The number of microbial species related to carbon fixation was 2779, nitrogen fixation was 1502, phosphorus metabolism was 1750.Compared with the original control soil, the bryophytes increased by 37%, 49% and 53% respectively after planting the rock surface. Compared with the original soil, the exposed rock surface increased by 20%, nitrogen fixation by 28% and phosphorus metabolism by 31%.Microbial species with significant differences between groups,Acidimimicrobia_bacterium,Acidimimicrobiaceae_bacterium,Acidimimicrobiales_bacterium, Iamiaceae_bacterium_SCSIO_58843 is significantly positively correlated with potassium content in soil,Microcoleus_Sp._PCC_7113 is a significant negative correlated with potassium content in soil.Alphaprotoobjective_bacterium, Solirubrobacteriales_bacterium, Betaproteobjective_bacterium is a significant positive correlated with succinic acid content in soil.Chloroflexi_bacterium is a significant positive correlated with oxalic acid content insoil.Acidobacteria_bacterium,Solirubrobacterales_bacterium,Acidimicrobiaceae_bacterium is a significant negative correlated with malic acid in soil.Gemmatimonadetes_bacterium is a significant negative correlated with oxalic acid.\\u003c/p\\u003e\\n\\u003cp\\u003eSmaragdicoccus_niigatensis,Gemmatimonadetes_bacterium,Nocardiaceae_bacterium_YC2-7 is significantly negatively correlated with succinic acid in soil.\\u003c/p\\u003e\\n\\u003cp\\u003eSolirubrobacterales_bacterium,Archangium_gephyra is a significant negative correlated with acetic acid in soil.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConclusions:\\u003c/strong\\u003eThe lithophytic bryophytes changed the microbial composition structure in the rock surface habitat, significantly increased the number of functional microorganisms, and then increased the accumulation of potassium, phosphorus, organic carbon and malic acid in the habitat, and promoted the positive development of the rock surface ecosystem.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Microbial differences in the habitats of lithophytic bryophytes and their relationship with soil nutrients\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-05-24 11:39:20\",\"doi\":\"10.21203/rs.3.rs-4417220/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"2aec8d91-c825-4bb5-9d54-037b9f88c238\",\"owner\":[],\"postedDate\":\"May 24th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-05-24T11:39:23+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-05-24 11:39:20\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4417220\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4417220\",\"identity\":\"rs-4417220\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}