PART 1 Qhia Txog Kev cuam tshuam ntawm Ib puag ncig ntawm Cistanche Salsa: Los ntawm Ntiaj Teb Ecological Regionalization rau Av Microbial Zej Zog Cov yam ntxwv

TSEEM CEEB:Kom nkag siab txog kev tswj hwm kev sib raug zoo ntawm ib puag ncig thiabCistanche salsa, ntawm no peb muab cov txheej txheem macro-thiab micro qhov ntev. Los ntawm macro foundations, tus qauv MaxEnt tau qhia tias lub teb chaws raws Txoj Kev Siv thiab Txoj Kev, xws li Tuam Tshoj, Egypt, thiab Libya, tshwj xeeb tshaj yog tsim nyog rau kev loj hlob ntawmC. salsatxij thaum ub (Last Glacial Maximum thiab mid-Holocene) mus rau yav tom ntej (2050 thiab 2070). Qhov kev sim Jackknife tau qhia tias nag lossis daus yog ib qho tseem ceeb ntawm ecological yam uas cuam tshuam rau C. salsa qhov kev faib tawm. Los ntawm macro foundations, 16S rRNA amplicon sequencing cov ntaub ntawv qhia tau hais tias cov av microbial zej zog ntawm peb ecotypes (desert-steppe, grassland, thiab gravel-desert) yog qhov txawv (p <0.001). cov="" tub="" ntxhais="" microbiome="" tsom="" xam="" pom="" tias="" cov="" kab="" mob="" genera="" arthrobacter,="" sphingomonas,="" thiab="" bacillus="" tau="" enriched="" core="" taxa="">C. salsa. LEfSe thiab random hav zoov tau siv los khawb Gillisia (desert-steppe), Flavisolibacter (grassland), thiab Variibacter(gravel-desert) raws li biomarkers uas muaj peev xwm paub qhov txawv ntawm cov zej zog microbial los ntawm peb ecotypes. Cov ntaub ntawv kwv yees qhia tau hais tias kev ua haujlwm hauv metabolic ntawm cov zej zog microbial tau txhawb nqa txoj hauv kev metabolic thiab kev ua cov ntaub ntawv ib puag ncig. Kev txheeb xyuas kev sib txheeb tau qhia tias qhov siab tshaj plaws, nag lossis daus ntawm lub quarter sov tshaj plaws (bio18), qhov nruab nrab ntawm diurnal range (bio2), thiab qhov ntsuas kub nruab nrab ntawm lub quarter warmest (bio10) yog ib qho tseem ceeb ntawm ecological yam uas cuam tshuam rau cov av microbial cov zej zog. Txoj haujlwm no tau muab kev nkag siab tshiab rau hauv kev tswj hwm kev sib raug zoo ntawm cov kev faib khoom tsim nyogC. salsa, av microbial zej zog, thiab ecological tsav tsheb. Tsis tas li ntawd, nws nkag siab qhov kev nkag siab ntawm kev sib cuam tshuam ntawm cov nroj tsuag suab puam thiab ecological yam nyob rau hauv arid ib puag ncig.

KEYWORDS:Cistanche salsa, MaxEnt, 16S rRNA amplicon sequencing, av microbial zej zog, kev cuam tshuam ib puag ncig.

Cistanche deserticola muaj ntau yam teebmeem, nyem qhov no kom paub ntau ntxiv

Yog xav paub ntxiv thov hu rau:Joanna.jia@wecistanche.com

1. Kev Taw Qhia

Kev lag luam tus nqi ntawm cov nroj tsuag tshuaj tau nce sai heev hauv xyoo tas los no nrog kev txhim kho thoob ntiaj teb thiab kev siv cov nroj tsuag ntuj. Lub qia ntawmCistanche salsaNws yog ib qho khoom noj khoom haus thiab tshuaj ntsuab uas feem ntau yog siv los ua lub raum tonic, tshuaj aphrodisiac, kev tiv thaiv kev laus thiab kev tiv thaiv oxidation, txoj hnyuv laxative, kev kho mob siab, thiab kev kho hluav taws xob tiv thaiv. Cov kev tshawb fawb phytochemical niaj hnub no ntawm C. salsa tau ua rau muaj kev txheeb xyuas thiab kev sib cais ntawm ntau qhov ntawm bioactive compounds, xws li ntau yam phenylethanoid glycosides, iridoids, alditols, thiab lignin, uas muaj qhov tseem ceeb ntawm kev kho mob thiab kev noj qab haus huv.1 Nrog rau kev loj hlob sai ntawm kev lag luam. rau cov tshuaj thiab cov zaub mov noj qab haus huv, qhov kev thov rauC. salsatau nce zuj zus thiab yog nrog los ntawm overexploitation thiab plunder ntawm cov tsiaj qus. Yog li ntawd, qhov loj-scale artificial taw qhia thiab cultivation ntawmC. salsatau dhau los ua ib qho kev ntsuas tseem ceeb los tiv thaiv cov peev txheej qus thiab qeeb ecological desertificationation. Txawm li cas los xij, kev tshawb fawb txog kev cog qoob loo ntawm artificialC. salsatseem txwv.

Ib puag ncig muaj kev cuam tshuam loj heev rau kev loj hlob thiab kev loj hlob ntawm C. salsa.2 Nroj tsuag loj hlob nyob rau hauv ib puag ncig sib txawv, tsim cov ecotypes sib txawv uas pom tias muaj kev hloov pauv tsis zoo ntawm cov khoom xyaw thiab cov noob qhia.3,4 Ntawm qib macro, kev tshawb fawb tau pom tias qhov kub thiab txias, Lub teeb, nag lossis daus, thiab hom av txhua yam muaj kev cuam tshuam rau cov nroj tsuag kev loj hlob thiab nws cov khoom xyaw nquag.5,6 Kev kawm txog kev sib cuam tshuam ntawm ib puag ncig thiab cov nroj tsuag thiab cov yam ntxwv ntawm cov ecotypes sib txawv ntawm cov nroj tsuag muaj txiaj ntsig zoo rau kev xaiv thiab cog qoob loo siab. -zoo ntau yam.7 Los ntawm kev pom macro, cov nroj tsuag tuaj yeem muaj kev cuam tshuam loj heev rau cov kab mob hauv av txawm tias lawv muaj me me lossis tsis muaj kev sib cuag nrog cov av los ntawm lawv cov hauv paus hniav.8,9 Txawm li cas los xij, tam sim no kev tshawb fawb txog kev sib raug zoo ntawm cov nroj tsuag thiab cov nroj tsuag. ib puag ncig, nrog rau cov xwm txheej huab cua thiab cov av microbial zej zog, tseem tsis paub meej.

Tus qauv faib hom (SDM) yog tus qauv kev txheeb cais uas tau tsim los siv cov ntaub ntawv faib cov tsiaj thiab ib puag ncig kev hloov pauv los txiav txim siab ecological xav tau ntawm cov tsiaj thiab ua haujlwm lawv cov peev txheej faib, 10,11 tshawb nrhiav thaj chaw tsim nyog los ntawm kev sib txuas ntawm huab cua thiab av, txiav txim siab qhov tsim nyog ib puag ncig rau cov nroj tsuag tshuaj, thiab kev tshawb fawb tiv thaiv thiab loj hlob cov nroj tsuag uas muaj kev puas tsuaj.12,13 SDMs, xws li MaxEnt thiab biomod2, tau ua tiav zoo rau cov kev kwv yees ntawm kev faib cov qauv tsim rau cov nroj tsuag muaj kev puas tsuaj thiab ecological thaum lub sij hawm hloov pauv huab cua.14,15

16S rRNA amplicon sequencing ntawm cov nroj tsuag rhizosphere av qauv tau ua los tshawb txog ntau haiv neeg ntawm microbial zej zog, muab kev nkag siab tshiab rau kev sib raug zoo ntawm cov nroj tsuag thiab av microbial zej zog.16,17

Hauv qhov kev tshawb fawb no, peb tshawb nrhiav kev sib raug zoo ntawm ib puag ncig thiabC. salsalos ntawm macro-thiab micro dimensions. Peb ua raws li nram no. (1) Peb siv tus qauv MaxEnt los kwv yees lub ntiaj teb tsim kev loj hlob thaj chaw ntawmC. salsalos ntawm lub sijhawm qub mus rau yav tom ntej (tsib lub sijhawm: Last Glacial Maximum [LGM], mid-Holocene [MH], tam sim no, 2050, thiab 2070) thiab suav cov cheeb tsam tsim nyog ntawm qib sib txawv thiab cov nyiaj pab thiab ntau yam ntawm bioclimatic variables cuam tshuam rau kev faib tawm ntawmC. salsa.(2) Ua ke nrog kev ua hauj lwm, peb tau sau peb ecotypes (desert-steppe, grassland, and gravel-desert) av samples los ntawm thaj chaw loj hlob zoo tshaj plaws (Tacheng thiab Xinjiang) ntawm

C. salsa. Peb tau ua 16S rRNA amplicon sequencing los tshawb txog cov yam ntxwv ntawm cov av microbial zej zog. Peb kuj tau sib piv qhov sib txawv ntawm cov av microbiomes hauv peb lub ecotypes thiab txiav txim siab cov tub ntxhais microbiomes thiab biomarkers uas tuaj yeem paub qhov txawv ntawm peb ecotypes. (3) Peb tau ua qhov kev txheeb xyuas kev sib raug zoo thiab kev txheeb xyuas rov qab los ntawm kev nplua nuj ntawm cov tub ntxhais microbiomes, biomarkers, thiab bioclimatic variables los tshawb txog kev tswj kev sib raug zoo ntawmC. salsathiab ib puag ncig.

Cistanchemuaj ntau yam cuam tshuam los tiv thaiv qaug zog

2. Cov khoom siv thiab cov txheej txheem

2.1. Ecological Niche Modeling.

Hauv kev tshawb fawb no, tsib lub peev txheej tau siv los tshawb nrhiav cov chaw tam sim no ntawmC. salsathoob ntiaj teb: (1) GBIF ; (3) National Specimen Information Infrastructure (4) luam tawm cov ntaub ntawv; thiab (5) kev ua haujlwm. Cov ntaub ntawv piv txwv yam tsis muaj latitude thiab longitude tau ua raws li daim ntawv qhia hauv online latitude thiab longitude query Cov txheej txheem buffer tsom xam tau siv los ua pov thawj thiab lim tau txais cov ntsiab lus faib tawm kom tshem tawm qhov cuam tshuam ntawm kev simulation ntau dhau los ntawm kev sib raug zoo loj. Lub spatial daws teeb meem ntawm bioclimatic variables yog 2.5 arc-min (kwv yees li 4.5 km2), thiab qhov buffer deb tau teem rau 3 km. Tsuas yog ib qho chaw faib khoom tau khaws cia thaum qhov kev ncua deb ntawm qhov chaw faib khoom tsawg dua 3 km. Tom qab cov ntsiab lus sib npaug raug tshem tawm, 76 qhov tshwm sim, tag nrho los ntawm qhov chaw tsim khoom tiag tiag ntawm C. salsa, tau sau thiab siv rau kev tshuaj xyuas (Daim duab 1a, Cov Ntaub Ntawv Ntxiv 1).

Peb siv 19 qhov hloov pauv bioclimatic ntawm WorldClim raws li kev kwv yees ib puag ncig. Cov ntaub ntawv hloov pauv bioclimatic tam sim no ntawm txoj kev tshawb no tau sau los ntawm kev soj ntsuam cov ntaub ntawv ntawm WorldClim version 1.4 database18 thaum xyoo 1960-1990 nrog kev daws teeb meem ntawm 2.5 arc-min. Cov qub (LGM thiab MH) thiab yav tom ntej (yav tom ntej 2050 thiab yav tom ntej 2070) bioclimatic variables muaj nyob rau hauv WorldClim version 1.4 ntawm 2.5 arc-min daws teeb meem tau ua hauj lwm raws li kev kwv yees ntawm hom kev faib qauv. Txhawm rau kom tsis txhob muaj ntau qhov sib txawv, peb tau khiav kev txheeb xyuas kev sib raug zoo ntawm cov ntsiab lus keeb kwm yav dhau los thiab tshem tawm ib qho ntawm qhov sib txawv hauv txhua khub nrog Pearson

correlation value > 0.819 (Daim duab S1). Yim qhov hloov pauv bioclimatic thaum kawg suav nrog hauv cov qauv yog qhov nruab nrab diurnal ntau yam (bio2), qhov ntsuas kub nruab nrab ntawm lub quarter wettest (bio8), qhov ntsuas kub nruab nrab ntawm lub quarter driest (bio9), qhov nruab nrab ntawm lub quarter warmest (bio10) , nag lossis daus txhua xyoo (bio12), nag lossis daus ntawm lub quarter qhuav (bio17), nag lossis daus ntawm lub quarter sov tshaj plaws (bio18), thiab nag lossis daus ntawm lub quarter txias tshaj plaws (bio19).

Peb siv qhov qhib software pob qhov siab tshaj plaws entropy qauv (MaxEnt v.3.4.0), 20 uas tuaj yeem rub tawm los tsim cov qauv faib rauC. salsa. Cov kev txwv hauv qab no tau siv rau hauv cov chaw yooj yim: cov noob random thiab qhov feem pua ​​​​ntawm 25 thiab 10 replicates. Los ntawm kev teeb tsa qhov feem pua ​​​​raug random mus rau 25 feem pua, peb xaiv 75 feem pua ​​​​ntawm cov ntsiab lus faib rau qhov kev cob qhia. Los ntawm kev teeb tsa tus naj npawb ntawm replicates rau 10, peb khiav tus qauv 10 zaug nrog tib qhov chaw thiab nruab nrab cov zis ntawm tag nrho cov khiav kom tau qhov kawg tshwm sim. Lub cheeb tsam nyob rau hauv qhov txais neeg ua haujlwm nkhaus (AUC) tau siv los ntsuas tus qauv kev ua tau zoo-ntawm-haum, thiab tus qauv nrog tus nqi siab tshaj AUC tau suav tias yog qhov ua tau zoo tshaj plaws. Cov txheej txheem jackknife tau siv los ntsuas qhov tseem ceeb ntawm qhov hloov pauv. Cov lus teb nkhaus tau siv los ua kom tau qhov ntau ntawm bioclimatic variables.

ArcGIS tau siv los txheeb xyuas cov kev hloov pauv bioclimatic uas cuam tshuam rau kev faib tawm ntawm C. salsa thiab txheeb xyuas thiab suav cov cheeb tsam tsim nyog rau kev tsim khoom.

2.2. Soil Sample Collection thiab Description. C. salsa

Nws yog ib txwm faib nyob rau hauv peb ecotypes ntawm gravel-desert, grassland, thiab suab puam-steppe. Nyob rau lub Plaub Hlis 2017, peb tau sau cov qia thiab cov qauv av uas sawv cev rau cov ecotypes loj ntawm C. salsa hauv Tacheng, Xinjiang, Suav teb (Daim duab 1b, Table 1). Gravel-desert samples tau sau los ntawm Hejiaoke, Toli County (HJ1, HJ2, thiab HJ3). Grassland qauv raug sau los ntawm Yumin County (YM1, YM2, YM3, thiab YM4). Cov qauv suab puam-steppe tau sau los ntawm Jiang Alhan (JA1, JA2, JA3, JA4, JA5, thiab JA6). Cov qauv av uas peb sau tau yog tag nrho los ntawm cov av nyob rau saum npoo ntawm C. salsa thiab nws tus tswv tsev parasitic site. Voucher qauv nrog

voucher naj npawb los ntawm 20170510079-DT mus rau 20170510091-DT tau muab tso rau hauv lub herbarium ntawm lub koom haum ntawm Medicinal Plant Development ntawm Tuam Tshoj Academy of Medical Sciences nyob rau hauv Beijing, Tuam Tshoj. Tom qab ntxuav, succulent qia cov ntaub so ntswg raug txiav mus rau hauv me me pieces, tam sim ntawd khov nyob rau hauv cov kua nitrogen, thiab ces muab cia rau ntawm -80 degree kom txog rau thaum ua ntxiv. Cov av cores tau muab coj los ntawm qhov tob ntawm 20 cm siv

stainless-steel cylindrical driller nrog ib txoj kab uas hla ntawm 5 cm thiab ces muab cia rau ntawm -20 degree nyob rau hauv ib tug portable tub yees. Tom qab thauj mus rau qhov chaw kuaj, cov qauv av tau dhau los ntawm 2 hli sieve kom tshem tawm cov ntaub so ntswg, cov hauv paus hniav, pob zeb, thiab lwm yam khib nyiab thiab tom qab ntawd muab khaws cia ntawm −20 degree hauv lub tub yees ua ntej kev sim ntxiv.

2.3. DNA Extraction thiab 16S rRNA Sequencing.

Av DNA tau muab rho tawm siv PowerSoil DNA Isolation Kit (MoBio Laboratories, Carlsbad, CA) raws li phau ntawv. Qhov purity thiab zoo ntawm genomic DNA tau kuaj xyuas ntawm 0.8 feem pua ​​agarose gels. V3-4 cheeb tsam hypervariable ntawm cov kab mob 16S rRNA noob tau nthuav tawm nrog cov primers 338F (ACTCCTACGGGAGGCAGCAG) thiab 806R (GGACTACHVGGGTWTCTAAT). ' xaus rau pem hauv ntej thiab rov qab primers (Allwegene Co., Beijing). PCR tau ua tiav ntawm Mastercycler Gradient (Eppendorf, Lub Tebchaws Yelemees) siv 25 μL cov tshuaj tiv thaiv ntim uas muaj 12.5 μL ntawm KAPA 2G Robust HotStart ReadyMix, 1 μL ntawm kev xa tawm primer (5 μM), 1 μL ntawm thim rov qab primer (5 μM), 5 μL ntawm DNA (tag nrho cov template kom muaj nuj nqis yog 30 ng), thiab 5.5 μL ntawm H2O. Cov kev caij tsheb kauj vab yog raws li nram no: 95 degree rau 5 min, ua raws li 28 lub voj voog ntawm 95 degree rau 45 s, 55 degree rau 50 s, thiab 72 degree rau 45 s, nrog rau qhov kawg ncua ntawm 72 degree rau 10 min. Peb cov khoom PCR ib qho piv txwv tau sib sau ua ke los txo qis cov tshuaj tiv thaiv-theem PCR kev tsis ncaj ncees. Cov khoom PCR raug ntxuav los ntawm kev siv QIAquick Gel Extraction Kit (QIAGEN, Lub teb chaws Yelemees) thiab tom qab ntawd suav nrog siv PCR lub sijhawm. Kev sib sib zog nqus tau ua tiav ntawm MiSeq platform los ntawm Allwegene Co. (Beijing). Tom qab kev khiav, kev txheeb xyuas duab, hu xov tooj, thiab kev kwv yees yuam kev tau ua tiav siv Illumina Analysis Pipeline Version 2.6.

2.4. Kev tshuaj xyuas ntawm 16S rRNA Amplicon Sequencing Data.

Tag nrho cov ntaub ntawv sib txuas tau xa mus rau NCBI Short Archive (SRA) raws li SRA xa SUB7456002. Cov ntaub ntawv nyoos tau raug tshuaj xyuas thawj zaug, thiab cov txheej txheem raug tshem tawm raws li cov kev xav hauv qab no: ntu luv dua 200 bp nrog cov qhab nia qis (tsawg dua lossis sib npaug li 20) thiab muaj cov hauv paus tsis meej lossis tsis sib piv cov txheej txheem primer thiab cov cim barcode. Cov ntawv nyeem tau raug muab sib cais siv cov qauv tshwj xeeb barcode ib ntus thiab txiav nrog Illumina Analysis Pipeline Version 2.6. Tom qab ntawd cov ntaub ntawv teev tau raug tshuaj xyuas siv QIIME. Cov kab ke tau muab tso rau hauv kev ua haujlwm taxonomic units (OTUs) ntawm qhov zoo sib xws ntawm 97 feem pua ​​22 los tsim cov curvature tsis tshua muaj thiab suav cov txiaj ntsig kev nplua nuj thiab ntau haiv neeg. Ribosomal Database Project Classifier cov cuab yeej tau siv los faib tag nrho cov kab ke mus rau hauv ntau pawg taxonomic, uas qhov kev ntseeg siab tau teem rau 0.7.23 Kev tshuaj xyuas pawg tau ua raws li OTU cov ntaub ntawv los ntawm txhua tus qauv los ntawm kev siv R 3.6.1 los tshuaj xyuas cov Qhov zoo sib xws ntawm cov qauv sib txawv.24 UniFrac nrug matrix ntawm microbial cov zej zog los ntawm txhua tus qauv raug xam los ntawm kev siv Tayc coefficient thiab sawv cev raws li ib tug unweighted khub-pab pawg txoj kev nrog lej txhais tau tias clustering tsob ntoo, uas piav txog qhov sib txawv (1- zoo sib xws) ntawm ntau cov qauv.25 Ib daim ntawv ntoo Newick-format kuj tau tsim los ntawm qhov kev tshuaj ntsuam no. Alpha diversity tau siv los txheeb xyuas qhov nyuaj ntawm hom kev sib txawv rau ib qho qauv siv plaub qhov ntsuas, xws li Chao1, pom hom, thiab Shannon thiab Fisher

kev sib txawv indexes. Cov kev ntsuas no tau suav nrog siv QIIME software (Boulder, CO, USA) hauv Python (v.1.8.0) (La Jolla, CA, USA).26 Beta diversity tsom xam tau siv los ntsuas qhov sib txawv hauv cov qauv hauv cov ntsiab lus ntawm hom complexity. Kev sib txawv ntawm Beta tau suav nrog kev txheeb xyuas tus thawj tswj hwm (PCoA) thiab kev tshuaj xyuas pawg hauv QIIME.27 Kev soj ntsuam ntawm molecular variance (AMOVA) tau ua los ntawm niam.28 Kev ntsuas Kruskal-Wallis tau siv los xam OTU qhov sib txawv ntawm pawg (p-value<0.05), and="" heat="" maps="" were="" drawn="" using="" pheatmap="" (r="" package).="" core="" microbiome="" analysis="" was="" adopted="" from="" the="" core="" function="" in="" the="" r="" package="" microbiome="" (sample="" prevalence="20%," relative="" abundance="0.01%)" by="" microbiomeanalyst29="" (https://www.="">

Kev txheeb cais tawm (LDA) thiab random hav zoov (RF) txoj hauv kev hauv MicrobiomeAnalyst Web site tau siv los txiav txim siab microbiome biomarkers. Ib qho nonparametric factorial Kruskal-Wallis sum-rank test tau ua thawj zaug los txheeb xyuas cov yam ntxwv nrog qhov tseem ceeb sib txawv ntawm kev xav txog qhov kev sim lossis chav kawm txaus siab. Tom ntej no, LDA (qhov pib yog teem rau 2) tau ua los xam cov txiaj ntsig loj ntawm txhua qhov sib txawv uas muaj ntau yam. Cov yam ntxwv tau suav tias yog qhov tseem ceeb ntawm lawv qhov kev hloov kho p-tus nqi. Lub neej ntawd hloov p-tus nqi txiav yog 0.05. RF tsom xam tau ua tiav siv randomForest pob5. Txoj kev no siv ib pawg ntawm cov ntoo cais, txhua tus uas yog cog los ntawm random feature xaiv los ntawm tus qauv bootstrap ntawm txhua ceg.

Tax4Fun (R pob, http://tax4fun.gobics.de/) tau siv los kwv yees cov microbial functional profiles ntawm microbiomes hauv cov qauv av. OTU Biom lub rooj ntawm cov av microbiome tau siv los ua cov ntaub ntawv nkag rau cov metagenome imputation ntawm C. salsa av cov qauv. Tom qab ntawd cov kev kwv yees ntawm cov gene class abundances tau soj ntsuam ntawm KEGG Orthology (KO) pawg theem 3. Cov txiaj ntsig tau los ntawm Tax4Fun tau txheeb xyuas hauv Doby (R pob).

2.5.Correlation Analysis of Key Microbial Communities and Bioclimatic Factors.

Peb siv ArcGIS los suav cov lej tseem ceeb ntawm bioclimatic yam tseem ceeb los ntawm 13 cov ntsiab lus hauv av. Kev txheeb xyuas qhov tsis txaus ntseeg ntawm cov genera tseem ceeb ntawm cov zej zog microbial (tsib biomarkers, tsib core microbiomes) thiab bioclimatic yam tau ua nrog

Canoco 5 software. Log2 cov ntaub ntawv hloov dua siab tshiab tau ua tib yam ua ntej kev tshuaj ntsuam. Spearman correlation coefficients raug xam rau ntau ntawm tsib biomarkers, tsib core microbiomes, thiab bioclimatic yam ntaub ntawv kev koom ua ke los ntawm kev siv SPSS. Cov txiaj ntsig kev txheeb xyuas kev sib raug zoo tau kos los ntawm pheatmap (R pob).

Cistanchetuaj yeem tiv taus kev laus

3. TSEEM CEEB

Kev twv ua ntej ntawm Ntiaj Teb Kev faib tawm ua haujlwm ntawm C. salsa nyob rau lub sijhawm sib txawv. Qhov xam ROC pom

uas tus nqi AUC yog {{0}}}.977 (Daim duab S2), qhia tias tus qauv pom zoo nrog cov ntaub ntawv.30 Habitat tsim nyog simulation tshwm sim rau tsib lub sijhawm (LGM, MH, tam sim no, 2050, thiab 2070 ) tau piav qhia nyob rau hauv daim duab 2 thiab Table 2. C. salsa-loj hlob haum tau muab faib ua tsib pawg raws li kev txheeb cais: tsis tsim nyog (chav kawm 5: 0-20 feem pua), marginal (chav kawm 4: 20-50 feem pua), ncaj ncees (chav kawm 3: 50-75 feem pua), zoo (chav kawm 2: 75−

90 feem pua), thiab zoo heev (chav kawm 1: 90-100 feem pua). Kab lus no tsuas yog tham txog cov chaw faib khoom tsim nyog rau chav kawm 1, chav kawm 2, thiab chav kawm 3. Tam sim no muaj peev xwm faib tawm ntawm C. salsa qhia tias hom kab no tau nthuav dav thoob plaws peb thaj chaw huab cua: huab cua huab cua huab cua sov, huab cua sov Mediterranean, thiab suab puam kub. thaj chaw huab cua. Txhua lub teb chaws, tshwj tsis yog Antarctica, muaj cov cheeb tsam uas tsim nyog rau C. salsa. Cov cheeb tsam no feem ntau tau faib rau hauv nruab nrab thiab sab hnub poob Asia thiab sab qaum teb Africa thiab tau tawg nyob rau hauv nruab nrab thiab sab hnub poob North America, Central South America, thiab sab hnub poob Oceania. Qhov zoo tshaj plaws (chav kawm 1) cov cheeb tsam tsim nyog tau nthuav dav tshaj plaws hauv tebchaws Iyiv (76 750 km2), qhov zoo (chav kawm 2) thiab kev ncaj ncees (chav kawm 3) cov cheeb tsam tsim nyog tau faib ntau tshaj plaws hauv Suav teb (chav kawm 2: {{ 13}} km2 thiab chav kawm 3: 1 024 600 km2). Cov cheeb tsam uas tsim nyog nyob rau hauv Asia feem ntau txwv rau Tuam Tshoj (chav kawm 1: 45 775 km2), Jordan (chav kawm 1: 16 075 km2), Israel (chav kawm 1: 14 975 km2), Saudi Arabia (chav kawm 1: 14 925 km2), thiab Iran (chav kawm 1: 12 600 km2). Qhov chaw tsim nyog ntawm C. salsa nyob rau hauv teb chaws Africa tau muab faib rau hauv Egypt, Libya (chav kawm 1: 34 400} km2), thiab Tunisia (chav kawm 1: 275 km2). Cov cheeb tsam tsim nyog rau C. salsa hauv South America feem ntau yog faib hauv Chile (chav kawm 1: 16 550 km2).

Los ntawm qhov kev xav ntawm spatial (Daim duab 2), qhov kev hloov pauv hauv cheeb tsam tsim nyog rau tsib lub sijhawm ua ntej tau nce thiab tom qab ntawd txo qis nrog thaj tsam loj tshaj plaws hauv xyoo 2050 (Daim duab 3a, Daim duab S3). Peb pom qhov txo qis hauv qhov tsim nyog C. salsa cheeb tsam nyob rau hauv lub LGM txheeb ze rau qhov ntawd nyob rau hauv plaub lub sij hawm. Piv nrog rau tam sim no (4 358 775 km2), thaj chaw tsim nyog tau txo los ntawm 50 feem pua ​​(2 160 975 km2) hauv lub sijhawm LGM, los ntawm 10 feem pua ​​(MH: 3 910 350 km2) hauv lub sijhawm MH, thiab los ntawm 1 feem pua ​​({13}} km2) hauv 2070 lub sijhawm thiab nce 2 feem pua ​​({17}} km2) hauv 2050 lub sijhawm. Qhov tseem ceeb, txij li tam sim no mus rau yav tom ntej (los ntawm 1960 txog 2080), thaj chaw zoo heev (chav kawm 1) rau C. salsa maj mam txo qis (tam sim no, 243 200 km2; 2050, 232 425 km2; 2070, { {29}}km2).

Cistancheextract muaj ntaunoj qab haus huvcov txiaj ntsig

3.2. Kev twv ua ntej ntawm thaj chaw tsim nyog faib ntawm C. salsa hauv Suav teb thiab ntau yam ntawm Bioclimatic variables.

Qhov chaw tsim nyog rau C. salsa kev loj hlob hauv Suav teb feem ntau txwv rau sab qaum teb sab hnub poob Xinjiang, central Inner Mongolia, sab qaum teb Shaanxi, sab qaum teb Shanxi, sab qaum teb Qinghai, sab qaum teb Gansu, thiab feem ntau ntawm Ningxia (Daim duab 2b). Cov cheeb tsam tsim nyog tshaj plaws hauv Suav teb feem ntau yog faib hauv Xinjiang, ntawm qhov chaw zoo heev (chav kawm 1) npog 23 650 km2, thaj chaw zoo (chav kawm

2) npog 220 950 km2, thiab thaj chaw ncaj ncees (chav kawm 3) npog 189 975 km2. Ntawm thaj chaw zoo (chav kawm 2), Xinjiang suav txog 60.99 feem pua, tom qab ntawd Inner Mongolia (86 300 km2) thiab Gansu ({11}} km2) nrog 23.82 feem pua ​​​​thiab 6.75 feem pua, raws li (Daim duab 3b). Cov txiaj ntsig ntawm MaxEnt qauv qhia tias Xinjiang tshwj xeeb yog tsim rau C. salsa kev loj hlob. Txawm li cas los xij, ntau qhov chaw nyob, xws li gravel-desert, grassland, and desert-steppe, tau pom nyob rau tib cheeb tsam thaum mus ncig ua si. Yog li ntawd, peb tau txaus siab rau cov yam ntxwv ntawm cov av microbial cov zej zog thiab lawv txoj kev sib raug zoo nrog ib puag ncig nyob rau hauv txawv ecotypes ntawm C. salsa nyob rau hauv Tacheng, Xinjiang, Suav teb.

Qhov tseem ceeb ntawm txhua qhov kev hloov pauv bioclimatic rau kev faib tawm ntawm C. salsa yog qhia hauv Table 2. Tag nrho cov txiaj ntsig tau txhais tau tias yog 10 replicate runs. Peb qhov kev hloov pauv bioclimatic saum toj kawg nkaus nrog kev cuam tshuam loj tshaj plaws ntawm C. salsa faib yog nag lossis daus ntawm lub quarter qhuav (bio17), nag lossis daus ntawm lub quarter sov tshaj plaws (bio18), thiab qhov nruab nrab diurnal range (bio2) nrog cov txheeb ze koom nrog 25.81. feem pua, 17.65 feem pua, thiab 13.70 feem pua, feem. Raws li qhia los ntawm cov lus teb nkhaus tau txais los ntawm MaxEnt qauv, qhov ntau ntawm cov khoom bio tuaj yeem suav tau thaum qhov kev kwv yees qhov tshwm sim tshaj 0.5. Piv txwv li, qhov ntau ntawm bio17 yog los ntawm −135.30 mus rau 11.34 mm, uas ntawm bio 18 yog los ntawm −206.60 mus rau 4.40 mm, thiab bio2 yog los ntawm 11.65 mus rau 12.50 degree.

3.3. Av Microbiomes Los Ntawm Peb Ecotypes ntawm C. salsa Exhibiting Distinct thiab Overlapping Microbial Zej Zog.

16S rRNA sequencing ua rau 518 217 nyeem ntawv nyoos, ntawm cov uas 441 576 raug tshuaj xyuas kom zoo thiab ntev. Cov ntaub ntawv teev muaj 11 818−{4}} (txhais tau tias: 20 150) ib ntus ib qho piv txwv. Cov ntawv nyeem tau zoo tau ua ke los ntawm lub hauv paus ntawm

>97 feem pua ​​​​qhia txog tus kheej rau hauv 2 788 microbial OTUs (Table 1).

Lub zej zog microbial tau muab faib ua 34 phyla thiab 321 genera. Nyob rau theem phylum (Daim duab 4a), Actinobacteria (DS, 31.94 feem pua; GD; 46.42 feem pua; GL, 33.33 feem pua), Proteobacteria (DS,

23.25% yog '; PIB, 22.53%; GL, 24.68 feem pua), thiab Gemmatimonadetes (DS, 17.77 feem pua; GD, 8.02 feem pua; GL, 8.36 feem pua) yog cov tseem ceeb nyob rau hauv peb ecotypes. Nyob rau theem genus (Daim duab 4b), cov suab puam- steppe tau dominated los ntawm Euzebya (4.82 feem pua) thiab Arthrobacter (1.74 feem pua), qhov tseem ceeb tshaj plaws genera nyob rau hauv gravel- suab puam yog Arthrobacter (8.35 feem pua) thiab Bacillus (4.95 feem pua) ). Bacillus (6.89 feem pua) thiab Arthrobacter (5.57 feem pua) yog cov tseem ceeb hauv cov nyom. Sab saum toj 10 cov zej zog microbial tshaj plaws nyob rau hauv cov suab puam-steppe tau muab faib ua yim phyla (Daim duab 4c), cov nyob hauv cov pob zeb- suab puam tau muab faib ua xya phyla (Daim duab 4d), thiab cov hauv hav zoov tau muab faib ua 10 phyla (Daim duab 4e). ).

Cov kev ntsuas ntawm cov qauv sib txawv (-diversity) qhia txog kev hloov pauv ntawm cov nyom mus rau gravel- suab puam thiab suab puam-steppe (Daim duab 4f). Qhov sib txawv ntawm cov av microbial cov zej zog nyob rau hauv txhua tus qauv raug soj ntsuam ntawm Shannon, Chao 1, Fisher, thiab soj ntsuam hom diversity indexes. Lub Shannon, Chao 1, pom hom, thiab Fisher indexes qhia tias - ntau haiv neeg ntawm cov nyom nyom av.

tau siab dua li ntawm ob lub zej zog ecotype. Cov txiaj ntsig ntawm qhov tsis tshua muaj qhov nkhaus (Daim duab S5) zoo ib yam li cov txiaj ntsig saum toj no, tshwj tsis yog YM1 thiab HJ3. AMOVA cov txiaj ntsig (Table S3) tau pom tias muaj qhov sib txawv tseem ceeb (p < 0.01)="" ntawm="" peb="" qhov="" ecotypes.="" cov="" txiaj="" ntsig="" ntawm="" pcoas="" uas="" tsis="" muaj="" kev="" txwv="" ntawm="" qhov="" tsis="" hnyav="" unifrac="" nyob="" deb="" 2d="" daim="" phiaj="" (daim="" duab="" 4g)="" tau="" qhia="" tias="" cov="" qauv="" av="" ntawm="" cov="" kab="" mob="" microbiomes="" los="" ntawm="" txawv="" ecotypes,="" tshwj="" tsis="" yog="" ym1="" thiab="" hj3,="" tau="" zoo="" clustered.="" lub="" bray="" nrug="" ntau="" haiv="" neeg="" pawg="" pawg="" raws="" li="" ib="" leeg-clustering="" algorithm="" tshwm="" sim="" (daim="" duab="" 4h)="" ntawm="" peb="" ecotypes="" av="" qauv="" qhia="" tau="" hais="" tias="" cov="" nyom="" nyom="" qauv,="" tshwj="" tsis="" yog="" ym2,="" tau="" nyob="" ze,="" cov="" suab="" puam-steppe="" av="" qauv,="" tshwj="" tsis="" yog="" ja1,="" tau="" ze="" ze,="" thiab="" cov="" gravel-desert="" av="" qauv="" tau="" zoo="" sib="">

3.4. Sib txawv Microbiome Kev Ntsuas ntawm Peb Ecotypes ntawm C. salsa.

Daim ntawv qhia tshav kub ntawm qhov kev nplua nuj ntawm ntau yam sib txawv (Daim duab 5a) qhia tau hais tias cov genus ntau ntawm cov qauv ntawm cov suab puam steppe txawv ntawm cov qauv ntawm ob lub ecotypes. Raws li pom nyob rau hauv cov kev tshwm sim ntawm

LEfSe (Daim duab 5c) thiab RF (Daim duab 5d), daim ntawv qhia hluav taws xob mini mus rau sab xis qhia txog kev nplua nuj ntawm cov kab mob microbial nyob rau hauv peb ecotypes ntawm qib genus. Cov genera uas tuaj yeem sawv cev rau cov suab puam-steppe ecotype muaj xws li Gillisia, Illumatobacter, Salegentibacter, Marinimicrobium, thiab lwm yam. Variibacter yog lub cim.

genus theem uas tuaj yeem sawv cev rau gravel-desert ecotype. Cov biomarkers nyob rau hauv cov nyom ecotype muaj Flavisolibacter thiab Agromyces. Tom qab cov txiaj ntsig ntawm ob txoj kev sib xyaw ua ke, 11 biomarkers raug xaiv (Table S4). Daim duab 5b qhia txog daim ntawv qhia tshav kub ntawm kev nplua nuj ntawm 11 biomarkers.

3.5. Core Microbiome Screening thiab Metabolic Function Prediction of the Three Ecotypes of C. salsa.

Daim duab Venn (Daim duab 5e) tau npaj rau OTUs tau txais los ntawm tag nrho cov qauv hauv av, thiab cov txiaj ntsig tau pom tias peb qhov ecotypes sib koom 1712 OTUs. Txoj kev persistence tau saws los ntawm cov tseem ceeb muaj nuj nqi hauv R pob microbiome los txheeb xyuas cov tub ntxhais microbiome nyob rau hauv peb ecotypes ntawm C. salsa. Cov kab mob tseem ceeb no muaj rau OTUs thiab sib npaug rau 19.64 feem pua ​​​​ntawm tag nrho cov microbiome. Nrog rau kev cais tawm ntawm cov tsis muaj npe thiab tsis sib xws, cov OTUs no tau muab faib ua rau 6 genera, thiab lawv cov kev nplua nuj tau kos rau hauv daim ntawv qhia tshav kub (Daim duab 5f).

Cov txiaj ntsig kev twv ua haujlwm tau zoo (Daim duab 6, Cov Ntaub Ntawv Ntxiv 2) tau qhia tias txoj hauv kev ua haujlwm metabolism hauv av microbiomes hauv peb ecotypes ntawm C. salsa zoo ib yam nyob rau hauv cov carbohydrates thiab cov amino acid metabolism yog ntau ntawm cov txheej txheem metabolic. Membrane kev thauj mus los thiab cov teeb liab hloov pauv kuj tseem muaj ntau hauv kev ua cov ntaub ntawv ib puag ncig.

3.6.Correlation Analysis ntawm Microbial Zej Zog thiab Bioclimatic Variables ntawm Peb Ecotypes ntawm C. salsa.

Kev soj ntsuam tsis tu ncua ntawm cov tub ntxhais kawm, biomarker microbiome abundance, thiab bioclimatic variables tau ua nyob rau hauv genus theem, thiab reanalysis tau ua raws li qhov tshwm sim. Qhov kev hloov pauv ntawm qhov sib txawv yog 32.5 feem pua. Los nag ntawm lub quarter sov tshaj plaws (bio18) piav qhia 23.9 feem pua ​​​​ntawm cov zej zog microbial (p=0.07). Qhov ntsuas kub nruab nrab ntawm lub quarter sov tshaj plaws (bio10) thiab qhov nruab nrab diurnal ntau yam (bio2) tau cuam tshuam zoo nrog Illumatobacter thiab cuam tshuam tsis zoo nrog Bacillus (Daim duab 7a).

Kev txheeb xyuas kev sib raug zoo tau ua rau lub hauv paus, biomarker microbiome abundance, thiab xya bioclimatic variables. Cov txiaj ntsig ntawm kev sib raug zoo (Daim duab 7b) tau qhia tias Illumatobacter thiab Salegentibacter (biomarkers nyob rau hauv suab puam-steppe) tau zoo sib xws nrog qhov nruab nrab diurnal range (bio2) thiab qhov nruab nrab kub ntawm lub caij sov tshaj plaws (bio10) tab sis tseem ceeb heev nrog qhov tsis zoo (correlated). alt) thiab nag lossis daus ntawm lub quarter sov tshaj plaws (bio18). Ntawm qhov tsis sib xws, Agromyces (biomarker hauv cov nyom) tau muaj txiaj ntsig zoo nrog qhov siab (alt) thiab nag lossis daus ntawm lub quarter sov tshaj plaws (bio18) tab sis tsis zoo cuam tshuam nrog qhov nruab nrab diurnal ntau (bio2) thiab qhov ntsuas kub ntawm lub quarter sov tshaj (bio10). Tsis tas li ntawd, Arthrobacter (core microbiome) yog qhov tsis zoo cuam tshuam nrog qhov nruab nrab diurnal range (bio2) thiab qhov ntsuas kub nruab nrab ntawm lub quarter warmest (bio10). Rubrobacter tau muaj txiaj ntsig zoo cuam tshuam nrog nag lossis daus txhua xyoo (bio12).