Phenolic Cov ntsiab lus thiab kev sib txawv ntawm cov noob caj noob ces nyob rau theem ntawm cov pej xeem thoob plaws Lub Ntuj Kev faib ntau yam ntawm Bearberry (Arctostaphylos Uva-ursi, Ericaceae) hauv Iberian Peninsula

Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com

Abstract:Bearberry (Arctostaphylos uva-ursi) yog cov nroj tsuag tshuaj ib txwm siv los kho cov kab mob urinary vim muaj cov ntsiab lus siab ntawmarbutin(hydroquinone -D-glucoside), uas tam sim no tsuas yog siv los ua cov tawv nqaij dawb-dawb hauv cov tshuaj pleev ib ce. Bearberry kuj tau pom zoo raws li ntuj tsimantioxidantNtxiv nrog rau cov ntsiab lus siab ntawm phenolic tebchaw hauv nplooj.Peb tau kawm txog qhov sib txawv ntawm phenolic tebchaw hauv 42 cov tsiaj qus ntawm bearberry, aiming toelucidate yog intrinsic biological, climatic, thiab/los yog geographic yam cuam tshuam rau cov ntsiab lus phenolic thoob plaws nwsnatural tis nyob rau hauv Iberian Peninsula. Cov nplooj Bearberry tau sau thaum lub caij nplooj zeeg dhau peb lub xyoos (2014-2016) hauv cov pej xeem thoob plaws ib cheeb tsam thiab qhov siab gradient. Methanolicextracts tau pom ntau qhov sib txawv ntawm tag nrho cov ntsiab lus phenols, thiab sib txawv phenolic profiles hais txogarbutin(qib ntawm cov ntsiab lus tseem ceeb no txawv ntawm 87 mus rau 232 mg / g dr wt), tab sis kuj muaj cov ntsiab lus catechinand myricetin, uas tau cuam tshuam los ntawm thaj chaw thiab huab cua. Cov theem nruab nrab ntawm kev hloov pauv ntawm cov genome loj - ntsuas los ntawm kev ntws cytometry - thiab ntawm ob thaj chaw plastid DNA kuj tau kuaj pom ntawm cov pej xeem. Cov noob caj noob ces thiab cytogenetic sib txawv ntawm cov pejxeem tsis muaj zog tab sis tseem ceeb cuam tshuam nrog kev sib txawv ntawm phytochemical. Cov neeg tseem ceeb bearberry genotypes nrog siab duaantioxidantmuaj peev xwm raug txheeb xyuas tom qab.

Ntsiab lus: arbutin; genetic thiab phytochemical variability; genome loj; haplotypes; ntujantioxidants

cistanche yog ntuj antioxidants

1. Taw qhia

Lub synthesis ntawm cov nroj tsuag tshwj xeeb metabolites sib txawv nyob rau hauv lub sij hawm (piv txwv li, ontogeny, phenology, thiab induceddefenses), thiab qhov chaw, raws li nws plays lub luag hauj lwm tseem ceeb nyob rau hauv cov nroj tsuag adaptation rau ib puag ncig tej yam kev mob, hos kuj genetic variation account rau chemodiversity [1]. Ntawm cov tebchaw no, phenolshave muaj ntau hom mono- thiab polymeric qauv uas ua tiav ntau yam ntawm physiologicalroles [2]. Lub biosynthesis ntawm cov metabolites tshwj xeeb cuam tshuam zoo heev los ntawm ib puag ncig xws li kub, nag lossis daus, lossis hnub ci hluav taws xob, uas feem ntau raug rau latitudinal, longitudinal, lossis altitudinal gradients. Tshwj xeeb, phenolic tebchaw tsub zuj zuj yog cov lus teb dav dav rau kev txhim kho qib ntawm UV-B (280-315 nm) hluav taws xob. Cov kev hloov pauv tshwj xeeb tau tshaj tawm hauv cov nroj tsuag loj hlob hauv thaj av Mediterranean thaum lub caij ntuj sov thiab ntawm qhov chaw siab, qhov twg muaj qhov tshwm sim ntau dua ntawm UV-B tshwm sim thiab cinnamic acids thiab flavonoids pom qhov siab tshaj UVabsorption [3].

Arbutin(hydroquinone -D-glucoside) yog ib qho yooj yim phenol compound nrog kev txwv tsis pub muaj nyob rau hauv nplooj ntawm qee hom genera xws li Arbutus, Arctostaphylos, Pyrus, los yog Vaccinium.Lub ntsiab ntuj tsim ntawm arbutin, lub bearberry (Arctostaphylos uva-ursi ( L.) Spreng.) tau siv rau ntau pua xyoo los kho cov kab mob urinary thiab lwm yam kab mob raum [4], thiab cov tshuaj ntsuab tseem npaj tau tam sim no [5]. Nyob rau hauv xyoo tas los no, spectrum ntawm kev siv ntawmarbutintau nthuav dav, feem ntau yog tus neeg sawv cev ntawm daim tawv nqaij dawb hauv kev lag luam kom zoo nkauj [6] thiab hauv kev kho mob vim nwsantioxidant, tshuaj tua kab mob, tshuaj tua kab mob, thiab tshuaj tua kab mob [7]. Yog li ntawd, muaj kev txaus siab ntau ntxiv hauv kev nrhiav cov peev txheej ntuj ntxiv ntawmarbutin, nrog rau cov txheej txheem biotechnological uas tuaj yeem hloov cov tshuaj synthesis [8,9]. Hauv cov ntsiab lus no, hauv vitro tsim cov arbutin sivDatura inoxia cell kab lis kev cai tau mus txog qhov ntsuas ntsuas [10].

Ntxiv rauarbutin, lwm cov phenolic tebchaw ua rau A. uva-ursi active zog, xws li flavonoids thiab tannins [11,12], uas catechin thiab corilagin yog cov feem ntau cuam tshuam inleaves, feem [13]. Hauv kev lag luam khoom noj khoom haus natural compounds raws li cov nroj tsuag phenols yog replacesyntheticantioxidantpreservatives [14]. A. uva-ursi tau siv los ua ib qho ntxiv, tshwj xeeb tshaj yog nyob rau hauv cov khoom noj nqaij [15–19] tab sis kuj muaj nyob rau hauv cov khoom ntim [20]. Thaum kawg, lub peev xwm ntawm A. uva-ursi raws li qhov chaw ntawm cov neeg sawv cev ntawm cov tawv nqaij tau raug npaj tsis ntev los no [21].

Cov "bearberry nplooj" monograph ntawm European Tshuaj Lub Koom Haum [5] tau thov kom tsawg kawg ntawm 7 feem puaarbutincov ntsiab lus nyob rau hauv qhuav nplooj raws li qhov yuav tsum tau rau kev npaj tshuaj ntsuab. Cov kev tshawb fawb tau ua dhau los xyoo dhau los tau piav qhia txog cov ntsiab lus ntawm arbutin hauv bearberry nplooj txawv ntawm 0 feem pua ​​​​rau 18 feem pua ​​, piav qhia cov txheej txheem los ntawm kev txheeb xyuas, kev hloov pauv ntuj, kev loj hlob, thiab hnub sau qoob [4]. Arbutin cov ntsiab lus feem ntau txiav txim siab los ntawm kev ua haujlwm siab ua kua chromatography [4,12,22–24], thiab cov kev tshawb fawb yav dhau los tau pom ntau duaarbutinCov ntsiab lus nyob rau hauv bearberry nroj tsuag sau nyob rau hauv lub caij nplooj ntoos zeeg tshaj li nyob rau hauv cov sau nyob rau hauv lub caij nplooj ntoos hlav [4] .Muaj ntau yam bearberry chemotypes tau piav, txuam nrog lub geographical distributionand rau intraspecific sib txawv nyob rau hauv subspecies ntawm A. uva-ursi. Qhov zoo tshaj plaws phytochemicalvariation tshaj tawm yog qhov tsis muaj arbutin hauv A. uva-ursi subsp. stipitata Packer thiab Denford, tab sis kuj muaj qhov sib txawv hauv methyl-arbutin, ellagic acid, lossis myricetin cov ntsiab lus tau raug xa mus. Quercetin kuj muaj nyob rau hauv bearberry nplooj, uas ob qho tib si aglycones, nrog rau lawv cov 3-O-glycosides yog cov feem ntau pom flavonoids [4]. Txawm li cas los xij, feem ntau ntawm cov kev tshawb fawb no tau ua tiav ntau xyoo dhau los, thiab tam sim no intraspecific variation hauv bearberry yog suav tias yog tas mus li. Lub 14 subspecies piav qhia, suav nrog ib qho los ntawm Spain (A. uva-ursi subsp. crassifolius (Braun-Blanq.) Rivas Mart. ex Torre, Alcaraz thiab MB Crespo), yog tam sim no tsis tau txais cov npe thiab sib txuas nrog A. uva-ursi [ 25].Cov kev tshawb nrhiav tshiab yog li ntawd yuav tsum tau txhim kho peb txoj kev paub txog cov khoom sib txuas ua ke hauv cov khoom siv raw uas tau sau los ntawm ntau qhov chaw rau kev lag luam txij li cov tshuaj profile ntawm A. uva-ursi nplooj tau piav qhia kom suav nrog ntau hom phenols, tannins, thiab flavonoidsin cov kev tshawb fawb tsis ntev los no tau ua los tshawb xyuas qhov sib txawv ntawm qib sib txawv [23,26,27].

Cov noob caj noob ces (xws li cytogenetic) ntau haiv neeg ntawm cov pej xeem theem txhawb kev hloov mus rau qhov sib txawv ntawm ib puag ncig [28]. Cov teebmeem ntawm polyploidization ntawm kev tsim cov metabolites tshwj xeeb hauv cov tshuaj thiab cov nroj tsuag muaj ntxhiab tsis ntev los no tau tshuaj xyuas [29]. Nyob rau hauv cov ntsiab lus no, genome size yog ib tug loj parameter nyob rau hauv cov nroj tsuag variability kev ntsuam xyuas, txij li thaum lub intraspecific variation yog feem ntau kuaj pom ([30], thiab cov ntaub ntawv nyob rau hauv), correlated nrog ntau lwm yam kab mob lom, thiab plays lub luag hauj lwm nyob rau hauv evolutionary txheej txheem [31-35. ]. Txawm li cas los xij, cov kev tshawb fawb hais txog kev sib koom ntawm cov nyiaj DNA ntawm nuclear (xws li genome loj) thiab phytochemical ntau haiv neeg tsawg heev [36,37]. Cov kev sib raug zoo ntawm phytochemical thiab genetic ntau haiv neeg tau raug soj ntsuam siv nuclear tab sis tshwj xeeb tshaj yog cov cim plastidDNA [38], tshwj xeeb tshaj yog nyob rau hauv tshuaj [39] thiab cultivated [40] hom nroj tsuag.

Bearberry yog dav faib thoob plaws hauv cheeb tsam circumboreal, tab sis feem ntau ntawm cov tsiaj qus sau nyob rau hauv cov teb chaws Europe yog nyob rau hauv lub Eastern lub teb chaws, Austria, Switzerland, Ltalis, thiab Spain. altitudes li ntawm 550 mus rau 2350 m saum toj no hiav txwv theem (asl). Nws muaj ntau dua nyob rau sab qaum teb vim cov pej xeem nyob hauv qhov chaw qis qis qis dua thiab feem ntau muaj cov tib neeg tsawg dua. Cov kev tshawb fawb uas twb muaj lawm hais txog cov ntsiab lus sib txawv ntawm arbutin hauv Spain tau xa mus rau qhov tseem ceeb xws li 8 feem pua ​​​​ntawm cov neeg nyob sab qaum teb sab hnub tuaj [41] thiab 19 feem pua ​​​​hauv Spanish cov khoom cog tau soj ntsuam hauv kev tshawb fawb hauv Tebchaws Yelemees [42]. Peb txoj kev tshawb fawb aims toelucidate yam, nrog rau qhov loj ntawm genome thiab genetic ntau haiv neeg, piav qhia phytochemical variationacross lub ntuj faib ntawm bearberry nyob Spain. Peb cov txiaj ntsig tuaj yeem pab txhawb rau kev xaiv cov khoom cog rau kev lag luam tshuaj, tshuaj pleev ib ce, thiab zaub mov.

daim tawv nqaij dawb zaub mov: CISATNCHE

2. Cov txiaj ntsig thiab kev sib tham

2.1. Chemical Diversity Analysis

Bearberry nplooj coj los ntawm tag nrho ntawm 249 cov nroj tsuag loj hlob hauv 42 qhov chaw Spanish, thiab thoob plaws peb lub xyoos, 2014–2016 (Daim duab 1, Table S1), pom qhov hnyav qhuav sib txawv (dr wt) feem pua, xws li 46.7 (LO) rau 55.1 feem pua ​​(CP), nrog rau qhov nruab nrab ntawm 50.1 ± 2.7 feem pua ​​(cov ntaub ntawv tsis qhia). Methanolicextracts npaj los ntawm cov qauv sau hauv 2014 thiab 2015 tau ua haujlwm rau tag nrho cov phenols thiab kev txiav txim siab arbutin ntsiab lus. Peb tau pom ntau yam kev hloov pauv tsis tu ncua rau ob qhov tsis muaj qhov sib txawv tseem ceeb (p <0.001) ntawm="" cov="" nroj="" tsuag.="" nplooj="" rho="" tawm="" ntawm="" 80="" nroj="" tsuag="" coj="" mus="" rau="" lub="" caij="" nplooj="" zeeg="" 2014="" (daim="" duab="" 2a)="" pom="" tag="" nrho="" cov="" ntsiab="" lus="" phenolic="" xws="" li="" 103.3="" ±="" 4.8="" mg="" gae/g="" dr="" wt="" (piv="" txwv="" li-4)="" to206.4="" ±="" 6.5="" mg="" gae/g="" dr="" wt="" (sr="" -2),="">arbutinCov ntsiab lus hloov pauv ntawm 92.0 ± 3.0 mg/g dr wt (AN-6) mus rau 194.2 ± 5.6 mg/g dr wt (SE-8). Kev soj ntsuam ntawm cov extracts npaj los ntawm nplooj sau nyob rau hauv 2015 los ntawm 94 nroj tsuag (Daim duab 2b) kuj pom ib tug dav phytochemical variation, los ntawm 110.5 ± 3.6 mg GAE/g dr wt(PI-4) mus rau 200.9 ± 9.8 mg GAE/g dr wt (LO-4) nyob rau hauv tag nrho cov ntsiab lus phenolic, thaum cov ntsiab lus arbutin ranged ntawm 87.1 ± 0.4 mg / g dr wt (ET-2) thiab 211.5 ± 5.9 mg / g dr wt (LI -2). Tsis tas li ntawd, tsis hais lub xyoo ntawm kev sau, qhov sib txawv tseem ceeb hauv tag nrho cov phenols thiabarbutinCov ntsiab lus tau kuaj pom ntawm cov nroj tsuag bearberry loj hlob nyob rau tib qhov chaw rau feem ntau ntawm cov pejxeem (cov ntaub ntawv tsis qhia).Arbutin qib tau cuam tshuam nrog tag nrho cov ntsiab lus phenols hauv bearberry nplooj extract txij thaum peb kwv yees qhov tseem ceeb Pearson coefficients ntawm {{0}} .332 (p=0.003) rau 2014, thiab 0.289 rau 2015data (p=0.005). Arbutin cov ntsiab lus ntawm 48 cov nroj tsuag coj mus rau hauv ob lub xyoos tsis txawv heev (p=0.380).

Txawm hais tias muaj qhov sib txawv nyob hauv cov neeg, thiab tsis hais txog lub xyoo sau qoob loo, kev txheeb xyuas qhov sib txawv tau pom tias cov ntsiab lus ntawm arbutin ntawm cov nplooj ntoo nplooj extract kuj tseem nyob ntawm (p < {{0}}="" 00="" 1)="" onpopulation="" qhov="" chaw="" (daim="" duab="" 3a,b),="" raws="" li="" tshwm="" sim="" rau="" tag="" nrho="" cov="" ntsiab="" lus="" phenols="" ntawm="" bearberry="" nplooj="" analyzedin="" 2015="" (p="">< 0.001,="" daim="" duab="" 3b).="" hauv="" qhov="" sib="" piv,="" qhov="" sib="" txawv="" tseem="" ceeb="" hauv="" tag="" nrho="" cov="" ntsiab="" lus="" phenols="" tsis="" tau="" pom="" (p="0.080)" ntawm="" 10="" tus="" neeg="" kuaj="" hauv="" xyoo="" 2014,="" uas="" tau="" nyob="" hauv="" thaj="" chaw="" me="" me,="" tab="" sis="" faib="" rau="" ntau="" qhov="" siab,="" los="" ntawm="" 424="" (ba.="" pejxeem)="" txog="" 1410="" m="" asl(pa="" pejxeem).="" qhov="" kev="" hloov="" pauv="" ntawm="" qhov="" siab="" no="" tau="" cuam="" tshuam="" rau="" qhov="" sib="" txawv="" ntawm="" kev="" nyab="" xeeb="" (table="" s1),="" txij="" li="" qhov="" chaw="" ntawm="" qhov="" chaw="" siab="" dua="" yog="" qhov="" ntsuas="" kub="" qis="" dua="" (pearson's="" coefficient="" of="" correlation,="" r="−0.666," p="">< 0.001)="" thiab="" ntau="" dua="" txhua="" xyoo="" nag="" lossis="" daus="" (="" r="0.485," p="">< 0.001),="" uas="" inturn="" ua="" rau="" lub="" ntiaj="" teb="" hluav="" taws="" xob="" qis="" qis="" (r="−0.390," p="">< 0.001).="" tsis="" tas="" li="" ntawd,="" los="" ntawm="" cov="" ntaub="" ntawv="" no="" tau="" kwv="" yees="" qhov="" qis="" tab="" sis="" tseem="" ceeb="" ntawm="" kev="" sib="" raug="" zoo="" ntawm="" cov="" dej="" nag="" txhua="" xyoo="" thiab="" tag="" nrho="" cov="" phenols="" cov="" ntsiab="" lus="" ntawm="" cov="" nroj="" tsuag="" bearberry="" (spearman's="" coefficient="" rho="0.256," p="0.022)," whereas="" los="" ntawm="" 2015="" cov="" ntaub="" ntawv="" teev="" thaum="" peb="" kuaj="" cov="" pej="" xeem="" nyob="" rau="" hauv="" ib="" cheeb="" tsam="" loj,="" peb="" tau="" kuaj="" pom="" muaj="" kev="" sib="" raug="" zoo="" ntawm="" cov="" dej="" nag="" txhua="" xyoo="" thiab="" cov="" ntsiab="" lus="" arbutin="" (rho="0.246," p="0.017)." qhov="" kev="" sib="" txheeb="" tseem="" ceeb="" no="" qhia="" tau="" hais="" tias="" cov="" nroj="" tsuag="" bearberry="" loj="" hlob="" nyob="" rau="" sab="" qaum="" teb="" qhov="" chaw="" thiab="" ntawm="" qhov="" chaw="" siab="" dua="" tau="" pom="" ntau="">arbutincov ntsiab lus (rho=0.217, p=0.035 thiab rho=0.269, p=0.009, feem). Raws li tau hais, cov ntsiab lus arbutin siab dua no cuam tshuam ntau dua rau cov ntsiab lus phenols ntau dua ntawm bearberryleaves, uas zoo sib xws (qhov nruab nrab 154.4 ± 19.6 mg GAE / g dr wt) rau cov pom hauv aqueousextracts los ntawm lwm hom nroj tsuag ua haujlwm raws liantioxidantadditives hauv kev lag luam zaub mov [43], xws li asrosemary (185.0 mg GAE/g dr wt), tshuaj yej (149.3 mg GAE/g dr wt), los yog guava (154.4 mg GAE/g dr wt). Cov txiaj ntsig no tau pom zoo nrog cov ntaub ntawv yav dhau los thiab tau lees paub hauv kev tshawb fawb ua los ntawm Wrona li al. [20] siv qee cov qauv Spanish bearberry nplooj sau hauv 2015, rau qhov siab duaantioxidantmuaj peev xwm tau txuam nrog ntau duaarbutincov ntsiab lus. Cov neeg tseem ceeb bearberry genotypes kuj tseem raug txheeb xyuas rau lub hom phiaj no, xws li cov tib neeg 1, 4, 7, thiab 8 los ntawm cov pejxeem LO, uas tau khaws cia nruab nrab ntawm 183.3 ± 16.4 mg GAE / g dr wt hauv ob xyoos ntawm kev kawm.

Txhawm rau kom paub ntxiv txog kev nyab xeeb thiab thaj chaw muaj feem cuam tshuam rau cov qauv ntawm phenolicvariation, peb tau ua ib qho piv txwv ntau dua nyob rau lub caij nplooj zeeg 2016: 140 bearberry nroj tsuag loj hlob hauv 29 qhov chaw, thiab peb suav tsib phenolic tebchaw. (Daim duab S1a) uas tau txheeb xyuas los ntawm kev sib txuas nrog cov qauv hauv acetonitrile / dej gradient (Daim duab S1b). Kev soj ntsuam tau pom ntau qhov sib txawv tsis tu ncua rau arbutin, caffeic acid, catechin, myricetin, thiab quercetin glucoside cov ntsiab lus, nrog qhov sib txawv tseem ceeb ntawm cov nroj tsuag uas feem ntau pom nyob rau hauv cov pej xeem (cov ntaub ntawv tsis qhia). Rau cov ntsiab lus tseem ceeb phenolic, peb pom tias cov ntsiab lus arbutin nyob ntawm 91.1 ± 5. }}), thaum cov ntsiab lus catechin nyob ntawm 4.1 ± 0.1 (AF{{20}}) txog 45.5 ± 1.4 mg / g dr wt (LB-5). Hais txog ob qho lwm yam flavonoids tau txiav txim siab, myricetin tsis raug kuaj pom hauv qee cov nroj tsuag los ntawm ntau tus neeg, thaum mus txog 21.2 ± 1.2 mg / g dr wt hauv PO{{30}}, thiab quercetin glucoside txawv ntawm 3.8 ± 0.1 (CO -3) rau 22.8 ± 0.8 mg/g dr wt (BT-3). Cov ntsiab lus qis ntawm caffeic acid tau txiav txim siab hauv bearberry nplooj extracts, uas txawv ntawm 1.8 ± 0.0 (IZ-1) txog 7.1 ± 0.3 mgPlants /g dr wt (SI-4).

Txawm hais tias muaj kev sib txawv ntawm cov tib neeg, qhov sib txawv tseem ceeb ntawm cov pej xeem kuj tau kwv yees rau tsib lub tebchaw no (Kruskal–Wallis tests, p < {{0}}.001,="" thiab="" p="0." 009="" forcaffeic="" acid="" cov="" ntsiab="" lus).="" cov="" ntsiab="" lus="" arbutin="" siab="" dua="" tau="" txiav="" txim="" siab,="" qhov="" nruab="" nrab,="" hauv="" cov="" nroj="" tsuag="" los="" ntawm="" ab,="" gu,="" ln,="" thiab="" lo="" natural="" pejxeem="" (daim="" duab="" 4a),="" uas="" txawv="" heev="" (tom="" qab="" bonferronicorrection)="" los="" ntawm="" cov="" ntsiab="" lus="" tsawg="" pom="" hauv="" cov="" nroj="" tsuag="" los="" ntawm="" lb="" pejxeem="" (186.7="" ±="" 22.3,="" 193.9="" ±="" 12.1,169.5="" ±="" 10.0,="" thiab="" 169.2="" ±="" 5.2="" mg="" g="" dr="" wt="" pem="" hauv="" ntej="" mus="" rau="" 111.8="" ±="" 16.0="" mg="" g="" dr="" wt,="" ntsig="" txog).="" ib="" zaug="">arbutincov ntsiab lus ntawm cov nroj tsuag uas tau coj mus kuaj kuj nyob rau hauv 2015 tsis sib txawv heev ntawm xyoo (p=0.821). Cov pej xeem LB tau pom nyob rau hauv lem siab dua txhais tau tias catechin theem (Daim duab 4b), uas txawv txav ntawm cov ntsiab lus qis ntawm cov flavonoid no pom hauv cov pejxeem AF thiab LC (32.8 ± 9.8 pem hauv ntej mus rau 5.7 ± 1.9 thiab 5.3 ± {{2{{62}). }}}.5 mg/g dr wt, ntsig). Txhais tau hais tias myricetin thiab quercetin glucosidecontents tau pom hauv daim duab 4c, nrog qhov sib txawv tseem ceeb ntawm cov pejxeem AB, CG, LB, thiab PO, nrog cov ntsiab lus myricetin ntau dua (15.1 ± 4.0, 14.6 ± 5.7, 10 .7 ± 2.8, thiab 17.2 ± 3.9 mg / g dr wt, ntsig txog), piv rau cov qib qis uas tau sau los ntawm cov nroj tsuag nruab nrab ntawm cov pejxeem CO thiab OD (1.2 ± 1.5and 2.4 ± 3.6 mg / g dr wt, raws li) , thiab tseem txhais tau hais tias quercetin glucoside cov ntsiab lus txiav txim siab cov neeg nyob hauv AA, BT, CP, IZ, thiab LB (15.2 ± 3.7, 15.9 ± 4.6, 11.8 ± 2.3, 15.7 ± 3.4, thiab 13.1 ± 4.6, mg / g rhiab heev) los ntawm qhov ntawd hauv cov pejxeem CO (4.0 ± 0.1 mg/g dr wt). Piv txwv li, cov ntsiab lus caffeic acid oscillated los ntawm 2.1 ± 0.2 mg / g dr wt hauv cov pejxeem AB, CE, PT, thiab SA, rau 3.5 ± 1.2 mg / g dr wt hauv cov pejxeem AF lossis 3.4 ± 2.2 mg / g dr wt hauv pejxeem SI; Yog li ntawd, qhov sib txawv tseem ceeb tsis tau pom vim qhov sib txawv ntawm intrapopulation (nruab nrab cov ntsiab lus 2.7 ± 0.8 mg / g dr wt).Cov txiaj ntsig no ntxiv qhov kev hloov pauv ntawm phenolic metabolites pom los ntawm Wrona li al. [20], rau yim ntawm qhov chaw bearberry coj mus kuaj hauv 2015 uas tau soj ntsuam siv UPLC®-ESI-Q-TOF nrog MSE thev naus laus zis.

Hauv kev sib piv rau cov txiaj ntsig tau txais hauv kev tshuaj ntsuam ua nrog cov qauv bearberry los ntawm 2014 thiab 2015,arbutincov ntsiab lus txiav txim siab hauv 2{{10}}16 cov qauv qhia pom qhov qis tab sis tseem ceeb ntawm kev sib raug zoo nrog hluav taws xob (rho=0.256, p=0.002), thiab qhov siab tshaj qhov ntsuas kub (rho{ {5}}.183, p=0.030), thiab yog li ntawd tau inversely correlated rau cov dej nag txhua xyoo (rho=−0.265, p=0.002). Cov kev sib txawv no kuj tseem cuam tshuam nrog kev hloov pauv hauv qhov siab (rho=−0.192, p=0.023), tab sis qhov kev sib raug zoo tsis zoo no tau piav qhia los ntawm qhov hais qis.arbutincov ntsiab lus ntawm cov nroj tsuag los ntawm LB cov pej xeem uas nyob ntawm 1720m. Ib qho piv txwv ntawm kev hloov pauv tau pom zoo rau cov ntsiab lus myricetin txij li qib siab ntawm qhov sib xyaw no tau cuam tshuam nrog cov nroj tsuag bearberry loj hlob nyob rau hauv qhov chaw nyob rau hauv cov hluav taws xob ntau dua (rho=0.226, p=0).{{1{{ 12}}}}07). Tus qauv rov qab rau ntawm arbutin tau pom zoo rau catechin, txij li qhov kev sib raug zoo tau kwv yees ntawm qhov flavonoid thiab dej nag txhua xyoo (rho=0.398,p < 0="" {21}}{{23="" }}1),="" thiab="" cov="" kev="" sib="" raug="" tsis="" zoo="" tau="" raug="" kuaj="" pom="" muaj="" hluav="" taws="" xob="" thiab="" nrog="" qhov="" ntsuas="" kub="" siab="" tshaj="" plaws="" (rho="−0.307," p="">< 0.001="" thiab="" rho="−0.470," p="">< 0.001,="" raws).="" qhov="" no="" variationin="" climatic="" yam="" piav="" qhia="" tias="" ob="" qhov="" chaw="" siab="" thiab="" latitude="" cuam="" tshuam="" cov="" ntsiab="" lus="" catechin="" ntawm="" bearberryleaves="" (rho="0.410" thiab="" 0.490,="" raws="" li,="" p=""><0.001). ib="" qho="" tseem="" ceeb="" ntawm="" kev="" tshuaj="" ntsuam="" xyuas="" tau="" ua="" nrog="" phytochemical,="" huab="" cua,="" thiab="" thaj="" chaw="" cov="" ntaub="" ntawv="" sib="" raug="" rau="" 29="" qhov="" chaw="" kuaj="" hauv="" xyoo="" 2016,="" uas="" ob="" qho="" tib="" si="" thawj="" zaug="" tau="" piav="" qhia="" 60="" feem="" pua="" ​​​​ntawm="" cov="" kev="" hloov="" pauv,="" tau="" lees="" paub="" cov="" txiaj="" ntsig="" no="" (daim="" duab="" 5).="" peb="" tsis="" pom="" muaj="" kev="" sib="" raug="" zoo="" ntawm="" caffeic="" acid="" los="" yog="" quercetin="" glucoside="" ntsiab="" lus="" ntawm="" cov="" nroj="" tsuag="" bearberry="" thiab="" cov="" huab="" cua="" lossis="" thaj="" chaw,="" txawm="" hais="" tias="" peb="" kwv="" yees="" me="" ntsis="" kev="" sib="" raug="" zoo="" ntawm="" myricetin="" thiab="" quercetin="" glucoside="" cov="" ntsiab="" lus="" (rho="0.184," p="" {{31}).="" }.030)="" thiab="" nruab="" nrab="" ntawm="" myricetin="" thiab="" caffeic="" acid="" cov="" ntsiab="" lus="" (rho="−0.176," p="0.037)" ntawm="" bearberry="" nroj="">

Qhov pom contrasting climatic qauv ntawm variation nyob rau hauvarbutinCov ntsiab lus ntawm cov nroj tsuag bearberry qhia tias, txawm hais tias biosynthesis ntawm cov metabolite no yuav nce ntxiv nyob rau hauv cov xwm txheej ntawm cov hluav taws xob ntau dua thiab kub, tej zaum dej tsis txaus yog qhov txwv tsis pub muaj rau A. uva-ursi hauv thaj tsam Mediterranean thiab yav qab teb. Del Valle et al. [44] tau tshaj tawm cov qauv latitudinal ntawm flavonoidsaccumulation ntawm Silene littorea (Caryophyllaceae) hauv Spain, qhov twg cov ntsiab lus flavonoids siab dua tau txiav txim siab nyob rau yav qab teb cov neeg, txij li latitude cuam tshuam tsis zoo nrog UV-B hluav taws xob thiab kub, thiab zoo nrog nag lossis daus. Cov hluav taws xob thoob ntiaj teb qhia pom qhov tseeb latitudinal gradient nrog qhov tshwm sim ntau dua nyob rau sab qab teb thiab sab hnub tuaj ntawm Iberian Peninsula, thiab tseem muaj qhov cuam tshuam tseem ceeb vim huab cua persistence modulates qhov xwm txheej ntawm cov hluav taws xob no ntawm qhov siab siab dua.A UV-radiation daim ntawv qhia ntawm Spain [45] xa mus rau kev sib raug zoo siab (r > 0.9) ntawm UV-B thiab cov ntaub ntawv hluav taws xob thoob ntiaj teb. Raws li qhov kev siv no, bearberry pejxeem uas pom tau tias siab duaarbutinCov ntsiab lus hauv peb txoj kev tshawb fawb no feem ntau nyob hauv thaj chaw hauv qab UV-B hluav taws xob theem ntawm 2403-2451 J / m2.

Peb cov txiaj ntsig rau catechin variation nyob rau hauv bearberry pom ib thaj chaw qauv (Daim duab 5) thiab pom zoo nrog cov kev tshawb fawb ua nyob rau hauv lwm hom nroj tsuag los ntawm sab qaum teb cov teb chaws Europe. Cov ntsiab lus siab dua ntawm cov metabolites tshwj xeeb, xws li flavonoids thiab anthocyanins, tau tshaj tawm nyob rau hauv cov nroj tsuag loj hlob ntawm qhov siab dua latitudes, tej zaum vim lub hnub ci ntev dua thiab qis dua hmo ntuj [46]. Yog li, cov ntsiab lus phenolic soluble nyob rau hauv Juniperus communis koob tau nce nrog latitude thiab qhov siab hauv Finlandpopulations [47], thiab kuj 10-19 feem pua ​​​​siab dua cov ntsiab lus phenolic tau pom nyob rau ntawm ib qho chaw siab dua hauv cov txiv hmab txiv ntoo ntawm peb Ribes spp. cultivars soj ntsuam nyob rau hauv ob qhov chaw Finnish [48]. Cov kws sau ntawv no hais txog cov ntsiab lus phenols qis dua nrog cov hluav taws xob ntau dua thiab kub. Flavonoid cov ntsiab lus nyob rau hauv cov txiv hmab txiv ntoo ntawm ob hom Vaccinium (tseem los ntawm tsev neeg Ericaceae) pom ib thaj chaw gradient, nrog ntau dua ntawm flavonoids nyob rau sab qaum teb latitudes [49,50].

Nyob rau hauv kev sib piv rau altitudinal variation, qib siab ntawm flavonoids tau txiav txim siab nyob rau hauv cov pej xeem ntawm lwm hom Ericaceae, Calluna vulgaris, loj hlob ntawm siab dua [51]. Altitudinal gradients kuj cuam tshuam rau cov metabolism hauv tshwj xeeb ntawm cov neeg Spanish ntawm Arnica montana [52] thiab Quercus robur [53].Cov haujlwm tom kawg tau xa mus rau qhov tseem ceeb hauv kev sib raug zoo ntawm cov concentration ntawm tag nrho cov nplooj phenolics thiab nce siab, ua gradient nyob ntawm tsuas yog flavonoids, yog li qhia tias cov no. compoundsdrove txoj kev sib raug zoo rau tag nrho phenolics. Cov txiaj ntsig no pom zoo nrog peb qhov kwv yees rau catechin contentpattern. Thaum kawg, cov dej nag gradient tau txuam nrog qhov sib txawv tseem ceeb hauv polyphenols muaj pes tsawg leeg hauv African tshuaj ntsuab tsob ntoo (Myrothamnus flabellifolia) uas qhov sib txawv ntawm cov metabolomic tau sib haum nrog cov qauv caj ces ntawm cov pejxeem [54].

CISTANCHE tuaj yeem tiv thaiv kev laus

2.2. Genome Size thiab Sequence Data Analysis

Genome loj 2C qhov tseem ceeb ntawm cov nroj tsuag bearberry ranged ntawm 2.50 txog 3.15 pg (Table 1). Qhov sib txawv me ntsis ntawm cov DNA DNA muaj nyob hauv cov pej xeem (Kruskal–Wallis χ2=87.639, df=36, p=3.39 × 10−6). Dunn qhov kev sim nrog Bonferroni kho tau qhia qhov sib txawv tseem ceeb ntawm qhov sib txawv: AL thiab SI pejxeem (Z=−3.972, p=0.024), LE thiab SI (Z=−4.196, p {{ 21}}.009), and SEand SI (Z=−4.236, p=0.008). Cov pejxeem ntawm Pontils (PO) yog hom cheeb tsam ntawm A. uva-ursi var.crassifolius, uas tau tom qab ua ke ua A. uva-ursi subsp. crassifolius, thiab tam sim no tsis suav tias yog taxonomically (saib cov lus qhia), muaj tus nqi nruab nrab (2.88 pg), nyob rau hauv qhov ntau ntawm cov txiaj ntsig tau txais hauv lwm haiv neeg ntawm hom. Qhov no yog thawj qhov kev kawm dav dav ntawm nuclear DNA ntau npaum li cas hauv hom thiab tag nrho cov genus txij li cov ntaub ntawv tsuas muaj nyob rau niaj hnub no los ntawm ib tus neeg Balkan ntawm tib taxon [55]. Tus nqi 2C muaj qhia, 2.49 pg, yog muab tso rau ntawm qhov qis qis ntawm qhov sib txawv ntawm qhov kev tshawb xyuas cov ntaub ntawv no. Tsis muaj kev sib raug zoo ntawm cov genome loj qhov tseem ceeb thiab cov pej xeem qhov siab tau pom. Tus nqi DNA ntawm cov tib neeg tau pom qhov tsis muaj txiaj ntsig zoo cuam tshuam nrog rauarbutincov ntsiab lus thaum cov ntaub ntawv los ntawm 158 bearberryplants tau muab piv (rho=0.187, p=0.018), thaum tsis muaj lwm yam tseem ceeb correlation ntawm genomesize thiab lwm yam sib txawv. Qhov txiaj ntsig no tau pom zoo nrog cov lus tshaj tawm nce ntxiv hauv kev tsim cov metabolites tshwj xeeb uas feem ntau pom hauv cov tshuaj thiab cov nroj tsuag muaj ntxhiab tsw qab polyploidization [29].

Cov khoom tsim tshiab tshiab rau rpl{{0}}trnL thiab psbE-petN intergenic spacers tau ua raws hauv obmatrices (feem ntau muaj 566 thiab 874 bp) uas tau muab sib xyaw rau hauv ib qho matrix ntawm 1440 bp.Moderate qib ntawm kev hloov pauv hauv cov cheeb tsam plastid no tau kuaj pom (rau lub nucleotide hloov pauv thiab plaub indels). Kaum qhov sib txawv haplotypes tau pom nrog tag nrho haplotype ntau haiv neeg (Hd) ntawm 0.468 (Table 1). Feem ntau cov pej xeem (21) tsuas muaj ib qho haplotype xwb, thaum tsib ntawm lawv pom muaj ntau haiv neeg ntawm cov tib neeg. Qhov ntau tshaj haplotype 1 pom qhov ntau zaus (71.4 feem pua), ua raws li haplotype 2 (15.2 feem pua), qhov seem ntawm lawv tau pom ntau zaus qis dua (0.04-0.01 feem pua). Lub geographic faib ntawm haplotypes yog qhia nyob rau hauv daim duab 6. Haplotype 1 tau pom nyob rau hauv 32 cov pejxeem - nws tau kho nyob rau hauv 18 ntawm lawv - faib thoob plaws tag nrho cov cheeb tsam ntawm cov qauv tshwj tsis yog rau cov cheeb tsam sab qab teb tshaj plaws. Haplotype 2 tau pom nyob rau hauv kaum tus neeg thiab yog qhov tshwj xeeb haplotype nyob rau hauv ob sab qab teb neeg coob (HU thiab LV) nrog rau hauv PR. Haplotype 4 yog ntiag tug los ntawm peb haiv neeg los ntawm Pyrenees, thaum haplotype 8 tshwm sim hauv ob pawg neeg los ntawm sab hnub tuaj IberianPeninsula. Hais txog kev sib raug zoo evolutionary uas qhia nyob rau hauv lub parsimony network (Daim duab 6), feem ntau ntawm cov haplotypes yog kev cob cog rua los ntawm ib tug los yog ob tug kev hloov pauv. Haplotype 1 nyob hauv nruab nrab, nrog xya haplotypes txuas nrog nws.

Kev ntsuam xyuas Mantel ntawm pDNA caj ces thiab tshuaj sib txuas sib txuas ntawm qhov matrices pom tias tsis muaj zog tab sis muaj kev sib raug zoo (r=0.301; p=0.048) ntawm kev sib txawv ntawm caj ces thiab qhov concentration ntawm tsib yam tshuaj ntsuas rau cov 2016 data set. Kev ntsuam xyuas Mantel tsis tau qhia txog kev sib raug zoo ntawm caj ces thiab thaj chaw nyob deb ntawm 25 tus neeg kuaj hauv 2016 (r=0.103, p=0.207) lossis nruab nrab ntawm cov tshuaj lom neeg thiab thaj chaw sib txawv ntawm cov no. 25 pejxeem (r=0.191, p=0.050). Lub phytochemical variability qhia ntawm no rau bearberry qus pej xeem thoob plaws Spain yog li ntawd complemented los ntawm nruab nrab theem ntawm pDNA haplotype variation, yog lub sab qaum teb-sab qab teb genetic sib txawv pom nyob rau hauv bearberry pejxeem piv rau phylogeographic qauv pom nyob rau hauv lwm yam nroj tsuag (xws li, [56–58]. Tag nrho cov no homogeneity yuav tsum tau nyob rau hauv cov nroj tsuag uas tsis muaj peev xwm yug me nyuam [59], raws li A. uva-ursi, uas nthuav tawm cov kab mob tsis zoo thiab cov neeg tuag siab ntawm cov tub ntxhais hluas seedlings [4]. Txawm li cas los xij, qhov sib txawv ntawm cov noob caj noob ces pom los ntawm pDNA cuam tshuam nrog biochemical. Pairwise nrug kwv yees los ntawm profiles ntawm tsib phenolic daim duab 6. Geographical tis ntawm pDNA haplotypes ntawm Iberian pejxeem ntawm Arctostaphylos uva-ursi.Population codes sib raug rau cov nyob rau hauv Table S1, thiab ncuav qab zib kab kos sawv cev rau feem pua ​​​​ntawm cov tib neeg uas qhia txhua haplotype hauv txhua tus pej xeem. daim duab plaub, qhov txheeb cais parsimony network qhia txog kev sib raug zoo ntawm te n plastid haplotypes pom nyob rau hauv A. uva-ursi populationsis sawv cev. Txhua kab txaij raws cov kab sib txuas cov haplotypes qhia ib kauj ruam hloov pauv hauv thaj tsam pDNA ua ntu zus hauv txoj kev tshawb no.A Mantel kev sim ntawm pDNA caj ces thiab tshuaj sib txuas sib txuas ntawm cov matrices pom qhov tsis muaj zog tab sis tseem ceeb correlation (r=0.301; p {{28. }}.048) ntawm kev sib txawv ntawm cov noob caj noob ces thiab qhov concentration ntawm tsib yam tshuaj uas ntsuas rau 2016 cov ntaub ntawv teev. Qhov kev sim Mantel tsis tau qhia txog qhov tseem ceeb ntawm kev sib raug zoo ntawm caj ces thiab thaj chaw nyob deb ntawm 25 tus neeg sampledin 2016 (r=0.103, p=0.207) lossis nruab nrab ntawm cov tshuaj lom neeg thiab thaj chaw sib txawv ntawm 25 tus neeg ( r=0.191, p=0.050). Lub phytochemical variability qhia ntawm no rau bearberrywild pej xeem thoob plaws Spain yog li ua tiav los ntawm theem nrab ntawm pDNA haplotypevariation, ua rau sab qaum teb-sab qab teb genetic sib txawv pom nyob rau hauv bearberry pejxeem piv rau phylogeographic qauv pom nyob rau hauv lwm yam nroj tsuag (xws li, [56–58]. Tag nrho cov no homogeneity yuav tsum inplants nrog ib tug tsawg reproductive muaj peev xwm [59], raws li A. uva-ursi, uas nthuav tawm tsis zoo germination tus nqi thiab siab tuag tus nqi ntawm cov tub ntxhais hluas seedlings [4]. Txawm li cas los xij, qhov sib txawv ntawm cov noob caj noob ces qhia los ntawm pDNA cuam tshuam nrog biochemical pairwise nrug kwv yees los ntawm profiles. Cov qauv no ntawm cov noob caj noob ces thiab phytochemical variation (uas, raws li tau hais, tsis yog txuam nrog thaj chaw deb), nrog rau cov tsis muaj zog tab sis tseem ceeb correlation ntawm genomesize thiab arbutin cov ntsiab lus, tej zaum yuav qhia tau hais tias ntuj genomic variability cuam tshuam cov yield ntawm phenoliccompounds nyob rau hauv A. uva-ursi. Txawm li cas los xij, hypervariable molecular m arkers nyob rau hauv ib qho piv txwv ntawm cov pej xeem ntau yog tsim nyog los txheeb xyuas cov qauv phylogeographic thiab kev sib koom ua ke ntawm cov noob caj noob ces thiab biochemical hloov pauv.

CISTANCHE muaj ANTI-OXIDATION EFFECTS

3. Cov ntaub ntawv thiab cov txheej txheem

3.1. Cov khoom cog

Rau phytochemical tsom xam, tag nrho ntawm 249 bearberry nroj tsuag loj hlob nyob rau hauv 42 pej xeem raug kuaj nyob rau lub caij nplooj zeeg 2014, 2015, thiab 2016, ua tus sawv cev ntawm lub ntuj faib ntawm no nroj tsuag nyob rau hauv Iberian Peninsula (Daim duab 1 thiab Table S1). Ua ntej, thaum lub caij nplooj zeeg 2014, peb tau sau cov terminalshoots los ntawm 80 nroj tsuag hauv kaum Spanish pejxeem (AG, AN, BA, LI, LO, PE, PÁ, SR, SC, thiab SE) nyob rau hauv ib cheeb tsam me me (80 × 50 km kwv yees) nyob rau sab qaum teb Spain tab sis nyob rau hauv ntau qhov chaw siab tshaj (424-1410 m asl). Thib ob, thaum lub caij nplooj zeeg xyoo 2015, peb tau sau 48 cov nroj tsuag los ntawm rau ntawm cov pejxeem (AG, LI, LO, PA, SE, thiab SR) thiab 46 bearberry nroj tsuag los ntawm rau cov pejxeem nyob rau hauv qis latitudes (AL, CH, ET, HU, LV. , thiab PI). Rau txhua tus pej xeem, peb tau sim yim tsob nroj sib cais tsawg kawg yog 5 m, tshwj tsis yog hauv HU, qhov chaw peb pom tsuas yog rau tus neeg. Peb, nyob rau lub caij nplooj zeeg 2016, tag nrho ntawm 140 cov nroj tsuag tau coj mus kuaj hauv 29 qhov chaw sib txawv (1-6 tus neeg los ntawm txhua tus) suav nrog 26 lub zos Spanish tshiab (IZ, CE, BT, MO, AA, GU, SI, AF, SA, PT, AB, CG, OD, PS, LB, CO, LE, MA, CP, PO, MZ, AV, LC, AY, LN, and ZU). Peb cov qauv ntsuas qhov siab ntawm 534 txog 1750 m. Ua ke, 42 tus neeg sawv cev rau ntau yam kev nyab xeeb (Table S1): qhov ntsuas qhov siab tshaj plaws pom tau tias 2-foldvariation (13–26 ◦C) thiab nag lossis daus txhua xyoo oscillated ntawm 399 (ZU) txog 1589 mm (PT). Cov hluav taws xob thoob ntiaj teb yog los ntawm 4.2 txog 5.1 kWh / m2d. Rau rau kaum lub davhlau ya nyob twg tua (15-20 cm ntev) tau sau rau txhua tsob ntoo kom tau 6-15 g ntawm cov nplooj noj qab haus huv, uas tau tom qab excised thiab qhuav ntawm 60 ◦C qhov hnyav tas li (3-4 hnub). Tom qab qhov kev txiav txim siab qhuav qhuav, nplooj tau ua homogenized hauv mortar thiab khaws cia ntawm 4 ◦C kom txog thaum kuaj xyuas. Cov khoom cog no tau ua haujlwm rau tag nrho cov phenols thiab HPLC kev txiav txim.

Cov ntaub ntawv nplooj ntoo rau kev ntsuas cytometric tau txais los ntawm nplooj tshiab ntawm 178 tus neeg koom nrog rau tag nrho 37 cov neeg (Table 1). Cov qauv tau txais los ntawm 33 ntawm cov neeg uas tau hais los saum toj no, thiab los ntawm plaub tus neeg tshiab (BE, JO, thiab PR hauv Spain, thiab EN hauv Andorra) qhov twg nplooj tau sau rau lub caij nplooj ntoo hlav 2017, thiab yog li ntawd tsis suav nrog phytochemicalanalysis (Daim duab 1 thiab Tab S1). Los ntawm ib mus rau rau tus neeg rau ib tug pej xeem raug ntsuas induplicates. Nplooj cov ntaub ntawv rau DNA rho tawm tau qhuav hauv silica gel thiab khaws cia ntawm chav tsev kub.DNA kev txheeb xyuas ntau haiv neeg tau ua nyob rau hauv tag nrho ntawm 105 nroj tsuag los ntawm 35 qhov chaw (Table 1).

Cog vouchers ntawm 46 bearberry pejxeem kawm tau muab tso rau hauv herbarium BC, ntawm lub koom haum Botànic de Barcelona, ​​hauv herbarium BCN, ntawm Center de Documentació de BiodiversitatVegetal, Universitat de Barcelona los yog hauv herbarium JACA, ntawm Instituto Pirenaico de Eco ( CSIC).

3.2. Kev npaj ntawm nplooj Extracts thiab tag nrho Phenols txiav txim

Peb replicates ntawm bearberry nplooj extract tau npaj los ntawm kev siv 50 mg ntawm cov qauv qhuav thiab 10 mL ntawm 80 feem pua ​​methanol. Cov hlab tau incubated rau 30 min nyob rau hauv ib tug ultrasonic da dej, thiab ces extractswere filtered (0.45 mm) thiab khaws cia ntawm 4 ◦C kom txog rau thaum kev soj ntsuam. Tag nrho cov ntsiab lus phenolic tau txiav txim siab nyob rau hauv cov extracts npaj los ntawm cov qauv sau hauv 2014 thiab 2015. Peb ua raws li Folin-Ciocalteumethod [60,61] nrog kev hloov me ntsis: mus rau 0.1 mL ntawm extract, 0.4 mL ntawm methanol (80 feem pua), 0.5 mLof Folin- Ciocalteu reagent thiab 8 mL ntawm ultrapure dej tau ntxiv. Tom qab 5 feeb hauv ib qho ultrasonicbath, 1 mL ntawm Na2CO3 20 feem pua ​​(w/v) tau ntxiv. Cov qauv raug tso rau hauv qhov tsaus ntuj rau 30 feeb ua ntej ntsuas qhov nqus ntawm 760 nm siv lub UV-pom pom spectrophotometer (CARY 50 BIO, Varian, AgilentTechnologies). Cov txiaj ntsig tau qhia nyob rau hauv qhov sib npaug gallic acid (GAE); uas yog, mg gallic acid / g dr wt, siv lub gallic acid tus qauv nkhaus (40–340 µg / g).

3.3. Arbutin thiab Lwm Phenolic Metabolite Kev Txiav Txim

Phenolic cov ntsiab lus ntawm bearberry nplooj methanolic extracts tau quantified los ntawm RP-HPLC sivan HPLC-UV/Vis system (LaChrom Merck Hitachi L-7400) nrog ib tug Kinetex 5 µm-EVO C18(250 hli × 4.6 hli ) kem thiab absorbance txiav txim siab ntawm 280 nm. Cov khoom siv los ntawm cov qauv sau tau hauv xyoo 2014 thiab 2015 tau diluted 1:10 (v/v) thiab txhaj nrog ib theem mobile ntawm methanol thiab acetonitrile.Cov kev pab cuam gradient muaj 0–5 min, 25 feem pua ​​ntawm methanol; 5-6 feeb, 100 feem pua; 6-10 min, 100 feem pua; thiab 10-20 min, dua 25 feem pua ​​​​ntawm methanol. Tus nqi ntws yog 1 mL / min thiab txhaj tshuaj yog 20 μL. Hauv cov xwm txheej no, lub sijhawm khaws cia ntawmarbutinyog 2.6 min, ua rau peb txiav txim siab qhov concentration ntawm qhov sib xyaw no siv lub calibration nkhaus tsim nrog 6 dilutions ntawm tus qauv, los ntawm 4 0 mus rau 340 µg / g, nrog rau kev sib txheeb coefficient ntawm r=0. ib 9997. Cov ntaub ntawv pov thawj los ntawm cov qauv sau hauv 2016 tau txheeb xyuas siv lub sijhawm txawb ntawm methanol thiab dej nrog cov txheej txheem gradient hauv qab no: 0-5 min, 10 feem pua ​​​​ntawm methanol; 5-6 feeb, 20 feem pua; 6-10 feeb, 20 feem pua; 10-11 feeb, 30 feem pua; 11-15 feeb, 30 feem pua; 15-16 feeb, 40 feem pua; 16-20 feeb, 40 feem pua; 20–21 feeb, 50 feem pua; 21–25 feeb, 50 feem pua; 25–26 feeb, 60 feem pua; 26-30 feeb, 60 feem pua; 30-31 feeb, 10 feem pua; thiab 31-36 min, dua 10 feem pua ​​​​ntawm methanol. Hauv cov xwm txheej no, peb txiav txim siabarbutin thiabLwm cov 4 phenolic tebchaw: caffeic acid, catechin, myricetin, thiab quercetin -3-O-glucopyranosideusing tus coj calibration nkhaus tsim nrog 5 dilutions ntawm cov qauv (25–500 µg/mL) , uas qhia txog kev sib txheeb coefficients ntawm {{10}}}.9992 rau catechin (lub sij hawm tuav 12.0 min), 0.9924 rau caffeicacid (lub sij hawm tuav 13.4 min), 0.9981 rau quercetin{{16} }O-glucoside (lub sijhawm tuav 23.4 min), thiab 0.9997 rau myricetin (lub sijhawm tuav 25.1 min).

Methanol thiab acetonitrile tau nyob hauv qib HPLC thiab yuav los ntawm Panreac (Barcelona, ​​Spain).Standards (purity Ntau dua lossis sib npaug li 98 feem pua) ntawmarbutin, caffeic acid, thiab gallic acid, nrog rau Folin-Ciocalteu reagent tau yuav los ntawm Sigma-Aldrich (Barcelona, ​​Spain). Catechin thiab quercetin-3-O-glucopyranosidestandards tau yuav los ntawm Extrasynthese (Genay Cedex, Fabkis), thaum myricetin tau yuav los ntawm Alfa Aesar (Karlsruhe, Lub teb chaws Yelemees).

Cov txiaj ntsig ntawm kev txiav txim siab tshuaj lom neeg tau txheeb xyuas siv SPSS v. 25 software thiab R 3.5.2.Tom qab kev sim cov ntaub ntawv rau qhov qub thiab homoscedasticity, peb tau ua qhov kev ntsuam xyuas ntawm qhov sib txawv ntawm cov ntaub ntawv sib txawv thiab txhais tau tias kev xeem sib cais (Tukey thiab Tamhane) lossis cov kev ntsuas tsis yog parametric. raws li Kruskal-Wallis, thiab Dunn ntau qhov kev sib piv nrog Bonferroni kho. Peb kuj kwv yees correlationcoefficients (Pearson los yog Spearman nyob ntawm seb cov ntaub ntawv ua raws li ib txwm faib) ntawm qhov sib txawv, suav nrog genome loj.

CISTANCHE muaj ANTI-OXIDATION EFFECTS

3.4. Genome Size kwv yees thiab DNA Sequencing

Lub genome loj ntawm cov nroj tsuag bearberry tau kwv yees los ntawm kev ntws cytometry ntawm Centers Científicsi Tecnològics, Universitat de Barcelona (CCiTUB) ua raws li cov txheej txheem piav qhia hauv Pellicer li al. [62].Petunia hybrida Vilm. 'PxPc6' (2C=2.85 pg) tau siv los ua tus qauv sab hauv. Cov noob ntawm tus qauv tau muab los ntawm Plateforme de cytométrie d'Imagerie-Gif, CNRS-I2BC (Gif-sur-Yvette, France).Nuclear DNA cov ntsiab lus (2C) tau suav los ntawm kev sib npaug ntawm cov ntsiab lus DNA paub ntawm tus qauv bythe quotient ntawm cov ncov txoj hauj lwm (hom) ntawm hom hom phiaj thiab tus qauv nyob rau hauv histogramof fluorescence intensities, piv txwv li ib tug linear correlation ntawm lub fluorescent teeb liab los ntawm thestained nuclei ntawm cov tsis paub cov qauv, paub nyob rau hauv tus qauv thiab cov DNA [63].

Kwv yees li 20 mg ntawm silica-dried nplooj cov ntaub so ntswg tau siv rau DNA rho tawm siv CTABprotocol [64] nrog kev hloov kho me me. Qhov zoo ntawm tag nrho DNA raug kuaj nrog NanoDrop 1000spectrophotometer (ThermoScientific, Wilmington, DE, USA). Cov cheeb tsam plastid intergenic rpl32-trnLand psbE-petN, nrog rau thaj tsam nuclear ribosomal DNA ITS, tau nthuav dav thiab ua raws li peb tus neeg hauv ib tus neeg. Ib ntus ntawm ITS tau nthuav tawm tsis muaj qhov sib txawv hauv 105 tus neeg txheeb cais, tshwj tsis yog rau ob peb txoj haujlwm uas qhia txog kev sib txawv ntawm cov nucleotide polymorphisms. Yog li ntawd, thaj av nuclear ribosomal DNA no raug tshem tawm los ntawm kev tshuaj xyuas ntxiv. Lwm thaj chaw plastid DNA (piv txwv li, ndhF; ndhF-rpl32; psbA-trnH; psbD-trnT; rps16; thiab trnL-trnF) kuj tau sim tab sis lawv raug muab pov tseg vim tsis tshua muaj kev sib txawv los yog cov teeb meem sequencing. Amplification txheej txheem tau ua raws li tau piav qhia hauv Vitales li al. [65]. Direct sequencing of the amplified DNA segments was performed with Big Dye Terminator Cycle Sequencing v 3.1 (PE Biosystems, Foster City, California, USA) ntawm Unitatde Genòmica, CCiTUB, ntawm ABI PRISM 3700 DNA analyzer (PE Biosystems). Cov rpl32-trnL thiabpsbE-petN ib ntus tau sib sau ua ke nrog BioEdit version 7.1.3.0 [66], ua raws li ClusalW MultipleAlignment v1.4 [67] thiab kho manually. GenBank tus lej nkag tau muab rau hauv Table S2.

3.5. Genome Size thiab Genetic Analysis

Kev kuaj tsis-parametric Kruskal-Wallis tau ua los kuaj xyuas qhov sib txawv tseem ceeb hauv 2C qhov tseem ceeb ntawm cov pejxeem. Dunn ntau qhov kev sib piv kuj tau ua los txiav txim seb cov neeg twg pom qhov sib txawv tseem ceeb ntawm lawv. Qhov kev kho Bonferroni tau siv los txo qhov kev ua yuam kev yam kuv (tsis muaj tseeb).

Kev sib txawv ntawm caj ces (polymorphic sites, parsimony cov chaw qhia paub, tus naj npawb ntawm haplotypes, thiab tag nrho haplotype ntau haiv neeg) tau kwv yees siv DnaSP v6 [68]. Plastid haplotypes tau txiav txim siab hauv cov ntaub ntawv sib koom ua ke uas suav nrog ob qho tib si rpl32-trnL thiab psbE-petN cheeb tsam.Indels tau codified nrog FastGap v.1.2 [69] thiab kho raws li ib qho kev hloov pauv. Cov evolutionaryrelationships ntawm haplotypes tau inferred raws li parsimony network tsim siv TCS [70] raws li siv nyob rau hauv PopArt [71]. Qhov siab tshaj plaws ntawm qhov sib txawv uas tshwm sim los ntawm singlesubstitutions ntawm haplotypes tau suav nrog 95 feem pua ​​​​kev ntseeg siab txwv.

Txhawm rau txheeb xyuas qhov kev sib raug zoo ntawm cov ntaub ntawv caj ces thiab tshuaj lom neeg ib qho kev sim Mantel tau ua. Ua ntej, peb kwv yees qhov phenotypic deb ntawm 25 cov neeg siv cov ntaub ntawv los ntawm cov nroj tsuag bearberry sau hauv 2016, uas tau ntsuas rau tsib yam phenolic (piv txwv li.arbutin, catechin, caffeic acid, quercetin, thiab myricetin). Peb siv R Commander los tsim cov ntaub ntawv no [72], thiab tom qab ntawd, los xam cov tshuaj nyob deb tau los ntawm cov qauv matrix nrog cov haujlwm "Dist" onR raws li Euclidean nrug. Thib ob, Nei cov pej xeem genetic pairwise nrug matrix tau suav nruab nrab ntawm 25 cov neeg siv DnaSP v6. Thaum kawg, peb suav cov geographicdistances pairwise ntawm cov pej xeem siv pob "geodist" hauv R. Pairwise correlations nruab nrab ntawm matrices tau suav nrog kev xeem Mantel nrog 10,000 permutations nrog functionmantel muaj nyob rau hauv R pob "vegan" [ 73] ib.

4. Cov lus xaus

Tag nrho cov txheeb xyuas bearberry nroj tsuag pomarbutincov ntsiab lus siab dua 7 feem pua; Yog li ntawd, cov khoom siv cog no tsim nyog rau kev npaj tshuaj ntsuab. Peb qhov kev soj ntsuam tau qhia ntau dua arbutin cov ntsiab lus ntau dua li cov (txog 9 feem pua) xa los ntawm Parejo li al. [40], uas tau txheeb xyuas plaub tus neeg nyob rau sab qaum teb sab hnub tuaj Pyrenees ntawm 1580-2030 m thiab kuaj pom qhov sib txawv ntawm cov ntsiab lus arbutin tsawg ntawm cov neeg. Tsis tas li ntawd, feem ntau ntawm cov txiaj ntsig tau txiav txim siab ntawm no yog nyob rau hauv thaj tsam ntawm cov khoom siv cog qoob loo uas tau xaiv rau kev cog qoob loo hauv tebchaws Poland [74] thiab hauv kev pom zoo nrog qhov siab.arbutinCov ntsiab lus ntawm Spanish bearberry nplooj qauv qhia los ntawm Sonnenschein thiab Tegtmeier [42] thaum xaiv cov nroj tsuag forcultivation hauv lub teb chaws Yelemees. Cov neeg tseem ceeb genotypes ntawm A. uva-ursi tuaj yeem nthuav tawm vegetatively, tab sis feem ntau ntawm cov clones tau ua tsis tiav thaum lub sij hawm tsim teb, uas txwv kev siv cov peev txheej no. Peb kuj tau piav qhia txog qhov kev hloov pauv uas twb muaj lawm hauv cov ntsiab lus ntawm peb flavonoids (catechin, myricetin, thiab quercetinglucoside), nrog rau caffeic acid, ntawm cov neeg Spanish bearberry, thiab cov xwm txheej huab cua (feem ntau yog lub ntiaj teb hluav taws xob thiab dej nag), uas cuam tshuam rau latitudinal thiab altitudinal. gradients, cuam tshuam qhov variation. Nyob rau tib lub sijhawm, txawm tias qhov qis qis ntawm haplotype thiab genome loj hloov pauv tau txiav txim siab hauv Iberian A. uva-ursi, genetic thiab cytogenetic sib txawv ntawm cov pejxeem tsis muaj zog tab sis cuam tshuam nrog ntau hom phytochemical. Zuag qhia tag nrho, cov txiaj ntsig no qhia tau hais tias cov txiaj ntsig ntawm ob qho tib si caj ces thiab cov kab mob abiotic yuav tsum tau txiav txim siab los piav qhia txog kev sib txawv ntawm phytochemical muaj nyob hauv cov nroj tsuag los ntawm cov tsiaj qus.

CISTANCHE EFFECTS

Cov ntaub ntawv

1. Moore, BD; Andrew, RL; Kulheim, C.; Foley, WJ Piav qhia txog kev sib txawv intraspecific hauv cov nroj tsuag theem nrab metabolites hauv cov ntsiab lus ecological. New Phytol. 2014, 201, 733–750. [CrossRef] [PubMed]

2. Cheynier, V.; Kom, G.; Davies, KM; Lattanzio, V. Plant phenolics: Kev nce qib tsis ntev los no ntawm lawv cov biosynthesis, noob caj noob ces, thiab ecophysiology. Nroj Physiol. Biochem. 2013, 72, 1–20. [CrossRef]

3. Bernal, M.; ib. Lus, L.; Julkunen-Tiitto, R.; Badosa, J.; Verdaguer, D. Altitudinal thiab raws caij nyoog hloov ntawm phenolic tebchaw hauv Buxus sempervirens nplooj thiab cuticles. Nroj Physiol. Biochem. 2013, 70, 471–482.[CrossRef] [PubMed]

4. Upton, R. (Ed.) Uva-ursi Nplooj. Arctostaphylos uva-ursi (L.) Spreng. Txuj kev soj ntsuam, Kev Tswj Xyuas Zoo, thiab Kev Kho Mob; American Herbal Pharmacopoeia: Scotts Valley, CA, USA, 2008; 30p ib.

5. European Medicines Agency. Daim ntawv ntsuam xyuas ntawm Arctostaphylos uva-ursi (L.) Spreng. Folium. EMA/HMPC/750266/2016; European Medicines Agency: Amsterdam, Lub Netherlands, 2018.6. Kanlayavattanakul, M.; Lourith, N. Nroj tsuag thiab cov khoom ntuj tsim rau kev kho mob ntawm daim tawv nqaij hyperpigmentation—Kev tshuaj xyuas. Planta Med. 2018, 84, 988–1006. [CrossRef]

7. Migas, P.; Krauze-Baranowska, M. Qhov tseem ceeb ntawm arbutin thiab nws cov derivatives hauv kev kho thiab tshuaj pleev ib ce.Phytochem. Lett. 2015, 13, 35–40. [CrossRef]

8. Seo, DH; Jung, JH; Li, JE; Jeon, EJ; Kim, W.; Park, CS Biotechnological zus tau tej cov arbutin (- thiab -arbutin), daim tawv nqaij-lightening agents, thiab lawv derivatives. Appl. Microbiol. Biotechnol. 2012, 95, 1417–1425 [CrossRef]

9. Xu, X. Tian, ​​Y.; Zhang, W. Zhang, T. Guang, C.; Mu, W. Tsis ntev los no kev nce qib ntawm cov khoom siv roj ntsha ntawm -arbutin. Appl. Microbiol. Biotechnol. 2018, 102, 8145–8152. [CrossRef] [PubMed]

10. Davey, M. Secondary metabolism hauv cov nroj tsuag kab lis kev cai. Hauv Encyclopedia of Applied Plant Sciences, 2nd ed.; Thomas, B., Murphy, DJ, Murray, BG, Eds.; Elsevier: London, UK, 2017; Phau 2, p. 462–467. [CrossRef]

11. Kurkin, VA; Ryazanova, TK; Daeva, ED; Kadentsev, VI Constituents of Arctostaphylos uva-ursi leaf.Chem. Nat. Compd. 2018, 54, 278–280. [CrossRef]

12. Panda, A.; Petrucci, R.; Marrosu, G.; Multari, G.; Romana-Gallo, F. HPLC-PDAESI-TOF/MS metabolicprofiling ntawm Arctostaphylos pungens thiab Arctostaphylos uva-ursi: Kev sib piv kev kawm ntawm phenolic compounds los ntawm nplooj nplooj metabolic extract. Phytochemistry 2015, 115, 79–88. [CrossRef]

13. Olennikov, DN; Chekhirova, GV 60-Galloylpicein thiab lwm yam phenolic tebchaw los ntawm Arctostaphylos uva-ursi.Chem. Nat. Compd. 2013, 49, 1–7. [CrossRef]

14. Brewer, MS Natural antioxidants: qhov chaw, tebchaw, mechanisms ntawm kev txiav txim, thiab tej yam kev siv.Compr. Rev. Food Sci. Food Safe. 2011, 10, 221–247. [CrossRef]

15. Amarowicz, R.; Peb, RB; Rahimi-Moghaddam, P.; Loj, B.; Weil, JA Dawb-radical scavenging muaj peev xwm thiab kev ua haujlwm antioxidant ntawm cov nroj tsuag xaiv los ntawm Canadian prairies. Khoom noj khoom haus Chem. 2004, 84, 551–562 [CrossRef]

16. Kws ntoo, R.; O'Grady, MN; O'Callaghan, YC; O'Brien, NM; Kerry, JP Kev Ntsuam Xyuas ntawm cov tshuaj antioxidant muaj peev xwm ntawm cov noob txiv hmab txiv ntoo thiab cov bearberry rho tawm hauv cov nqaij npuas nyoos thiab siav. Nqaij Sci. 2007, 76, 604–610. [CrossRef][PubMed]

17. Mekini'c, IG; Scroza, D.; Ljubenkov, ib.; Katalini'c, V.; Šimat, V. Antioxidant, thiab antimicrobial peev xwm ntawm phenolic metabolites los ntawm ib txwm siv Mediterranean tshuaj ntsuab thiab txuj lom. Khoom noj khoom haus 2019, 8, 579. [CrossRef]

18. Mohd Azman, NA; Gallego, MG; Segovia, F.; Abdullah, S.; Shaarani, SM; Almajano, MP Kawm txog cov khoom ntawm bearberry nplooj extract raws li ntuj antioxidant hauv cov qauv zaub mov. Antioxidants 2016, 5, 11.[CrossRef] [PubMed]

19. Peg, RB; Amarowicz, R. Naczk, M. Antioxidant kev ua ntawm polyphenolics los ntawm bearberry nplooj (Arctostaphylos uva-ursi L. Sprengel) extract nyob rau hauv cov nqaij systems. Nyob rau hauv Phenolic Cheebtsam nyob rau hauv cov zaub mov thiab Natural HealthProducts; Shahidi, F., Ho, CT, Eds.; American Chemical Society: Washington, DC, USA, 2005; pp. 67–82 [CrossRef]

20. Wrona, M.; Blasco, S.; Becerril, R. Nerín, C.; Kev muag khoom, E.; Asensio, E. Antioxidant thiab antimicrobial markers byUPLC®–ESI-Q-TOF-MSE ntawm ib tug tshiab multilayer active ntim raws li Arctostaphylos uva-ursi. Xyoo 2019, 196, 498–509. [CrossRef]

21. Maier, M.; Olbermann, UA; Renner, M.; ib. Weidner, E. Kev tshuaj xyuas cov tshuaj European tshuaj ntsuab ntawm lawv cov ntsiab lus tannin-Tshiab muaj peev xwm tanning tus neeg sawv cev rau kev lag luam tawv. Ind. qoob loo. Prod. 2017, 99, 19–26. [CrossRef]

22. Boros, B.; Jakabová, S.; Madarász, T.; Molnár, R. Galambos, IB; Kilár, F.; Fellinger, UA; Farkas, A. ValidatedHPLC txoj kev rau ib txhij kom muaj nuj nqis ntawm bergenin, arbutin, thiab gallic acid nyob rau hauv nplooj ntawm txawv Bergenia hom. Chromatographia 2014, 77, 1129–1135. [CrossRef]

23. Miaw-Ling, C.; Chur-Min, C. Simultaneous HPLC kev txiav txim siab ntawm hydrophilic whitening tus neeg sawv cev hauv cov khoom kom zoo nkauj. J. Pharm. Biomed. Qhov quav. 2003, 33, 617–626. [CrossRef]

24. Paejo, I.; Viladomat, F.; Bastida, J.; Codina, C. Ib kauj ruam extraction nyob rau hauv lub quantitative tsom xam ntawm arbutinin bearberry (Arctostaphylos uva-ursi) tawm los ntawm high-performance kua chromatography. Phytochem. Anal.2001, 12, 336–339. [CrossRef]

25. Hwj chim. Nroj tsuag ntawm Ntiaj Teb Online. Muaj nyob online: http://www.plantsoftheworldonline.org/ ( nkag mus rau 29 Plaub Hlis 2020).

26. Saleem, A.; Harris, NWS; Asim, M.; Cuerrier, UA; Martineau, L.; ib. Hadad, PS; Arnason, JTRP-HPLC-DAD-APCI/MSD txoj kev rau tus cwj pwm ntawm cov tshuaj Ericaceae siv los ntawm Eeyou IstcheeCree First Nations. Phytochem. Qhov quav. 2010, 21, 328–339. [CrossRef] [PubMed]

27. Stefanescu, BE; Saib, K.; Mocan, A.; Crisan, G. Phenolic tebchaw los ntawm tsib hom Ericaceae nplooj thiab lawv cov bioavailability thiab cov txiaj ntsig kev noj qab haus huv. Molecules 2019, 24, 2046. [CrossRef] [PubMed]

28. Onda, Y.; Mochida, K. Tshawb nrhiav kev sib txawv ntawm cov noob caj noob ces hauv cov nroj tsuag uas siv cov txheej txheem sib txuas ua ke. Genom. 2016, 17, 358–367. [CrossRef] [PubMed]

29. Iannicelli, J.; Guariniello, J.; Tossib, VE; Regalado, JJ; Di Ciaccioa, L.; van Barene, CM; Pitta Álvarez, SI; Escandón, AS Lub "polyploid effect" hauv kev yug me nyuam ntawm hom tshuaj tsw qab thiab tshuaj. Sci. Hortic. 2020, 260, 108854. [CrossRef]

30. Leitch, IJ; Leitch, AR Genome loj ntau haiv neeg thiab evolution hauv av nroj tsuag. Nyob rau hauv-Nroj genome DiversityVolume 2, Lub cev qauv, tus cwj pwm thiab evolution ntawm cog genomes; Leitch, IJ, Greilhuber, J., Doležel, J., Wendel, JF, Eds.; Springer: Vienna, Austria, 2013; Ib., 307–322. [CrossRef]

31. Bennett, MD; Leitch, IJ Nuclear DNA ntau npaum li cas hauv angiosperms—Kev nce qib, teeb meem, thiab kev cia siab.Ann. Bot. 2005, 95, 45–90. [CrossRef]

32. Gregory, TR (Ed.) Lub Evolution ntawm Genome; Elsevier: San Diego, CA, USA, 2005.

33. Hoang, PN; Schubert, V.; Meister, UA; Fuchs, J.; Shubert, I. Variation nyob rau hauv genome loj, cell thiab nucleus ntim, chromosome tooj thiab rDNA loci ntawm duckweeds. Sci. Rep. 2019, 9, 3234. [CrossRef]

34. Knight, CA; Molinari, N.; Petrov, DA Qhov loj genome txwv hypothesis: Evolution, ecology, thiab phenotype. Ann. Bot. 2005, 95, 177–190. [CrossRef]

35. Pellicer, J.; Hidalgo, UA; Dodsworth, S.; Leitch, IJ Genome loj ntau haiv neeg thiab nws cov kev cuam tshuam rau cov evolution ofland nroj tsuag. Genes 2018, 9, 88. [CrossRef]

36. Kev, I.; Rušˇci´c, M.; Skoˇcibuši'c, M.; Maravi'c, UA; Siljak-Yakovlev, S.; Politico, O. Phytochemical thiab cytogeneticcharacterization ntawm Centaurea solstitialis (Asteraceae) los ntawm Croatia. Chem. Biodivers. 2017, 14, e1600213.[CrossRef]

37. Cole, IB; Cai, J.; Alan, RA; Saxena, PK; Murch, SJ Sib piv ntawm Scutellaria baicalensis, Scutellarialateriflora thiab Scutellaria racemosa: Genome loj, muaj peev xwm antioxidant thiab phytochemistry. Planta Med. 2008, 74, 474–481. [CrossRef]

38. Carvalho, YGS; Vitorino, LC; de Souza, UJB; Bessa, LA Tsis ntev los no cov qauv hauv kev tshawb fawb txog cov noob caj noob ces ntawm cov nroj tsuag: Qhov cuam tshuam rau kev txuag. Diversity 2019, 11, 62. [CrossRef]

39. Wei, S.; Yang, W.; Wang, X.; Hou, Y. Cov noob caj noob ces muaj ntau haiv neeg nyob rau hauv cov nroj tsuag tshuaj ntsuab, Saussureainvolucrata (Saussurea, Asteraceae), nyob rau sab hnub poob Tianshan Roob, Suav. Txuag. Genet. Xyoo 2017, 18, 1435–1447. [CrossRef]

40. Chang, YJ; Cao, YF; Zhang, JM; Tian, ​​LM; Dong, XG; Zhang, YJ; Qi, D.; Zhang, XS Kawm txog chloroplastDNA ntau haiv neeg ntawm kev cog qoob loo thiab qus pears (Pyrus L.) nyob rau sab qaum teb Tuam Tshoj. Tsob Ntoo Genet. Genomes 2017, 13, 44.[CrossRef]

41. Paejo, I.; Viladomat, F.; Bastida, J.; Codina, C. Variation ntawm arbutin cov ntsiab lus nyob rau hauv txawv qus pejxeem ntawm Arctostaphylos uva-ursi nyob rau hauv Catalonia, Spain. J. Herbs Spices Med. Paj 2002, 9, 329–333. [CrossRef]

42. Sonnenschein, M.; Tegtmeier, M. Kev sim nyob rau hauv domestication ntawm bearberry (Arctostaphylos uva-ursi (L.)Spreng.). J. Med. Spices Plants 2012, 17, 124–128.

43. Chen, HY; Lin, YC; Hsieh, CL Kev Ntsuam Xyuas ntawm cov tshuaj tiv thaiv antioxidant ntawm aqueous extract ntawm qee cov tshuaj ntsuab xaiv. Khoom noj khoom haus Chem. 2008, 104, 1418–1424. [CrossRef]

44. Del Valle, JC; Buide, ML; Casimiro-Soriguer, I.; Whittall, JB; Narbona, E. Ntawm flavonoid tsub zuj zuj ntawm cov nroj tsuag sib txawv: Cov qauv sib txawv ntawm cov tib neeg thiab cov neeg nyob hauv ntug dej hiav txwv campion (Silene littorea). Pem hauv ntej. Tsob Ntoo Sci. 2015, 6, 939. [CrossRef]

45. Martínez-Cadenas, C.; Lopez, S.; Ribas, G.; Flores, C.; García, UA; Sevilla, UA; Smith-Zubiaga, I.; Ibarrola-Villaba, M.; Pino-Yanes, MM; Gardeazabal, J.; ua al. Simultaneous purifying xaiv ntawm cov poj koob yawm txwv MC1R allele thiab zoo xaiv ntawm melanoma-risk allele v60l nyob rau sab qab teb Europes.Mol. Biol. Evol. 2013, 30, 2654–2665. [CrossRef]

46. ​​Jakaaola, L.; Hohtola, A. Cov nyhuv ntawm latitude ntawm flavonoid biosynthesis hauv cov nroj tsuag. Plant Cell Environ. 2010, 33, 1239–1247. [CrossRef] 47. Martz, F.; Peltola, R.; Fontana, S.; Duval, RE; Julkunen-Tiitto, R.; Stark, S. Cov nyhuv ntawm latitude thiab qhov siab ntawm qhov terpenoid thiab soluble phenolic muaj pes tsawg leeg ntawm juniper (Juniperus communis) koob thiab kev ntsuam xyuas ntawm lawv cov tshuaj tua kab mob hauv thaj chaw boreal. J. Agric. Khoom noj khoom haus Chem. Xyoo 2009, 57, 9575–9584. [CrossRef]

48. Yaj, B.; Zheng, J.; Laaksonen, O.; Tauv, R.; Kallio, H. Cov teebmeem ntawm latitude thiab huab cua tej yam kev mob onphenolic tebchaw nyob rau hauv currant (Ribes spp.) cultivars. J. Agric. Khoom noj khoom haus Chem. 2013, 61, 3517–3532. [CrossRef][PubMed]

49. Lätti, AK; Yakola, L.; Riihien, KR; Kainulainen, PS Anthocyanin thiab flavonol variation hauv bogbil berries (Vaccinium uliginosum L.) hauv Finland. J. Agric. Khoom noj khoom haus Chem. Xyoo 2010, 58, 427–433. [CrossRef] [PubMed]

50. Lätti, AK; Riihien, KR; Kainulainen, PS Kev tshuaj xyuas ntawm anthocyanin variation nyob rau hauv cov tsiaj qus ntawm bilberry (Vaccinium myrtillus L.) hauv Finland. J. Agric. Khoom noj khoom haus Chem. 2008, 56, 190–196. [CrossRef]

51. Monschein, M.; Iglesias, J.; Koj, O.; Bucar, F. Phytochemistry ntawm heather (Calluna vulgaris (L.) Hull) thiab nws qhov kev hloov pauv hloov pauv. Phytochem. Rev. 2010, 9, 205–215. [CrossRef]

52. Perry, NB; Burgess, EJ; Rodríguez Guitián, MA; Romero Franco, R. López Mosquera, E.; Smallfield, BM; Joyce, NI; Littlejohn, RP Sesquiterpene lactones hauv Arnica montana: Helenalin thiab dihydrohelenalinchemotypes hauv Spain. Planta Med. Xyoo 2009, 75, 660–666. [CrossRef] [PubMed]

53. Abdala-Roberts, L.; Assmann, S.; Berny-Mier, Y.; Terán, JC; Covelo, F.; Glauser, G.; Moreira, X. Biotic andabiotic yam cuam tshuam nrog kev hloov pauv ntawm cov nroj tsuag thiab herbivory nyob rau hauv hom ntoo qhib hom.Am. J. Bot. 2016, 103, 2070–2078. [CrossRef] [PubMed]

54. Bentley, J.; ib. Moore, JP; Farrant, JM Metabolomics ua ib qho ntxiv rau phylogenetics rau kev ntsuam xyuas intraspecific ciam teb nyob rau hauv lub desiccation-tolerant medicinal shrub Myrothamnus flabellifolia (Myrothamnaceae).Phytochemistry 2019, 159, 127-136. [CrossRef] [PubMed]

55. Siljak-Yakovlev, S.; Pustahija, F.; Solic, EM; Boguni'c, F.; Muratov'c, E.; Basi'c, N.; Catrice, UA; Brown, CSTowards ib database ntawm genome loj thiab chromosome tus naj npawb ntawm Balkan flora: C-tus nqi nyob rau hauv 343 taxa withnovel qhov tseem ceeb rau 252. Adv. Sci. Lett. 2010, 3, 190–213. [CrossRef]

56. Garrido, JL; Alcántara, JM; Rey, PJ; Medrano, M.; Guitián, J.; Castellanos, MC; Bastida, JM; Jaime, R.; Herrera, CM Geographic genetic qauv ntawm Iberian columbines (gen. Aquilegia). Plant Syst. Evol. 2017, 303, 1145–1160. [CrossRef

57. Lis, D.; Poschlod, P.; Reisch, C. Phylogeography ntawm ib tug tawv pob zeb ciaj sia nyob rau hauv European qhuav grasslands.PLoS ib 2017, 12, e0179961. [CrossRef]

58. Olalde, M.; ib. Herrán, UA; Espinel, S.; Goicoechea, P. Dawb oaks phylogeography hauv Iberian Peninsula.Ecol. Manag. 2002, 156, 89–102. [CrossRef]

59. Barrett, SCH Kev cuam tshuam ntawm clonality ntawm cov nroj tsuag kev sib deev. Proc. Natl. Acad. Sci. USA 2015, 112,8859–8866. [CrossRef] [PubMed]

60. Skowyra, M.; Falguera, V.; Gallego, G.; Peiro, S.; Almajano, MP Cov nyhuv ntawm Perilla frutescens extracts ntawm oxidative stability ntawm cov qauv zaub mov emulsions. Antioxidants 2014, 3, 38–54. [CrossRef] [PubMed]

61. Pascoal, A.; Quirantes-Piné, R.; Fernando, UA; Alexopoulo, E.; Segura-Carretero, A. Phenolic compositionand antioxidant kev ua ntawm kenaf nplooj. Ind. qoob loo. Prod. 2015, 78, 116–123. [CrossRef]

62. Pellicer, J.; Garnatje, T.; Molero, J.; Pustahija, F.; Siljak-Yakovlev, S.; Vallès, J. Keeb Kwm, thiab evolution ntawm SouthAmerican Artemisia hom (Asteraceae); pov thawj los ntawm molecular phylogeny, ribosomal DNA thiab genomesize cov ntaub ntawv. Aust. J. Bot. 2010, 58, 605–616. [CrossRef]

63. Doležel, J. Flow cytometric tsom xam ntawm nuclear DNA cov ntsiab lus hauv cov nroj tsuag siab dua. Phytochem. Qhov quav. 1991, 2, 143–154, ib. [CrossRef]

64. Doyle, JJ; Doyle, JL Ib txoj kev cais DNA ceev ceev rau me me ntawm cov ntaub so ntswg tshiab. Phytochem. Bull.Bot. Soc. Am. 1987, 19, 11–15.

65. Vitales, D.; Feliner, NPE; Valles, J.; Garnatje, T.; Firat, M.; Álvarez, I. Ib tug tshiab circumscription ntawm Mediterranean genus Anacyclus (Anthemideae, Asteraceae) raws li plastid thiab nuclear DNA markers.Phytotaxa 2018, 349, 1-17. [CrossRef]

66. Hall, TA BioEdit: Ib tug neeg siv-phooj ywg kab lis kev cai kev sib raug zoo thiab kev tsom xam cov kev pab cuam rau Windows95/98/NT. Nucleic Acids Symp. Ser. 1999, 41, 95–98.

67. Thompson, JD; Higgins, DG; Gibson, TJ CLUSTAL W: Txhim kho qhov rhiab heev ntawm kev nce qib ntawm ntau qhov sib txawv los ntawm kev sib tw hnyav, kev nplua rau txoj haujlwm tshwj xeeb, thiab qhov hnyav matrix xaiv.Nucleic Acids Res. 1994, 22, 4673–4680. [CrossRef]

68. Rozas, J.; Ferrer-Mata, UA; Sánchez-DelBarrio, JC; Guirao-Rico, S.; Librado, P.; Ramos-Onsins, SE;Sánchez-Gracia, A. DnaSP 6: DNA Sequence Polymorphism Analysis of Large Datasets. Mol. Biol. Evol.2017, 34, 3299–3302. [CrossRef] [PubMed]

69. Borchsenius, F. FastGap, Version 1.2; Department of Biosciences, Aarhus University: Aarhus, Denmark, 2009; Muaj nyob hauv online: http://www.aubot.dk/FastGap_home.htm ( nkag mus rau 1 Lub Xya Hli 2019).

70. Clement, M.; Posada, DCKA; Crandall, KA TCS: Ib qho khoos phis tawj los kwv yees cov noob caj noob ces.Mol. Ecol. 2000, 9, 1657–1659. [CrossRef] [PubMed]

71. Leeg, JW; Bryant, D. Popart: Tag nrho-feature software rau haplotype network tsim. Cov txheej txheem Ecol. Evol.2015, 6, 1110–1116. [CrossRef]

72. Fox, J. Pib nrog tus thawj coj R: Ib qho yooj yim-statistics graphical user interface rau RJ Stat. Softw.2005, 14, 1–42. [CrossRef]

73. Oksanen, J.; Blanchet, FG; Kev, R.; Legendre, P.; O'Hara, RB; Simpson, GL; Solymos, P.; Stevens, M.; Wagner, H. Vegan: Community Ecology Package. R pob Version 1.17-4. 2010. Muaj nyob online:http://CRAN.R-project.org/package=vegan (accessed on 19 September 2010).74. Malinowska, H. Cov xaiv ntawm bearberry (Arctostaphylos uva-ursi (L.) Spreng.) los ntawm cov neeg hauv tebchaws Poland. Acta Soc. Bot. Pol. 1995, 64, 91–96. [CrossRef]