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Fungicides inowanzoshandiswa panguva yekutumbuka kwemuti uye inogona kutyisidzira mapollinator ezvipembenene.Zvakadaro, zvishoma zvinozivikanwa pamusoro pemabatirwo emukume asiri nyuchi (semuenzaniso, nyuchi dzega, Osmia cornifrons) dzinopindura pakubata uye systemic fungicides inowanzoshandiswa pamaapuro panguva yemaruva.Iyi gaka rezivo inodzikamisa sarudzo dzekutonga dzinotaridza kuchengetedzeka kwakachengeteka uye nguva yekupfapfaidza fungicide.Takaongorora mhedzisiro yezviviri zvinobata fungicides (captan uye mancozeb) uye ina interlayer/phytosystem fungicides (ciprocycline, myclobutanil, pyrostrobin uye trifloxystrobin).Mhedzisiro pahuremu hwehuremu, kurarama, bonde reshiyo uye kusiyana kwebhakitiriya.Ongororo iyi yakaitwa pachishandiswa chisingaperi chemuromo bioassay umo mukume waibatwa mumiyero mitatu zvichibva pane yazvino inokurudzirwa dhigirii rekushandisa mumunda (1X), hafu yedosi (0.5X) uye yakaderera dose (0.1X).Ese madosi e mancozeb uye pyritisoline akadzikisa zvakanyanya huremu hwemuviri uye kurarama kwelarval.Isu takazoteedzera iyo 16S gene kuratidza iyo larval bacteriome ye mancozeb, fungicide inokonzera kufa kwakanyanya.Takaona kuti kusiyana kwemabhakitiriya uye kuwanda kwakaderedzwa zvakanyanya mumakonye akadyiswa pamukume wakarapwa mancozeb.Mhedzisiro yedu yemurabhoritari inoratidza kuti kumwaya mamwe efungicides aya panguva yekutumbuka kunonyanya kukuvadza hutano hweO. cornifrons.Idzi ruzivo rwakakosha kune ramangwana manejimendi sarudzo maererano nekushandiswa kwakasimba kwemiti yemichero inodzivirira zvigadzirwa uye inoshanda sehwaro hwemaitiro ekudzora ane chinangwa chekudzivirira mapollinator.
Iyo nyuchi yega yega Osmia cornifrons (Hymenoptera: Megachilidae) yakaunzwa kuUnited States kubva kuJapan mukupera kwema1970 uye kutanga kwema1980s, uye mhuka iyi yakaita basa rakakosha remukume mukuchengetedza zvipenyu kubva ipapo.Huwandu hwevanhu verudzi rwenyuchi iyi chikamu chemhando dzinosvika makumi mashanu dzenyuchi dzemusango dzinoenderana nenyuchi dzinosvibisa maarmond nemaapuro minda yemiti muUnited States2,3.Mason nyuchi dzinosangana nematambudziko mazhinji, kusanganisira kupatsanurwa kwenzvimbo, utachiona, uye mishonga yezvipembenene3,4.Pakati pemishonga yezvipembenene, fungicides inoderedza simba rekuwana, kutsvaga5 uye kugadzirisa muviri6,7.Kunyangwe tsvagiridzo ichangoburwa inoratidza kuti hutano hweMason nyuchi hunokanganiswa zvakananga necommensal uye ectobactic microorganisms, 8,9 nekuti mabhakitiriya nefungi zvinogona kukanganisa hutano uye mhinduro dzekudzivirira zvirwere, mhedzisiro yekufumurwa kwefungicide pane hutachiona hwakasiyana hweMason nyuchi huchangotanga kuva. akadzidza.
Fungicides emhando dzakasiyana-siyana (contact and systemic) inopfapfaidzwa muminda yemichero isati yatumbuka uye panguva yemaruva kurapa zvirwere zvakaita seapuro scab, kuora kunorwadza, brown rot uye powdery mildew10,11.Fungicides anoonekwa seasina ngozi kune mapollinator, saka anokurudzirwa kuvarimi panguva yekutumbuka;Kufumurwa nekupinzwa kwemafungicides aya nenyuchi kunonyanya kuzivikanwa, sezvo chiri chikamu chekunyoresa chezvipembenene neUS Environmental Protection Agency nemamwe akawanda masangano anodzora nyika12,13,14.Zvisinei, zvinokonzerwa nefungicides kune vasiri nyuchi hazvizivikanwi nokuti hazvidikanwi pasi pezvibvumirano zvemvumo yekutengeserana muUnited States15.Pamusoro pezvo, kazhinji hapana maprotocol akamisikidzwa ekuyedza nyuchi imwe chete16,17, uye kuchengetedza makoroni anopa nyuchi kuti dziedze zvinonetsa18.Miedzo yenyuchi dzakasiyana-siyana dziri kuramba dzichiitwa muEurope nekuUSA kuti dziongorore zvinokonzerwa nemishonga yezvipembenene panyuchi dzemusango, uye maprotocol akaenzana achangobva kugadzirwa eO. cornifrons19.
Nyuchi dzine nyanga dzine mamonocytes uye dzinoshandiswa mukutengeserana muzvirimwa zvecarp sekuwedzera kana kutsiva nyuchi.Nyuchi idzi dzinobuda pakati paKurume naKubvumbi, nehono dzisati dzakura dzichibuda mazuva matatu kana mana hadzi dzisati dzasvika.Mushure mekusangana, hadzi inoshingaira kuunganidza pollen nenekita kuti ipe nhevedzano yemasero ebrood mukati metubular dendere mhango (yakasikwa kana yekugadzira)1,20.Mazai anokandwa pamukume mukati memasero;hadzi inobva yavaka madziro evhu isati yagadzirira sero rinotevera.Mapere ekutanga akavharirwa muchorion uye anodya pamvura yeembryo.Kubva pane yechipiri kusvika kune yechishanu instar (prepupa), makonye anodya pollen22.Kana mukume wapera zvachose, makonye anoita zvikoko, pupate uye anobuda sevakuru mukamuri imwe chete yevana, kazhinji mukupera kwezhizha20,23.Vakuru vanobuda muchirimo chinotevera.Kurarama kwevanhu vakuru kunobatanidzwa net net energy gain (kurema kuwedzera) zvichienderana nekudya kwekudya.Nokudaro, hutano hwehutano hwehukume, pamwe chete nezvimwe zvinhu zvakadai semamiriro ekunze kana kutarisana nemishonga yezvipembenene, zvinogadzirisa kurarama uye hutano24.
Mishonga yezvipembenene uye fungicides yakashandiswa isati yatumbuka inokwanisa kufamba mukati mechirimwa vasculature kusvika kune madhigirii akasiyana-siyana, kubva kune translaminar (semuenzaniso, inokwanisa kufamba kubva kumusoro kwemashizha kusvika kune yakadzika pasi, semamwe fungicides) 25 kune zvechokwadi systemic madhara., iyo inogona kupinda korona kubva mumidzi, inogona kupinda muchero wemaruva eapuro26, iyo inogona kuuraya munhu mukuru O. cornifrons27.Mimwe mishonga inopinzawo mukume, izvo zvinokanganisa kukura kwegonye rechibage zvichikonzera kufa kwavo19.Zvimwe zvidzidzo zvakaratidza kuti mamwe mafungicides anogona kusandura zvakanyanya maitiro ekuchengeta matendere emarudzi ane hukama O. lignaria28.Pamusoro pezvo, ongororo dzemurabhoritari nedzemumunda dzinotevedzera zviitiko zvekuuraya zvipembenene (kusanganisira mishonga yefungidzi) zvakaratidza kuti mishonga yezvipembenene inokanganisa physiology 22 morphology 29 uye kurarama kwenyuchi nedzimwe nyuchi dziri dzoga.Kupfapfaidza kwakasiyana-siyana kwefungicidal kunoshandiswa zvakananga kumaruva akazaruka panguva yekutumbuka kunogona kusvibisa mukume inounganidzwa nevakuru kuti mhute isimudzirwe, mhedzisiro yacho inoramba ichidzidzwa30.
Zviri kuwedzera kucherechedzwa kuti kukura kwemakonye kunokonzerwa nekukume uye nharaunda dzehutachiona hwekudya.Iyo nyuchi microbiome inopesvedzera paramita senge body mass31, metabolic shanduko22 uye kubatwa nehutachiona32.Zvidzidzo zvakapfuura zvakaongorora pesvedzero yedanho rekusimudzira, chikafu, uye nharaunda pane microbiome yenyuchi dziri dzoga.Zvidzidzo izvi zvakaratidza kufanana muchimiro uye kuwanda kwelarval uye pollen microbiomes33, pamwe neiyo yakajairika mabhakitiriya genera Pseudomonas neDelftia, pakati pemhando dzenyuchi dzega.Zvisinei, kunyange zvazvo fungicides yakabatanidzwa nemaitiro ekudzivirira hutano hwenyuchi, migumisiro yefungicides pa larval microbiota kuburikidza nemuromo wakananga kuratidzwa inoramba isina kuongororwa.
Chidzidzo ichi chakaedza mhedzisiro yedosi chaiyo yepasi rose yezvitanhatu zvinowanzo shandiswa fungicides akanyoreswa kushandiswa pamuchero wemuti muUnited States, kusanganisira kubata uye systemic fungicides inopihwa nemuromo kuchibage hornworm moth makonye kubva kuchikafu chine utachiona.Isu takaona kuti kubata uye systemic fungicides kwakaderedza huremu hwenyuchi hwemuviri uye kuwedzera kufa, nemhedzisiro yakaipisisa ine chekuita nemancozeb uye pyrithiopide.Takazoenzanisa hutachiona hwakasiyana-siyana hwemakonye anodyiswa pane mancozeb-yakagadziriswa pollen kudya neaya anodyiswa pakudya kwekutonga.Isu tinokurukura nzira dzingangodaro dziri pasi pekufa uye zvinokonzeresa kune integrated pest and pollinator management (IPPM)36 zvirongwa.
Mukuru O. cornifrons overwintering in cocoons akawanikwa kubva kuFruit Research Center, Biglerville, PA, uye akachengetwa pa -3 kusvika 2 ° C (± 0.3 ° C).Pamberi pekuedza (600 makokoni muhuwandu).Muna Chivabvu 2022, 100 O. cornifrons makokoni aiendeswa zuva nezuva mumakapu epurasitiki (50 makoko pamukombe, DI 50 cm × 15 cm kureba) uye zvekupukuta zvakaiswa mukati memakombe kukurudzira kuvhurwa uye kupa chinotsenga substrate, kuderedza kushushikana padombo. nyuchi37 .Isa makapu maviri epurasitiki ane makoko mukeji yezvipembenene (30 × 30 × 30 cm, BugDorm MegaView Science Co.23 ° C, humidity 60%, photoperiod 10 l (yakaderera intensity): mazuva gumi nemana.100 vakadzi vakasangana nevarume vakasunungurwa mangwanani ega ega kwemazuva matanhatu (100 pazuva) mumatendere maviri ekugadzira panguva yepamusoro peapuro maruva (musungo dendere: upamhi 33.66 × kureba 30.48 × kureba 46.99 cm; Supplementary Figure 1).Yakaiswa paPennsylvania State Arboretum, pedyo necherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), peach (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), pear (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), coronaria apple tree (Malus coronaria) uye akawanda marudzi emiti yemaapuro (Malus coronaria, Malus), muti wemuapuro wepamba 'Co-op 30' Enterprise™, Malus apple tree 'Co- Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy').Imwe neimwe yebhuruu plastiki imba yeshiri inokwana pamusoro pemabhokisi maviri emapuranga.Bhokisi rega rega redendere raive ne800 isina chinhu kraft mapepa machubhu (spiral yakavhurika, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) yakaiswa mune opaque cellophane machubhu (0.7 OD ona Plastic plugs (T-1X plugs) inopa nzvimbo dzekugara. .
Mabhokisi ese edendere akatarisana nechekumabvazuva uye aive akafukidzwa negirinhi yebindu repurasitiki refence (Everbilt modhi #889250EB12, kuvhura saizi 5 × 5 cm, 0.95 m × 100 m) kudzivirira makonzo neshiri kupinda uye kuiswa pamusoro pevhu padivi pebhokisi redendere ivhu. mabhokisi.Nest bhokisi (Supplementary Figure 1a).Mazai echibage aiunganidzwa zuva nezuva nekuunganidza machubhu makumi matatu kubva mumatendere uye kuaendesa kurabhoritari.Uchishandisa chigero, cheka kumagumo kwechubhu, wobva wabvisa spiral chubhu kuratidza mabrood cells.Mazai ega nemukume zvawo zvakabviswa pachishandiswa spatula yakakombama (Microslide tool kit, BioQuip Products Inc., California).Mazai akaiswa papepa resefa yakanyorova uye akaiswa muPetri dish kwemaawa maviri asati ashandiswa mukuedza kwedu (Supplementary Figure 1b-d).
Murabhoritari, takaongorora huturu hwemukanwa hwemafungicides matanhatu akaiswa pamberi uye panguva yekutumbuka kweapuro pazvikamu zvitatu (0.1X, 0.5X, uye 1X, apo 1X ndiyo mucherechedzo unoiswa pamarita zana emvura/eka. mumunda)., Tafura 1).Kusungwa kwega kwega kwakadzokororwa 16 nguva (n = 16).Mishonga miviri yefungicides (Table S1: mancozeb 2696.14 ppm uye captan 2875.88 ppm) uye mana systemic fungicides (Table S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobupp5 5 cyprodinil 8; 8 azole 5ppm2 m) chepfu kune michero, miriwo uye zvirimwa zvekushongedza .Isu takaita homogenized pollen tichishandisa grinder, takatamisa 0.20 g kutsime (24-tsime Falcon Plate), uye yakawedzera uye yakasanganiswa 1 μL yefungicide solution kuti iite pyramidal pollen nematsime akadzika 1 mm umo mazai akaiswa.Isa nzvimbo uchishandisa mini spatula (Supplementary Figure 1c,d).Falcon mahwendefa akachengetwa mukamuri tembiricha (25 ° C) uye 70% hama humidity.Takazvienzanisa nemakonye ekudzora akadyiswa homogeneous pollen kudya yakabatwa nemvura yakachena.Takarekodha kufa uye kuyera huremu hwegonye zuva rega rega kusvika makonye asvika zera rekutanga kushandisa chiyero chekuongorora (Fisher Sainzi, kunyatsojeka = 0.0001 g).Pakupedzisira, chiyero chepabonde chakaongororwa nekuvhura cocoon mushure memwedzi 2.5.
DNA yakatorwa kubva muhonye yeO. cornifrons (n = 3 pamamiriro ekurapa, mancozeb-yakarapwa uye mukume isina kurapwa) uye takaita ongororo yemhando dzakasiyana-siyana pamasampuli aya, kunyanya nekuti mu mancozeb kufa kwakanyanya kwakaonekwa mumakonye.kugamuchira MnZn.DNA yakakwidziridzwa, ikanatswa pachishandiswa DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), uye yakatevedzana (600 cycles) paIllumina® MiSeq™ ichishandisa v3 kit.Kutevedzana kwakanangwa kwebhakitiriya 16S ribosomal RNA genes yakaitwa pachishandiswa Quick-16S ™ NGS Library Prep Kit (Zymo Research, Irvine, CA) vachishandisa maprimers akananga V3-V4 dunhu re16S rRNA gene.Pamusoro pezvo, kutevedzana kwe18S kwakaitwa pachishandiswa 10% PhiX kuiswa, uye kukwidziridzwa kwakaitwa pachishandiswa primer pair 18S001 uye NS4.
Ngenisa uye maitirwo akabatanidzwa kuverenga39 uchishandisa QIIME2 pombi (v2022.11.1).Aya maverengero akachekwa uye akabatanidzwa, uye chimeric sequences yakabviswa uchishandisa iyo DADA2 plugin muQIIME2 (qiime dada2 ruzha pairing)40.Iwo 16S uye 18S makirasi ekuita akaitwa pachishandiswa chinhu classifier plugin Classify-sklearn uye pre-yakadzidziswa artifact silva-138-99-nb-classifier.
Yese data yekuyedza yakaongororwa kuti yakajairwa (Shapiro-Wilks) uye homogeneity yekusiyana (bvunzo yaLevene).Nekuti iyo data seti haina kusangana nekufungidzira kwekuongororwa kweparametric uye shanduko yakatadza kuenzanisa zvakasara, takaita nonparametric nzira mbiri ANOVA (Kruskal-Wallis) ine zvinhu zviviri [nguva (nhatu-chikamu 2, 5, uye 8 zuva. nguva mapoinzi) uye fungicide] kuongorora maitiro ekurapa pahuremu hutsva huremu, ipapo post hoc nonparametric pairwise kuenzanisa kwakaitwa pachishandiswa bvunzo yeWilcoxon.Takashandisa generalized linear model (GLM) ine Poisson kugoverwa kuenzanisa migumisiro yefungicides pakurarama kune matatu fungicide concentrations41,42.Nekusiyanisa kuwanda kuongororwa, nhamba yeamplicon sequence variants (ASVs) yakadonha padanho re genus.Kuenzanisa kwekuwanda kwekusiyanisa pakati pemapoka anoshandisa 16S (genus level) uye 18S hukama huzhinji hwakaitwa pachishandiswa generalized additive model for position, scale, and shape (GAMLSS) ine beta zero-inflated (BEZI) kugoverwa kwemhuri, iyo yakafananidzirwa pane macro. .muMicrobiome R43 (v1.1).1).Bvisa mitochondrial uye chloroplast mhando pamberi pekusiyanisa ongororo.Nekuda kweakasiyana taxonomic mazinga e18S, chete yakaderera mwero yemutero wega wega wakashandiswa pakuongorora kwakasiyana.Zvose zvinongororwa zvenhamba zvakaitwa uchishandisa R (v. 3.4.3., CRAN project) (Team 2013).
Kuratidzwa kune mancozeb, pyrithiostrobin, uye trifloxystrobin zvakanyanya kuderedza uremu hwemuviri muO. cornifrons (Fig. 1).Migumisiro iyi yaigara ichionekwa kune ese matatu madosi akaongororwa (Fig. 1a-c).Cyclostrobin uye myclobutanil hazvina kunyanya kuderedza uremu hwemakonye.
Avhareji yehuremu hutsva hwe stem borer larvae yakayerwa panguva nhatu pasi pemishonga ina yekudya (homogeneous pollen feed + fungicide: control, 0.1X, 0.5X uye 1X doses).(a) Mushonga wakaderera (0.1X): nguva yekutanga (zuva 1): χ2: 30.99, DF = 6;P <0.0001, yechipiri nguva pfungwa (zuva 5): 22.83, DF = 0.0009;kechitatu;pfungwa (zuva 8): χ2: 28.39, DF = 6;(b) hafu yehafu (0.5X): nguva yekutanga (zuva 1): χ2: 35.67, DF = 6;P <0.0001, yechipiri nguva poindi (zuva rekutanga).): χ2: 15.98, DF = 6;P = 0.0090;nguva yechitatu (zuva 8) χ2: 16.47, DF = 6;(c) Nzvimbo kana chiyero chakazara (1X): nguva yekutanga (zuva 1) χ2: 20.64, P = 6;P = 0.0326, nguva yechipiri (zuva 5): χ2: 22.83, DF = 6;P = 0.0009;nguva yechitatu (zuva 8): χ2: 28.39, DF = 6;nonparametric kuongororwa kwekusiyana.Mabhawa anomiririra zvinoreva ± SE yekuenzanisa paviri (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Pamushonga wakaderera (0.1X), uremu hwemakonye hwakaderedzwa ne60% ne trifloxystrobin, 49% ine mancozeb, 48% ine myclobutanil, uye 46% ine pyrithistrobin (Fig. 1a).Kana yaiswa hafu yemumunda (0.5X), huremu hwemancozeb huremu hwakaderedzwa ne86%, pyrithiostrobin ne52% uye trifloxystrobin ne50% (Fig. 1b).Mushonga wakazara wemumunda (1X) wemancozeb wakaderedza huremu hwemakonye ne82%, pyrithiostrobin ne70%, uye trifloxystrobin, myclobutanil uye sangard neinenge 30% (Fig. 1c).
Kufa kwaive kwakanyanya pakati pemakonye akadyiswa mancozeb-akarapwa mukume, achiteverwa nepyrithiostrobin uye trifloxystrobin.Kufa kwakawedzera nekuwedzera kwemaitiro e mancozeb uye pyritisoline (Fig. 2; Tafura 2).Zvisinei, kufa kwechibage kwakawedzera zvishoma sezvo trifloxystrobin yakawedzera;cyprodinil uye captan hazvina kuwedzera zvakanyanya kufa kana zvichienzaniswa nekurapa kurapwa.
Kufa kwegonye renhunzi kwakafananidzwa mushure mekumedzwa kwemukume mumwe nemumwe akarapwa nemishonga yefungicide matanhatu.Mancozeb uye pentopyramide zvainyanya kunzwisiswa nemuromo kune honye dzechibage (GLM: χ = 29.45, DF = 20, P = 0.0059) (mutsara, mutserendende = 0.29, P <0.001; slope = 0.24, P) <0.00).
Paavhareji, pakurapa kwese, 39.05% yevarwere vaive vakadzi uye 60.95% vaive varume.Pakati pemishonga yekudzora, chikamu chevakadzi chaive 40% mune zvese zvakaderera-dose (0.1X) uye hafu-dose (0.5X) zvidzidzo, uye 30% muminda-dose (1X) zvidzidzo.Pa 0.1X dose, pakati pezvipembenene zvakadyiswa mukume zvakabatwa ne mancozeb uye myclobutanil, 33.33% yevakuru vaiva vakadzi, 22% yevakuru vaiva vakadzi, 44% yemakondohwe makuru aiva echikadzi, 44% emakondohwe makuru aiva vakadzi.chikadzi, 41% yehonye huru dzaive hadzi, uye zvidzori zvaive 31% (Fig. 3a).Panguva ye0.5 dosi, 33% yehonye dzevakuru muboka remancozeb nepyrithiostrobin vaive vakadzi, 36% muboka retrifloxystrobin, 41% muboka remyclobutanil, uye 46% muboka recyprostrobin.Nhamba iyi yaive 53% muboka.muboka re captan uye 38% muboka rekutonga (Fig. 3b).Pa1X dose, 30% yeboka remancozeb vaiva vakadzi, 36% yeboka repyrithiostrobin, 44% yeboka re trifloxystrobin, 38% yeboka re myclobutanil, 50% yeboka rekutonga vaiva vakadzi - 38.5% (Fig. 3c) .
Mazana emabhunu echikadzi neechirume mushure mekuratidzwa nefungicide.(a) Mushonga wakaderera (0.1X).(b) Half dose (0.5X).(c) Munda wemumunda kana chiyero chakazara (1X).
16S kutevedzana kwekuongorora kwakaratidza kuti boka rebhakitiriya rakasiyana pakati pemakonye anodyiswa nemancozeb-yakagadziriswa pollen uye magonye anodyiswa nemafuta asina kurapwa (Fig. 4a).Indekisi yemicrobial yemakonye asina kurapwa akadyiswa mukume yaive yakakwira kupfuura iyo yegonye inodyiwa nemancozeb-treated pollen (Fig. 4b).Kunyange zvazvo kusiyana kwakaonekwa muhupfumi pakati pemapoka kwakanga kusingaverengeki zvakanyanya, kwaive kuderera zvakanyanya kudarika izvo zvakaonekwa kuti zvipembenene zvinodya pane pollen isina kurapwa (Fig. 4c).Huwandu huzhinji hwakaratidza kuti microbiota yemakonye akadyiswa pamukume wekudzora aive akasiyana-siyana pane emakonye aidyiwa nemakonye anorapwa mancozeb (Fig. 5a).Kuongorora kunotsanangurwa kwakaratidza kuvapo kwe28 genera mukutonga uye mancozeb-treated samples (Fig. 5b).c Kuongorora uchishandisa 18S kutevedzana kwakaratidza hapana misiyano yakakosha (Supplementary Figure 2).
SAV profiles yakavakirwa pa16S kutevedzana kwakaenzaniswa neShannon hupfumi uye yakacherechedzwa hupfumi padanho refilum.(a) Principal coordination analysis (PCoA) yakavakirwa paruzhinji utachiona nharaunda chimiro mune isina kurapwa mukume-inodyiswa kana kudzora (bhuruu) uye mancozeb-akadyiswa mhute (orenji).Imwe neimwe data point inomiririra yakaparadzana sampuli.PCoA yakaverengerwa uchishandisa iyo Bray-Curtis chinhambwe chekugovera multivariate t.Ovals inomiririra 80% yekuvimba level.(b) Boxplot, mbishi Shannon hupfumi data (mapoinzi) uye c.Pfuma inooneka.Mabhokisi anoratidza mabhokisi emutsara wepakati, interquartile range (IQR), uye 1.5 × IQR (n = 3).
Kuumbwa kwenharaunda dzeutachiona hwemakonye anodyiswa pamancozeb-yakarapwa uye isina kurapwa pollen.(a) Hukama huzhinji hwehutachiona hwejenara hunoverengerwa mumakonye.(b) Mepu inopisa yenharaunda dzakazivikanwa dzehutachiona.Delftia (odd ratios (OR) = 0.67, P = 0.0030) uye Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060);Mitsetse yemepu yekupisa inounganidzwa uchishandisa correlation daro uye avhareji yekubatanidza.
Zvatinowana zvinoratidza kuti kubatwa nemumuromo (mancozeb) uye systemic (pyrostrobin and trifloxystrobin) fungicides, inoshandiswa zvakanyanya panguva yekutumbuka, inoderedza kurema zvakanyanya uye kuwedzera kufa kwegonye rechibage.Pamusoro pezvo, mancozeb yakaderedza zvakanyanya kusiyana uye kupfuma kweiyo microbiome panguva yeprepupal nhanho.Myclobutanil, imwe systemic fungicide, yakaderedza zvakanyanya kuwanda kwehuremu hwemuviri pamadosi ese matatu.Izvi zvakaonekwa panguva yechipiri (zuva 5) uye yechitatu (zuva 8) nguva.Kusiyana neizvi, cyprodinil uye captan hazvina kunyanya kuderedza uremu kana kupona kana zvichienzaniswa neboka rinodzora.Sekuziva kwedu, basa iri ndiro rekutanga kuona mhedzisiro yemitengo yeminda yemhando dzakasiyana dzefungicides dzinoshandiswa kuchengetedza zvirimwa zvechibage kuburikidza nekufumurwa kwepollen.
Ese marapirwo efungicide akanyanya kuderedza kuwanda kwehuremu uchienzaniswa nekurapa kwekudzora.Mancozeb yakave nemhedzisiro yakakura pakukura kwehuremu hwemuviri neavhareji kudzikiswa ne51%, ichiteverwa nepyrithiostrobin.Nekudaro, zvimwe zvidzidzo hazvina kutaura zvakaipa zvinokonzeresa zvemumunda madhizi efungicides pamatanho emalarval44.Kunyange zvazvo dithiocarbamate biocides yakaratidza kuve ine low acute toxicity45, ethylene bisdithiocarbamates (EBDCS) yakaita semancozeb inogona kusvibisa kuita urea ethylene sulfide.Tichifunga nezvayo mutagenic mhedzisiro mune dzimwe mhuka, ichi chinosvibisa chigadzirwa chinogona kunge chine mutoro wezvakaonekwa mhedzisiro46,47.Zvidzidzo zvakapfuura zvakaratidza kuti kuumbwa kwe ethylene thiourea kunokonzerwa nezvinhu zvakaita seyepamusoro tembiricha48, humidity level49 uye kureba kwechigadzirwa chekuchengetedza50.Mamiriro ekuchengetedza akakodzera emabiocides anogona kudzikisira izvi mhedzisiro.Pamusoro pezvo, European Food Safety Authority yakaratidza kushushikana nezvehupfu hwepyrithiopide, iyo yakaratidza kuve carcinogenic kune digestive masisitimu edzimwe mhuka51.
Kudzorwa nemuromo kwemancozeb, pyrithiostrobin, uye trifloxystrobin kunowedzera kufa kwemakonye echibage.Kusiyana neizvi, myclobutanil, ciprocycline uye captan hazvina kukanganisa kufa.Migumisiro iyi yakasiyana neyaLadurner et al.52, uyo akaratidza kuti captan yakaderedza zvakanyanya kurarama kwevanhu vakuru O. lignaria uye Apis mellifera L. (Hymenoptera, Apisidae).Pamusoro pezvo, mishonga yefungidziro yakadai secaptan uye boscalid yakawanikwa ichikonzera kufa kwerukova52,53,54 kana kushandura hunhu hwekudyisa55.Shanduko idzi, zvakare, dzinogona kukanganisa hutano hwehukume uye pakupedzisira kuwana simba kwechikamu chelarval.Kufa kwakaonekwa muboka rekutonga kwaienderana nezvimwe zvidzidzo 56,57.
Huwandu hwepabonde hwevarume hunocherechedzwa mubasa redu hunogona kutsanangurwa nezvinhu zvakadai sokusakwana kukwirana uye kushata kwemamiriro ekunze panguva yemaruva, sezvakambotaurwa kuO. cornuta naVicents naBosch.Kunyange zvazvo vanhukadzi nevarume muchidzidzo chedu vaiva nemazuva mana ekuroorana (nguva inowanzonzi yakakwana kuti ibudirire kuroorana), takaita nemaune kuderedza chiedza kuti kuderedze kushungurudzika.Nekudaro, shanduko iyi inogona kukanganisa nekusaziva nzira yekubatana61.Pamusoro pezvo, nyuchi dzinowana mazuva akati wandei emamiriro ekunze asina kunaka, anosanganisira kunaya kwemvura uye tembiricha yakaderera (<5°C), izvo zvinogonawo kukanganisa kusvirana kubudirira4,23.
Kunyangwe chidzidzo chedu chakanangana neiyo larval microbiome yese, mhedzisiro yedu inopa nzwisiso muhukama hunogona kuitika pakati penharaunda dzebhakitiriya dzinogona kuve dzakakosha kune chikafu chenyuchi uye kuratidzwa kwefungicide.Semuyenzaniso, magonye akadyiswa mukume ane mancozeb akanga aderedza zvikuru chimiro chenharaunda uye kuwanda kuenzaniswa nemakonye akadyiswa mukume usina kurapwa.Mumakonye aidya mukume asina kurapwa, mapoka emabhakitiriya eProteobacteria neActinobacteria ndiwo ainyanya uye ainyanya kuita aerobic kana facultatively aerobic.Mabhakitiriya eDelft, anowanzo sangana nemhando dzenyuchi dzega, anozivikanwa kuva nemishonga inorwisa mabhakitiriya, zvichiratidza basa rinogona kudzivirira kurwisa zvirwere.Imwe mhando yebhakitiriya, Pseudomonas, yaive yakawanda mumakonye akadyiswa mukume asina kurapwa, asi akaderedzwa zvakanyanya mumakonye ane mancozeb.Mibairo yedu inotsigira zvidzidzo zvakapfuura zvinozivisa Pseudomonas seimwe yegenera rakawanda muO. bicornis35 nemamwe mavumbu ari oga34.Kunyange zvazvo uchapupu hwekuedza kwebasa rePseudomonas muhutano hweO. cornifrons husati hwadzidzwa, bhakitiriya iri rakaratidzwa kuti rinokurudzira kuumbwa kwehupfu hunodzivirira mubeetle Paederus fuscipes uye kukurudzira arginine metabolism in vitro 35, 65. Izvi zvinoratidzira zvinoratidza basa rinogona kuita mukudzivirira kwehutachiona uye bhakitiriya panguva yekukura kweO. cornifrons.Microbacterium imwe genus yakawanikwa muchidzidzo chedu inonzi iripo muhuwandu hwemakonye enhunzi yevarwi vatema pasi penzara66.MuO. cornifrons larvae, microbacteria inogona kubatsira pakuenzanisa uye kusimba kwegumbo microbiome pasi pemamiriro ekunetseka.Pamusoro pazvo, Rhodococcus inowanikwa muO. cornifrons larvae uye inozivikanwa nekugona kubvisa huturu67.Iyi genus inowanikwawo muura hweA. florea, asi mune yakaderera zvakanyanya68.Mhedzisiro yedu inoratidza kuvepo kweakawanda genetic misiyano kune akati wandei microbial taxa iyo inogona kushandura metabolic maitiro mumakonye.Zvisinei, kunzwisisa kuri nani kwekushanda kwakasiyana-siyana kweO. cornifron kunodiwa.
Muchidimbu, zvakabuda zvinoratidza kuti mancozeb, pyrithiostrobin, uye trifloxystrobin zvakaderedza kuwanda kwehuremu hwemuviri uye kuwedzera kufa kwemakonye echibage.Kunyangwe paine kushushikana kuri kukura pamusoro pezvinokonzeresa fungicides pamapollinator, pane kudikanwa kwekunzwisisa zvirinani mhedzisiro yekusara kwemametabolites emakomponi aya.Mhedzisiro iyi inogona kuiswa muzvirumbidzo zvezvirongwa zvakabatanidzwa zvekuchengetedza mukume zvinobatsira varimi kuti vadzivise kushandisa mamwe mafungicides muti wemichero usati waita maruva nekusarudza mishonga yefungidzi uye kusiyanisa nguva yekushandiswa, kana nekukurudzira kushandiswa kwezvimwe zvinokuvadza zvishoma 36. Ruzivo urwu. zvakakosha pakugadzira mazano.pakushandisa mishonga yezvipembenene, sekugadzirisa zvirongwa zvekupfapfaidza zviripo uye kushandura nguva yekupfapfaidza pakusarudza mishonga yefungidziro kana kukurudzira kushandiswa kwedzimwe nzira dzisina ngozi.Kumwe kutsvagurudza kunodiwa mumhedzisiro yakaipa yefungicides pachiyero chebonde, hunhu hwekudyisa, gut microbiome, uye mamorekuru maitiro ari pasi pechibage borer kuonda uye kufa.
Kwakabva data 1, 2 uye 3 muFigure 1 uye 2 zvakaiswa mufigshare data repository DOI: https://doi.org/10.6084/m9.figshare.24996245 uye https://doi.org/10.6084/m9.figshare.24996233.Kutevedzana kwakaongororwa muchidzidzo chezvino (Figs. 4, 5) inowanikwa muNCBI SRA repository pasi pekuwedzera nhamba PRJNA1023565.
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