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Mishonga yefungicide inowanzo shandiswa panguva yekutumbuka kwemiti uye inogona kutyisidzira zvipembenene zvinoparadzira zvipembenene. Zvisinei, zvishoma zvinozivikanwa nezvekuti zvipembenene zvisiri zvenyuchi (semuenzaniso, nyuchi dzega, Osmia cornifrons) zvinopindura sei kana zvasangana nezvipembenene uye mishonga yefungicide inoshandiswa pamaapuro panguva yekutumbuka. Musiyano uyu weruzivo unodzikamisa sarudzo dzemutemo dzinoona huwandu hwakachengeteka uye nguva yekupfapfaidza fungicide. Takaongorora mhedzisiro yemishonga miviri yefungicide inoparadzira fungicide (captan ne mancozeb) uye mishonga mina ye interlayer/phytosystem fungicides (ciprocycline, myclobutanil, pyrostrobin ne trifloxystrobin). Mhedzisiro pakuwedzera huremu hwemakonye, kurarama, huwandu hwebonde uye kusiyana kwemabhakitiriya. Ongororo iyi yakaitwa uchishandisa nzira yekurapa zvipembenene nemuromo umo pollen yakarapwa muzvikamu zvitatu zvichibva pane dose inokurudzirwa pari zvino yekushandiswa mumunda (1X), hafu dose (0.5X) uye dose yakaderera (0.1X). Dose dzese dze mancozeb ne pyritisoline dzakaderedza zvakanyanya huremu hwemuviri uye kupona kwemakonye. Takazoongorora jini re16S kuti tiratidze bhakitiriya remakonye remancozeb, mushonga wefungi unokonzera kufa kwakanyanya. Takaona kuti kusiyana kwemabhakitiriya nekuwanda kwawo kwakadzikira zvakanyanya muhonye dzinodyiswa nemukume wakarapwa nemancozeb. Zvakabuda murabhoritari yedu zvinoratidza kuti kupfapfaidza mimwe yemishonga iyi yefungi panguva yemaruva kunonyanya kukuvadza hutano hweO. cornifrons. Ruzivo urwu rwakakosha pakusarudza kweramangwana maererano nekushandiswa kwenguva refu kwezvigadzirwa zvekuchengetedza miti yemichero uye kunoshanda sehwaro hwemaitiro ekutonga akagadzirirwa kuchengetedza zvinoparadzira mukume.
Nyuchi dzemason dzinonzi Osmia cornifrons (Hymenoptera: Megachilidae) dzakaunzwa kuUnited States dzichibva kuJapan mukupera kwema1970 nekutanga kwema1980, uye mhando iyi yakaita basa guru rekuunza mukumedza mukuchengetedza nharaunda kubvira ipapo. Kuwanda kwenyuchi iyi kwakasikwa kuri chikamu chemhando dzinosvika makumi mashanu dzenyuchi dzemusango dzinobatsira nyuchi dzinobereka mukumedza minda yemichero yemaarumondi nemaapuro muUnited States2,3. Nyuchi dzeMason dzinosangana nematambudziko akawanda, kusanganisira kupatsanurwa kwenzvimbo dzekugara, utachiona, nemishonga yekuuraya zvipembenene3,4. Pakati pemishonga yekuuraya zvipembenene, mishonga yekuuraya zvipembenene inoderedza simba, kutsvaga chikafu chemhuka5 uye kugadzirisa muviri6,7. Kunyangwe tsvakiridzo yazvino ichiratidza kuti hutano hwenyuchi dzeMason hunokanganiswa zvakananga nehutachiona hwe commensal uye ectobactic, 8,9 nekuti mabhakitiriya nefungus zvinogona kukanganisa kudya uye mhinduro dzemuviri, mhedzisiro yekusangana nefungicide pakusiyana-siyana kwenyuchi dzeMason iri kutanga kudzidzwa.
Mishonga yefungicides ine migumisiro yakasiyana-siyana (kusangana uye systemic) inopfapfaidzwa muminda yemichero isati yatumbuka uye panguva yekurapa zvirwere zvakaita se apple scab, bitter rot, brown rot uye powdery mildew10,11. Mishonga yefungicides inoonekwa seisina njodzi kune vanofudza zvirimwa, saka inokurudzirwa kuvarimi munguva yekutumbuka; Kuonekwa uye kutorwa kwemishonga iyi yefungicides nenyuchi kunozivikanwa zvakanyanya, sezvo iri chikamu chekunyoreswa kwemishonga yezvipembenene neUS Environmental Protection Agency nemamwe masangano mazhinji emutemo wenyika12,13,14. Zvisinei, migumisiro yemishonga yefungicides kune vasiri nyuchi hainyanyozivikanwa nekuti haidiwi pasi pezvibvumirano zvemvumo yekushambadzira muUnited States15. Pamusoro pezvo, kazhinji hapana nzira dzakajairwa dzekuedza nyuchi imwe chete16,17, uye kuchengetedza makoroni anopa nyuchi kuti dziongororwe kwakaoma18. Kuedzwa kwenyuchi dzakasiyana-siyana dziri kutarisirwa kuri kuitwa zvakanyanya muEurope neUSA kuti vadzidze mhedzisiro yemishonga yezvipembenene panyuchi dzesango, uye nzira dzakajairwa dzakagadzirwa munguva pfupi yapfuura dzeO. cornifrons19.
Nyuchi dzine nyanga dzinonzi monocytes uye dzinoshandiswa muzvirimwa zvecarp sekuwedzera kana kutsiva nyuchi dzeuchi. Nyuchi idzi dzinobuda pakati paKurume naKubvumbi, varume vasati vasvika mazuva matatu kusvika mana vasati vasvika. Mushure mekusangana, hadzi inounganidza mukume nenectar kuti ipe masero evana mukati mehunde yechubhu (yechisikigo kana yekugadzira)1,20. Mazai anoiswa pamukume mukati memasero; hadzi inozovaka rusvingo rwevhu isati yagadzirira sero rinotevera. Makume ekutanga anovharirwa muchorion uye anodya mvura yemucheche. Kubva padanho rechipiri kusvika padanho rechishanu (prepupa), makume anodya mukume22. Kana mukume wapera zvachose, makume anoumba makoko, mapupate uye anobuda seakura mukamuri rimwe chete revana, kazhinji mukupera kwezhizha20,23. Vakuru vanobuda muchirimo chinotevera. Kurarama kwevakuru kunobatanidzwa nekuwedzera simba (kuwedzera uremu) zvichibva pakudya. Saka, hutano hwemukume, pamwe nezvimwe zvinhu zvakaita semamiriro ekunze kana kusangana nemishonga inouraya zvipembenene, ndizvo zvinosarudza kupona uye hutano24.
Mishonga yezvipembenene nefungicides inoshandiswa isati yatumbuka inokwanisa kufamba mukati memitsipa yemiti kusvika padanho rakasiyana, kubva pakushandura (semuenzaniso, inokwanisa kufamba kubva pamusoro pemashizha kuenda pasi, semamwe mishonga yefungicides) 25 kusvika kumhedzisiro chaiyo yesystemic. , iyo inogona kupinda mumusoro kubva pamidzi, inogona kupinda mu nectar yemaruva eapuro26, kwainogona kuuraya O. cornifrons yakakura27. Mimwe mishonga yezvipembenene inosvetukirawo mukume, ichikanganisa kukura kwemakonye echibage uye ichikonzera kufa kwawo19. Zvimwe zvidzidzo zvakaratidza kuti mimwe mishonga yefungicides inogona kuchinja zvakanyanya maitiro ekugara kwematendere emhando dzakabatana O. lignaria28. Pamusoro pezvo, zvidzidzo zverabhoritari uye zvemunda zvinotevedzera mamiriro ekushandiswa kwemishonga yezvipembenene (kusanganisira fungicides) zvakaratidza kuti mishonga yezvipembenene inokanganisa physiology22 morphology29 uye kupona kwenyuchi dzeuchi nedzimwe nyuchi dziri dzoga. Mishonga yakasiyana-siyana yefungicidal inoshandiswa zvakananga kumaruva akavhurika panguva yemaruva inogona kusvibisa mukume unounganidzwa nevakuru kuti ukure makonye, mhedzisiro yacho ichiri kudzidzwa30.
Zviri kuramba zvichizivikanwa kuti kukura kwemakonye kunokanganiswa nemaruva nemapoka ezvipembenene mugadziriro yekugaya. Huchi hwenyuchi hunokanganisa zvinhu zvakaita sehuremu hwemuviri31, shanduko dzemetabolism22 uye mukana wekutapukirwa nezvirwere32. Zvidzidzo zvekare zvakaongorora simba rekukura, zvinovaka muviri, uye nharaunda pahutachiona hwenyuchi dziri dzoga. Zvidzidzo izvi zvakaratidza kufanana muchimiro uye kuwanda kwemakonye nemaruva33, pamwe nerudzi rwemabhakitiriya rwakajairika rwePseudomonas neDelftia, pakati pemhando dzenyuchi dziri dzoga. Zvisinei, kunyangwe mishonga yefungicides yakabatana nemaitiro ekuchengetedza hutano hwenyuchi, mhedzisiro yemishonga yefungicides pahutachiona hwemakonye kuburikidza nekubuda zvakananga mumuromo haisati yaongororwa.
Chidzidzo ichi chakaedza mhedzisiro yemishonga mitanhatu inoshandiswa yefungicides yakanyoreswa kushandiswa pamichero yemiti muUnited States, kusanganisira mishonga yefungicides inobatwa nemuromo uye inopihwa nemuromo kuzvipembenene zvechibage zvechibage kubva muchikafu chakasvibiswa. Takaona kuti mishonga yefungicides inobatwa nemuviri uye inobatwa nemuviri inoderedza kuwedzera kwehuremu hwemuviri wenyuchi uye kuwedzera kufa, nemhedzisiro yakanyanya inobatanidzwa ne mancozeb ne pyrithiopide. Takazoenzanisa kusiyana kwehutachiona hwezvipembenene zvinopihwa pakudya kwemupollen kwakarapwa ne mancozeb neizvo zvinopihwa pakudya kwekudzora. Tinokurukura nzira dzinogona kukonzera kufa uye mhedzisiro yehurongwa hwekutarisira zvipembenene nezvipembenene (IPPM)36.
Makonzo eO. cornifrons akura aichengetwa munguva yechando mukonzoni akawanikwa kubva kuFruit Research Center, Biglerville, PA, uye akachengetwa pa −3 kusvika 2°C (± 0.3°C). Pamberi pekuyedza (makonzoni mazana matanhatu ese). Muna Chivabvu 2022, makonzoni zana eO. cornifrons aiendeswa zuva nezuva mumakapu epurasitiki (makonzoni makumi mashanu pakapu imwe neimwe, DI 5 cm × 15 cm kureba) uye machira akaiswa mukati memakapu kuti avhurike uye ape substrate inotsengwa, zvichideredza kushushikana nyuchi dzine matombo37. Isa makapu maviri epurasitiki ane makonzoni mukeji yezvipembenene (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) nema feeders e10 ml ane 50% sucrose solution uye chengeta kwemazuva mana kuti uve nechokwadi chekuvharwa uye kusangana. 23°C, hunyoro hwe60%, photoperiod 10 l (hushoma simba): mazuva gumi nemana. Hadzi nevarume zana dzakaberekeswa vaisunungurwa mangwanani ega ega kwemazuva matanhatu (100 pazuva) mumatendere maviri ekugadzira panguva yekutumbuka kwemaapuro (dendere remusungo: upamhi 33.66 × kureba 30.48 × kureba 46.99 cm; Mufananidzo Wekuwedzera 1). Yakaiswa paPennsylvania State Arboretum, pedyo necherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), peach (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), peya (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), coronaria apple tree (Malus coronaria) uye mhando dzakasiyana dzemiti yemaapuro (Malus coronaria, Malus), domestic apple tree 'Co-op 30′ Enterprise™, Malus apple tree 'Co-Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy'). Imba yega yega yepurasitiki yepurasitiki inokwana pamusoro pemabhokisi maviri emapuranga. Bhokisi rega rega redendere raiva nemachubhu epepa asina chinhu mazana masere (akavhurika, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) akaiswa mumachubhu ecellophane asina kujeka (0.7 OD ona maplug epurasitiki (T-1X plugs) anopa nzvimbo dzekuvakira matendere.
Mabhokisi ese ezvitunha akatarisa kumabvazuva uye akafukidzwa nefenzi yegirini yepurasitiki (Everbilt model #889250EB12, saizi yekuvhura 5 × 5 cm, 0.95 m × 100 m) kudzivirira kupinda kwembeva neshiri uye akaiswa pamusoro pevhu pedyo nemabhokisi evhu ezvitunha zvezvitunha. Bhokisi rezvitunha (Mufananidzo wekuwedzera 1a). Mazai ezvitunha zvechibage aiunganidzwa zuva nezuva nekuunganidza machubhu makumi matatu kubva mumatendere uye kuatakura kuenda kurabhoritari. Uchishandisa chigero, cheka pamucheto wechubhu, wobva wabvisa chubhu yespiral kuti uone masero evana. Mazai ega ega negoridhe rawo zvakabviswa uchishandisa spatula yakakombama (Microslide tool kit, BioQuip Products Inc., California). Mazai akaiswa papepa rekusefa rakanyorova ndokuiswa mudhishi rePetri kwemaawa maviri asati ashandiswa mukuyedza kwedu (Mufananidzo wekuwedzera 1b-d).
Murabhoritari, takaongorora chepfu yemumuromo yemishonga mitanhatu yefungicides yakashandiswa isati yatanga uye panguva yemaruva emuapuro pazvikamu zvitatu (0.1X, 0.5X, uye 1X, uko 1X iri chiratidzo chakashandiswa pa 100 galoni dzemvura/eka. Dosi yakawanda mumunda = huwandu mumunda). , Tafura 1). Huwandu hwega hwega hwakadzokororwa ka16 (n = 16). Mishonga miviri yefungicides (Tafura S1: mancozeb 2696.14 ppm uye captan 2875.88 ppm) uye mishonga mina yefungicides inogadzirwa mumuviri (Tafura S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75 .12 ppm; cyprodinil 280.845 ppm) chepfu kumichero, miriwo nezvirimwa zvekushongedza. Takaisa mukume muhombe tichishandisa muchina wekukuya, tikaisa 0.20 g mugomba (24-well Falcon Plate), ndokuwedzera nekusanganisa 1 μL yemushonga wefungicide kuti tigadzire mukume wepiramidhi nematsime akadzika 1 mm umo mazai akaiswa. Isa uchishandisa mini spatula (Supplementary Picture 1c, d). Maplate eFalcon akachengetwa patembiricha yemukamuri (25°C) uye 70% hunyoro. Takaaenzanisa nemapundu anodyisa akapihwa chikafu chemukume chakafanana chakarapwa nemvura yakachena. Takanyora kufa uye takayera huremu hwemapundu mazuva ese kusvika mapundu asvika zera rekutanga tichishandisa analytical balance (Fisher Scientific, accuracy = 0.0001 g). Pakupedzisira, huwandu hwepabonde hwakaongororwa nekuvhura cocoon mushure memwedzi 2.5.
DNA yakatorwa kubva muhonye dzeO. cornifrons dzese (n = 3 pakurapwa kwese, pollen yakarapwa ne mancozeb uye isina kurapwa) uye takaita ongororo ye microbial diversity pane aya masampuli, kunyanya nekuti mu mancozeb kufa kwakanyanya kwakaonekwa muhonye. Kugamuchira MnZn. DNA yakawedzerwa, yakacheneswa uchishandisa DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), uye yakatevedzana (600 cycles) paIllumina® MiSeq™ uchishandisa v3 kit. Kutevedzana kwakanangana kwemajini eribosomal RNA ebhakitiriya 16S kwakaitwa uchishandisa Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) uchishandisa ma primers akananga kunharaunda yeV3-V4 ye16S rRNA gene. Pamusoro pezvo, 18S sequencing yakaitwa uchishandisa 10% PhiX inclusion, uye amplification yakaitwa uchishandisa primer pair 18S001 neNS4.
Import uye process paired reads39 uchishandisa QIIME2 pipeline (v2022.11.1). Aya mareads akachekwa uye akabatanidzwa, uye chimeric sequences dzakabviswa uchishandisa DADA2 plugin muQIIME2 (qiime dada2 noise pairing)40. 16S ne18S class assignments dzakaitwa uchishandisa object classifier plugin Classify-sklearn uye pre-trained artifact silva-138-99-nb-classifier.
Data rese rekuyedza rakaongororwa kuti rionekwe kuti rakajairwa here (Shapiro-Wilks) uye kuti rakafanana here kana kuti kwete (bvunzo yaLevene). Nekuti data racho harina kusangana nepfungwa dzekuongorora parametric uye shanduko yacho yakatadza kugadzirisa zvasara, takaita nonparametric two-way ANOVA (Kruskal-Wallis) nezvinhu zviviri [nguva (three-phase 2, 5, uye 8 day time points) uye fungicide] kuti tiongorore mhedzisiro yekurapa pahuremu hutsva hwemakonzo, ipapo kuenzanisa kwe post hoc nonparametric pairiwise kwakaitwa tichishandisa Wilcoxon test. Takashandisa generalized linear model (GLM) ine Poisson distribution kuti tienzanise mhedzisiro yefungicides pakurarama muhuwandu hutatu hwefungicide41,42. Kuti tiongorore huwandu hwakasiyana, huwandu hwe amplicon sequence variants (ASVs) hwakaderedzwa padanho re genus. Kuenzanisa kwehuwandu hwakasiyana pakati pemapoka achishandisa 16S (genus level) uye 18S relative abundance kwakaitwa uchishandisa generalized additive model yenzvimbo, chiyero, uye chimiro (GAMLSS) nekugoverwa kwemhuri kwe beta zero-inflated (BEZI), izvo zvakaenzanisirwa pa macro . mu Microbiome R43 (v1.1). 1). Bvisa mitochondrial ne chloroplast species usati waongorora musiyano. Nekuda kwemazinga akasiyana etaxonomic e18S, nhanho yakaderera chete yetaxon yega yega ndiyo yakashandiswa pakuongorora musiyano. Kuongorora kwese kwestatistiki kwakaitwa uchishandisa R (v. 3.4.3., CRAN project) (Team 2013).
Kusangana ne mancozeb, pyrithiostrobin, uye trifloxystrobin zvakaderedza zvakanyanya kuwedzera kwehuremu hwemuviri muO. cornifrons (Mufananidzo 1). Mhedzisiro iyi yakaonekwa nguva dzose pamatanho ese matatu akaongororwa (Mufananidzo 1a–c). Cyclostrobin ne myclobutanil hazvina kuderedza zvakanyanya huremu hwemakonye.
Avhareji yehuremu hutsva hwemabukira emuborera anoyerwa panguva nhatu pasi pemishonga mina yekudya (homogeneous pollen feed + fungicide: control, 0.1X, 0.5X uye 1X doses). (a) Dose shoma (0.1X): nguva yekutanga (zuva 1): χ2: 30.99, DF = 6; P < 0.0001, nguva yechipiri (zuva 5): 22.83, DF = 0.0009; nguva yechitatu; nguva (zuva 8): χ2: 28.39, DF = 6; (b) hafu yedose (0.5X): nguva yekutanga (zuva 1): χ2: 35.67, DF = 6; P < 0.0001, nguva yechipiri (zuva rekutanga). ): χ2: 15.98, DF = 6; P = 0.0090; poindi yechitatu (zuva rechi8) χ2: 16.47, DF = 6; (c) Nzvimbo kana dose yakazara (1X): poindi yekutanga (zuva rechi1) χ2: 20.64, P = 6; P = 0.0326, poindi yechipiri (zuva rechi5): χ2: 22.83, DF = 6; P = 0.0009; poindi yechitatu (zuva rechi8): χ2: 28.39, DF = 6; ongororo isina parameter yevariance. Mabhawa anomiririra avhareji ± SE yekuenzanisa kwepairise (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Pamuyero wakaderera (0.1X), huremu hwemuviri wemakonye hwakaderedzwa ne60% ne trifloxystrobin, 49% ne mancozeb, 48% ne myclobutanil, uye 46% ne pyrithistrobin (Mufananidzo 1a). Pakaiswa hafu yemuyero wemunda (0.5X), huremu hwemuviri wemakonye e mancozeb hwakaderedzwa ne86%, pyrithiostrobin ne52% uye trifloxystrobin ne50% (Mufananidzo 1b). Muyero wakazara wemunda (1X) we mancozeb wakaderedza huremu hwemakonye ne82%, pyrithiostrobin ne70%, uye trifloxystrobin, myclobutanil nesangard neanenge 30% (Mufananidzo 1c).
Huwandu hwevanhu vakafa hwakanyanya pakati pemakonye akadyiswa mukume wakagadziriswa mancozeb, zvichiteverwa nepyrithiostrobin netrifloxystrobin. Huwandu hwevanhu vakafa hwakawedzera nekuwedzera kwehuwandu hwemancozeb nepyritisoline (Mufananidzo 2; Tafura 2). Zvisinei, kufa kwevane borer kwakangowedzera zvishoma sezvo huwandu hwetrifloxystrobin hwakakwira; cyprodinil necaptan hazvina kuwedzera zvakanyanya kufa zvichienzaniswa nekurapwa kwekudzivirira.
Kufa kwevana vezvipembenene vane fungus kwakaenzaniswa mushure mekunwa mupunga wakarapwa nemishonga mitanhatu yakasiyana yefungicide. Mancozeb nepentopyramide zvainyanya kubatwa nehonye dzechibage nemuromo (GLM: χ = 29.45, DF = 20, P = 0.0059) (mutsetse, slope = 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 pekurapa kwekudzivirira, huwandu hwevakadzi hwaive 40% muzvidzidzo zvedosi shoma (0.1X) nehafu (0.5X), uye 30% muzvidzidzo zvedosi yemunda (1X). Padosi ye0.1X, pakati pemakonye akadyiswa mupureni akarapwa ne mancozeb ne myclobutanil, 33.33% yevanhu vakuru vaive vakadzi, 22% yevanhu vakuru vaive vakadzi, 44% yemakonye makuru aive vakadzi, 44% yemakonye makuru aive vakadzi. vakadzi, 41% yemakonye makuru aive vakadzi, uye kudzora kwaive 31% (Mufananidzo 3a). Pakapetwa ka0.5 kubva pamushonga, 33% yehonye huru muboka remancozeb nepyrithiostrobin dzaive dzechikadzi, 36% muboka retrifloxystrobin, 41% muboka remyclobutanil, uye 46% muboka recyprostrobin. Nhamba iyi yaive 53% muboka. muboka recaptan uye 38% muboka rekutonga (Mufananidzo 3b). Pamushonga we1X, 30% yeboka remancozeb vaiva vakadzi, 36% yeboka repyrithiostrobin, 44% yeboka retrifloxystrobin, 38% yeboka remyclobutanil, 50% yeboka rekutonga vaiva vakadzi - 38.5% (Mufananidzo 3c).
Huwandu hwezvipembenene zvechikadzi nezvechirume mushure mekubatwa nefungus padanho remakonzo. (a) Dose shoma (0.1X). (b) Hafu yedose (0.5X). (c) Dose yemunda kana dose yakazara (1X).
Kuongororwa kwe16S sequence kwakaratidza kuti boka remabhakitiriya rakasiyana pakati pemakonye akadyiswa nemupore wakarapwa ne mancozeb nemakonye akadyiswa nemupore usina kurapwa (Mufananidzo 4a). Nhamba ye microbial yemakonye asina kurapwa akadyiswa mumupore waive wakakwira kupfuura wemakonye akadyiswa mumupore wakarapwa ne mancozeb (Mufananidzo 4b). Kunyangwe musiyano wakaonekwa mukupfuma pakati pemapoka wakanga usina kukosha, waive wakaderera zvakanyanya pane wakaonekwa kune makonye akadyiswa mumupore usina kurapwa (Mufananidzo 4c). Kuwanda kwakaratidza kuti microbiota yemakonye akadyiswa mumupore wakadzora waive wakasiyana zvakanyanya kupfuura wemakonye akadyiswa mumupore wakarapwa ne mancozeb (Mufananidzo 5a). Ongororo inotsanangura yakaratidza kuvapo kwemajena makumi maviri nemasere muzviyero zvekudzora uye zvakarapwa ne mancozeb (Mufananidzo 5b). c Ongororo ichishandisa 18S sequencing yakaratidza kuti hapana musiyano wakakosha (Mufananidzo Wekuwedzera 2).
Mapurofayi eSAV akavakirwa pa16S sequences akaenzaniswa neShannon richness uye akaona richness pa phylum level. (a) Principal coordinate analysis (PCoA) yakavakirwa pachimiro che microbial community structure mu pollen-fed kana control (blue) uye mancozeb-fed larvae (orenji). Poindi yega yega yedata inomiririra sampuro yakasiyana. PCoA yakaverengerwa uchishandisa daro reBray-Curtis rekugoverwa kwe multivariate t. Maovals anomiririra 80% confidence level. (b) Boxplot, raw Shannon wealth data (points) uye c. Observable wealth. Boxplots inoratidza mabhokisi e median line, interquartile range (IQR), uye 1.5 × IQR (n = 3).
Kuumbwa kwemapoka ezvipembenene zvemakonye anodyiswa mukume wakarapwa newasina kurapwa ne mancozeb. (a) Kuwanda kwemarudzi ezvipembenene mumapoka. (b) Mepu yekupisa yemapoka ezvipembenene anozivikanwa. Delftia (odds ratio (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 yakakamurwa uchishandisa kureba kwekubatana uye avhareji yekubatana.
Zvakabuda muongororo yedu zvinoratidza kuti kushandisa mushonga we "contact" (mancozeb) uye "systemic (pyrostrobin and trifloxystrobin), wakashandiswa zvakanyanya panguva yekutumbuka, zvakaderedza zvakanyanya kuwedzera kwehuremu uye kuwedzera kufa kwe "cornea". Pamusoro pezvo, mancozeb yakaderedza zvakanyanya kusiyana-siyana uye kupfuma kwe "microbiome" panguva yekugadzirira. Myclobutanil, imwe "systemic fungicide", yakaderedza zvakanyanya kuwedzera kwehuremu hwemuviri we "larval" pamatanho ese matatu. Mhedzisiro iyi yakaonekwa panguva yechipiri (yezuva rechi5) neyechitatu (yezuva rechi8). Kusiyana neizvi, cyprodinil ne "captan" hazvina kuderedza zvakanyanya kuwedzera kwehuremu kana kurarama zvichienzaniswa neboka rekutonga. Sekuziva kwedu, basa iri ndiro rekutanga kuona mhedzisiro yehuwandu hwe "field fungicides" dzakasiyana dzinoshandiswa kuchengetedza zvirimwa zvechibage kuburikidza nekuratidzwa zvakananga kwe "pollen".
Mishonga yese yefungicide yakaderedza zvakanyanya kuwedzera kwehuremu hwemuviri zvichienzaniswa nekurapa kwekudzora. Mancozeb yakava nemhedzisiro yakakura pakuwedzera kwehuremu hwemuviri wemakonzo nekudzikira kweavhareji ye51%, ichiteverwa nepyrithiostrobin. Zvisinei, zvimwe zvidzidzo hazvina kutaura nezvemigumisiro yakaipa yemishonga yefungicide mumunda pamatanho emakonzo44. Kunyangwe dithiocarbamate biocides yakaratidzwa kuti ine chepfu shoma45, ethylene bisdithiocarbamates (EBDCS) yakadai semancozeb inogona kuora kuita urea ethylene sulfide. Zvichienderana nemhedzisiro yayo yekuchinja mune dzimwe mhuka, chigadzirwa ichi chekuora chinogona kunge chiri icho chinokonzera mhedzisiro yakaonekwa46,47. Zvidzidzo zvekare zvakaratidza kuti kuumbwa kwe ethylene thiourea kunokanganiswa nezvinhu zvakaita sekupisa kwakanyanya48, mwero wehunyoro49 uye kureba kwekuchengetwa kwechigadzirwa50. Mamiriro akakodzera ekuchengetera biocides anogona kuderedza mhedzisiro iyi. Pamusoro pezvo, European Food Safety Authority yakaratidza kunetseka nezve chepfu ye pyrithiopide, iyo yakaratidzwa kuti inokonzera kenza kumasisitimu ekugaya emamwe mhuka51.
Kunwa mancozeb, pyrithiostrobin, uye trifloxystrobin nemuromo kunowedzera kufa kwehonye dzechibage. Kusiyana neizvi, myclobutanil, ciprocycline uye captan hazvina kukanganisa kufa. Izvi zvakasiyana nezvaLadurner et al.52, avo vakaratidza kuti captan yakaderedza zvakanyanya kupona kweO. lignaria yevakuru naApis mellifera L. (Hymenoptera, Apisidae). Pamusoro pezvo, mishonga yefungicide yakaita secaptan neboscalid yakawanikwa ichikonzera kufa kwehonye dzechibage52,53,54 kana kuchinja maitiro ekudyisa55. Shanduko idzi, dzinogona kukanganisa kunaka kwekudya kwemukume uye pakupedzisira kuwana simba kwechikamu chemakonye. Kufa kwakaonekwa muboka rekutonga kwaienderana nedzimwe ongororo 56,57.
Kuwanda kwevarume nevakadzi kwakaonekwa mubasa redu kunogona kutsanangurwa nezvinhu zvakaita sekusakwana kwekusangana kwemhuka uye mamiriro ekunze asina kunaka panguva yekutumbuka, sezvakambotaurwa kare nezveO. cornuta naVicens naBosch. Kunyangwe hadzi nevarume muchidzidzo chedu vaive nemazuva mana ekusangana (nguva inowanzoonekwa seyakakwana kuti kusangana kubudirire), takaderedza simba rechiedza nemaune kuti tideredze kushushikana. Zvisinei, shanduko iyi inogona kukanganisa maitiro ekusangana kwemhuka nemaune61. Pamusoro pezvo, nyuchi dzinosangana nemamiriro ekunze asina kunaka kwemazuva akati wandei, kusanganisira mvura uye tembiricha yakaderera (<5°C), izvo zvinogonawo kukanganisa kubudirira kwekusangana kwemhuka4,23.
Kunyangwe chidzidzo chedu chakatarisana ne microbiome yese yemakonye, zvatakawana zvinopa ruzivo rwehukama hunogona kuitika pakati pemabhakitiriya hunogona kuve hwakakosha pakudya kunovaka muviri uye kusangana nefungicide. Semuenzaniso, mupure wakarapwa ne mancozeb wakadyiswa mankozeb wakaderedza zvakanyanya chimiro chemunharaunda yemakonye uye kuwanda kwawo zvichienzaniswa ne mupure usina kurapwa. Mumakonye akadya mupure usina kurapwa, mapoka emabhakitiriya eProteobacteria neActinobacteria ndiwo aive akapararira uye ainyanya aerobic kana facultatively aerobic. Mabhakitiriya eDelft, anowanzobatanidzwa nemhando yenyuchi yega, anozivikanwa kuti ane basa rekuuraya mabhakitiriya, zvichiratidza basa rekudzivirira kubva kuzvirwere. Imwe mhando yemabhakitiriya, Pseudomonas, yaive yakawanda mu mupure usina kurapwa, asi yakaderedzwa zvakanyanya mu mupure wakarapwa ne mancozeb. Zvatakawana zvinotsigira zvidzidzo zvekare zvinoratidza Pseudomonas seimwe yemarudzi akawanda muO. bicornis35 nedzimwe magonye ega ega34. Kunyangwe humbowo hwekuyedza hwebasa rePseudomonas muhutano hweO. cornifrons husati hwadzidzwa, bhakitiriya iyi yakaratidzwa kuti inokurudzira kugadzirwa kwechepfu dzinodzivirira muPaederus fuscipes yebeetle uye inokurudzira metabolism yearginine mu vitro 35, 65. Izvi zvinoratidza basa rinogona kuitwa mukudzivirira mavhairasi nemabhakitiriya panguva yekukura kweO. cornifrons larvae. Microbacterium imwe mhando yakawanikwa muchidzidzo chedu inonzi inowanikwa muhuwandu hwakawanda muBlack Soldier Flight Flight Larvae pasi penzara66. MuO. cornifrons larvae, microbacteria inogona kubatsira mukuenzana uye kusimba kwe microbiome yemudumbu pasi pemamiriro ekumanikidzwa. Pamusoro pezvo, Rhodococcus inowanikwa muO. cornifrons larvae uye inozivikanwa nekugona kwayo kubvisa chepfu67. Rudzi urwu runowanikwawo mudumbu reA. florea, asi muhuwandu hushoma kwazvo68. Mhedzisiro yedu inoratidza kuvapo kwemarudzi akasiyana-siyana emajini pamarudzi akawanda ema microbial anogona kuchinja maitiro emetabolism mumhungu. Zvisinei, kunzwisisa kuri nani kwekusiyana kwebasa reO. cornifrons kunodiwa.
Muchidimbu, mhedzisiro inoratidza kuti mancozeb, pyrithiostrobin, uye trifloxystrobin zvakaderedza kuwedzera kwehuremu hwemuviri uye kuwedzera kufa kwehonye dzechibage. Kunyangwe paine kunetseka kuri kukura pamusoro pemhedzisiro yefungicides pazvipfukuto, pane chikonzero chekunzwisisa zviri nani mhedzisiro yemetabolites yasara yemakemikari aya. Mhedzisiro iyi inogona kuiswa muzvirongwa zvehurongwa hwakabatanidzwa hwekutarisira fungicides zvinobatsira varimi kudzivirira kushandiswa kwemamwe fungicides usati watumbuka miti yemichero nekusarudza fungicides uye kuchinja nguva yekushandiswa, kana nekukurudzira kushandiswa kwemamwe masuru asina kukuvadza 36. Ruzivo urwu rwakakosha pakugadzira mazano ekushandisa zvipembenene, zvakaita sekugadzirisa mapurogiramu aripo ekupfapfaidza uye kuchinja nguva yekupfapfaidza pakusarudza fungicides kana kukurudzira kushandiswa kwemamwe masuru asina njodzi. Tsvagiridzo yakawedzerwa inodiwa mumhedzisiro yakaipa yefungicides pahuwandu hwepabonde, maitiro ekudyisa, gut microbiome, uye mamorekuru maitiro ari kukonzera kurasikirwa nehuremu nerufu rwechibage chechibage.
Data remanyuko 1, 2 ne3 muMifananidzo 1 ne2 rakaiswa mufigshare data repository DOI: https://doi.org/10.6084/m9.figshare.24996245 uye https://doi.org/10.6084/m9. figshare.24996233. Ma sequences akaongororwa muchidzidzo chazvino (Mifananidzo 4, 5) anowanikwa muNCBI SRA repository pasi penhamba yePRJNA1023565.
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