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Imichiza yokungunda isoloko isetyenziswa ngexesha lokudumba kweziqhamo zomthi kwaye inokoyikisa izinambuzane.Nangona kunjalo, kuncinci okwaziwayo malunga nendlela i-pollinators ezingezizo iinyosi (umzekelo, iinyosi ezihlala zodwa, i-Osmia cornifrons) zisabela njani kuqhagamshelwano kunye nezibulali zifo zokungunda ezidla ngokusetyenziswa kwii-apile ngexesha lokudubula kweentyatyambo.Lo msantsa wolwazi unciphisa izigqibo zolawulo ezimisela ukugxila okukhuselekileyo kunye nexesha lokutshiza ngemichiza yokungunda.Siye savavanya iziphumo zemichiza yokungunda yoqhagamshelwano (i-captan ne-mancozeb) kunye ne-interlayer/phytosystem fungicides ezine (ciprocycline, myclobutanil, pyrostrobin kunye ne-trifloxystrobin).Iziphumo ekufumaneni ubunzima bombungu, ukusinda, umlinganiselo wesondo kunye nokwahlukana kwebhaktiriya.Uvavanyo lwenziwe kusetyenziswa i-bioassay yomlomo engapheliyo apho i-pollen iphathwe kwiidosi ezintathu ngokusekelwe kwi-dose ekhuthazwayo ngoku yokusetyenziswa kwentsimi (1X), i-dose yesiqingatha (0.5X) kunye ne-dose ephantsi (0.1X).Zonke iidosi ze-mancozeb kunye ne-pyritisoline zinciphise kakhulu ubunzima bomzimba kunye nokusinda kombungu.Emva koko salandelelanisa ijini ye-16S ukubonisa i-larval bacteriome ye-mancozeb, isibulala-zinambuzane esinoxanduva lokufa okuphezulu.Sifumanise ukuba iyantlukwano yebhaktiriya kunye nobuninzi bancitshiswe kakhulu kwimibungu eyondliwe kwipollen ene-mancozeb.Iziphumo zethu zaselabhorethri zibonisa ukuba ukutshiza ezinye zezi zibulala-ntsholongwane zokungunda ngexesha lokudubula kweentyatyambo kuyingozi kakhulu kwimpilo ye-O. cornifrons.Olu lwazi lufanelekile kwizigqibo zolawulo lwexesha elizayo malunga nokusetyenziswa ngokuzinzileyo kwemveliso yokhuselo lwemithi yeziqhamo kwaye isebenza njengesiseko seenkqubo zolawulo ezijolise ekukhuseleni abavuzi.
Inyosi ehlala yodwa i-Osmia cornifrons (i-Hymenoptera: i-Megachilidae) yaziswa e-United States isuka e-Japan ekupheleni kweminyaka yoo-1970 kunye nasekuqaleni koo-1980, kwaye uhlobo ludlale indima ebalulekileyo yomvumvuzelwano kwi-ecosystem elawulwayo ukususela ngoko.Abemi bolu hlobo lwenyosi bayinxalenye yeentlobo ezingama-50 zeenyosi zasendle ezincedisa iinyosi ezisasaza ipoleni yealmondi kunye nama-apile e-United States2,3.Iinyosi zeMason zijongene nemingeni emininzi, kubandakanya ukuqhekeka kwendawo yokuhlala, iintsholongwane, kunye nezitshabalalisi3,4.Phakathi kwezibulala-zinambuzane, imichiza yokungunda inciphisa ukufumana amandla, ukutya5 kunye nokulungisa umzimba6,7.Nangona uphando lwakutsha nje lubonisa ukuba impilo yeenyosi zeMason ichaphazeleka ngokuthe ngqo yi-commensal kunye ne-ectobactic microorganisms, i-8,9 kuba iibhaktheriya kunye nefungi zinokuchaphazela isondlo kunye neempendulo ze-immune, iziphumo zokuvezwa kwe-fungicide kwi-microbial diversity of Mason bees ziqala ukuba wafunda.
Imichiza yokungunda yeziphumo ezahlukeneyo (zonxibelelwano kunye nenkqubo) zitshizwa kumabhoma ngaphambi nangexesha lokuntshula kweentyatyambo ukunyanga izifo ezifana nokhoko lwama-apile, ukubola okukrakrayo, ukubola okumdaka kunye nomgubo we-powdery mildew10,11.I-fungicides ithathwa njengengenabungozi kwi-pollinators, ngoko ke iyacetyiswa kubalimi ngexesha leentyantyambo;Ukuvezwa kunye nokungeniswa kwezi fungicides ziinyosi ziyaziwa kakuhle, njengoko kuyinxalenye yenkqubo yokubhalisa i-pesticide yi-Arhente yoKhuseleko lweNdalo yase-US kunye nezinye ii-arhente ezilawulayo zelizwe12,13,14.Nangona kunjalo, iziphumo zemichiza yokungunda kwizinto ezingezizo iinyosi zaziwa kancinci kuba azifunwa phantsi kwezivumelwano zogunyaziso lwentengiso e-United States15.Ukongeza, akukho ziprothokholi eziqhelekileyo zokuvavanya iinyosi ezingatshatanga16,17, kunye nokugcina iikholoni ezibonelela ngeenyosi zovavanyo mngeni18.Ulingo lweenyosi ezahlukeneyo ezilawulwayo ziya ziqhutywa ngakumbi eYurophu nase-USA ukufunda iziphumo zezitshabalalisi kwiinyosi zasendle, kwaye iiprothokholi ezisemgangathweni ziye zaphuhliswa kutsha nje ze-O. cornifrons19.
Iinyosi ezineempondo ziyi-monocytes kwaye zisetyenziselwa urhwebo kwizityalo ze-carp njengesongezelelo okanye ukutshintshwa kweenyosi zobusi.Ezi nyosi zivela phakathi kuka-Matshi no-Epreli, kunye neenkunzi ezingaphambi kwexesha zivela kwiintsuku ezintathu ukuya kwezine phambi kweemazi.Emva kokukhwelana, imazi iqokelela i-pollen kunye nencindi ukuze ibonelele ngoluhlu lweeseli ze-brood ngaphakathi kwendlwane ye-tubular (yendalo okanye yokwenziwa)1,20.Amaqanda abekwe kwi-pollen ngaphakathi kweeseli;imazi ngoko yakha udonga lodongwe ngaphambi kokuba ilungise iseli elandelayo.Iimbungu zokuqala ze-instar zifakwe kwi-chorion kwaye zondla ulwelo lwe-embryonic.Ukususela kwisibini ukuya kweyesihlanu instar (prepupa), imibungu itya i-pollen22.Nje ukuba unikezelo lomungu luphele ngokupheleleyo, imibungu yenza amaqomboloko, intshontsho kwaye ivele ibe ngabantu abadala kwigumbi elinye lenzala, ngokuqhelekileyo ekupheleni kwehlobo20,23.Abantu abadala baphuma kwintwasahlobo elandelayo.Ukuphila kwabantu abadala kuhambelana nokufumana amandla e-net (ukuzuza ubunzima) ngokusekelwe ekutyeni ukutya.Ngaloo ndlela, umgangatho wezondlo we-pollen, kunye nezinye izinto ezifana nemozulu okanye ukuvezwa kwi-pesticides, zizigqibo zokusinda kunye nempilo24.
Amayeza okubulala izinambuzane kunye nemichiza yokungunda efakwe phambi kokudubula kweentyatyambo ayakwazi ukuhamba ngaphakathi kwevasculature yesityalo ukuya kumaqondo ahlukeneyo, ukusuka kwi-translaminar (umzekelo, ekwazi ukusuka kumphezulu wamagqabi ukuya kumphezulu osezantsi, njengamanye amachiza okungunda) 25 ukuya kwiziphumo zenkqubo yenyani., enokungena kwisithsaba ukusuka kwiingcambu, inokungena kwi-nectar yeentyantyambo ze-apula26, apho inokubulala umntu omdala u-O. cornifrons27.Ezinye izitshabalalisi ziphumela kwipollen, nto leyo echaphazela ukukhula kwemibungu yombona kwaye ibangele ukufa kwayo19.Olunye uphononongo lubonise ukuba eminye imichiza yokungunda inokuguqula ngokubonakalayo indlela yokuzalela yeentlobo ezinxulumene ne-O. lignaria28.Ukongeza, iilabhoratri kunye nezifundo zentsimi ezilinganisa iimeko zokuvezwa kwezitshabalalisi (kubandakanywa nemichiza yokungunda) zibonise ukuba izibulala-zinambuzane zichaphazela kakubi i-physiology 22 morphology 29 kunye nokusinda kweenyosi zobusi kunye neenyosi ezizimeleyo.Izitshizi ezahlukahlukeneyo zokubulala umngundo ezisetyenziswa ngokuthe ngqo kwiintyatyambo ezivulekileyo ngexesha lokudubula kweentyatyambo zinokungcolisa umungu oqokelelwe ngabantu abadala ukuze kuphuhliswe imibungu, iziphumo zayo kusafuneka zifundwe30.
Kuya kuqondwa ngakumbi ukuba ukuphuhliswa kombungu kuphenjelelwa yi-pollen kunye ne-microbial community of the digestive system.I-honeybee microbiome ichaphazela iiparameters ezifana nobunzima bomzimba31, utshintsho lwe-metabolic22 kunye nokuchaphazeleka kwi-pathogens32.Izifundo zangaphambili ziye zavavanya impembelelo yenqanaba lophuhliso, izondlo, kunye nokusingqongileyo kwi-microbiome yeenyosi ezizimeleyo.Ezi zifundo zibonise ukufana kwisakhiwo kunye nobuninzi be-larval kunye ne-pollen microbiomes33, kunye neyona genera yebhaktheriya eqhelekileyo iPseudomonas kunye neDelftia, phakathi kweentlobo zeenyosi ezizimeleyo.Nangona kunjalo, nangona i-fungicides idibene nezicwangciso zokukhusela impilo yezinyosi, imiphumo ye-fungicides kwi-microbiota ye-larval ngokusebenzisa ukuvezwa komlomo ngokuthe ngqo ihlala ingahlolisiswa.
Olu phononongo luvavanye iziphumo zeedosi zehlabathi lokwenyani zemichiza yokungunda esetyenziswa rhoqo emithandathu ebhaliselwe ukusetyenziswa kwisiqhamo somthi e-United States, kubandakanywa ukudibana kunye nezibulali-kungunda ezilawulwa ngomlomo kwimibungu ye-corn hornworm moth ephuma kukutya okungcolisekileyo.Siye safumanisa ukuba ukudibana kunye nenkqubo yefungicides yanciphisa ubunzima bomzimba wenyosi kunye nokunyuka kokufa, kunye neziphumo ezibi kakhulu ezinxulumene ne-mancozeb kunye ne-pyrithiopide.Emva koko siye sathelekisa i-microbial diversity ye-larvae etyiswa kukutya kwe-pollen ene-mancozeb kunye nezo zondliwa kulawulo lokutya.Sixoxa ngeendlela ezinokubakho ezikhokelela ekufeni kunye nefuthe kulawulo oludibeneyo lwezinambuzane kunye ne-pollinator (IPPM)36 iinkqubo.
Ii-cornifron zabantu abadala ezigqithisa ubusika kwi-cocoons zafunyanwa kwi-Fruit Research Centre, e-Biglerville, e-PA, kwaye zigcinwe ku-−3 ukuya ku-2 °C (±0.3°C).Ngaphambi kovavanyo (ama-cocoons angama-600 ewonke).NgoMeyi ka-2022, i-100 O. cornifrons cocoons yathunyelwa yonke imihla kwiikomityi zeplastiki (ama-cocoons angama-50 ngekomityi nganye, i-DI 5 cm × 15 cm ubude) kunye nokucima kwafakwa ngaphakathi kweekomityi ukukhuthaza ukuvulwa kunye nokubonelela nge-substrate ehlafunwayo, ukunciphisa uxinzelelo kwi-stony. iinyosi37 .Beka iikomityi ezimbini zeplastiki eziqulethe ama-cocoons kwi-cage yezinambuzane (30 × 30 × 30 cm, i-BugDorm MegaView Science Co. Ltd., eTaiwan) kunye ne-10 ml feeders equlethe i-50% yesisombululo se-sucrose kunye nokugcina iintsuku ezine ukuqinisekisa ukuvala kunye nokukhwelana .I-23 ° C, umswakama ohambelana ne-60%, i-photoperiod 10 l (ubukhulu obuphantsi): iintsuku eziyi-14.I-100 leemazi kunye neenkunzi ezitshatileyo zakhululwa rhoqo kusasa ngeentsuku ezintandathu (i-100 ngosuku) kwiindlwana zokwenziwa ezimbini ngexesha leentyatyambo ze-apile eziphezulu (indlwane yomgibe: ububanzi 33.66 × ubude 30.48 × ubude 46.99 cm; Umfanekiso owongezelelweyo 1).Ibekwe ePennsylvania State Arboretum, kufutshane necherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), ipesika (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), ipear (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), i-coronaria apple tree (Malus coronaria) kunye neentlobo ezininzi zemithi yama-apile (Malus coronaria, Malus), umthi wama-apile wasekhaya 'Co-op 30′ Enterprise™, Malus apple tree' Co- Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy').Indlu nganye yentaka yeplastiki eluhlaza ingena phezu kweebhokisi ezimbini zomthi.Ibhokisi yendlwane nganye iqulathe iityhubhu zephepha zekraft ezingama-800 (i-spiral open, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) efakwe kwiityhubhu ze-opaque ze-cellophane (0.7 OD bona iiplagi zePlastiki (iiplagi ze-T-1X) zibonelela ngeendawo zokuhlala. .
Zombini iibhokisi zendlwane zijonge ngasempuma kwaye zagqunywa ngocingo lwegadi yeplastiki eluhlaza (imodeli ye-Everbilt #889250EB12, ubukhulu bokuvula 5 × 5 cm, 0.95 m × 100 m) ukuthintela ukufikelela kweempuku kunye neentaka kwaye zibekwe kumphezulu womhlaba ecaleni komhlaba webhokisi yendlwane. iibhokisi.Ibhokisi ye-Nest (Umfanekiso owongezelelweyo 1a).Amaqanda e-corn borer ayeqokelelwa yonke imihla ngokuqokelela iityhubhu ezingama-30 kwiindlwane aze azithuthe ukuya kwilebhu.Usebenzisa isikere, yenza ukusika ekupheleni kombhobho, emva koko uqhaqhe ityhubhu ejikelezayo ukuze uveze iiseli zebrood.Amaqanda ngamnye kunye ne-pollen yawo yasuswa kusetyenziswa i-spatula egobileyo (i-Microslide tool kit, i-BioQuip Products Inc., eCalifornia).Amaqanda afukanywa kwiphepha lokucoca elifumileyo kwaye abekwe kwisitya sePetri iiyure ezi-2 ngaphambi kokuba asetyenziswe kuvavanyo lwethu (Umfanekiso owongezelelweyo 1b-d).
Kwilabhoratri, siye savavanya ubutyhefu bomlomo bemichiza yokungunda emithandathu esetyenziswa ngaphambi nangexesha lokutyatyamba kweapile kwiinkalo ezintathu (0.1X, 0.5X, kunye ne-1X, apho i-1X iluphawu olusetyenziswa kwiilitha ezili-100 zamanzi/ehektare. ebaleni)., Uluhlu 1).Ugxininiso ngalunye luphindwe ngamaxesha e-16 (n = 16).Izibulala-zinambuzane ezimbini zoqhagamshelwano (Itheyibhile S1: mancozeb 2696.14 ppm kunye ne-captan 2875.88 ppm) kunye ne-systemic fungicides ezine (Itheyibhile S1: i-pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobupp5m8 2 azolecypro.2 azole m) ubutyhefu kwiziqhamo, imifuno kunye nezityalo zokuhombisa .Thina i-homogenized i-pollen ngokusebenzisa i-grinder, idluliselwe i-0.20 g kwiqula (i-24-well Falcon Plate), kwaye yongezwa kwaye ixutywe i-1 μL yesisombululo se-fungicide ukwenza i-pyramidal pollen kunye ne-1 mm yemithombo enzulu apho amaqanda abekwe khona.Beka usebenzisa i-spatula encinci (umzobo owongezelelweyo 1c, d).Iipleyiti ze-Falcon zigcinwe kwindawo yokushisa (25 ° C) kunye ne-70% yomswakama ohambelanayo.Sazithelekisa kunye nemibungu yokulawula eyondla ukutya kwe-pollen exutywe ngamanzi acocekileyo.Sirekhode ukufa kunye nokulinganisa ubunzima bombungu yonke imihla de imibungu ifikelele kwiminyaka yangaphambi kwe-prepupal isebenzisa ibhalansi yohlalutyo (Fisher Scientific, accuracy = 0.0001 g).Ekugqibeleni, umlinganiselo wesondo wavavanywa ngokuvula i-cocoon emva kweenyanga ezi-2.5.
I-DNA yatsalwa kuyo yonke imibungu ye-O. cornifrons (n = 3 ngokwemeko yonyango, i-mancozeb-enyangwayo kunye ne-pollen enganyangwanga) kwaye siye senza uhlalutyo lokwahlukana kwe-microbial kwezi sampuli, ngakumbi kuba kwi-mancozeb eyona nto iphezulu yokufa yabonwa kwi-larvae.ukufumana uMnZn.I-DNA yandiswa, yahlanjululwa kusetyenziswa i-DNAZymoBIOMICS®-96 MagBead DNA kit (i-Zymo Research, Irvine, CA), kwaye ilandelelaniswe (imijikelo engama-600) kwi-Illumina® MiSeq™ isebenzisa i-v3 kit.Ulandelelwano olujoliswe kuyo lwebhaktiriya ye-16S ribosomal RNA yofuzo lwenziwa kusetyenziswa i-Quick-16S™ NGS Library Prep Kit (iZymo Research, Irvine, CA) kusetyenziswa iiprimers ezijolise kwingingqi ye-V3-V4 ye-16S rRNA gene.Ukongezelela, ukulandelelana kwe-18S kwenziwa ngokusebenzisa i-10% yokufakwa kwe-PhiX, kwaye ukukhulisa kwenziwa ngokusebenzisa i-primer pair 18S001 kunye ne-NS4.
Ngenisa kunye nenkqubo idityanisiwe kufundwa39 usebenzisa umbhobho we-QIIME2 (v2022.11.1).Ezi zifundo zilungiswe kwaye zidityanisiwe, kwaye ukulandelelana kwe-chimeric kwasuswa kusetyenziswa iplagin ye-DADA2 kwi-QIIME2 (qiime dada2 ukubhanqa ingxolo)40.Izabelo zeklasi ze-16S kunye ne-18S zenziwe kusetyenziswa iplagi yohlelo lodidi lwento Classify-sklearn kunye ne-artifact eqeqeshwe kwangaphambili i-silva-138-99-nb-classifier.
Zonke iinkcukacha zovavanyo zihlolwe ukuba ziqhelekile (i-Shapiro-Wilks) kunye ne-homogeneity of variances (uvavanyo lukaLevene).Ngenxa yokuba isethi yedatha ayizange ihlangabezane neengcinga zokuhlalutya i-parametric kwaye ukuguqulwa kwahluleka ukulungelelanisa i-residuals, senze i-nonparametric yeendlela ezimbini ze-ANOVA (Kruskal-Wallis) kunye nezinto ezimbini [ixesha (isigaba sesithathu se-2, i-5, kunye ne-8 ngosuku). amanqaku exesha) kunye nesichiza sokungunda] ukuvavanya isiphumo sonyango kubunzima obutsha bombungu, emva koko uthelekiso lwe-post hoc nonparametric pairwise lwenziwa kusetyenziswa uvavanyo lweWilcoxon.Sasebenzisa imodeli yomgca ngokubanzi (GLM) kunye nokusabalalisa kwePoisson ukuthelekisa imiphumo ye-fungicides ekusindeni kuzo zonke iintlobo ezintathu ze-fungicide concentrations41,42.Ukuhlalutya ubuninzi obuhlukeneyo, inani le-amplicon sequence variants (ASVs) yawa phantsi kwinqanaba le-genus.Ukuthelekiswa kobuninzi bokwahlukana phakathi kwamaqela asebenzisa i-16S (inqanaba le-genus) kunye ne-18S ubuninzi obunxulumene nobuninzi benziwa kusetyenziswa imodeli eyongezelelweyo ngokubanzi kwisithuba, isikali, kunye nokuma (GAMLSS) kunye ne-beta zero-inflated (BEZI) yosasazo lwentsapho, oluyimodeli kwi-macro. .kwiMicrobiome R43 (v1.1).1).Susa iintlobo ze-mitochondrial kunye ne-chloroplast ngaphambi kohlalutyo oluhlukeneyo.Ngenxa yamanqanaba ahlukeneyo etaxonomic ye-18S, kuphela elona nqanaba lisezantsi letaxon nganye lisetyenziselwe uhlalutyo olwahlukileyo.Lonke uhlalutyo lwamanani lwenziwa kusetyenziswa i-R (v. 3.4.3., iprojekthi ye-CRAN) (Iqela le-2013).
Ukubonakaliswa kwi-mancozeb, i-pyrithiostrobin, kunye ne-trifloxystrobin kunciphisa kakhulu ubunzima bomzimba kwi-O. cornifrons (Umfanekiso 1).Ezi ziphumo bezijongwa rhoqo kuzo zontathu iidosi ezivavanyiweyo (Umfanekiso 1a–c).I-Cyclostrobin kunye ne-myclobutanil ayizange inciphise kakhulu ubunzima beembungu.
I-avareji yobunzima obutsha bombungu we-stem borer olinganiswa ngamaxesha amathathu phantsi konyango lwe-dietary ezine (i-homogeneous pollen feed + fungicide: control, 0.1X, 0.5X kunye ne-1X iidosi).(a) Idosi ephantsi (0.1X): indawo yokuqala (usuku 1): χ2: 30.99, DF = 6;P <0.0001, inqaku lesibini (usuku 5): 22.83, DF = 0.0009;okwesithathu;ingongoma (usuku 8): χ2: 28.39, DF = 6;(b) isiqingatha sedosi (0.5X): indawo yokuqala (usuku 1): χ2: 35.67, DF = 6;P <0.0001, inqaku lesibini (usuku lokuqala).): χ2: 15.98, DF = 6;P = 0.0090;indawo yesithathu (usuku 8) χ2: 16.47, DF = 6;(c) Isiza okanye idosi epheleleyo (1X): indawo yokuqala (usuku 1) χ2: 20.64, P = 6;P = 0.0326, inqaku lesibini (usuku 5): χ2: 22.83, DF = 6;P = 0.0009;inqaku lesithathu (usuku 8): χ2: 28.39, DF = 6;uhlalutyo lwe-nonparametric yokwahluka.Imivalo imele intsingiselo ± SE yokuthelekisa ngababini (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Kwidosi ephantsi (0.1X), ubunzima bomzimba we-larval buncitshiswe nge-60% kunye ne-trifloxystrobin, i-49% ne-mancozeb, i-48% ne-myclobutanil, kunye ne-46% nge-pyrithistrobin (umzobo 1a).Xa ubonakaliswe kwisiqingatha sedosi yentsimi (0.5X), ubunzima bomzimba we-mancozeb larvae buncitshiswe ngama-86%, i-pyrithiostrobin ngama-52% kunye ne-trifloxystrobin ngama-50% (umzobo 1b).Idosi egcweleyo yasendle (1X) ye-mancozeb yehlise ubunzima bombungu ngama-82%, ipyrithiostrobin ngama-70%, kunye ne-trifloxystrobin, myclobutanil kunye ne-sangard malunga nama-30% (Fig. 1c).
Ukubhubha kwabaphezulu phakathi kwemibungu etyiswe umungu one-mancozeb, ilandelwa yipyrithiostrobin kunye ne-trifloxystrobin.Ukufa kwanda ngokunyuka kwamanani e-mancozeb kunye ne-pyritisoline (umzobo 2; iThebhile 2).Nangona kunjalo, ukufa kwe-corn borer kuye kwanda nje kancinci njengoko ukugxila kwe-trifloxystrobin kusanda;I-cyprodinil kunye ne-captan ayizange ikhulise kakhulu ukufa xa kuthelekiswa nonyango lokulawula.
Ukufa kwemibungu yeempukane kuthelekiswe emva kokufunyaniswa komungu ngamnye kunyangwa ngemichiza emithandathu eyahlukeneyo yokungunda.I-Mancozeb kunye ne-pentopyramide babenovelwano ngakumbi ekuvezweni komlomo kwiimpethu zombona (i-GLM: χ = 29.45, DF = 20, P = 0.0059) (umgca, i-slope = 0.29, P <0.001; i-slope = 0.24, P) <0.00).
Ngokomyinge, kulo lonke unyango, i-39.05% yezigulane yayingabasetyhini kwaye i-60.95% yayingamadoda.Phakathi kwezonyango zolawulo, umlinganiselo wabasetyhini wawungama-40% kuzo zombini idosi ephantsi (0.1X) kunye nesiqingatha sedosi (0.5X) izifundo, kunye ne-30% kwizifundo zedosi yasendle (1X).Kwidosi ye-0.1X, phakathi kwemibungu eyondliwe i-pollen enyangwa nge-mancozeb kunye ne-myclobutanil, i-33.33% yabantu abadala yayingabantu basetyhini, i-22% yabantu abadala yayingabasetyhini, i-44% ye-larvae yabantu abadala yayingabasetyhini, i-44% ye-larvae yabantu abadala yayingabasetyhini.imazi, i-41% yemibungu yabantu abadala yayingabafazi, kwaye ulawulo lwalungama-31% (Umfanekiso 3a).Ngamaxesha angama-0.5 ethamo, i-33% yeentshulube zabantu abadala kwiqela le-mancozeb kunye ne-pyrithiostrobin yayingabantu basetyhini, i-36% kwiqela le-trifloxystrobin, i-41% kwiqela le-myclobutanil, kunye ne-46% kwiqela le-cyprostrobin.Eli nani belingama-53% kwiqela.kwiqela le-captan kunye ne-38% kwiqela lolawulo (Umfanekiso 3b).Kwi-dose ye-1X, i-30% yeqela le-mancozeb yayingabasetyhini, i-36% yeqela le-pyrithiostrobin, i-44% yeqela le-trifloxystrobin, i-38% yeqela le-myclobutanil, i-50% yeqela lokulawula yayingabasetyhini - 38.5% (Umfanekiso 3c) .
Ipesenti yemazi nenkunzi emva kokuba sesichengeni sokubulala umngundo.(a) Idosi ephantsi (0.1X).(b) Isiqingatha sedosi (0.5X).(c) Idosi yasendle okanye idosi epheleleyo (1X).
Uhlalutyo lokulandelelana kwe-16S lubonise ukuba iqela lebhaktheriya lahlukile phakathi kwemibungu eyondliwe nge-pollen enyangwa yi-mancozeb kunye ne-larvae ezondliwa nge-pollen engaphathwanga (Fig. 4a).Isalathiso semicrobial semibungu enganyangwanga eyondliwe kwipollen yayiphezulu kunelo leemibungu ezondliwa kwipollen enyangwa yimancozeb (Fig. 4b).Nangona umahluko oboniweyo kubutyebi phakathi kwamaqela wawungabalulekanga ngokweenkcukacha-manani, wawusezantsi kakhulu kunoko ujongwe ukuba imibungu isitya kwipollen enganyangwanga (Fig. 4c).Ubuninzi obunxulumeneyo bubonise ukuba i-microbiota ye-larvae eyondliwe kwi-pollen yokulawula yayihluke kakhulu kuneembungu ezondliwe kwi-mancozeb-treated larvae (Fig. 5a).Uhlalutyo oluchazayo lubonakalise ubukho be-28 genera ekulawuleni kunye neesampuli eziphathwe nge-mancozeb (umzobo 5b).c Uhlahlelo olusebenzisa ulandelelwano lwe-18S alubonakalisi mahluko ubalulekileyo (Umfanekiso owongezelelweyo 2).
Iiprofayili ze-SAV ezisekelwe kulandelelwano lwe-16S zafaniswa nobutyebi be-Shannon kwaye zaqaphela ubutyebi kwinqanaba le-phylum.(a) Uhlalutyo oluphambili lolungelelwaniso (PCoA) olusekwe kulwakhiwo loluntu jikelele lwemicrobial kwi-pollen-fed or control (blue) kunye ne-mancozeb-fed (orenji).Inqaku ngalinye ledatha limele isampuli eyahlukileyo.I-PCoA ibalwe kusetyenziswa umgama weBray-Curtis we-multivariate t yokusabalalisa.I-ovals imele i-80% yezinga lokuzithemba.(b) I-Boxplot, idatha ekrwada yobutyebi bukaShannon (amanqaku) kunye c.Ubutyebi obubonakalayo.Iibhokisi zebhokisi zibonisa iibhokisi ze-median line, i-interquartile range (IQR), kunye ne-1.5 × IQR (n = 3).
Ukudityaniswa koluntu lwemicrobial lwemibungu etyiswe kwi-mancozeb-enyangweni kunye nepollen enganyangwanga.(a) Ubuninzi obunxulumeneyo bemicrobial genera bufundeka kwilarvae.(b) Imephu yobushushu yeentsholongwane ezichongiweyo.I-Delftia (i-odd ratio (OR) = 0.67, P = 0.0030) kunye ne-Pseudomonas (OKANYE = 0.3, P = 0.0074), i-Microbacterium (OKANYE = 0.75, P = 0.0617) (OKANYE = 1.5, P = 0.0060);Imiqolo yemephu yobushushu idityanisiwe kusetyenziswa umgama wolungelelwaniso kunye nomndilili woqhagamshelo.
Iziphumo zethu zibonisa ukuba ukuba sesichengeni somlomo kunxibelelwano (mancozeb) kunye ne-systemic (pyrostrobin kunye ne-trifloxystrobin) imichiza yokungunda, esetyenziswa kakhulu ngexesha leentyantyambo, inciphise kakhulu ukutyeba kunye nokunyuka kokusweleka kwemibungu yombona.Ukongeza, i-mancozeb yanciphisa kakhulu iyantlukwano kunye nobutyebi be-microbiome ngexesha lenqanaba le-prepupal.I-Myclobutanil, enye i-systemic fungicide, yanciphisa ngokuphawulekayo ukufumana ubunzima bomzimba wombungu kuzo zontathu iidosi.Esi siphumo sabonakala kwinqanaba lesibini (usuku lwe-5) kunye neyesithathu (usuku lwe-8).Ngokwahlukileyo, i-cyprodinil kunye ne-captan ayizange inciphise kakhulu ukufumana ubunzima okanye ukusinda xa kuthelekiswa neqela lokulawula.Ngokolwazi lwethu, lo msebenzi ngowokuqala ukumisela ifuthe lemilinganiselo yentsimi yezibulali-kungunda ezahlukeneyo ezisetyenziselwa ukukhusela izityalo zombona ngokutyhileka ngokuthe ngqo kwepollen.
Lonke unyango lokungunda lunciphise kakhulu ubunzima bomzimba xa kuthelekiswa nonyango lolawulo.IMancozeb ibe neyona mpembelelo inkulu ekufumaneni ubunzima bombungu ngokuncipha komndilili wama-51%, ilandelwa yipyrithiostrobin.Nangona kunjalo, olunye uphononongo alukhange luchaze iziphumo ezibi zedosi yasendle yemichiza yokungunda kumanqanaba ombungu44.Nangona i-dithiocarbamate biocides ibonakaliswe inetyhefu ephantsi ye-acute45, i-ethylene bisdithiocarbamates (EBDCS) efana ne-mancozeb inokwehlisela kwi-urea ethylene sulfide.Ukunikezelwa kwemiphumo ye-mutagenic kwezinye izilwanyana, le mveliso yokuthotywa ingaba noxanduva lweziphumo ezibonwayo46,47.Izifundo zangaphambili zibonise ukuba ukubunjwa kwe-ethylene thiourea kuphenjelelwa zizinto ezifana neqondo lokushisa eliphakamileyo48, amanqanaba okufuma49 kunye nobude bokugcinwa kwemveliso50.Iimeko ezifanelekileyo zokugcina ii-biocides zinokuthomalalisa ezi ziphumo bezingalindelekanga.Ukongezelela, i-European Food Safety Authority ibonise inkxalabo malunga netyhefu yepyrithiopide, ebonakaliswe ukuba i-carcinogenic kwiinkqubo zokugaya ezinye izilwanyana51.
Ukulawulwa ngomlomo kwe-mancozeb, i-pyrithiostrobin, kunye ne-trifloxystrobin kwandisa ukusweleka kwemibungu yombona.Ngokwahlukileyo, i-myclobutanil, i-ciprocycline kunye ne-captan yayingenayo impembelelo ekufeni.Ezi ziphumo zihluke kwezo zikaLadurner et al.52, obonise ukuba i-captan yanciphisa kakhulu ukusinda kwabantu abadala i-O. lignaria kunye ne-Apis mellifera L. (Hymenoptera, Apisidae).Ukongeza, imichiza yokungunda efana ne-captan kunye ne-boscalid ifunyenwe ibangela ukufa kombungu52,53,54 okanye ukuguqula indlela yokuziphatha55.Olu tshintsho, lunokuchaphazela umgangatho wesondlo se-pollen kwaye ekugqibeleni ukuzuza kwamandla kwinqanaba le-larval.Ukufa okubonwe kwiqela lolawulo kwakuhambelana nezinye izifundo ze-56,57.
Umlinganiselo wesondo othanda indoda ebonwayo kumsebenzi wethu unokuchazwa ngezinto ezifana nokungonelanga kokukhwelana kunye neemeko ezimbi zemozulu ngexesha leentyantyambo, njengoko kwakucetyiswe ngaphambili kwi-O. cornuta nguVicents noBosch.Nangona iimazi kunye namadoda kuphononongo lwethu zineentsuku ezine zokutshata (ixesha elithathwa njengelikwaneleyo ukukhwelana ngempumelelo), siye sanciphisa ngamabom ukukhanya ukuze sinciphise uxinzelelo.Nangona kunjalo, olu tshintsho lungaphazamisana nenkqubo yokukhwelana ngokungeyonjongo61.Ukongeza, iinyosi zifumana iintsuku ezininzi zemozulu embi, kuquka imvula kunye namaqondo obushushu aphantsi (<5°C), nto leyo enokuchaphazela kakubi impumelelo4,23 yokukhwelana.
Nangona uphononongo lwethu lujolise kuyo yonke i-larval microbiome, iziphumo zethu zibonelela ngokuqonda kubudlelwane obunokubakho phakathi koluntu lwebhaktiriya olunokubaluleka kwisondlo seenyosi kunye nokuvezwa kwefungicide.Umzekelo, imibungu etyiswe umungu enyangwa yi-mancozeb inciphise kakhulu ubume bentsholongwane yoluntu kunye nobuninzi xa kuthelekiswa nemibungu etyiswe umungu onganyangwanga.Kwimibungu etya umungu onganyangwanga, amaqela ebhaktiriya iProteobacteria kunye ne-Actinobacteria ayengamele kwaye ubukhulu becala yayi-aerobic okanye i-facultatively aerobic.Ibhaktheriya yeDelft, edla ngokunxulunyaniswa neentlobo zeenyosi ezizimeleyo, zaziwa ngokuba nomsebenzi wokubulala iintsholongwane, nto leyo ebonisa indima enokubakho yokukhusela kwiintsholongwane.Olunye uhlobo lwebhaktiriya, iPseudomonas, yayininzi kwimibungu eyondla umungu onganyangwanga, kodwa yancitshiswa kakhulu kwimibungu enyangwa yimancozeb.Iziphumo zethu zixhasa izifundo zangaphambili ezichonga i-Pseudomonas njengenye yezona genera zininzi kwi-O. bicornis35 kunye nabanye oomeva ababodwa34.Nangona ubungqina bovavanyo lwendima ye-Pseudomonas kwimpilo ye-O. cornifrons ayizange ifundwe, le bhaktiriya ibonakaliswe ukukhuthaza ukuhlanganiswa kwee-toxins ezikhuselayo kwi-beetle Paederus fuscipes kunye nokukhuthaza i-arginine metabolism in vitro 35, 65. Ezi ngqalelo zibonisa indima enokubakho kukhuselo lwentsholongwane kunye neebhaktheriya ngexesha lophuhliso lwe-O. cornifrons larvae.I-Microbacterium lolunye uhlobo oluchongiweyo kuphononongo lwethu ekuxelwe ukuba lukhona ngamanani aphezulu kwimibungu yeempukane zejoni elimnyama phantsi kweemeko zendlala66.Kwi-O. cornifrons larvae, i-microbacteria ingaba negalelo kwibhalansi kunye nokuqina kwe-gut microbiome phantsi kweemeko zoxinzelelo.Ukongeza, i-Rhodococcus ifumaneka kwi-O. cornifrons larvae kwaye iyaziwa ngezakhono zayo zokukhupha i-toxification67.Olu hlobo lukwafumaneka emathunjini e-A. florea, kodwa ngobuninzi obuphantsi kakhulu68.Iziphumo zethu zibonisa ubukho bokwahluka kofuzo oluninzi kwii-tax ezininzi zemicrobial ezinokuguqula iinkqubo zemetabolism kwimibungu.Nangona kunjalo, ukuqonda okungcono kokungafani okusebenzayo kwe-O. cornifrons kuyadingeka.
Ngamafutshane, iziphumo zibonisa ukuba i-mancozeb, i-pyrithiostrobin, kunye ne-trifloxystrobin zinciphise ukufumana ubunzima bomzimba kunye nokunyuka kokusweleka kwemibungu yombona.Nangona kukho inkxalabo ekhulayo malunga nemiphumo ye-fungicides kwi-pollinators, kukho imfuneko yokuqonda ngcono imiphumo ye-metabolites eseleyo yale mixube.Ezi ziphumo zinokudityaniswa kwiingcebiso zeenkqubo ezihlangeneyo zolawulo lwe-pollinator ezinceda amafama ukuba aphephe ukusebenzisa imichiza ethile yokungunda ngaphambi nangexesha lokudumba kwemithi yeziqhamo ngokukhetha imichiza yokungunda kunye nokwahluka kwexesha lokufakwa kwayo, okanye ngokukhuthaza ukusetyenziswa kweendlela ezingeyongozi kangako 36. Olu lwazi ibalulekile ekuphuhliseni iingcebiso.ekusetyenzisweni kwezitshabalalisi, ezifana nokulungelelanisa iinkqubo ezikhoyo zokutshiza kunye nokutshintsha ixesha lokutshiza xa ukhetha imichiza yokungunda okanye ukukhuthaza ukusetyenziswa kwezinye iindlela ezingenabungozi kangako.Uphando olongezelelweyo luyafuneka kwiziphumo ezibi zokubulala umngundo kumlinganiselo wesondo, indlela yokuziphatha, i-gut microbiome, kunye neendlela zemolekyuli eziphantsi kokuncipha kobunzima bombona kunye nokufa.
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