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Ama-fungicides avame ukusetshenziswa ngesikhathi sokuqhakaza kwezithelo zesihlahla futhi angasongela ama-pollinator ezinambuzane.Kodwa-ke, kuncane okwaziwayo mayelana nendlela impova ezingezona izinyosi (isb., izinyosi ezihlala zodwa, i-Osmia cornifrons) ezisabela ngayo ekuthinteni kanye nezibulala-fungi ezihleliwe ezivame ukusetshenziswa kuma-apula ngesikhathi sokuqhakaza.Leli gebe lolwazi likhawulela izinqumo zokulawula ezinquma ukugxila okuphephile kanye nesikhathi sokufafaza ngesikhunta.Sihlole imiphumela yezibulala-sikhunta ezimbili ezithintana naye (i-captan ne-mancozeb) kanye nama-interlayer/phytosystem fungicides amane (i-ciprocycline, i-myclobutanil, i-pyrostrobin ne-trifloxystrobin).Imithelela ekuzuzeni isisindo sezibungu, ukusinda, isilinganiso sobulili nokuhlukahluka kwamagciwane.Ukuhlola kwenziwe kusetshenziswa i-bioassay yomlomo engapheli lapho impova yelashwa ngemithamo emithathu ngokusekelwe kumthamo onconywayo okwamanje wokusetshenziswa ensimini (1X), umthamo oyingxenye (0.5X) kanye nedosi ephansi (0.1X).Yonke imithamo ye-mancozeb ne-pyritisoline inciphise kakhulu isisindo somzimba kanye nokusinda kwezibungu.Sabe sesilandelanisa isakhi sofuzo se-16S ukuze sibonise i-bacteriome yesibungu ye-mancozeb, isibulala-sikhunta esibangela ukufa okuphezulu kakhulu.Sithole ukuthi ukuhlukahluka nobuningi bamagciwane kwehliswe kakhulu ezibungwini ezifunzwa impova efakwe i-mancozeb.Imiphumela yethu yaselabhorethri ikhombisa ukuthi ukufafaza amanye alawa ma-fungicides ngesikhathi sokuqhakaza kuyingozi kakhulu empilweni ye-O. cornifrons.Lolu lwazi lufanelekile ezinqumweni zokuphatha zesikhathi esizayo mayelana nokusetshenziswa okuqhubekayo kwemikhiqizo yokuvikela izihlahla zezithelo futhi lusebenza njengesisekelo sezinqubo zokulawula okuhloswe ngazo ukuvikela abathumbi bempova.
Inyosi ewumakhi yodwa i-Osmia cornifrons (Hymenoptera: Megachilidae) yethulwa e-United States isuka e-Japan ngasekupheleni kwawo-1970 kanye nasekuqaleni kwawo-1980, futhi lolu hlobo seludlale indima ebalulekile yokuthuthwa kwempova ku-ecosystem ephethwe kusukela ngaleso sikhathi.Izibalo ezingokwemvelo zale nyosi ziyingxenye yezinhlobo ezingaba ngu-50 zezinyosi zasendle ezihambisana nezinyosi ezithulisa impova ezisimini zama-alimondi nama-apula e-United States2,3.Izinyosi zamaMason zibhekana nezinselelo eziningi, okuhlanganisa ukuhlukana kwezindawo zokuhlala, amagciwane, nezibulala-zinambuzane3,4.Phakathi kwama-insecticides, ama-fungicides anciphisa ukuzuza kwamandla, ukuphakela ukudla5 kanye nokulungisa umzimba6,7.Nakuba ucwaningo lwakamuva luphakamisa ukuthi impilo yezinyosi zaseMason ithonywa ngokuqondile ama-commensal kanye ne-ectobactic microorganisms, i-8,9 ngoba amagciwane kanye nesikhunta singathonya umsoco kanye nezimpendulo zokuzivikela komzimba, imiphumela yokuchayeka kwe-fungicide ezinhlobonhlobo ze-microbial ye-Mason bee wafunda.
Ama-fungicides anemiphumela ehlukahlukene (othintana nawo kanye nesistimu) afuthwa ezinsimini zezithelo ngaphambi nangesikhathi sokuqhakaza ukuze kwelaphe izifo ezinjengoqweqwe lwama-apula, ukubola okubabayo, ukubola okunsundu kanye nesikhutha10,11.Ama-fungicides abhekwa njengengozi kuma-pollinator, ngakho-ke anconywa kubalimi ngesikhathi sokuqhakaza;Ukuchayeka kanye nokumunca lezi zibulala-sikhunta yizinyosi kwaziwa kakhulu, njengoba kuyingxenye yenqubo yokubhaliswa kwezibulala-zinambuzane yi-US Environmental Protection Agency kanye nezinye izikhungo eziningi zokulawula zikazwelonke12,13,14.Kodwa-ke, imiphumela yemithi ebulala isikhunta kwabangezona izinyosi ayaziwa kancane ngoba ayidingeki ngaphansi kwezivumelwano zokugunyazwa kokumaketha e-United States15.Ukwengeza, ngokuvamile azikho izimiso ezijwayelekile zokuhlola izinyosi ezingabodwa16,17, futhi ukugcina amakoloni ahlinzeka ngezinyosi ukuze zihlolwe kuyinselele18.Ukuhlolwa kwezinyosi ezehlukene ezilawulwayo kuya ngokuya kwenziwa e-Europe nase-USA ukuze kufundwe imiphumela yezibulala-zinambuzane ezinyosini zasendle, futhi izivumelwano ezijwayelekile zisanda kwakhiwa ze-O. cornifrons19.
Izinyosi ezinezimpondo ziyi-monocyte futhi zisetshenziselwa ukuhweba izitshalo ze-carp njengesengezo noma esikhundleni sezinyosi zoju.Lezi zinyosi zivela phakathi kukaNdasa kanye no-April, kanti eyeduna engaphambi kwesikhathi iphuma ezinsukwini ezintathu kuya kwezine ngaphambi kwezinsikazi.Ngemva kokukhwelana, insikazi iqoqa ngenkuthalo impova kanye nompe ukuze kuhlinzekwe uchungechunge lwamangqamuzana ama-brood ngaphakathi kwesidleke se-tubular (imvelo noma yokwenziwa)1,20.Amaqanda abekwa phezu kwempova ngaphakathi kwamaseli;insikazi bese yakha udonga lobumba ngaphambi kokulungisa ingqamuzana elilandelayo.Izibungu zokuqala ze-instar zivalelwe ku-chorion futhi zidla uketshezi lwe-embryonic.Kusukela kweyesibili kuya kweyesihlanu (prepupa), izibungu zidla impova22.Lapho impova isiphelile ngokuphelele, izibungu zakha imifece, izibungu futhi ziphuma sezikhulile ekamelweni elilodwa lezingane, ngokuvamile ekupheleni kwehlobo20,23.Abantu abadala bayavela entwasahlobo elandelayo.Ukusinda kwabantu abadala kuhlotshaniswa nokuzuza kwamandla ensalela (ukuzuza isisindo) okusekelwe ekudleni.Ngakho, izinga lokudla okunomsoco wempova, kanye nezinye izici ezifana nesimo sezulu noma ukuchayeka ezibulala-zinambuzane, yizici zokusinda kanye nempilo24.
Ama-insecticide nama-fungicides asetshenziswa ngaphambi kokuqhakaza ayakwazi ukuhamba ngaphakathi kwe-vasculature yesitshalo kuye kumazinga ahlukahlukene, ukusuka ku-translaminar (isb., ekwazi ukusuka phezulu kwamaqabunga ukuya endaweni engezansi, njengamanye ama-fungicides) 25 kuya emiphumeleni yesistimu yangempela., engangena emqheleni isuka ezimpandeni, ingangena umpe wezimbali ze-apula26, lapho zingabulala khona umuntu omdala u-O. cornifrons27.Ezinye izibulala-zinambuzane ziphinde zingene empovani, ziphazamise ukukhula kwezibungu zommbila futhi zibangele ukufa kwazo19.Olunye ucwaningo lubonise ukuthi amanye ama-fungicides angashintsha kakhulu ukuziphatha kokuzalela kwezinhlobo ezihlobene ze-O. lignaria28.Ngaphezu kwalokho, ucwaningo lwaselabhorethri nolwensimu olulingisa izimo zokuchayeka kwezibulala-zinambuzane (kuhlanganise nezibulala-sikhunta) lubonise ukuthi izibulala-zinambuzane zithinta kabi i-physiology 22 morphology 29 nokuphila kwezinyosi zoju nezinye izinyosi ezihlala zodwa.Izifutho ezihlukahlukene zokubulala isikhunta ezisetshenziswa ngokuqondile ezimbalini ezivulekile ngesikhathi sokuqhakaza zingase zingcolise impova eqoqwe abantu abadala ukuze kuthuthukiswe izibungu, imiphumela yayo okusamele ifundwe30.
Kuya ngokuya kuqashelwa ukuthi ukuthuthukiswa kwezibungu kuthonywa impova kanye nemiphakathi yama-microbial ohlelweni lokugaya ukudla.I-honeybee microbiome ithonya imingcele efana nesisindo somzimba31, izinguquko ze-metabolic22 kanye nokuba sengozini kumagciwane32.Ucwaningo lwangaphambilini luye lwahlola ithonya lesigaba sokuthuthuka, izakhi, kanye nemvelo ku-microbiome yezinyosi ezizimele.Lezi zifundo zembule ukufana ekwakhekeni nasekuchichimeni kwe-larval ne-pollen microbiomes33, kanye nesizukulwane sebhaktheriya esivame kakhulu i-Pseudomonas ne-Delftia, phakathi kwezinhlobo zezinyosi ezizimele zodwa.Kodwa-ke, nakuba ama-fungicides ahlotshaniswa namasu okuvikela impilo yezinyosi, imiphumela ye-fungicides ku-larval microbiota ngokusebenzisa ukuchayeka komlomo okuqondile ihlala ingahloliwe.
Lolu cwaningo luhlole imiphumela yemithamo yomhlaba wangempela yezibulala-sikhunta eziyisithupha ezivame ukusetshenziswa ezibhaliselwe ukusetshenziswa esithelweni sesihlahla e-United States, okuhlanganisa ukuthintana nezibulala-sikhunta ezihleliwe ezilawulwa ngomlomo ezibungwini zenundu lommbila ezivela ekudleni okungcolile.Sithole ukuthi ukuthintana kanye nama-systemic fungicides anciphisa ukuzuza kwesisindo somzimba wezinyosi kanye nokunyuka kokufa, nemiphumela emibi kakhulu ehambisana ne-mancozeb ne-pyrithiopide.Sabe sesiqhathanisa ukuhlukahluka kwamagciwane ezibungu ezidla ukudla kwempova okulashwe nge-mancozeb nalezo ezidla ukudla okulawulwayo.Sixoxa ngezindlela ezingase zibe khona zokushona kanye nemithelela yokulawulwa kwezinambuzane ezididiyelwe kanye ne-pollinator (IPPM)36 izinhlelo.
Ama-O. cornifrons ama-cornifron ama-overwintering emifeceni atholwe e-Fruit Research Center, e-Biglerville, e-PA, futhi agcinwa ku-−3 kuya ku-2°C (±0.3°C).Ngaphambi kokuhlolwa (ama-cocoons angama-600 esewonke).NgoMeyi 2022, imifece engu-100 O. cornifrons yayidluliselwa nsuku zonke ezinkomishini zepulasitiki (imifece engu-50 inkomishi ngayinye, i-DI 5 cm × 15 cm ubude) futhi izinto zokusula zafakwa ngaphakathi kwezinkomishi ukuze kuthuthukiswe ukuvuleka nokuhlinzeka ngendawo ehlafunwayo, ukunciphisa ingcindezi etsheni. izinyosi37 .Beka izinkomishi ezimbili zepulasitiki eziqukethe imifece ekhejini lezinambuzane (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) neziphakeli ezingu-10 ml eziqukethe isixazululo se-sucrose esingu-50% futhi uzigcine izinsuku ezine ukuze uqinisekise ukuvala nokukhwelana.23°C, umswakama ohlobene 60%, photoperiod 10 l (amandla aphansi): 14 izinsuku.Izinsikazi eziyi-100 eziganiwe nezinduna zazidedelwa njalo ekuseni izinsuku eziyisithupha (i-100 ngosuku) zibe izidleke zokwenziwa ezimbili ngesikhathi sokuqhakaza kwezimbali ze-apula (isidleke somgibe: ububanzi 33.66 × ubude 30.48 × ubude 46.99 cm; Umfanekiso Owengeziwe 1).Kubekwe e-Pennsylvania State Arboretum, eduze kwe-cherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), ipentshisi (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), ipear (Pyrus perifolia 'Olympic', Pyrus i-perifolia 'Shinko', i-Pyrus perifolia 'Shinseiki'), isihlahla sama-apula se-coronaria (i-Malus coronaria) nezinhlobo eziningi zezihlahla zama-apula (i-Malus coronaria, i-Malus), isihlahla sama-apula sasekhaya 'Co-op 30' Enterprise™, i-apula i-Malus 'Co- Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy').Indlu ngayinye yezinyoni yepulasitiki eluhlaza okwesibhakabhaka ilingana phezu kwamabhokisi amabili okhuni.Ibhokisi lesidleke ngalinye linamashubhu ephepha e-kraft angu-800 angenalutho (evuliwe, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) afakwe kumashubhu e-cellophane opaque (0.7 OD bona amapulagi ePlastiki (T-1X plugs) ahlinzeka ngezindawo zokuzalela .
Womabili amabhokisi esidleke abheke ngasempumalanga futhi embozwe ngothango lwengadi yepulasitiki eluhlaza (imodeli ye-Everbilt #889250EB12, usayizi wokuvula 5 × 5 cm, 0.95 m × 100 m) ukuze kuvinjelwe ukungena kwamagundane nezinyoni futhi kubekwe phezu komhlabathi eduze kwebhokisi lesidleke senhlabathi. amabhokisi.Ibhokisi le-Nest (Umfanekiso Owengeziwe 1a).Amaqanda e-corn borer ayeqoqwa nsuku zonke ngokuqoqa amashubhu angu-30 ezidlekeni futhi awahambise endaweni yokucwaninga.Usebenzisa isikelo, yenza ukusikeka ekugcineni kweshubhu, bese uqaqa ishubhu elizungezayo ukuze uveze ama-brood cells.Amaqanda ngamanye kanye nempova yawo kwasuswa kusetshenziswa i-spatula egobile (ikhithi yamathuluzi e-Microslide, i-BioQuip Products Inc., California).Amaqanda afukanyelwa ephepheni lokuhlunga elimanzi futhi afakwa esitsheni sePetri amahora ama-2 ngaphambi kokuba asetshenziswe ekuhloleni kwethu (Umfanekiso Owengeziwe 1b-d).
Elabhorethri, sihlole ubuthi bomlomo bemithi ebulala isikhunta eyisithupha esetshenziswa ngaphambi nangesikhathi sokuqhakaza kwe-apula ezindaweni ezintathu (0.1X, 0.5X, kanye no-1X, lapho u-1X kuwuphawu olusetshenziswa ngamalitha ayi-100 amanzi/ihektare. Umthamo wensimu ephezulu = ukugxilisa ingqondo enkundleni)., Ithebula 1).Ukuhlushwa ngakunye kwaphindwa izikhathi ze-16 (n = 16).Ama-fungicides amabili okuxhumana (Ithebula S1: i-mancozeb 2696.14 ppm ne-captan 2875.88 ppm) kanye nezibulala-sikhunta ezine zesistimu (Ithebula S1: i-pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobupp5m8 cyprodinil 2; 2 azole m) ubuthi ezithelweni, imifino kanye nezitshalo zokuhlobisa .Senza impova ibe yi-homogenized sisebenzisa umshini wokugaya, sidlulisele u-0.20 g emthonjeni (24-well Falcon Plate), futhi sengeza futhi saxuba i-1 μL yesisombululo sesikhunta ukuze senze impova ye-pyramidal nemithombo ejulile engu-1 mm lapho amaqanda abekwe khona.Beka usebenzisa i-spatula encane (Umfanekiso Owengeziwe 1c,d).Amapuleti e-Falcon agcinwa ekamelweni lokushisa (25°C) kanye nomswakama ohlobene no-70%.Saziqhathanisa nezibungu zokulawula eziphakelwa ukudla kwempova okuxubile okuphathwe ngamanzi ahlanzekile.Sarekhoda ukufa futhi salinganisa isisindo sezibungu zonke izinsuku kuze kube yilapho izibungu zifinyelela iminyaka yangaphambi kokuzalwa zisebenzisa ibhalansi yokuhlaziya (I-Fisher Scientific, ukunemba = 0.0001 g).Ekugcineni, isilinganiso sobulili sahlolwa ngokuvula umfece ngemva kwezinyanga ezingu-2.5.
I-DNA ikhishwe kuzibungu eziphelele ze-O. cornifrons (n = 3 esimweni ngasinye sokwelashwa, impova elashwe i-mancozeb nengaphathwanga) futhi senza ukuhlaziya kokuhlukahluka kwamagciwane kulawa masampuli, ikakhulukazi ngenxa yokuthi ku-mancozeb ukufa okuphezulu kakhulu kwabonwa ezibungwini.ukuthola uMnZn.I-DNA iye yakhuliswa, yahlanzwa kusetshenziswa i-DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), futhi yahlelwa (imijikelezo engu-600) ku-Illumina® MiSeq™ kusetshenziswa ikhithi ye-v3.Ukulandelana okuhlosiwe kwezakhi zofuzo zebhaktheriya ze-16S ribosomal RNA kwenziwe kusetshenziswa i-Quick-16S™ NGS Library Prep Kit (i-Zymo Research, Irvine, CA) kusetshenziswa ama-primers aqondise isifunda se-V3-V4 sofuzo lwe-16S rRNA.Ukwengeza, ukulandelana kwe-18S kwenziwa kusetshenziswa i-10% yokufakwa kwe-PhiX, futhi ukukhulisa kwenziwa kusetshenziswa i-primer pair 18S001 kanye ne-NS4.
Ngenisa futhi ucubungule ukufunda39 usebenzisa ipayipi le-QIIME2 (v2022.11.1).Lokhu kufundwa kuye kwasikwa futhi kwahlanganiswa, futhi ukulandelana kwe-chimeric kwasuswa kusetshenziswa i-plugin ye-DADA2 ku-QIIME2 (qiime dada2 noise pairing)40.Izabelo zekilasi le-16S nele-18S zenziwe kusetshenziswa i-plugin ye-classify classify-sklearn kanye ne-artifact eqeqeshwe ngaphambilini i-silva-138-99-nb-classifier.
Yonke idatha yokuhlola ihlolelwe ukujwayelekile (i-Shapiro-Wilks) kanye ne-homogeneity yokuhluka (ukuhlolwa kuka-Levene).Ngenxa yokuthi isethi yedatha ayizange ihlangabezane nokucatshangelwa kokuhlaziywa kwepharamitha futhi ukuguqulwa kwehlulekile ukwenza izinsalela zifane, senze i-ANOVA (Kruskal-Wallis) enezindlela ezimbili ezingezona parametric enezici ezimbili [isikhathi (isigaba sesithathu, 5, nosuku olungu-8). amaphuzu esikhathi) nesibulala-sikhunta] ukuhlola umthelela wokwelashwa esisindweni esisha sezibungu, kwase kuba ukuqhathaniswa kwe-post hoc nonparametric pairwise kwenziwa kusetshenziswa ukuhlolwa kwe-Wilcoxon.Sisebenzise imodeli yomugqa ojwayelekile (i-GLM) ngokusatshalaliswa kwe-Poisson ukuze siqhathanise imiphumela yemithi yokubulala isikhunta ekusindeni kuzo zonke izici ezintathu zesikhunta41,42.Ngokuhlaziywa kobuningi obuhlukile, inani lokuhlukahluka kokulandelana kwe-amplicon (ASVs) liye lagoqwa ezingeni lohlobo.Ukuqhathaniswa kobuningi bokwahlukana phakathi kwamaqembu asebenzisa i-16S (izinga lohlobo lohlobo) kanye nobuningi obuhlobene obungu-18S kwenziwa kusetshenziswa imodeli yokwengeza evamile yesikhundla, isikali, nokuma (i-GAMLSS) enokusabalalisa komndeni kwe-beta zero-inflated (BEZI), okufaniswe ku-macro. .ku-Microbiome R43 (v1.1).1).Susa izinhlobo ze-mitochondrial ne-chloroplast ngaphambi kokuhlaziya okuhlukile.Ngenxa yamazinga e-taxonomic ahlukene we-18S, izinga eliphansi kuphela le-taxon ngayinye lisetshenziswe ekuhlaziyeni okuhlukile.Konke ukuhlaziya kwezibalo kwenziwa kusetshenziswa i-R (v. 3.4.3., iphrojekthi ye-CRAN) (Ithimba lango-2013).
Ukuchayeka ku-mancozeb, i-pyrithiostrobin, ne-trifloxystrobin kunciphisa kakhulu ukuzuza kwesisindo somzimba ku-O. cornifrons (Umfanekiso 1).Le miphumela yaqashelwa njalo kuyo yomithathu imithamo ehloliwe (Fig. 1a–c).I-Cyclostrobin ne-myclobutanil ayizange inciphise kakhulu isisindo sezibungu.
Isilinganiso sesisindo esisha sezibungu ze-stem borer esikalwa izikhathi ezintathu ngaphansi kokwelashwa okune okudliwayo (i-homogeneous pollen feed + fungicide: control, 0.1X, 0.5X kanye nemithamo engu-1X).(a) Umthamo ophansi (0.1X): iphuzu lokuqala (usuku 1): χ2: 30.99, DF = 6;P <0.0001, iphoyinti lesibili (usuku 5): 22.83, DF = 0.0009;okwesithathu;iphuzu (usuku 8): χ2: 28.39, DF = 6;(b) umthamo oyingxenye (0.5X): iphuzu lokuqala (usuku 1): χ2: 35.67, DF = 6;P <0.0001, iphuzu lesibili (usuku lokuqala).): χ2: 15.98, DF = 6;P = 0.0090;iphuzu lesithathu (usuku 8) χ2: 16.47, DF = 6;(c) Isizinda noma umthamo ogcwele (1X): iphuzu lokuqala (usuku 1) χ2: 20.64, P = 6;P = 0.0326, iphuzu lesibili (usuku 5): χ2: 22.83, DF = 6;P = 0.0009;iphuzu lesithathu (usuku 8): χ2: 28.39, DF = 6;ukuhlaziywa kwe-nonparametric kokuhluka.Amabha amele incazelo ± SE yokuqhathanisa ngakubili (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Ngesilinganiso esiphansi kakhulu (0.1X), isisindo somzimba wezibungu sehliswe ngo-60% nge-trifloxystrobin, 49% nge-mancozeb, 48% nge-myclobutanil, kanye nama-46% nge-pyrithistrobin (Fig. 1a).Lapho kuvezwa uhhafu womthamo wensimu (0.5X), isisindo somzimba wezibungu ze-mancozeb sehliswe ngo-86%, i-pyrithiostrobin ngo-52% kanye ne-trifloxystrobin ngo-50% (Fig. 1b).Umthamo wenkundla ogcwele (1X) we-mancozeb wehlise isisindo sobungu ngo-82%, i-pyrithiostrobin ngo-70%, kanye ne-trifloxystrobin, myclobutanil ne-sangard cishe ngama-30% (Fig. 1c).
Ukufa bekuphakeme kakhulu phakathi kwempova efakwe izibungu efakwe i-mancozeb, kulandele i-pyrithiostrobin ne-trifloxystrobin.Ukufa kwanda ngemithamo ekhulayo ye-mancozeb ne-pyritisoline (Fig. 2; Ithebula 2).Kodwa-ke, ukufa kwe-corn borer kukhuphuke kancane kuphela njengoba ukugxila kwe-trifloxystrobin kwanda;I-cyprodinil ne-captan ayizange ikhulise kakhulu ukufa uma kuqhathaniswa nemithi yokulawula.
Ukufa kwezibungu zempukane kwaqhathaniswa ngemva kokumunca impova ngayinye ephathwe ngama-fungicides ahlukene ayisithupha.I-Mancozeb ne-pentopyramide bekuzwela kakhulu ekuchayekeni komlomo ezimpethuni (i-GLM: χ = 29.45, DF = 20, P = 0.0059) (umugqa, ukuthambeka = 0.29, P <0.001; ukuthambeka = 0.24, P) <0.0.
Ngokwesilinganiso, kukho konke ukwelashwa, ama-39.05% eziguli kwakungabesifazane kanti ama-60.95% kwakungabesilisa.Phakathi kwemithi yokulawula, ingxenye yabesifazane ibingu-40% kuzo zombili izifundo zomthamo ophansi (0.1X) nohhafu wedosi (0.5X), kanye nama-30% ocwaningweni lwedosi yasensimini (1X).Ngomthamo ongu-0.1X, phakathi kwezibungu ezifunzwe impova eziphathwe nge-mancozeb ne-myclobutanil, u-33.33% wabantu abadala kwakungabantu besifazane, u-22% wabantu abadala kwakungabesifazane, u-44% wezibungu ezindala kwakuyizinsikazi, u-44% wezibungu ezindala kwakuyizinsikazi.zesifazane, ama-41% ezibungu ezindala kwakungezesifazane, futhi izilawuli zazingama-31% (Fig. 3a).Ezikhathini ezingu-0.5 zomthamo, ama-33% ezikelemu ezindala eqenjini le-mancozeb ne-pyrithiostrobin kwakungabesifazane, ama-36% eqenjini le-trifloxystrobin, ama-41% eqenjini le-myclobutanil, nama-46% eqenjini le-cyprostrobin.Lesi sibalo besingu-53% eqenjini.eqenjini lika-captan kanye ne-38% eqenjini lokulawula (Fig. 3b).Ngomthamo we-1X, i-30% yeqembu le-mancozeb kwakungabesifazane, i-36% yeqembu le-pyrithiostrobin, i-44% yeqembu le-trifloxystrobin, i-38% yeqembu le-myclobutanil, i-50% yeqembu lokulawula kwakungabesifazane - 38.5% (Fig. 3c) .
Iphesenti lezinsikazi nezinduna ngemva kokuchayeka kwesibulala-sikhunta.(a) Umthamo ophansi (0.1X).(b) Uhhafu wethamo (0.5X).(c) Umthamo wenkundla noma umthamo ogcwele (1X).
Ukuhlaziywa kokulandelana kwe-16S kubonise ukuthi iqembu lebhaktheriya lihlukile phakathi kwezibungu eziphakelwa impova ephethwe yi-mancozeb nezibungu eziphakelwa impova engalashwa (Fig. 4a).Inkomba ye-microbial yezibungu ezingalashwanga ezidliwe ngempova yayiphezulu kunaleyo yezibungu ezidla impova efakwe i-mancozeb (Fig. 4b).Nakuba umehluko obonwe ekucebeni phakathi kwamaqembu wawungabalulekile ngokwezibalo, wawuphansi kakhulu kunalowo obonwa izibungu ezidla impova engalashwa (Fig. 4c).Ubuningi obuhlobene bubonise ukuthi i-microbiota yezibungu ezidla impova yokulawula yayihluke kakhulu kunezibungu ezidla izibungu ezilashwe nge-mancozeb (Fig. 5a).Ukuhlaziywa okuchazayo kwembula ukuba khona kwesizukulwane se-28 ekulawuleni kanye namasampula enziwe nge-mancozeb (Fig. 5b).c Ukuhlaziya kusetshenziswa ukulandelana kwe-18S akuvezanga umehluko obalulekile (Umfanekiso Owengeziwe 2).
Amaphrofayela e-SAV asekelwe ekulandeleni kwe-16S aqhathaniswe nokunotha kwe-Shannon futhi kwabonwa ukunotha ezingeni le-phylum.(a) Ukuhlaziywa kwe-Principal coordinate (PCoA) okusekelwe kusakhiwo somphakathi we-microbial wonkana ku-pollen-feed or control (blue) kanye nezibungu ezifunzwa i-mancozeb (iwolintshi).Iphoyinti ledatha ngalinye limelela isampula ehlukile.I-PCoA ibalwe kusetshenziswa ibanga le-Bray-Curtis lokusabalalisa kuka-t okuxubile.Ama-oval amele izinga lokuzethemba elingu-80%.(b) I-Boxplot, idatha yengcebo ye-Shannon eluhlaza (amaphuzu) kanye no-c.Umcebo obonakalayo.Amabhokisi abonisa amabhokisi omugqa omaphakathi, ububanzi be-interquartile (IQR), kanye no-1.5 × IQR (n = 3).
Ukwakheka kwemiphakathi ye-microbial yezibungu ezidla impova eselashwe nge-mancozeb futhi engelashiwe.(a) Ubuningi obuhlobene bezakhi ze-microbial bufundeka ezibungwini.(b) Imephu yokushisa yemiphakathi ekhonjiwe yamagciwane.I-Delftia (isilinganiso sokungezwani (NOMA) = 0.67, P = 0.0030) kanye ne-Pseudomonas (NOMA = 0.3, P = 0.0074), i-Microbacterium (NOMA = 0.75, P = 0.0617) (NOMA = 1.5, P = 0.0060);Imigqa yemephu yokushisa ihlanganiswe kusetshenziswa ibanga lokuxhumanisa nokuxhumana okumaphakathi.
Imiphumela yethu ikhombisa ukuthi ukuchayeka ngomlomo ekuthinteni (i-mancozeb) kanye nezibulala-fungi ze-systemic (pyrostrobin kanye ne-trifloxystrobin), ezisetshenziswa kakhulu ngesikhathi sokuqhakaza, kunciphisa kakhulu ukukhuluphala kanye nokwanda kokufa kwezibungu zommbila.Ngaphezu kwalokho, i-mancozeb inciphise kakhulu ukuhlukahluka kanye nokunotha kwe-microbiome ngesikhathi sesigaba sangaphambi kokuzalwa.I-Myclobutanil, enye i-systemic fungicide, yehlisa kakhulu ukuzuza kwesisindo somzimba wezibungu kuyo yonke imithamo emithathu.Lo mphumela ubonakale ngesikhathi sesibili (usuku 5) nolwesithathu (usuku 8).Ngokuphambene, i-cyprodinil ne-captan ayizange inciphise kakhulu ukuzuza kwesisindo noma ukusinda uma kuqhathaniswa neqembu lokulawula.Ngokwazi kwethu, lo msebenzi ungowokuqala ukunquma imiphumela yamazinga ensimu emithi ehlukahlukene yokubulala isikhunta esetshenziselwa ukuvikela izitshalo zommbila ngokuchayeka kwempova eqondile.
Yonke imithi yokwelapha isikhunta yanciphisa kakhulu ukukhuphuka kwesisindo somzimba uma kuqhathaniswa nokwelapha okulawula.I-Mancozeb ibe nomthelela omkhulu ekuzuzeni isisindo somzimba wezibungu ngokuncipha okumaphakathi okungama-51%, kulandele i-pyrithiostrobin.Kodwa-ke, olunye ucwaningo aluzange lubike imiphumela emibi yemithamo yasensimini yezibulala-sikhunta ezigabeni zezibungu44.Nakuba i-dithiocarbamate biocides iboniswe inobuthi obuphansi obuphansi45, i-ethylene bisdithiocarbamates (EBDCS) efana ne-mancozeb ingakwazi ukwehlisela izinga le-urea ethylene sulfide.Uma kubhekwa imiphumela yawo eguquguqukayo kwezinye izilwane, lo mkhiqizo wokuwohloka ungase ube nesibopho semiphumela ebonwayo46,47.Ucwaningo lwangaphambilini lubonise ukuthi ukwakheka kwe-ethylene thiourea kuthonywa izici ezifana nezinga lokushisa eliphakeme48, amazinga womswakamo49 kanye nobude bokugcinwa komkhiqizo50.Izimo ezifanele zokugcina ama-biocides zinganciphisa le miphumela engemihle.Ngaphezu kwalokho, i-European Food Safety Authority izwakalise ukukhathazeka ngobuthi be-pyrithiopide, okuboniswe ukuthi i-carcinogenic ezinhlelweni zokugaya zezinye izilwane51.
Ukuphathwa ngomlomo kwe-mancozeb, i-pyrithiostrobin, ne-trifloxystrobin kwandisa ukufa kwezibungu ze-corn borer.Ngokuphambene, i-myclobutanil, i-ciprocycline ne-captan ayizange ibe nomthelela ekufeni.Le miphumela ihlukile kuleyo kaLadurner et al.52, owabonisa ukuthi u-captan wanciphisa kakhulu ukusinda komuntu omdala u-O. lignaria no-Apis mellifera L. (Hymenoptera, Apisidae).Ukwengeza, izibulala-sikhunta ezifana ne-captan ne-boscalid zitholwe zidala ukufa kwezibungu52,53,54 noma ziguqula indlela yokuziphatha55.Lezi zinguquko, nazo, zingathinta ikhwalithi yomsoco we-pollen futhi ekugcineni ukuzuza kwamandla esiteji sezibungu.Ukufa okubonwe eqenjini lokulawula kwakuhambisana nezinye izifundo ze-56,57.
Isilinganiso sobulili obuvuna abesilisa esibonwa emsebenzini wethu singachazwa ngezinto ezifana nokunganeli kokukhwelana nezimo zezulu ezimbi ngesikhathi sokuqhakaza, njengoba kwakusikiselwe ngaphambilini ku-O. cornuta nguVicents noBosch.Nakuba abesifazane nabesilisa ocwaningweni lwethu babenezinsuku ezine zokushada (isikhathi ngokuvamile esibhekwa njengesanele ukukhwelana okuphumelelayo), sehlise ngamabomu ukukhanya ukuze sinciphise ingcindezi.Nokho, lokhu kuguqulwa kungase kuphazamise inqubo yokukhwelana ngokungenhloso61.Ukwengeza, izinyosi ziba nezinsuku ezimbalwa zesimo sezulu esibi, okuhlanganisa imvula namazinga okushisa aphansi (<5°C), okungase futhi kube nomthelela omubi empumelelweni yokukhwelana4,23.
Nakuba ucwaningo lwethu lugxile kuyo yonke i-larval microbiome, imiphumela yethu inikeza ukuqonda ebudlelwaneni obungase bube phakathi kwemiphakathi yamagciwane engase ibaluleke ekondlekeni kwezinyosi kanye nokuchayeka kwesikhunta.Isibonelo, izibungu ezifunzwe impova ene-mancozeb zinciphise kakhulu ukwakheka komphakathi wamagciwane kanye nensada uma kuqhathaniswa nezibungu eziphakelwa impova engelashiwe.Ezibungwini ezidla impova engelashiwe, amaqembu ebhaktheriya i-Proteobacteria ne-Actinobacteria ayegqamile futhi ayesebenzisa kakhulu i-aerobic noma i-facultatively aerobic.Amagciwane e-Delft, ngokuvamile ahlotshaniswa nezinhlobo zezinyosi ezizimele zodwa, aziwa ngokuba nomsebenzi wama-antibiotic, okubonisa indima engaba khona yokuvikela emagciwaneni.Olunye uhlobo lwebhaktheriya, i-Pseudomonas, lwalugcwele izibungu ezidla impova engelashiwe, kodwa zancipha kakhulu ezibungwini ezilashwe nge-mancozeb.Imiphumela yethu isekela izifundo zangaphambilini ezikhomba i-Pseudomonas njengenye yesizukulwane esivame kakhulu ku-O. bicornis35 kanye neminye iminyovu ehlala yodwa34.Nakuba ubufakazi bokuhlola bendima ye-Pseudomonas empilweni ye-O. cornifrons abuzange bufundwe, leli bhaktheriya liye laboniswa ukukhuthaza ukuhlanganiswa kobuthi obuvikelayo ku-beetle Paederus fuscipes futhi kukhuthaze i-arginine metabolism in vitro 35, 65. Lokhu kuhlola kusikisela indima engaba khona ekuvikelweni kwegciwane kanye nebhaktheriya ngesikhathi sokukhula kwezibungu ze-O. cornifrons.I-Microbacterium ngolunye uhlobo oluhlonzwe ocwaningweni lwethu olubikwa ukuthi lukhona ngamanani aphezulu ezibunguni zempukane yamasosha amnyama ngaphansi kwezimo zendlala66.Ku-O. cornifrons izibungu, i-microbacteria ingase ibe nomthelela ekulinganiseni nasekuqineni kwe-gut microbiome ngaphansi kwezimo zokucindezeleka.Ukwengeza, i-Rhodococcus itholakala ezibungwini ze-O. cornifrons futhi yaziwa ngamakhono ayo okukhipha ubuthi obuyingozi67.Lolu hlobo luyatholakala futhi emathunjini e-A. florea, kodwa ngobuningi obuphansi kakhulu68.Imiphumela yethu ikhombisa ukuba khona kokuhlukahluka kwezakhi zofuzo eziningi kuzo zonke i-microbial taxa ezingashintsha izinqubo ze-metabolic ezibungwini.Nokho, ukuqonda okungcono kokuhlukahluka kokusebenza kwe-O. cornifron kuyadingeka.
Kafushane, imiphumela ibonisa ukuthi i-mancozeb, i-pyrithiostrobin, ne-trifloxystrobin yehlisa ukuzuzwa kwesisindo somzimba kanye nokunyuka kokushona kwezibungu ze-corn borer.Nakuba kunokukhathazeka okukhulayo mayelana nemiphumela yemithi ebulala isikhunta kuma-pollinator, kunesidingo sokuqonda kangcono imiphumela yensalela yama-metabolites alezi zinhlanganisela.Le miphumela ingafakwa ezincomweni zezinhlelo ezididiyelwe zokuphatha impova ezisiza abalimi ukuthi bagweme ukusetshenziswa kwemithi ethile yokubulala isikhunta ngaphambi nangesikhathi sokuqhakaza kwesihlahla sezithelo ngokukhetha ama-fungicides kanye nokushintsha isikhathi sokufakwa kwawo, noma ngokukhuthaza ukusetshenziswa kwezinye izindlela ezingalimazi kangako 36. Lolu lwazi kubalulekile ekuthuthukiseni izincomo.ekusetshenzisweni kwezibulala-zinambuzane, njengokulungisa izinhlelo ezikhona zokufutha kanye nokushintsha isikhathi sokufuthwa lapho kukhethwa izibulala-sikhunta noma ukukhuthaza ukusetshenziswa kwezinye izindlela ezingenabungozi kangako.Ucwaningo olwengeziwe luyadingeka emiphumeleni emibi yemithi yokubulala isikhunta esilinganisweni sobulili, indlela yokudla, i-gut microbiome, kanye nezindlela zamangqamuzana ezingaphansi kokuncipha kwesisindo se-corn borer nokufa.
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