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Ama-fungicides avame ukusetshenziswa ngesikhathi sokuqhakaza kwezithelo zesihlahla futhi angasongela izinambuzane ezithutha impova. Kodwa-ke, kuncane okwaziwayo ngendlela ama-pollinator angewona ama-bee (isb. izinyosi ezizimele, ama-Osmia cornifrons) asabela ngayo ekuthinteni kanye ne-systemic fungicides evame ukusetshenziswa kuma-apula ngesikhathi sokuqhakaza. Leli gebe lolwazi linciphisa izinqumo zomthetho ezinquma amazinga aphephile kanye nesikhathi sokufafaza i-fungicide. Sihlole imiphumela yama-fungicide amabili okuthintana (i-captan ne-mancozeb) kanye nama-fungicide amane aphakathi kwezingqimba/phytosystem (i-ciprocycline, i-myclobutanil, i-pyrostrobin kanye ne-trifloxystrobin). Imiphumela ekuzuzeni kwesisindo sama-larval, ukusinda, isilinganiso sobulili kanye nokuhlukahluka kwamagciwane. Ukuhlolwa kwenziwe kusetshenziswa i-bioassay yomlomo engapheli lapho impova yaphathwa khona ngemithamo emithathu ngokusekelwe kumthamo onconywayo njengamanje wokusetshenziswa ensimini (1X), uhhafu womthamo (0.5X) kanye nomthamo ophansi (0.1X). Yonke imithamo ye-mancozeb kanye ne-pyritisoline yehlise kakhulu isisindo somzimba kanye nokusinda kwama-larval. Sabe sesihlela i-gene ye-16S ukuze sichaze i-bacteriome yama-larval ye-mancozeb, i-fungicide ebangela ukufa okuphezulu kakhulu. Sithole ukuthi ukuhlukahluka kwamagciwane kanye nobuningi bawo kuncishiswe kakhulu ezimpungusheni ezidla impova ephathwe yi-mancozeb. Imiphumela yethu yelabhorethri ikhombisa ukuthi ukufafaza amanye ala ma-fungicide ngesikhathi sokuqhakaza kuyingozi kakhulu empilweni ye-O. cornifrons. Lolu lwazi lubalulekile ezinqumweni zokuphatha zesikhathi esizayo maqondana nokusetshenziswa okuqhubekayo kwemikhiqizo yokuvikela izihlahla zezithelo futhi lusebenza njengesisekelo sezinqubo zokulawula ezihlose ukuvikela ama-pollinator.
Inyosi eyodwa yama-mason i-Osmia cornifrons (Hymenoptera: Megachilidae) yethulwa e-United States isuka eJapan ngasekupheleni kwawo-1970 nasekuqaleni kwawo-1980, futhi lolu hlobo ludlale indima ebalulekile ekuthuthweni kwempova ezindaweni zemvelo eziphethwe kusukela ngaleso sikhathi. Izinhlobo zale nyosi ezizimele ziyingxenye yezinhlobo ezingaba ngu-50 zezinyosi zasendle ezihambisana nezinyosi ezithutha impova ezivandeni zama-alimondi nama-apula e-United States2,3. Izinyosi zeMason zibhekene nezinselele eziningi, okuhlanganisa ukuhlukana kwendawo yokuhlala, amagciwane, kanye nezibulala-zinambuzane3,4. Phakathi kwezibulala-zinambuzane, izibulala-zinambuzane zinciphisa ukutholakala kwamandla, ukudla ukudla5 kanye nokulungisa umzimba6,7. Nakuba ucwaningo lwamuva lubonisa ukuthi impilo yezinyosi zeMason ithonywa ngqo yizinambuzane ezincane ezihlala njalo kanye neziphilayo, 8,9 ngoba amabhaktheriya kanye nezikhunta kungathonya ukudla okunomsoco kanye nezimpendulo zomzimba, imiphumela yokuchayeka kwezikhunta ekuhlukeni kwamagciwane ezinyosi zeMason isanda kuqala ukufundwa.
Imithi yokubulala isikhunta enemiphumela eyahlukahlukene (ukuthintana kanye neyomzimba) ifuthwa ezivandeni zezithelo ngaphambi nangesikhathi sokuqhakaza ukuze kwelashwe izifo ezifana ne-apula scab, ukubola okubabayo, ukubola okunsundu kanye ne-powdery mildew10,11. Imithi yokubulala isikhunta ibhekwa njengengenangozi kubathuthi bezinambuzane, ngakho-ke inconywa kubalimi ngesikhathi sokuqhakaza; Ukuchayeka kanye nokugwinywa kwalezi mithi yokubulala isikhunta yizinyosi kwaziwa kahle, njengoba kuyingxenye yenqubo yokubhalisa izibulala-zinambuzane yi-US Environmental Protection Agency kanye nezinye izinhlangano eziningi zokulawula zikazwelonke12,13,14. Kodwa-ke, imiphumela yemithi yokubulala isikhunta kwabangebona izinyosi ayaziwa kangako ngoba ayidingeki ngaphansi kwezivumelwano zokugunyazwa kokumaketha e-United States15. Ngaphezu kwalokho, ngokuvamile azikho izindlela zokuhlola izinyosi ezilodwa16,17, futhi ukugcina amakoloni ahlinzeka ngezinyosi ukuze zihlolwe kuyinselele18. Ukuhlolwa kwezinyosi ezahlukene ezilawulwayo kuqhutshwa kabanzi eYurophu nase-USA ukuze kufundwe imiphumela yemithi yokubulala izinambuzane ezinyosini zasendle, futhi izindlela zokuhlola ezijwayelekile zisanda kuthuthukiswa ze-O. cornifrons19.
Izinyosi ezinezimpondo ziyi-monocytes futhi zisetshenziswa kwezentengiselwano ezitshalweni ze-carp njengesengezo noma esikhundleni sezinyosi zoju. Lezi zinyosi ziphuma phakathi kukaMashi no-Ephreli, kanti iziduna ezikhula ngokushesha ziphuma ezinsukwini ezintathu kuya kwezine ngaphambi kwezinsikazi. Ngemva kokuhlangana, insikazi iqoqa impova kanye ne-nectar ukuze inikeze uchungechunge lwamaseli ezinsana ngaphakathi kwesidleke se-tubular (semvelo noma sokwenziwa)1,20. Amaqanda abekwa phezu kwempova ngaphakathi kwamaseli; insikazi yakha udonga lobumba ngaphambi kokulungiselela iseli elilandelayo. Izibungu zokuqala ezingaphakathi zivalelwe ku-chorion futhi zondla uketshezi lwe-embryonic. Kusukela ku-instar yesibili kuya kweyesihlanu (prepupa), izibungu zondla impova22. Uma ukutholakala kwempova sekuphelile ngokuphelele, izibungu zakha ama-cocoon, ama-pupate futhi ziphuma njengabantu abadala ekamelweni elifanayo lezibungu, ngokuvamile ekupheleni kwehlobo20,23. Abantu abadala baphuma entwasahlobo elandelayo. Ukusinda kwabantu abadala kuhlotshaniswa nokuzuza kwamandla aphelele (ukuzuza isisindo) ngokusekelwe ekudleni. Ngakho-ke, ikhwalithi yokudla okunempilo yempova, kanye nezinye izici ezifana nesimo sezulu noma ukuchayeka ezibulala-zinambuzane, kuyizinto ezinquma ukusinda kanye nempilo24.
Izibulala-zinambuzane kanye nezibulala-fungi ezisetshenziswa ngaphambi kokuqhakaza ziyakwazi ukuhamba ngaphakathi kwemithambo yezitshalo ngamazinga ahlukahlukene, kusukela ku-translaminar (isb., ukukwazi ukuhamba kusuka phezulu kwamaqabunga kuya phansi, njengamanye ama-fungicides) 25 kuya emiphumeleni yesistimu yangempela. , engangena emqheleni kusukela ezimpandeni, ingangena ku-nectar yezimbali ze-apula 26, lapho ingabulala khona ama-o. cornishfrons amadala 27. Ezinye izibulala-zinambuzane nazo zingena empoveni, zithinta ukukhula kwezibungu zommbila futhi zibangele ukufa kwazo 19. Ezinye izifundo zikhombisile ukuthi ezinye izibulala-fungi zingashintsha kakhulu ukuziphatha kokuzalela kwezinhlobo ezihlobene ne-o. lignaria 28. Ngaphezu kwalokho, izifundo zelebhu kanye nezifundo zasensimini ezilingisa izimo zokuchayeka kwezibulala-zinambuzane (kufaka phakathi ama-fungicides) zikhombisile ukuthi izibulala-zinambuzane zithinta kabi i-physiology 22 morphology 29 kanye nokusinda kwezinyosi zoju kanye nezinye izinyosi ezizimele. Izifutho ezahlukahlukene zokubulala isikhunta ezisetshenziswa ngqo ezimbalini ezivulekile ngesikhathi sokuqhakaza zingangcolisa impova eqoqwe ngabantu abadala ukuze kuthuthukiswe izibungu, imiphumela yazo esazofundwa 30.
Kuyaziwa kabanzi ukuthi ukuthuthukiswa kwama-larval kuthonywa yi-pollen kanye nemiphakathi yama-microbial ohlelweni lokugaya ukudla. I-honeybee microbiome ithonya imingcele efana nobunzima bomzimba31, izinguquko ze-metabolic22 kanye nokuthambekela kwamagciwane32. Izifundo zangaphambilini zihlole ithonya lesigaba sokukhula, izakhamzimba, kanye nendawo ezungezile kuma-microbiome ezinyosi ezizimele. Lezi zifundo zembule ukufana esakhiweni kanye nobuningi bama-larval kanye nama-pollen microbiomes33, kanye nohlobo lwebhaktheriya oluvame kakhulu i-Pseudomonas ne-Delftia, phakathi kwezinhlobo zezinyosi ezizimele. Kodwa-ke, yize ama-fungicides ehlotshaniswa namasu okuvikela impilo yezinyosi, imiphumela yama-fungicides kuma-larval microbiota ngokuchayeka ngqo emlonyeni ayikahlolwa.
Lolu cwaningo luhlole imiphumela yemithamo yangempela yemithi eyisithupha evame ukusetshenziswa yokubulala izinambuzane ebhaliswe ukusetshenziswa ezithelweni zezihlahla e-United States, okuhlanganisa imithi yokubulala izinambuzane ethintanayo kanye neyohlelo enikezwa ngomlomo ezibungwini ze-corn hornworm moth ezivela ekudleni okungcolile. Sithole ukuthi imithi yokubulala izinambuzane ethintanayo kanye neyohlelo inciphisa ukukhuluphala komzimba wezinyosi kanye nokufa okwandisiwe, kanye nemiphumela emibi kakhulu ehlotshaniswa ne-mancozeb kanye ne-pyrithiopide. Sabe sesiqhathanisa ukuhlukahluka kwamagciwane ezibungwini ezidla ukudla kwempova okuphathwe yi-mancozeb kanye nalezo ezidla ukudla okulawulayo. Sixoxa ngezindlela ezingaba khona ezibangela ukufa kanye nemiphumela yezinhlelo zokuphathwa kwezinambuzane kanye nempova ezihlanganisiwe (IPPM)36.
Ama-O. cornifron amadala ahlala ebusika kuma-cocoon atholakale e-Fruit Research Center, eBiglerville, PA, futhi agcinwa ku-−3 kuya ku-2°C (±0.3°C). Ngaphambi kokuhlolwa (ama-cocoon angu-600 esewonke). NgoMeyi 2022, ama-cocoon angu-100 e-O. cornifron ayedluliselwa nsuku zonke ezinkomishini zepulasitiki (ama-cocoon angu-50 ngendebe, i-DI 5 cm × 15 cm ubude) futhi ama-wipe afakwa ngaphakathi kwezinkomishi ukukhuthaza ukuvulwa nokuhlinzeka nge-substrate ehlafunwayo, ukunciphisa ukucindezeleka ezinyosini ezinamatshe. Beka izinkomishi ezimbili zepulasitiki eziqukethe ama-cocoon ekhejini lezinambuzane (30 × 30 × 30 cm, i-BugDorm MegaView Science Co. Ltd., eTaiwan) kanye ne-feeders engu-10 ml equkethe isisombululo se-sucrose esingu-50% bese ugcina izinsuku ezine ukuqinisekisa ukuvalwa nokuhlangana. 23°C, umswakama ohlobene 60%, isikhathi se-photoperiod 10 l (ukuqina okuphansi): izinsuku ezingu-14. Izinsikazi eziyi-100 nezinduna ezizalwe ndawonye zazikhululwa njalo ekuseni izinsuku eziyisithupha (100 ngosuku) zifakwe ezidlekeni ezimbili zokwenziwa ngesikhathi sokuqhakaza kwama-apula aphezulu (isidleke somgibe: ububanzi 33.66 × ukuphakama 30.48 × ubude 46.99 cm; Isithombe Esengeziwe 1). Zibekwe ePennsylvania State Arboretum, eduze kwe-cherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), ipentshisi (Prunus persica 'Contender'), iPrunus persica 'PF 27A' Flamin Fury®), ipentshisi (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), coronaria apple tree (Malus coronaria) kanye nezinhlobo eziningi zezihlahla zama-apula (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'). Indlu ngayinye yezinyoni yepulasitiki eluhlaza okwesibhakabhaka ingena phezu kwamabhokisi amabili okhuni. Ibhokisi ngalinye lesidleke laliqukethe amashubhu ephepha le-kraft angenalutho angu-800 (avulekile ngokuzungeza, angu-0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) afakwe kumashubhu e-cellophane angacacile (0.7 OD bheka ama-plug epulasitiki (ama-plug e-T-1X) ahlinzeka ngezindawo zokuzalela.
Womabili amabhokisi ezidleke abheke empumalanga futhi ambozwe ngocingo lwengadi oluluhlaza lwepulasitiki (imodeli ye-Everbilt #889250EB12, usayizi wokuvula ongu-5 × 5 cm, 0.95 m × 100 m) ukuvimbela ukufinyelela kwamagundane nezinyoni futhi abekwa ebusweni bomhlabathi eduze kwamabhokisi omhlabathi webhokisi lezidleke. Ibhokisi lezidleke (Isithombe Esingeziwe 1a). Amaqanda e-corn borer ayeqoqwa nsuku zonke ngokuqoqa amashubhu angu-30 ezidlekeni bese ewathutha aye elabhorethri. Usebenzisa isikele, sika ekugcineni kwethubhu, bese uhlukanisa ithubhu elijikelezayo ukuze uveze amaseli ezinsana. Amaqanda ngamanye kanye nempova yawo kwasuswa kusetshenziswa i-spatula egobile (ikhithi yamathuluzi e-Microslide, i-BioQuip Products Inc., eCalifornia). Amaqanda afakwa ephepheni lokuhlunga elimanzi futhi afakwa esitsheni se-Petri amahora ama-2 ngaphambi kokusetshenziswa ekuhlolweni kwethu (Isithombe Esingeziwe 1b-d).
Elabhorethri, sihlole ubuthi bomlomo bemithi yokubulala isikhunta eyisithupha esetshenziswe ngaphambi nangesikhathi sokuqhakaza kwe-apula ngamanani amathathu (0.1X, 0.5X, kanye no-1X, lapho u-1X kuwuphawu olusetshenziswa ngamalitha ayi-100 amanzi/ihektare. Umthamo omkhulu wensimu = ukugcwala ensimini). , Ithebula 1). Ukugcwala ngakunye kuphindwe izikhathi eziyi-16 (n = 16). Ama-fungicide amabili okuthintana (Ithebula S1: mancozeb 2696.14 ppm kanye ne-captan 2875.88 ppm) kanye nama-fungicide amane ahlelekile (Ithebula S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75 .12 ppm; cyprodinil 280.845 ppm) ubuthi ezithelweni, emifino nasezitshalweni zokuhlobisa. Sihlanganise impova ngendlela efanayo sisebenzisa umshini wokugaya, sadlulisela u-0.20 g emthonjeni (i-Falcon Plate enama-well angu-24), sabe sesengeza futhi saxuba i-1 μL yesisombululo se-fungicide ukuze sakhe impova ye-pyramidal enemigodi ejulile engu-1 mm lapho kwafakwa khona amaqanda. Beka usebenzisa i-spatula encane (Isithombe Esingeziwe 1c, d). Amapuleti e-Falcon agcinwe ekushiseni kwegumbi (25°C) kanye nomswakama ohlobene ongu-70%. Siwaqhathanise nezibungu ezilawulayo eziphakelwa ukudla kwempova okufanayo okuphathwe ngamanzi ahlanzekile. Saqopha ukufa futhi salinganisa isisindo sezibungu njalo ngemva kwezinsuku ezimbili kwaze kwaba yilapho izibungu zifinyelela eminyakeni yobudala sisebenzisa ibhalansi yokuhlaziya (Fisher Scientific, ukunemba = 0.0001 g). Ekugcineni, isilinganiso sobulili sahlolwa ngokuvula i-cocoon ngemva kwezinyanga ezingu-2.5.
I-DNA ikhishwe ezibungwini ze-O. cornifrons eziphelele (n = 3 ngesimo ngasinye sokwelashwa, impova ephathwe nge-mancozeb nengaphathwanga) futhi senze ukuhlaziywa kokuhlukahluka kwamagciwane kulezi zibonelo, ikakhulukazi ngoba ku-mancozeb ukufa okuphezulu kakhulu kwabonwa ezibungwini. kuthola i-MnZn. I-DNA yandiswa, yahlanzwa kusetshenziswa i-DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), futhi yahlelwa ngokulandelana (imijikelezo engu-600) ku-Illumina® MiSeq™ kusetshenziswa i-v3 kit. Ukulandelana okuqondiwe kwezakhi zofuzo ze-ribosomal RNA ze-bacterial 16S kwenziwa kusetshenziswa i-Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) kusetshenziswa ama-primer aqondise esifundeni se-V3-V4 sezakhi zofuzo ze-16S rRNA. Ngaphezu kwalokho, ukulandelana kwe-18S kwenziwa kusetshenziswa i-10% PhiX inclusion, futhi ukukhulisa kwenziwa kusetshenziswa i-primer pair 18S001 kanye ne-NS4.
Ngenisa futhi ucubungule ama-reads abhangqiwe39 usebenzisa ipayipi le-QIIME2 (v2022.11.1). Lawa ma-reads ancishisiwe futhi ahlanganiswa, futhi ama-sequence e-chimeric asusiwe kusetshenziswa i-plugin ye-DADA2 ku-QIIME2 (ukubhangqa komsindo we-qiime dada2)40. Izabelo zekilasi le-16S kanye ne-18S zenziwe kusetshenziswa i-plugin ye-object classifier Classify-sklearn kanye ne-artifact silva-138-99-nb-classifier eqeqeshwe kusengaphambili.
Yonke idatha yokuhlola ihlolwe ukuthi ijwayelekile (uShapiro-Wilks) kanye nokulingana kokuhlukahluka (ukuhlolwa kukaLevene). Ngenxa yokuthi isethi yedatha ayizange ihlangabezane nokucatshangwa kokuhlaziywa kwe-parametric futhi ukuguqulwa kwehlulekile ukulinganisa ama-residues, senze i-ANOVA (Kruskal-Wallis) engeyona i-parametric enezindlela ezimbili ezinezici ezimbili [isikhathi (amaphuzu esikhathi esigaba sesithathu sesi-2, sesi-5, kanye nesi-8) kanye ne-fungicide] ukuze sihlole umphumela wokwelashwa esisindweni esisha sama-larval, kwabe sekwenziwa ukuqhathaniswa kwe-post hoc nonparametric pairwise kusetshenziswa ukuhlolwa kwe-Wilcoxon. Sisebenzise imodeli eqondile ejwayelekile (i-GLM) ene-Poisson distribution ukuze siqhathanise imiphumela ye-fungicide ekusindeni kuwo wonke amazinga amathathu e-fungicide41,42. Ukuze kuhlaziywe ukuchichima okuhlukile, inani le-amplicon sequence variants (ama-ASV) lawa ezingeni lohlobo. Ukuqhathaniswa kobuningi bokuhluka phakathi kwamaqembu kusetshenziswa i-16S (izinga lohlobo) kanye nobuningi obuhlobene be-18S kwenziwe kusetshenziswa imodeli yokwengeza ejwayelekile yesikhundla, isikali, kanye nesimo (GAMLSS) ngokusatshalaliswa komndeni we-beta zero-inflated (BEZI), okwaklanywa ku-macro. ku-Microbiome R43 (v1.1). 1). Susa izinhlobo ze-mitochondrial kanye ne-chloroplast ngaphambi kokuhlaziywa kokungafani. Ngenxa yamazinga ahlukene e-taxonomic e-18S, kwasetshenziswa izinga eliphansi kakhulu le-taxon ngayinye ekuhlaziyweni kokungafani. Zonke izibalo zokuhlaziya zenziwa kusetshenziswa i-R (v. 3.4.3., iphrojekthi ye-CRAN) (Ithimba 2013).
Ukuchayeka ku-mancozeb, i-pyrithiostrobin, kanye ne-trifloxystrobin kunciphisa kakhulu ukutholakala kwesisindo somzimba ku-O. cornifrons (Isithombe 1). Le miphumela ibonwe njalo kuzo zonke izilinganiso ezintathu ezihlolwe (Isithombe 1a–c). I-Cyclostrobin kanye ne-myclobutanil azizange zinciphise kakhulu isisindo sezibungu.
Isisindo esimaphakathi sesibungu se-stem borer silinganiswa ngezikhathi ezintathu ngaphansi kokwelashwa okune kokudla (ukudla kwempova okufanayo + isikhunta: ukulawula, imithamo engu-0.1X, 0.5X kanye ne-1X). (a) Isilinganiso esiphansi (0.1X): iphuzu lokuqala (usuku 1): χ2: 30.99, DF = 6; P < 0.0001, iphuzu lesibili (usuku 5): 22.83, DF = 0.0009; okwesithathu; iphuzu (usuku 8): χ2: 28.39, DF = 6; (b) isigamu sesilinganiso (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 lesikhathi lesithathu (usuku 8) χ2: 16.47, DF = 6; (c) Isiza noma umthamo ogcwele (1X): iphuzu lesikhathi sokuqala (usuku 1) χ2: 20.64, P = 6; P = 0.0326, iphuzu lesikhathi lesibili (usuku 5): χ2: 22.83, DF = 6; P = 0.0009; iphuzu lesikhathi lesithathu (usuku 8): χ2: 28.39, DF = 6; ukuhlaziywa okungeyona ipharamitha kokuhlukahluka. Amabha amelela isilinganiso ± SE sokuqhathaniswa okubili (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Ngesilinganiso esiphansi kakhulu (0.1X), isisindo somzimba wama-larval sehlisiwe ngo-60% nge-trifloxystrobin, 49% nge-mancozeb, 48% nge-myclobutanil, kanye no-46% nge-pyrithistrobin (Isithombe 1a). Lapho kuvezwa isigamu sesilinganiso sensimu (0.5X), isisindo somzimba sama-larvae e-mancozeb sehlisiwe ngo-86%, i-pyrithiostrobin ngo-52% kanye ne-trifloxystrobin ngo-50% (Isithombe 1b). Isilinganiso esigcwele sensimu (1X) se-mancozeb sehlise isisindo sama-larval ngo-82%, i-pyrithiostrobin ngo-70%, kanye ne-trifloxystrobin, i-myclobutanil kanye ne-sangard cishe ngo-30% (Isithombe 1c).
Ukufa bekuphezulu kakhulu phakathi kwezibungu ezidliwe impova ephathwe nge-mancozeb, kulandelwe yi-pyrithiostrobin kanye ne-trifloxystrobin. Ukufa kwanda ngokwandisa imithamo ye-mancozeb kanye ne-pyritisoline (Isithombe 2; Ithebula 2). Kodwa-ke, ukufa kwe-corn borer kwanda kancane njengoba amazinga e-trifloxystrobin anda; i-cyprodinil kanye ne-captan akuzange kukhulise kakhulu ukufa uma kuqhathaniswa nokwelashwa kokulawula.
Ukufa kwezibungu ze-borer fly kwaqhathaniswa ngemva kokungenisa impova ngayinye elashwe ngemithi eyisithupha ehlukene yokubulala isikhunta. I-Mancozeb ne-pentopyramide zazizwela kakhulu ekuvezweni ngomlomo yizimpethu zommbila (GLM: χ = 29.45, DF = 20, P = 0.0059) (umugqa, umthambeka = 0.29, P < 0.001; umthambeka = 0.24, P <0.00)).
Ngokwesilinganiso, kuzo zonke izindlela zokwelapha, ama-39.05% eziguli kwakungabesifazane kanti ama-60.95% kwakungabesilisa. Phakathi kwezindlela zokwelapha zokulawula, isilinganiso sabesifazane sasingu-40% kokubili ezifundweni zomthamo ophansi (0.1X) kanye nowesigamu (0.5X), kanye nama-30% ezifundweni zomthamo wasensimini (1X). Kumthamo we-0.1X, phakathi kwezibungu ezidla impova eziphathwe nge-mancozeb kanye ne-myclobutanil, ama-33.33% abantu abadala kwakungabesifazane, ama-22% abantu abadala kwakungabesifazane, ama-44% ezibungu ezindala kwakungabesifazane, ama-44% ezibungu ezindala kwakungabesifazane. abesifazane, ama-41% ezibungu ezindala kwakungabesifazane, kanti abalawulayo babengu-31% (Isithombe 3a). Ngesikhathi somthamo ophindwe izikhathi ezingu-0.5, ama-33% ezikelemu ezindala eqenjini le-mancozeb kanye ne-pyrithiostrobin kwakungabesifazane, ama-36% eqenjini le-trifloxystrobin, ama-41% eqenjini le-myclobutanil, kanye nama-46% eqenjini le-cyprostrobin. Lesi sibalo sasingu-53% eqenjini. eqenjini le-captan kanye nama-38% eqenjini lokulawula (Isithombe 3b). Kumthamo we-1X, ama-30% eqenjini le-mancozeb kwakungabesifazane, ama-36% eqenjini le-pyrithiostrobin, ama-44% eqenjini le-trifloxystrobin, ama-38% eqenjini le-myclobutanil, ama-50% eqenjini lokulawula kwakungabesifazane - ama-38.5% (Isithombe 3c).
Iphesenti lezimbobo zesifazane nesesilisa ngemva kokuchayeka ku-fungicide esigabeni sama-larval. (a) Isilinganiso esiphansi (0.1X). (b) Isilinganiso esiyingxenye (0.5X). (c) Isilinganiso sasensimini noma isilinganiso esigcwele (1X).
Ukuhlaziywa kokulandelana kwe-16S kubonise ukuthi iqembu lamagciwane lihlukile phakathi kwezibungu eziphakelwa impova ephathwe nge-mancozeb kanye nezibungu eziphakelwa impova engelashwanga (Isithombe 4a). Inkomba yamagciwane ezibungu ezingalashwanga eziphakelwa impova yayiphezulu kuneyezibungu eziphakelwa impova ephathwe nge-mancozeb (Isithombe 4b). Nakuba umehluko obonwe ekucebeni phakathi kwamaqembu wawungabalulekile ngokwezibalo, wawuphansi kakhulu kunalowo obonwe yizibungu eziphakelwa impova engelashwanga (Isithombe 4c). Ubuningi obuhlobene bubonise ukuthi i-microbiota yezibungu eziphakelwa impova elawulwayo yayihluke kakhulu kuneyezibungu eziphakelwa izibungu eziphathwe nge-mancozeb (Isithombe 5a). Ukuhlaziywa okuchazayo kwembule ukuthi kukhona izinhlobo ezingu-28 ezilawulwayo kanye namasampula aphathwe nge-mancozeb (Isithombe 5b). c Ukuhlaziywa kusetshenziswa ukulandelana kwe-18S akuvezi mehluko obalulekile (Isithombe Esingeziwe 2).
Amaphrofayili e-SAV asekelwe ku-sequences ye-16S aqhathaniswa nokuceba kwe-Shannon futhi aqaphela ukuceba ezingeni le-phylum. (a) Ukuhlaziywa kwe-Principal coordinate (PCoA) okusekelwe esakhiweni somphakathi we-microbial jikelele ku-pollen-fed noma control (blue) kanye ne-mancozeb-fed larvae (orange). Iphuzu ngalinye ledatha limelela isampula ehlukile. I-PCoA ibalwe kusetshenziswa ibanga le-Bray-Curtis lokusatshalaliswa kwe-t multivariate. Ama-oval amelela izinga lokuzethemba elingu-80%. (b) I-Boxplot, idatha yengcebo ye-Shannon eluhlaza (amaphuzu) kanye no-c. Ingcebo ebonakalayo. Ama-boxplot abonisa amabhokisi omugqa ophakathi, ububanzi be-interquartile (IQR), kanye no-1.5 × IQR (n = 3).
Ukwakheka kwemiphakathi yamagciwane ezibungu ezidla impova ephathwe nge-mancozeb nengaphathwanga. (a) Ubuningi bezinhlobo zamagciwane bufundeka ezibunguni. (b) Imephu yokushisa yemiphakathi yamagciwane etholakele. I-Delftia (isilinganiso samathuba (OR) = 0.67, P = 0.0030) kanye ne-Pseudomonas (OR = 0.3, P = 0.0074), i-Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060); Imigqa yemephu yokushisa ihlanganiswe ngokusebenzisa ibanga lokuhlobana kanye nokuxhumana okumaphakathi.
Imiphumela yethu ikhombisa ukuthi ukuchayeka ngomlomo kuma-fungicides okuthintana (i-mancozeb) kanye nama-fungicides asebenzisa i-systemic (pyrostrobin kanye ne-trifloxystrobin), asetshenziswa kabanzi ngesikhathi sokuqhakaza, kunciphisa kakhulu ukukhuluphala kanye nokwanda kokufa kwama-larvae ommbila. Ngaphezu kwalokho, i-mancozeb inciphise kakhulu ukuhlukahluka kanye nokuceba kwe-microbiome ngesikhathi sokulungiselela. I-Myclobutanil, enye i-fungicide esebenzisa i-systemic, inciphise kakhulu ukukhuluphala komzimba wama-larvae kuzo zonke izilinganiso ezintathu. Lo mphumela wabonakala ezikhathini zesibili (usuku 5) kanye nesithathu (usuku 8). Ngokuphambene nalokho, i-cyprodinil kanye ne-captan azizange zinciphise kakhulu ukukhuluphala noma ukusinda uma kuqhathaniswa neqembu lokulawula. Ngokwazi kwethu, lo msebenzi ungowokuqala ukunquma imiphumela yamazinga ensimu yama-fungicide ahlukene asetshenziswa ukuvikela izitshalo zommbila ngokuchayeka ngqo empova.
Zonke izindlela zokwelapha nge-fungicide zinciphise kakhulu ukukhuluphala komzimba uma kuqhathaniswa nezindlela zokwelapha zokulawula. I-Mancozeb ibe nomthelela omkhulu ekukhuleni kwesisindo somzimba we-larval ngokunciphisa okumaphakathi okungu-51%, kulandelwe yi-pyrithiostrobin. Kodwa-ke, ezinye izifundo azizange zibike imiphumela emibi yemithamo yasensimini ye-fungicides ezigabeni ze-larval44. Nakuba ama-biocides e-dithiocarbamate aboniswe ukuthi anobuthi obuphansi kakhulu45, i-ethylene bisdithiocarbamates (EBDCS) njenge-mancozeb ingonakalisa ibe yi-urea ethylene sulfide. Njengoba ibangelwa yimiphumela yayo yokuguquka kwezinye izilwane, lo mkhiqizo wokonakala ungase ube yimbangela yemiphumela ebonwe46,47. Izifundo zangaphambilini zibonise ukuthi ukwakheka kwe-ethylene thiourea kuthonywa yizici ezifana nokushisa okuphezulu48, amazinga omswakama49 kanye nobude bokugcinwa komkhiqizo50. Izimo ezifanele zokugcina ama-biocides zinganciphisa le miphumela emibi. Ngaphezu kwalokho, i-European Food Safety Authority iveze ukukhathazeka ngobuthi be-pyrithiopide, okuye kwabonakala ukuthi ibangela umdlavuza ezinhlelweni zokugaya ukudla zezinye izilwane51.
Ukuphathwa ngomlomo kwe-mancozeb, i-pyrithiostrobin, kanye ne-trifloxystrobin kwandisa ukufa kwezibungu ze-corn borer. Ngokuphambene nalokho, i-myclobutanil, i-ciprocycline kanye ne-captan azizange zibe nomthelela ekufeni. Le miphumela ihlukile kwekaLadurner et al.52, ababonise ukuthi i-captan inciphise kakhulu ukusinda kwe-O. lignaria yabantu abadala kanye ne-Apis mellifera L. (Hymenoptera, Apisidae). Ngaphezu kwalokho, ama-fungicide afana ne-captan kanye ne-boscalid atholakale ebangela ukufa kwezibungu52,53,54 noma ashintshe indlela yokudla55. Lezi zinguquko, nazo, zingathinta ikhwalithi yokudla kwempova futhi ekugcineni kuzuze amandla esigaba sezibungu. Ukufa okubonwe eqenjini lokulawula kwakuhambisana nezinye izifundo 56,57.
Isilinganiso sobulili esithandwa ngabesilisa esibonwe emsebenzini wethu singachazwa yizici ezifana nokuzalana okunganele kanye nezimo zezulu ezimbi ngesikhathi sokuqhakaza, njengoba kwaphakanyiswa ngaphambili nge-O. cornuta nguVicens noBosch. Nakuba abesifazane nabesilisa ocwaningweni lwethu babenezinsuku ezine zokuzalana (isikhathi ngokuvamile esibhekwa njengesanele ukuzalana okuphumelelayo), sinciphise ngamabomu ukukhanya ukuze sinciphise ukucindezeleka. Kodwa-ke, lokhu kuguqulwa kungaphazamisa inqubo yokuzalana ngengozi61. Ngaphezu kwalokho, izinyosi zithola izinsuku eziningana zesimo sezulu esibi, okuhlanganisa imvula kanye namazinga okushisa aphansi (<5°C), okungathinta kabi impumelelo yokuzalana4,23.
Nakuba ucwaningo lwethu lugxile kuyo yonke i-microbiome yama-larval, imiphumela yethu inikeza ukuqonda ngobudlelwano obungaba khona phakathi kwemiphakathi yamagciwane obungaba yinto ebalulekile ekudleni kwezinyosi kanye nokuchayeka ku-fungicide. Isibonelo, impova ephethwe yizibungu ezidla i-mancozeb yayinciphise kakhulu isakhiwo somphakathi wamagciwane kanye nobuningi uma kuqhathaniswa nempova ephethwe yizibungu ezidla impova engelashwanga. Ezibunguni ezidla impova engelashwanga, amaqembu amabhaktheriya i-Proteobacteria ne-Actinobacteria ayenamandla futhi ayevame kakhulu i-aerobic noma i-facultatively aerobic. Amabhaktheriya ase-Delft, avame ukuhlotshaniswa nezinhlobo zezinyosi ezizimele, ayaziwa ukuthi anomsebenzi wama-antibiotic, okubonisa indima engaba khona yokuvikela ngokumelene namagciwane. Olunye uhlobo lwamabhaktheriya, i-Pseudomonas, lwaluningi ezibunguni ezidla impova engelashwanga, kodwa lwancipha kakhulu ezibunguni eziphathwa yi-mancozeb. Imiphumela yethu isekela izifundo zangaphambilini ezikhomba i-Pseudomonas njengenye yezinhlobo eziningi kakhulu ku-O. bicornis35 kanye nezinye izinyo ezizimele. Nakuba ubufakazi bokuhlola ngendima ye-Pseudomonas empilweni ye-O. cornifrons bungakafundwa, leli gciwane liboniswe ukuthi likhuthaza ukwakheka kobuthi obuvikelayo kuma-beetle Paederus fuscipes futhi likhuthaza i-arginine metabolism in vitro 35, 65. Lokhu okubonwe kuphakamisa indima engaba khona ekuvikeleni amagciwane namagciwane ngesikhathi sokuthuthukiswa kwezibungu ze-O. cornifrons. I-Microbacterium ingenye uhlobo olukhonjwe ocwaningweni lwethu okubikwa ukuthi lukhona ngamanani aphezulu ezibungu ze-black soldier fly ngaphansi kwezimo zendlala66. Kuzibungu ze-O. cornifrons, ama-microbacteria angafaka isandla ekulinganisweni nasekuqineni kwe-microbiome yamathumbu ngaphansi kwezimo zokucindezeleka. Ngaphezu kwalokho, i-Rhodococcus itholakala ezibungu ze-O. cornifrons futhi yaziwa ngamakhono ayo okususa ubuthi67. Lolu hlobo lutholakala futhi emathunjini e-A. florea, kodwa ngobuningi obuphansi kakhulu68. Imiphumela yethu ikhombisa ukuba khona kwezinguquko eziningi zofuzo kuzo zonke izinhlobo eziningi zamagciwane ezingashintsha izinqubo ze-metabolic ezibunguni. Kodwa-ke, ukuqonda kangcono ukuhlukahluka kokusebenza kwe-O. cornifrons kuyadingeka.
Ngamafuphi, imiphumela ikhombisa ukuthi i-mancozeb, i-pyrithiostrobin, kanye ne-trifloxystrobin kunciphisa ukukhuluphala komzimba kanye nokwanda kokufa kwezibungu ze-corn borer. Nakuba kunokukhathazeka okukhulayo ngemiphumela ye-fungicides kuma-pollinator, kunesidingo sokuqonda kangcono imiphumela ye-metabolites esele yalezi zinhlanganisela. Le miphumela ingafakwa ezincomweni zezinhlelo zokuphathwa kwe-pollinator ezihlanganisiwe ezisiza abalimi ukugwema ukusebenzisa ama-fungicides athile ngaphambi nangesikhathi sokuqhakaza kwezihlahla zezithelo ngokukhetha ama-fungicides nokushintsha isikhathi sokusetshenziswa, noma ngokukhuthaza ukusetshenziswa kwezinye izindlela ezingalimazi kakhulu 36. Lolu lwazi lubalulekile ekuthuthukiseni izincomo. ngokusetshenziswa kwezibulala-zinambuzane, njengokulungisa izinhlelo ezikhona zokufafaza nokushintsha isikhathi sokufafaza lapho ukhetha ama-fungicides noma ukukhuthaza ukusetshenziswa kwezinye izindlela ezingalimazi kakhulu. Ucwaningo olwengeziwe luyadingeka ngemiphumela emibi yama-fungicides kusilinganiso sobulili, ukuziphatha kokudla, i-gut microbiome, kanye nezinqubo zama-molecule ezisekela ukwehla kwesisindo nokufa kwe-corn borer.
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