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Likokonyana tse bolaeang dikokwana-hloko hangata di sebediswa nakong ya dipalesa tsa ditholwana tsa sefate mme di ka sokela dikokonyana tse silafatsang dikokwana-hloko. Leha ho le jwalo, ho tsejwa hanyane ka moo dikokonyana tse sa silafatseng dinotshi (mohlala, dinotshi tse ikemetseng, Osmia cornifrons) di arabelang ka ho kopana le dikokonyana tse sireletsang dikokwana-hloko tse sebediswang hangata diapoleng nakong ya dipalesa. Lekhalo lena la tsebo le fokotsa diqeto tsa taolo tse etsang hore ho be le maemo a bolokehileng le nako ya ho fafatsa dikokonyana. Re hlahlobile ditlamorao tsa dikokonyana tse pedi tse sireletsang dikokwana-hloko (captan le mancozeb) le dikokonyana tse nne tse sireletsang dikokwana-hloko tse fapaneng/phytosystem (ciprocycline, myclobutanil, pyrostrobin le trifloxystrobin). Ditlamorao hodima boima ba dikokonyana, ho phela, karolelano ya bong le mefutafuta ya dibaktheria. Tlhahlobo e entswe ka ho sebedisa tlhahlobo ya bioassay e sa foleng ya molomo moo peo e phofshoang ka ditekanyo tse tharo ho latela tekanyo e kgothalletswang hajwale bakeng sa tshebediso ya tshimo (1X), tekanyo e halofo (0.5X) le tekanyo e tlase (0.1X). Ditekanyo tsohle tsa mancozeb le pyritisoline di fokoditse boima ba mmele haholo le ho phela ha dikokonyana. Eaba re hlophisa liphatsa tsa lefutso tsa 16S ho hlalosa baktheria ea larval ea mancozeb, e leng fungicide e ikarabellang bakeng sa lefu le phahameng ka ho fetisisa. Re fumane hore mefuta-futa ea libaktheria le bongata ba tsona li fokotsehile haholo ho liboko tse fepuoang ke peo e phofshoana e phekotsoeng ke mancozeb. Liphetho tsa rona tsa laboratori li bontša hore ho fafatsa tse ling tsa fungicide tsena nakong ea lipalesa ho kotsi haholo bophelong ba limela tsa O. cornifrons. Tlhahisoleseling ena e bohlokoa bakeng sa liqeto tsa tsamaiso ea nakong e tlang mabapi le tšebeliso e tsitsitseng ea lihlahisoa tsa tšireletso ea lifate tsa litholoana 'me e sebetsa e le motheo oa lits'ebetso tsa taolo tse reretsoeng ho sireletsa li-pollinator.
Notši e le 'ngoe ea mason ea Osmia cornifrons (Hymenoptera: Megachilidae) e ile ea tlisoa United States ho tsoa Japane bofelong ba lilemo tsa bo-1970 le mathoasong a bo-1980, 'me mofuta ona o bapetse karolo ea bohlokoa ea ho tsamaisa peo ea limela tikolohong e laoloang ho tloha ka nako eo. Mekhatlo ea linotši tsena e nang le tlhaho ke karolo ea mefuta e ka bang 50 ea linotši tsa naha tse tlatsanang le linotši tse tsamaisang peo ea limela tsa almonde le apole United States2,3. Linotši tsa Mason li tobana le liphephetso tse ngata, ho kenyeletsoa ho arohana ha libaka tsa bolulo, likokoana-hloko le chefo e bolaeang likokoanyana3,4. Har'a likokoanyana tse bolaeang likokoanyana, li-fungicides li fokotsa ho fumana matla, ho batla lijo tse ling5 le ho hlokomela 'mele6,7. Le hoja lipatlisiso tsa morao tjena li bontša hore bophelo bo botle ba linotši tsa Mason bo susumetsoa ka ho toba ke likokoana-hloko tsa commensal le ectobactic, 8,9 hobane libaktheria le li-fungus li ka susumetsa phepo e nepahetseng le likarabo tsa 'mele oa ho itšireletsa mafung, liphello tsa ho pepesehela fungicide ho mefuta-futa ea likokoana-hloko tsa linotši tsa Mason li sa tsoa qala ho ithutoa.
Likokonyana tse bolaeang dikokwana-hloko tse nang le ditlamorao tse fapaneng (ho kopana le ho kopana le mmele) di fafatswa dirapeng tsa difate pele le nakong ya dipalesa ho phekola mafu a kang lehlaka la apole, ho bola ho babang, ho bola ho sootho le hlobo e phofshoana10,11. Likokonyana tse bolaeang dikokwana-hloko di nkwa di se kotsi ho di-pollinator, kahoo di kgothalletswa ho balemi ba dirapa nakong ya dipalesa; Ho pepesehela le ho ja dikokonyana tsena ke dinotshi ho tsebahala haholo, kaha ke karolo ya tshebetso ya ngodiso ya dikokonyana tse bolayang dikokwana-hloko ke US Environmental Protection Agency le ditheo tse ding tse ngata tsa naha tsa taolo12,13,14. Leha ho le jwalo, ditlamorao tsa dikokonyana tse bolaeang dikokwana-hloko ho bao e seng dinotshi ha di tsejwe hakaalo hobane ha di hlokehe tlasa ditumellano tsa tumello ya papatso United States15. Ho phaella moo, ka kakaretso ha ho na diprotokoto tse tloaelehileng tsa ho leka dinotshi tse le nngwe16,17, mme ho boloka dikolone tse fanang ka dinotshi bakeng sa teko ho thata18. Diteko tsa dinotshi tse fapaneng tse laolwang di ntse di etswa ka ho eketsehileng Europe le USA ho ithuta ditlamorao tsa dikokonyana tse bolayang dikokwana-hloko dinotshing tsa naha, mme diprotokoto tse tloaelehileng di sa tswa ntshetswa pele bakeng sa O. cornifrons19.
Linotši tse nang le manaka ke li-monocyte 'me li sebelisoa khoebong lijalong tsa carp e le tlatsetso kapa sebaka sa linotši tsa mahe a linotši. Linotši tsena li hlaha pakeng tsa Hlakubele le Mmesa, 'me tse tona tse qalang li hlaha matsatsi a mararo ho isa ho a mane pele ho tse tšehali. Kamora ho kopana, e tšehali e bokella peō le lero ka mafolofolo ho fana ka letoto la lisele tsa bana ka har'a sehlaha sa tubular (sa tlhaho kapa sa maiketsetso)1,20. Mahe a beoa peōng ka har'a lisele; e tšehali ebe e haha lebota la letsopa pele e lokisa sele e latelang. Li-larvae tsa pele tse ka hare li koaletsoe ka har'a chorion 'me li ja metsi a embryonic. Ho tloha ho ea bobeli ho ea bohlano (prepupa), li-larvae li ja peō22. Hang ha phepelo ea peō e felile ka botlalo, li-larvae li etsa li-cocoon, li-pupate 'me li hlaha e le batho ba baholo ka kamoreng e le' ngoe ea bana, hangata qetellong ea lehlabula20,23. Batho ba baholo ba hlaha selemo se latelang. Ho phela ha batho ba baholo ho amahanngoa le ho eketseha ha matla a tlhaho (ho eketseha ha boima ba 'mele) ho latela lijo tse jeoang. Kahoo, boleng ba phepo ea peō, hammoho le lintlha tse ling tse kang boemo ba leholimo kapa ho pepesehela chefo e bolaeang likokoanyana, ke lintho tse etsang hore ho phele le bophelo bo botle24.
Libolaya-dikokwanyana le dibolaya-dikokwanyana tse sebediswang pele di thunya di kgona ho tsamaya ka hara methapo ya dimela ho ya maemong a fapaneng, ho tloha ho translaminar (mohlala, ho kgona ho tsamaya ho tloha hodima bokahodimo ba makhasi ho ya bokahodimo bo ka tlase, jwalo ka dibolaya-dikokwanyana tse ding) 25 ho ya ditlamoraong tsa sebele tsa tsamaiso. , e ka kenang moqhaka ho tloha metsong, e ka kena lero la dipalesa tsa apole26, moo di ka bolayang dibolaya-dikokwanyana tse kgolo tsa O. cornifrons27. Dibolaya-dikokwanyana tse ding le tsona di kena peong, di ama ntshetsopele ya diboko tsa poone mme tsa baka lefu la tsona19. Diphuputso tse ding di bontshitse hore dibolaya-dikokwanyana tse ding di ka fetola haholo boitshwaro ba ho behela dihlaha ba mofuta o amanang le tsona wa O. lignaria28. Ho phaella moo, dithuto tsa laboratori le tsa tshimo tse etsisang maemo a ho pepesehela dibolaya-dikokwanyana (ho kenyeletswa le dibolaya-dikokwanyana) di bontshitse hore dibolaya-dikokwanyana di ama hampe physiology22 morphology29 le ho phela ha dinotshi tsa mahe a dinotshi le dinotshi tse ding tse ikemetseng. Di-spray tse fapaneng tse bolayang dikokwanyana tse sebediswang ka ho toba dipaleng tse bulehileng nakong ya dipalesa di ka silafatsa peo e bokellwang ke batho ba baholo bakeng sa ntshetsopele ya diboko, tseo ditlamorao tsa tsona di sa ntseng di lokela ho ithutwa30.
Ho ntse ho lemohuoa haholoanyane hore ntshetsopele ya diboko e susumetswa ke peo e phofshoana le dikokwana-hloko tsa tsamaiso ya tshilo ya dijo. Dikokoana-hloko tsa dinotshi di susumetsa diparamente tse kang boima ba mmele31, diphetoho tsa metabolism22 le ho pepesehela dikokwana-hloko32. Diphuputso tse fetileng di hlahlobile tshusumetso ya mohato wa ntshetsopele, dikotla, le tikoloho hodima dikokoana-hloko tsa dinotshi tse ikemetseng. Diphuputso tsena di senotse ho tshwana sebopehong le bongata ba dikokoana-hloko tsa diboko le tsa pollen33, hammoho le mofuta o tlwaelehileng wa baktheria wa Pseudomonas le Delftia, hara mefuta ya dinotshi tse ikemetseng. Leha ho le jwalo, le hoja dikokoana-hloko di amana le mawa a ho sireletsa bophelo bo botle ba dinotshi, ditlamorao tsa dikokoana-hloko hodima dikokoana-hloko tsa diboko ka ho pepesehela molomo ka ho toba di ntse di sa hlahlojwe.
Phuputso ena e lekile liphello tsa litekanyetso tsa 'nete tsa li-fungicide tse tšeletseng tse sebelisoang hangata tse ngolisitsoeng bakeng sa ho sebelisoa litholoana tsa lifate United States, ho kenyeletsoa le li-fungicide tsa ho kopana le tsa tsamaiso tse fuoang ka molomo li-larvae tsa corn hornworm moth tse tsoang lijong tse silafetseng. Re fumane hore li-fungicide tsa ho kopana le tsa tsamaiso li fokotsa boima ba 'mele oa linotši le ho eketseha ha lefu, ka liphello tse mpe ka ho fetisisa tse amanang le mancozeb le pyrithiopide. Eaba re bapisa mefuta e fapaneng ea likokoana-hloko tsa li-fungus tse fepuoang lijong tsa peo e phofshoang e phekotsoeng ke mancozeb le tse fepuoang lijong tse laoloang. Re buisana ka mekhoa e ka bang teng e bakang lefu le litlamorao bakeng sa mananeo a kopaneng a taolo ea likokoanyana le peo e phofshoang (IPPM).
Li-cornifron tse kholo tsa O. cornifron tse ntseng li hola mariha ka har'a li-cocoon li fumanoe Setsing sa Lipatlisiso tsa Litholoana, Biglerville, PA, 'me li bolokiloe mochesong oa −3 ho isa ho 2°C (± 0.3°C). Pele ho teko (li-cocoon tse 600 ka kakaretso). Ka Mots'eanong 2022, li-cocoon tse 100 tsa O. cornifron li ne li fetisetsoa letsatsi le letsatsi ka har'a likopi tsa polasetiki (li-cocoon tse 50 ka kopi, DI 5 cm × 15 cm bolelele) 'me masela a ne a kenngoa ka har'a likopi ho khothaletsa ho buleha le ho fana ka substrate e ka hlafunoang, ho fokotsa khatello ea linotši tse majoe37. Beha likopi tse peli tsa polasetiki tse nang le li-cocoon ka har'a lesaka la likokoanyana (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) ka li-feeder tse 10 ml tse nang le tharollo ea sucrose ea 50% 'me u li boloke matsatsi a mane ho netefatsa hore lia koaloa le ho kopana. 23°C, mongobo o lekanyelitsoeng 60%, photoperiod 10 l (matla a tlase): matsatsi a 14. Tse tshehadi tse 100 tse nyetsweng le tse tona di ne di lokollwa hoseng ho hong le ho hong ka matsatsi a tsheletseng (100 ka letsatsi) ka hara dihlaha tse pedi tsa maiketsetso nakong ya ho thunya ha apole ka sehlohlolong (sehlaha sa leraba: bophara 33.66 × bolelele 30.48 × bolelele 46.99 cm; Setšoantšo se Tlatsetsang 1). Di beilwe Arboretum ya Naha ya Pennsylvania, haufi le ciliegia (Prunus cerasus 'Eubank' Sweet Cherry Pie™), perekisi (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), pere (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), sefate sa apole sa coronaria (Malus coronaria) le mefuta e mengata ya difate tsa apole (Malus coronaria, Malus), sefate sa apole sa lapeng 'Co-op 30′ Enterprise™, sefate sa apole sa Malus 'Co-Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy'). Ntlo e 'ngoe le e 'ngoe ea linonyana ea polasetiki e putsoa e lekana holim'a mabokose a mabeli a lehong. Lebokose le leng le le leng la sehlaha le ne le e-na le lipeipi tse 800 tse se nang letho tsa pampiri ea kraft (e bulehileng ka lebelo, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) e kentsoe ka har'a lipeipi tsa cellophane tse sa bonahaleng (0.7 OD bona lipeipi tsa polasetiki (lipeipi tsa T-1X) li fana ka libaka tsa ho behela lihlaha.
Mabokose a sehlaha ka bobedi a ne a shebile bochabela mme a kwahetswe ka terata ya serapa sa polasetiki e tala (mohlala wa Everbilt #889250EB12, boholo ba ho bula 5 × 5 cm, 0.95 m × 100 m) ho thibela phihlello ya ditoeba le dinonyana mme a bewa hodima mobu pela mabokose a mobu a lebokose la sehlaha. Lebokose la sehlaha (Setshwantsho se Tlatsetsang sa 1a). Mahe a poone a ne a bokellwa letsatsi le letsatsi ka ho bokella dipeipi tse 30 ho tswa dihlaheng le ho di isa laboratoring. O sebedisa sekere, etsa seha qetellong ya peipi, ebe o qhaqha tube ya spiral ho pepesa disele tsa bana. Mahe ka bomong le peo ya wona a ile a tloswa ho sebediswa spatula e kobehileng (Kit ya disebediswa tsa Microslide, BioQuip Products Inc., California). Mahe a ile a fuama pampiring ya sefe e mongobo mme a bewa ka sejaneng sa Petri ka dihora tse 2 pele a sebediswa ditekong tsa rona (Setshwantsho se Tlatsetsang sa 1b-d).
Laboratoring, re hlahlobile chefo ea li-fungicide tse tšeletseng tse sebelisitsoeng pele le nakong ea lipalesa tsa apole ka maemo a mararo (0.1X, 0.5X, le 1X, moo 1X e leng letšoao le sebelisoang ka lilithara tse 100 tsa metsi/hekthere. Tekanyo e phahameng ea tšimo = mahloriso tšimong). , Tafole ea 1). Hlobo ka 'ngoe e phetiloe ka makhetlo a 16 (n = 16). Li-fungicide tse peli tse amang (Tafole ea S1: mancozeb 2696.14 ppm le captan 2875.88 ppm) le li-fungicide tse 'ne tsa systemic (Tafole ea S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75 .12 ppm; cyprodinil 280.845 ppm) chefo ho litholoana, meroho le lijalo tsa mekhabiso. Re ile ra kopanya peo e phofshoana re sebedisa grinder, ra fetisetsa 0.20 g selibeng (Falcon Plate ya diliba tse 24), mme ra eketsa le ho kopanya 1 μL ya tharollo ya fungicide ho etsa peo e phofshoana ya pyramidal ka diliba tse tebileng tsa 1 mm tseo mahe a neng a kentswe ho tsona. Beha ka ho sebedisa spatula e nyane (Setshwantsho se Tlatsetsang sa 1c, d). Dipoleiti tsa Falcon di ne di bolokilwe mochesong wa kamore (25°C) le mongobo wa 70%. Re di bapisitse le diboko tse laolang tse fepuwang dijo tse tshwanang tsa peo e phofshoana tse phekotsweng ka metsi a hlwekileng. Re tlalehile lefu mme ra lekanya boima ba diboko letsatsi le leng le le leng ho fihlela diboko di fihla dilemong tsa pele ho hola re sebedisa tekanyo ya tlhahlobo (Fisher Scientific, nepahalo = 0.0001 g). Qetellong, karolelano ya bong e ile ya lekolwa ka ho bula kokone kamora dikgwedi tse 2.5.
DNA e ntšitsoe ho li-larvae tsa O. cornifrons kaofela (n = 3 ka boemo ba kalafo, peo e phofshoana e phekotsoeng ke mancozeb le e sa phekoloang) 'me re entse litlhahlobo tsa mefuta-futa ea likokoana-hloko lisampoleng tsena, haholo-holo hobane ho mancozeb lefu le phahameng ka ho fetisisa le ile la bonoa ho li-larvae. Ho amohela MnZn. DNA e ile ea atolosoa, ea hloekisoa ho sebelisoa DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), 'me ea aroloa ka tatellano (lipotoloho tse 600) ho Illumina® MiSeq™ ho sebelisoa v3 kit. Tatellano e reretsoeng ea liphatsa tsa lefutso tsa ribosomal RNA tsa baktheria ea 16S e entsoe ho sebelisoa Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) ho sebelisoa li-primer tse shebaneng le sebaka sa V3-V4 sa gene ea 16S rRNA. Ho phaella moo, tatellano ea 18S e entsoe ho sebelisoa 10% PhiX inclusion, 'me amplification e entsoe ho sebelisoa primer pair 18S001 le NS4.
Kenya le ho sebetsana le di-read39 tse kopantsweng ho sebediswa phaephe ya QIIME2 (v2022.11.1). Di-read tsena di ile tsa kgaolwa mme tsa kopanngwa, mme di-chatellano tsa chimeric di ile tsa tloswa ho sebediswa plugin ya DADA2 ho QIIME2 (qiime dada2 lerata la ho kopanya)40. Dikabelo tsa sehlopha sa 16S le 18S di entswe ho sebediswa plugin ya object classifier Classify-sklearn le artifact silva-138-99-nb-classifier e koetlisitsweng esale pele.
Lintlha tsohle tsa liteko li hlahlobiloe bakeng sa ho tloaeleha (Shapiro-Wilks) le ho tšoana ha liphapang (teko ea Levene). Hobane sete ea data ha ea ka ea fihlela likhopolo tsa tlhahlobo ea parametric 'me phetoho e hlōlehile ho etsa hore masala a be maemong a tloaelehileng, re entse ANOVA (Kruskal-Wallis) e seng ea parametric ka lintlha tse peli [nako (lintlha tsa nako tsa mohato oa boraro oa 2, 5, le 8) le fungicide] ho lekola phello ea kalafo boima ba larval fresh, ebe ho bapisoa ha post hoc nonparametric pairiwise ho entsoe ho sebelisoa teko ea Wilcoxon. Re sebelisitse mohlala o akaretsang oa linear (GLM) o nang le kabo ea Poisson ho bapisa liphello tsa fungicide ho pholoheng ho pholletsa le lihlopha tse tharo tsa fungicide41,42. Bakeng sa tlhahlobo ea bongata bo fapaneng, palo ea mefuta-futa ea tatellano ea amplicon (ASVs) e ile ea putlama boemong ba mofuta. Lipapiso tsa bongata bo fapaneng pakeng tsa lihlopha tse sebelisang 16S (boemo ba mofuta) le bongata bo amanang le 18S li entsoe ho sebelisoa mohlala oa tlatsetso o akaretsang bakeng sa boemo, sekala le sebopeho (GAMLSS) ka likabo tsa lelapa la beta zero-inflated (BEZI), tse entsoeng mohlala ho macro . ho Microbiome R43 (v1.1). 1). Tlosa mefuta ea mitochondrial le chloroplast pele ho tlhahlobo ea phapang. Ka lebaka la maemo a fapaneng a taxonomic a 18S, ke boemo bo tlase ka ho fetisisa ba taxon ka 'ngoe bo sebelisitsoeng bakeng sa tlhahlobo ea phapang. Litlhahlobo tsohle tsa lipalo-palo li entsoe ho sebelisoa R (v. 3.4.3., projeke ea CRAN) (Sehlopha sa 2013).
Ho pepesehela mancozeb, pyrithiostrobin, le trifloxystrobin ho fokoditse haholo keketseho ya boima ba mmele ho O. cornifrons (Setshwantsho sa 1). Ditlamorao tsena di ile tsa bonwa ka ho tswela pele bakeng sa ditekanyo tsohle tse tharo tse lekotsweng (Setshwantsho sa 1a–c). Cyclostrobin le myclobutanil ha di a ka tsa fokotsa boima ba diboko haholo.
Boima bo tloaelehileng ba foreshe ba diboko tsa diboko tsa dimela bo lekanngwa ka dinako tse tharo tlasa kalafo tse nne tsa dijo (phepo ya peo e phofshoana e tshwanang + fungicide: taolo, ditekanyo tsa 0.1X, 0.5X le 1X). (a) Tekanyo e tlase (0.1X): ntlha ya pele (letsatsi la 1): χ2: 30.99, DF = 6; P < 0.0001, ntlha ya bobedi (letsatsi la 5): 22.83, DF = 0.0009; nako ya boraro; ntlha (letsatsi la 8): χ2: 28.39, DF = 6; (b) tekanyo e halofo (0.5X): ntlha ya pele (letsatsi la 1): χ2: 35.67, DF = 6; P < 0.0001, ntlha ya bobedi (letsatsi la pele). ): χ2: 15.98, DF = 6; P = 0.0090; ntlha ea boraro (letsatsi la 8) χ2: 16.47, DF = 6; (c) Sebaka kapa tekanyo e felletseng (1X): ntlha ea pele (letsatsi la 1) χ2: 20.64, P = 6; P = 0.0326, ntlha ea bobeli (letsatsi la 5): χ2: 22.83, DF = 6; P = 0.0009; ntlha ea boraro (letsatsi la 8): χ2: 28.39, DF = 6; tlhahlobo e seng ea parametric ea phapang. Li-bar li emela karolelano ea ± SE ea lipapiso tse peli (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Ka tekanyo e tlase ka ho fetisisa (0.1X), boima ba 'mele ba liboko bo fokotsehile ka 60% ka trifloxystrobin, 49% ka mancozeb, 48% ka myclobutanil, le 46% ka pyrithistrobin (Setšoantšo sa 1a). Ha li pepesetsoa halofo ea tekanyo ea tšimo (0.5X), boima ba 'mele ba liboko tsa mancozeb bo fokotsehile ka 86%, pyrithiostrobin ka 52% le trifloxystrobin ka 50% (Setšoantšo sa 1b). Tekanyo e felletseng ea tšimo (1X) ea mancozeb e fokolitse boima ba liboko ka 82%, pyrithiostrobin ka 70%, le trifloxystrobin, myclobutanil le sangard ka hoo e ka bang 30% (Setšoantšo sa 1c).
Palo ea batho ba shoang e ne e le holimo ka ho fetisisa har'a peō e phofshoana e fepuoang ke mancozeb, e lateloa ke pyrithiostrobin le trifloxystrobin. Palo ea batho ba shoang e ile ea eketseha ka ho eketseha ha litekanyetso tsa mancozeb le pyritisoline (Setšoantšo sa 2; Tafole ea 2). Leha ho le joalo, lefu la batho ba bolaeang poone le ile la eketseha hanyane feela ha mahloriso a trifloxystrobin a ntse a eketseha; cyprodinil le captan ha lia ka tsa eketsa lefu haholo ha li bapisoa le mekhoa ea phekolo ea taolo.
Ho shoa ha liboko tsa li-borer fly ho bapisitsoe ka mor'a ho ja peo e phofshoana ka bonngoe e phekotsoeng ka meriana e tšeletseng e fapaneng ea fungicide. Mancozeb le pentopyramide li ne li le bonolo ho pepesehela liboko tsa poone ka molomo (GLM: χ = 29.45, DF = 20, P = 0.0059) (mola, leralla = 0.29, P < 0.001; leralla = 0.24, P <0.00)).
Ka karolelano, ho pholletsa le mekhoa eohle ea phekolo, bakuli ba 39.05% e ne e le basali 'me 60.95% e ne e le banna. Har'a mekhoa ea phekolo ea taolo, karolo ea basali e ne e le 40% lithutong tsa tekanyo e tlase (0.1X) le halofo (0.5X), le 30% lithutong tsa tekanyo ea tšimo (1X). Ka tekanyo ea 0.1X, har'a liboko tse fepuoang ke peō tse phekotsoeng ka mancozeb le myclobutanil, 33.33% ea batho ba baholo e ne e le basali, 22% ea batho ba baholo e ne e le basali, 44% ea liboko tse kholo e ne e le basali, 44% ea liboko tse kholo e ne e le basali. tse tšehali, 41% ea liboko tse kholo e ne e le basali, 'me tse laoloang e ne e le 31% (Setšoantšo sa 3a). Ka makhetlo a 0.5 a tekanyo, 33% ea liboko tse kholo sehlopheng sa mancozeb le pyrithiostrobin e ne e le tsa basali, 36% sehlopheng sa trifloxystrobin, 41% sehlopheng sa myclobutanil, le 46% sehlopheng sa cyprostrobin. Palo ena e ne e le 53% sehlopheng. sehlopheng sa captan le 38% sehlopheng sa taolo (Setšoantšo sa 3b). Ka tekanyo ea 1X, 30% ea sehlopha sa mancozeb e ne e le basali, 36% ea sehlopha sa pyrithiostrobin, 44% ea sehlopha sa trifloxystrobin, 38% ea sehlopha sa myclobutanil, 50% ea sehlopha sa taolo e ne e le basali - 38.5% (Setšoantšo sa 3c).
Peresente ea li-bore tse tšehali le tse tona ka mor'a ho pepesehela fungicide mohatong oa larval. (a) Tekanyo e tlase (0.1X). (b) Tekanyo e halofo (0.5X). (c) Tekanyo ea tšimo kapa tekanyo e felletseng (1X).
Tlhahlobo ea tatellano ea 16S e bontšitse hore sehlopha sa baktheria se fapane pakeng tsa liboko tse fepuoang ka peo e phofshoana e phekotsoeng ke mancozeb le liboko tse fepuoang ka peo e phofshoana e sa phekoloang (Setšoantšo sa 4a). Palo ea likokoana-hloko tsa liboko tse sa phekoloang tse fepuoang peo e phofshoana e ne e le holimo ho feta ea liboko tse fepuoang peo e phofshoana e phekotsoeng ke mancozeb (Setšoantšo sa 4b). Leha phapang e bonoang leruong lipakeng tsa lihlopha e ne e se ea bohlokoa ka lipalo-palo, e ne e le tlase haholo ho feta e bonoang bakeng sa liboko tse jang peo e phofshoana e sa phekoloang (Setšoantšo sa 4c). Bongata bo itseng bo bontšitse hore microbiota ea liboko tse fepuoang peo e laoloang e ne e fapane haholo ho feta ea liboko tse fepuoang peo e phekotsoeng ke mancozeb (Setšoantšo sa 5a). Tlhahlobo e hlalosang e senotse boteng ba mefuta e 28 lisampoleng tse laoloang le tse phekotsoeng ke mancozeb (Setšoantšo sa 5b). c Tlhahlobo e sebelisang tatellano ea 18S ha ea ka ea senola liphapang tse kholo (Setšoantšo se Tlatsetsang sa 2).
Liprofaele tsa SAV tse thehiloeng ho tatellano ea 16S li bapisitsoe le leruo la Shannon 'me tsa bona leruo boemong ba phylum. (a) Tlhahlobo ea likhokahano tse kholo (PCoA) e thehiloeng sebopehong sa sechaba sa likokoana-hloko tse sa phekoloang tse fepuoang kapa tse laoloang (putsoa) le li-larvae tse fepuoang ke mancozeb (lamunu). Ntlha e 'ngoe le e 'ngoe ea data e emela sampole e arohaneng. PCoA e baliloe ho sebelisoa sebaka sa Bray-Curtis sa kabo ea t ea multivariate. Li-oval li emela boemo ba kholiseho ba 80%. (b) Boxplot, data ea leruo la Shannon e tala (lintlha) le c. Leruo le bonoang. Boxplot li bontša mabokose a mola o bohareng, sebaka sa interquartile (IQR), le 1.5 × IQR (n = 3).
Sebopeho sa dihlopha tsa dikokwana-hloko tsa diboko tse fepuwang ke peo e phofshwang ke mancozeb le e sa phekolwang. (a) Bongata bo itseng ba dikokonyana bo balwa ka hara diboko. (b) Mmapa wa mocheso wa dihlopha tsa dikokwana-hloko tse hlwailweng. Delftia (karolelano ya menyetla (OR) = 0.67, P = 0.0030) le Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060); Mela ya dimapo tsa mocheso e arolwa ka ho sebedisa sebaka sa kamano le kgokelo e tloaelehileng.
Liphetho tsa rona li bontša hore ho pepesehela li-fungicide tse amang (mancozeb) le tsa 'mele (pyrostrobin le trifloxystrobin) ka molomo, tse sebelisoang haholo nakong ea lipalesa, ho fokolitse haholo keketseho ea boima ba 'mele le keketseho ea lefu la liboko tsa poone. Ho phaella moo, mancozeb e fokolitse haholo mefuta-futa le leruo la microbiome nakong ea mohato oa pele ho limela. Myclobutanil, e 'ngoe ea fungicide ea 'mele, e fokolitse haholo keketseho ea boima ba 'mele ba liboko ka tekanyo eohle e meraro. Phello ena e ile ea bonahala haholo linakong tsa bobeli (letsatsi la 5) le la boraro (letsatsi la 8). Ka lehlakoreng le leng, cyprodinil le captan ha lia ka tsa fokotsa keketseho ea boima ba 'mele kapa ho phela ha tsona haholo ha li bapisoa le sehlopha sa taolo. Ho ea ka tsebo ea rona, mosebetsi ona ke oa pele ho fumana liphello tsa sekhahla sa masimo a li-fungicide tse fapaneng tse sebelisoang ho sireletsa lijalo tsa poone ka ho pepesehela peo e phofshoana ka ho toba.
Kalafo eohle ea fungicide e fokolitse keketseho ea boima ba 'mele haholo ha e bapisoa le kalafo ea taolo. Mancozeb e bile le tšusumetso e kholo ho keketseho ea boima ba 'mele ea liboko ka phokotso e tloaelehileng ea 51%, e lateloa ke pyrithiostrobin. Leha ho le joalo, lithuto tse ling ha lia tlaleha litlamorao tse mpe tsa tekanyo ea masimo ea li-fungicide mehatong ea liboko44. Leha li-biocide tsa dithiocarbamate li bontšitsoe li na le chefo e tlase e matla45, ethylene bisdithiocarbamate (EBDCS) tse kang mancozeb li ka senyeha ho ba urea ethylene sulfide. Ka lebaka la litlamorao tsa eona tsa phetoho liphoofolong tse ling, sehlahisoa sena sa ho senyeha se ka 'na sa ikarabella bakeng sa litlamorao tse bonoang46,47. Lithuto tse fetileng li bontšitse hore ho thehoa ha ethylene thiourea ho susumetsoa ke lintlha tse kang mocheso o phahameng48, maemo a mongobo49 le bolelele ba polokelo ea sehlahisoa50. Maemo a nepahetseng a polokelo bakeng sa liboko a ka fokotsa litla-morao tsena. Ho phaella moo, Bolaoli ba Polokeho ea Lijo ba Europe bo bontšitse matšoenyeho ka chefo ea pyrithiopide, e bontšitsoeng e baka kankere lits'ebetsong tsa tšilo ea lijo tsa liphoofolo tse ling51.
Ho nooa ha mancozeb, pyrithiostrobin, le trifloxystrobin ho eketsa lefu la liboko tsa poone. Ka lehlakoreng le leng, myclobutanil, ciprocycline le captan ha lia ka tsa ama lefu. Liphetho tsena li fapana le tsa Ladurner et al.52, ba bontšitseng hore captan e fokolitse haholo ho phela ha O. lignaria ea batho ba baholo le Apis mellifera L. (Hymenoptera, Apisidae). Ho phaella moo, li-fungicide tse kang captan le boscalid li fumanoe li baka lefu la liboko52,53,54 kapa li fetola boitšoaro ba ho ja55. Liphetoho tsena, ka lehlakoreng le leng, li ka ama boleng ba phepo ea peo e phofshoana 'me qetellong matla a ho fumana boemo ba liboko. Ho shoa ho bonoeng sehlopheng sa taolo ho ne ho lumellana le lithuto tse ling 56,57.
Karolelano ea bong e ratang banna e bonoang mosebetsing oa rona e ka hlalosoa ke lintlha tse kang ho se lekane ha ho kopana le maemo a leholimo a mabe nakong ea lipalesa, joalo ka ha ho ile ha etsoa tlhahiso ea pele bakeng sa O. cornuta ke Vicens le Bosch. Le hoja tse tšehali le tse tona thutong ea rona li ne li e-na le matsatsi a mane a ho kopana (nako eo ka kakaretso e nkoang e lekane bakeng sa ho kopana ka katleho), re ile ra fokotsa matla a khanya ka boomo ho fokotsa khatello ea maikutlo. Leha ho le joalo, phetoho ena e ka sitisa ts'ebetso ea ho kopana ka phoso61. Ho phaella moo, linotši li ba le matsatsi a 'maloa a boemo ba leholimo bo bobe, ho kenyeletsoa pula le mocheso o tlase (<5°C), e leng se ka boela sa ama katleho ea ho kopana ka tsela e mpe4,23.
Leha thuto ea rona e ne e shebane le microbiome eohle ea larval, liphetho tsa rona li fana ka temohisiso mabapi le likamano tse ka bang teng har'a lichaba tsa baktheria tse ka bang tsa bohlokoa bakeng sa phepo e nepahetseng ea linotši le ho pepesehela fungicide. Mohlala, peō e phofshoana e fepuoang ke mancozeb e ne e fokolitse sebopeho sa sechaba sa likokoana-hloko le bongata ba tsona haholo ha li bapisoa le peō e phofshoana e fepuoang ke liboko. Libokong tse jang peō e phofshoana e sa phekoloang, lihlopha tsa baktheria tsa Proteobacteria le Actinobacteria li ne li le matla 'me li ne li le matla haholo ho feta aerobic kapa facultatively aerobic. Libaktheria tsa Delft, hangata li amahanngoa le mefuta ea linotši tse ikemetseng, li tsejoa li na le ts'ebetso ea lithibela-mafu, e leng se bontšang karolo e ka bang teng ea tšireletso khahlanong le likokoana-hloko. Mofuta o mong oa baktheria, Pseudomonas, o ne o le mongata ho peō e phofshoana e fepuoang ke liboko, empa o ne o fokotsehile haholo ho peō e phekotsoeng ke mancozeb. Liphetho tsa rona li tšehetsa lithuto tsa pejana tse khethollang Pseudomonas e le e 'ngoe ea mefuta e mengata ka ho fetisisa ho O. bicornis35 le bobi bo bong bo ikemetseng34. Leha bopaki ba liteko ba karolo ea Pseudomonas bophelong bo botle ba O. cornifrons bo so ithutoe, baktheria ena e bontšitsoe e khothalletsa tlhahiso ea chefo e sireletsang ho Paederus fuscipes ea kokoanyana le ho khothaletsa metabolism ea arginine in vitro 35, 65. Litlhahlobo tsena li bontša karolo e ka bang teng ts'ireletsong ea vaerase le ea baktheria nakong ea nts'etsopele ea li-larvae tsa O. cornifrons. Microbacterium ke mofuta o mong o khethiloeng thutong ea rona o tlalehiloeng o le teng ka bongata ho li-larvae tsa black soldier fly tlas'a maemo a tlala66. Ho li-larvae tsa O. cornifrons, li-microbacteria li ka kenya letsoho ho leka-lekana le ho tiea ha microbiome ea mala tlas'a maemo a khatello. Ho feta moo, Rhodococcus e fumanoa ho li-larvae tsa O. cornifrons 'me e tsejoa ka bokhoni ba eona ba ho ntša chefo67. Mofuta ona o boetse o fumanoa ka mpeng ea A. florea, empa ka bongata bo fokolang haholo68. Liphetho tsa rona li bontša boteng ba mefuta e mengata ea liphatsa tsa lefutso ho pholletsa le mefuta e mengata ea likokoana-hloko tse ka fetolang lits'ebetso tsa metabolism ho li-larvae. Leha ho le joalo, ho hlokahala kutloisiso e betere ea mefuta-futa ea ts'ebetso ea O. cornifrons.
Ka bokhutšoanyane, liphetho li bontša hore mancozeb, pyrithiostrobin, le trifloxystrobin li fokolitse boima ba 'mele le ho eketseha ha lefu la liboko tsa poone. Le hoja ho na le matšoenyeho a ntseng a hola mabapi le liphello tsa li-fungicides ho li-pollinator, ho na le tlhoko ea ho utloisisa hamolemo liphello tsa metabolites tse setseng tsa metsoako ena. Liphetho tsena li ka kenyelletsoa likhothaletsong tsa mananeo a kopaneng a taolo ea li-fungicides tse thusang balemi ho qoba tšebeliso ea li-fungicides tse itseng pele le nakong ea lipalesa tsa lifate tsa litholoana ka ho khetha li-fungicides le ho fetola nako ea ts'ebeliso, kapa ka ho khothaletsa tšebeliso ea mekhoa e meng e seng kotsi haholo. Tlhahisoleseling ena e bohlokoa bakeng sa ho nts'etsapele likhothaletso mabapi le tšebeliso ea chefo e bolaeang likokoanyana, joalo ka ho fetola mananeo a teng a ho fafatsa le ho fetola nako ea ho fafatsa ha ho khethoa li-fungicides kapa ho khothaletsa tšebeliso ea mekhoa e meng e seng kotsi haholo. Ho hlokahala lipatlisiso tse ling mabapi le liphello tse mpe tsa li-fungicides ka karolelano ea bong, boitšoaro ba ho fepa, microbiome ea mala, le mekhoa ea limolek'hule e bakang tahlehelo ea boima le lefu la li-poone borer.
Lintlha tsa mohloli 1, 2 le 3 ho Setšoantšo sa 1 le sa 2 li kentsoe polokelong ea data ea figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 le https://doi.org/10.6084/m9. figshare.24996233. Tatellano e sekasekiloeng thutong ea hajoale (Setšoantšo sa 4, 5) e fumaneha polokelong ea NCBI SRA tlas'a nomoro ea kamohelo ea PRJNA1023565.
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