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Hangata li-fungicides li sebelisoa nakong ea lipalesa tsa lifate 'me li ka tšosa likokoanyana tse tsamaisang peō ea limela.Leha ho le joalo, ha ho tsejoe hakaalo ka hore na likokoana-hloko tseo e seng tsa linotsi (mohlala, linotsi tse ikemetseng, Osmia cornifrons) li arabela joang ha li kopana le li-fungicides tse sebelisoang hangata liapoleng nakong ea lipalesa.Lekhalo lena la tsebo le fokotsa liqeto tsa taolo tse khethollang libaka tse bolokehileng le nako ea ho fafatsa ka fungicide.Re ile ra hlahloba liphello tsa li-fungicides tse peli tsa ho kopana (captan le mancozeb) le li-fungicides tse 'nè tsa interlayer/phytosystem (ciprocycline, myclobutanil, pyrostrobin le trifloxystrobin).Liphello ho boima ba 'mele, ho phela, karolelano ea thobalano le mefuta-futa ea libaktheria.Tlhahlobo e entsoe ho sebelisoa tlhahlobo e sa foleng ea molomo moo peō e phofshoana e ileng ea phekoloa ka litekanyetso tse tharo ho latela tekanyo e khothalletsoang hona joale bakeng sa tšebeliso ea tšimo (1X), halofo ea tekanyo (0.5X) le tekanyo e tlaase (0.1X).Litekanyetso tsohle tsa mancozeb le pyritisoline li fokolitse boima ba 'mele haholo le ho phela ha larval.Ka mor'a moo re ile ra hlophisa mofuta oa 16S ho tšoaea bacteriome ea larval ea mancozeb, 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 li-larvae tse feptjoang ka peo e phofshoana e hloekisitsoeng ke mancozeb.Liphetho tsa rona tsa laboratori li bontša hore ho fafatsa tse ling tsa li-fungicides tsena nakong ea lipalesa ho kotsi haholo ho bophelo bo botle ba O. cornifrons.Tlhahisoleseding ena e bohlokoa bakeng sa liqeto tsa tsamaiso ea nako e tlang mabapi le tšebeliso e tsitsitseng ea lihlahisoa tsa tšireletso ea lifate tsa litholoana 'me e sebetsa e le motheo oa mekhoa ea taolo e reretsoeng ho sireletsa likokoana-hloko.
Nonyana e ikemetseng ea mason bee Osmia cornifrons (Hymenoptera: Megachilidae) e ile ea tsebisoa United States ho tsoa Japane bofelong ba lilemo tsa bo-1970 le mathoasong a lilemo tsa bo-1980, 'me mofuta ona o bile le karolo ea bohlokoa ea ho tsamaisa peō ea limela tikolohong e laoloang ho tloha ka nako eo.Baahi ba tlhaho ba linotsi tsena ke karolo ea mefuta e ka bang 50 ea linotši tse hlaha tse tlatsanang le linotši tse tsamaisang peō ea lirapa tsa almonde le liapole United States2,3.Linotši tsa Mason li tobane le mathata a mangata, ho kenyelletsa le ho arohana ha libaka, likokoana-hloko le chefo e bolaeang likokoanyana3,4.Har'a meriana e bolaeang likokoanyana, li-fungicides li fokotsa ho fumana matla, ho batla lijo5 le ho lokisa 'mele6,7.Leha lipatlisiso tsa morao-rao li fana ka maikutlo a hore bophelo bo botle ba linotši tsa Mason bo susumetsoa ka kotloloho ke likokoana-hloko tsa commensal le ectobactic, 8,9 hobane libaktheria le li-fungus li ka ama phepo e nepahetseng le karabelo ea 'mele ea ho itšireletsa mafung, litlamorao tsa ho pepeseha ha fungicide ho mefuta e fapaneng ea likokoana-hloko tsa Mason linotsi li qala ho ba teng. ithutiloe.
Likokoana-hloko tsa liphello tse fapa-fapaneng (ho kopana le tsamaiso) li fafatsoa lirapeng tsa lifate tsa litholoana pele le nakong ea lipalesa ho phekola maloetse a kang lekhopho la apole, bola bo babang, bola bo sootho le phori10,11.Li-fungicides li nkuoa li se kotsi ho li-pollinator, ka hona li khothaletsoa ho balemi ba lirapa nakong ea lipalesa;Ho pepeseha le ho jeoa ha li-fungicides tsena ke linotsi ho tsebahala haholo, kaha ke karolo ea ts'ebetso ea ngoliso ea chefo e bolaeang likokoanyana ke Setsi sa Ts'ireletso ea Tikoloho sa US le mekhatlo e meng e mengata ea taolo ea naha12,13,14.Leha ho le joalo, litlamorao tsa li-fungicides ho bao e seng linotsi ha li tsejoe haholo hobane ha li hlokehe tlasa litumellano tsa tumello ea ho bapatsa United States15.Ho feta moo, ka kakaretso ha ho na liprothokholo tse emeng bakeng sa tlhahlobo ea linotsi tse le 'ngoe16,17, 'me ho boloka likolone tse fanang ka linotsi bakeng sa tlhahlobo ho thata18.Liteko tsa linotsi tse laoloang tse fapaneng li ntse li tsoela pele ho etsoa Europe le USA ho ithuta litlamorao tsa chefo e bolaeang likokoanyana linotsing tse hlaha, 'me haufinyane tjena ho entsoe liprothokholo tsa O. cornifrons19.
Linotsi tse nang le manaka ke li-monocyte 'me li sebelisoa khoebong ea lijalo tsa carp e le tlatsetso kapa sebaka sa linotsi tsa mahe a linotši.Linotši tsena li hlaha pakeng tsa Tlhakubele le Mmesa, tse tona tse sa tsoa feta nako li hlaha matsatsi a mararo ho isa ho a mane pele ho tse tšehali.Ka mor'a ho kopana, e tšehali e bokella peō le lero ka mafolofolo ho fana ka letoto la lisele tsa brood ka har'a sehlaha sa tubular (tlhaho kapa maiketsetso)1,20.Mahe a behoa holim'a peo e phofo ka har'a lisele;e tšehali ebe e haha lebota la letsopa pele e lokisa sele e latelang.Li-larvae tsa pele tsa instar li koaletsoe ka har'a chorion 'me li fepa ka maro a embryonic.Ho tloha ho ea bobeli ho isa ho ea bohlano (prepupa), li-larvae li ja peō e phofshoana22.Hang ha peō e phofshoana e felile ka ho feletseng, li-larvae li etsa li-cocoons, pupate 'me li hlahella e le batho ba baholo ka kamoreng e le 'ngoe ea bana, hangata qetellong ea lehlabula20,23.Batho ba baholo ba hlaha nakong ea selemo e latelang.Ho phela ha batho ba baholo ho amahanngoa le phaello ea matla a marang-rang (ho fumana boima ba 'mele) ho latela ho ja lijo.Ka hona, boleng ba phepo ea peō e phofshoana, hammoho le lintlha tse ling tse kang boemo ba leholimo kapa ho pepesehela chefo e bolaeang likokoanyana, ke lintlha tsa ho phela le bophelo bo botle24.
Likokoana-hloko le li-fungicides tse sebelisoang pele li thunya li khona ho tsamaea ka har'a "vasculature" ea semela ho ea ho tse fapa-fapaneng, ho tloha ho translaminar (mohlala, ho khona ho tloha holimo holimo ho makhasi ho ea sebakeng se ka tlaase, joalo ka likokoana-hloko tse ling) 25 ho liphello tsa tsamaiso ea 'nete., e ka kenang moqhaka ho tloha metso, e ka kena lero la lipalesa tsa apole26, moo li ka bolaeang motho e moholo O. cornifrons27.Likokoana-hloko tse ling le tsona li kena peōng ea limela, tse amang nts'etsopele ea li-larvae tsa poone le ho baka lefu la tsona19.Liphuputso tse ling li bonts'itse hore li-fungicides tse ling li ka fetola haholo mokhoa oa ho haha sehlaha oa mofuta o amanang le O. lignaria28.Ho feta moo, liphuputso tsa laboratori le tsa masimong tse etsisang maemo a ho pepesehela chefo e bolaeang likokoanyana (ho kenyeletsoa li-fungicides) li bontšitse hore chefo e bolaeang likokoanyana e ama physiology hampe 22 morphology 29 le ho phela ha mahe a linotsi le linotši tse ling tse ikemetseng.Li-sprays tse fapaneng tsa fungicidal tse sebelisoang ka kotloloho lipalesa tse bulehileng nakong ea lipalesa li ka silafatsa peo e phofshoana e bokeletsoeng ke batho ba baholo bakeng sa nts'etsopele ea larval, e leng litlamorao tsa eona tse lokelang ho ithutoa30.
Ho ntse ho hlokomeloa ka ho eketsehileng hore tsoelo-pele ea larval e susumetsoa ke peō ea limela le likokoana-hloko tsa tsamaiso ea lijo.The honeybee microbiome e susumetsa mekhoa e kang boima ba 'mele31, liphetoho tsa metabolism22 le ho ba bonolo ho likokoana-hloko32.Lithuto tse fetileng li hlahlobile tšusumetso ea sethala sa nts'etsopele, limatlafatsi le tikoloho ho microbiome ea linotsi tse ikemetseng.Liphuputso tsena li senotse ho tšoana ka sebopeho le bongata ba li-microbiomes tsa larval le pollen33, hammoho le mofuta o tloaelehileng oa baktheria Pseudomonas le Delftia, har'a mefuta ea linotsi tse ikemetseng.Leha ho le joalo, le hoja li-fungicides li 'nile tsa amahanngoa le mekhoa ea ho sireletsa bophelo bo botle ba linotsi, liphello tsa fungicides ho larval microbiota ka ho pepeseha ka ho toba ka molomo li lula li sa tsejoe.
Boithuto bona bo lekile litlamorao tsa litekanyetso tsa lefats'e tsa 'nete tsa li-fungicides tse tšeletseng tse sebelisoang hangata tse ngolisitsoeng ho sebelisoa litholoana tsa lifate United States, ho kenyeletsoa ho kopana le li-fungicides tsa systemic tse fanoang ka molomo ho li-corn hornworm moth larvae ho tsoa lijong tse silafetseng.Re fumane hore ho kopana le li-fungicides tsa tsamaiso li fokolitse boima ba 'mele oa linotsi le ho eketseha ha lefu, ka liphello tse matla ka ho fetisisa tse amanang le mancozeb le pyrithiopide.Ka mor'a moo re ile ra bapisa mefuta-futa ea likokoana-hloko tsa li-larvae tse feptjoang lijong tsa phofshoana e entsoeng ka mancozeb le tse feptjoang ka lijo tse laoloang.Re tšohla mekhoa e ka bang teng e bakang lefu le litlamorao bakeng sa taolo e kopaneng ea likokoanyana le pollinator (IPPM)36.
Batho ba baholo ba O. cornifrons overwintering in cocoons ba ile ba fumanoa Fruit Research Center, Biglerville, PA, 'me ba bolokoa ho −3 ho 2°C (±0.3°C).Pele ho teko (li-cocoons tse 600 ka kakaretso).Ka May 2022, li-cocoons tse 100 tsa O. cornifrons li ne li fetisetsoa letsatsi le leng le le leng ka har'a linoelo tsa polasetiki (50 cocoons ka senoelo, DI 5 cm × 15 cm ka bolelele) 'me li hlakola li ne li kenngoa ka har'a linoelo ho khothalletsa ho bula le ho fana ka substrate e chewable, ho fokotsa khatello ea kelello holim'a lejoe. linotsi37 .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-feeders tse 10 ml tse nang le 50% ea tharollo ea sucrose 'me u li boloke matsatsi a mane ho netefatsa ho koala le ho nyalana .23 ° C, mongobo o lekanyelitsoeng 60%, photoperiod 10 l (tlaase matla): matsatsi a 14.Tse 100 tse mated tse tšehali le tse tona li ne li lokolloa hoseng ho hong le ho hong ka matsatsi a tšeletseng (100 ka letsatsi) lihlaheng tse peli tsa maiketsetso nakong ea lipalesa tsa apole tse phahameng (sehlaha sa sefi: bophara 33.66 × bophahamo 30.48 × bolelele 46.99 cm; Tlatsetso Figure 1).E behiloe Pennsylvania State Arboretum, haufi le cheri (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 ea lifate tsa liapole (Malus coronaria, Malus), sefate sa liapole 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 polasetiki e putsoa ea linonyana e lekana ka holim'a mabokose a mabeli a lehong.Lebokose le leng le le leng la sehlaha le ne le na le li-tubes tsa pampiri tsa kraft tse 800 tse se nang letho (spiral open, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) e kentsoeng ka har'a li-opaque cellophane tubes (0.7 OD bona Li-plug tsa Plastic (T-1X plugs) li fana ka libaka tsa ho haha .
Mabokose a mabeli a lihlaha a ne a shebane le bochabela 'me a ne a koahetsoe ka terata ea serapa sa polasetiki e tala (mohlala oa Everbilt #889250EB12, boholo ba ho bula 5 × 5 cm, 0.95 m × 100 m) ho thibela litoeba le linonyana ho kena le ho behoa holim'a mobu haufi le mobu oa lebokose la sehlaha. mabokose.Nest box (Setshwantsho sa Tlatsetso 1a).Mahe a poone a ne a bokelloa letsatsi le leng le le leng ka ho bokella li-tubes tse 30 lihlaheng ebe o li isa laboratoring.U sebelisa sekere, etsa seha qetellong ea tube, ebe u qhaqholla spiral tube ho pepesa lisele tsa brood.Mahe a motho ka mong le peo e phofo ea ’ona a ile a ntšoa ka spatula e kobehileng (Microslide tool kit, BioQuip Products Inc., California).Mahe a ne a kenngoa pampiring e mongobo 'me a kenngoa ka sejana sa Petri bakeng sa lihora tsa 2 pele a sebelisoa litekong tsa rona (Setšoantšo sa Tlatsetso 1b-d).
Ka laboratoring, re ile ra lekola chefo ea molomo ea li-fungicides tse tšeletseng tse sebelisitsoeng pele le nakong ea palesa ea apole maemong a mararo (0.1X, 0.5X, le 1X, moo 1X e leng letšoao le sebelisoang ka lilithara tse 100 tsa metsi/acre. tšimong)., Lethathamo la 1).Khatiso e 'ngoe le e' ngoe e phetoa makhetlo a 16 (n = 16).Li-fungicides tse peli tsa ho kopana (Letlapa la S1: mancozeb 2696.14 ppm le captan 2875.88 ppm) le li-fungicides tse 'nè tsa systemic (Letlapa la S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobupp5 5 cyprodinil 2.1 m) chefo ho litholoana, meroho le lijalo tsa mekhabiso .Re ile ra etsa homogenized peo e phofshoana ka grinder, ra fetisetsa 0.20 g ho seliba (24-seliba Falcon Plate), 'me re eketsa le ho kopanya 1 μL ea tharollo ea fungicide ho etsa peō ea pyramidal ka liliba tse tebileng tsa 1 mm tseo mahe a neng a behoa ho tsona.Beha ho sebelisa spatula e nyane (Setšoantšo sa Tlatsetso 1c,d).Lipoleiti tsa phakoe li ne li bolokiloe mocheso oa kamore (25 ° C) le 70% ea mongobo o lekanyelitsoeng.Re ile ra li bapisa le li-larvae tsa taolo tse feptjoang ka lijo tse nang le peō ea homogeneous e tšetsoeng ka metsi a hloekileng.Re ile ra tlaleha lefu le ho lekanya boima ba li-larval letsatsi le leng le le leng ho fihlela li-larvae li fihla lilemong tsa pele ho pupal li sebelisa tekanyo ea tlhahlobo (Fisher Scientific, accuracy = 0.0001 g).Qetellong, karo-karolelano ea thobalano e ile ea hlahlojoa ka ho bula cocoon ka mor'a likhoeli tse 2,5.
DNA e ntšitsoe ho li-larvae tsa O. cornifrons (n = 3 ka boemo ba phekolo, peō e phekotsoeng ea mancozeb le e sa phekoloang) 'me ra etsa litlhahlobo tsa mefuta-futa ea likokoana-hloko lisampoleng tsena, haholo hobane ho mancozeb ho ile ha hlokomeloa lefu le phahameng ka ho fetisisa ho li-larvae.ho amohela MnZn.DNA e ile ea holisoa, ea hloekisoa ka ho sebelisa DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), 'me ea hlophisoa (li-cycles tse 600) ho Illumina® MiSeq™ ho sebelisa lisebelisoa tsa v3.Tatelano e reriloeng ea liphatsa tsa lefutso tsa 16S ribosomal RNA ea baktheria e ile ea etsoa ho sebelisoa Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) ho sebelisoa li-primers tse lebisang sebakeng sa V3-V4 sa 16S rRNA gene.Ho feta moo, tatellano ea 18S e ile ea etsoa ho sebelisoa 10% PhiX kenyeletso, 'me ho hōlisa ho ile ha etsoa ho sebelisa primer pair 18S001 le NS4.
Ho kenya le ho sebetsa ka bobeli ho bala39 o sebelisa phaephe ea QIIME2 (v2022.11.1).Litemana tsena li ile tsa hloekisoa 'me tsa kopanngoa,' me tatelano ea chimeric e ile ea tlosoa ho sebelisoa plugin ea DADA2 ho QIIME2 (qiime dada2 noise pairing)40.Likabelo tsa sehlopha sa 16S le 18S li entsoe ho sebelisoa plugin ea classifier Classify-sklearn le artifact e koetlisitsoeng esale pele silva-138-99-nb-classifier.
Lintlha tsohle tsa liteko li ile tsa hlahlojoa bakeng sa ho tloaeleha (Shapiro-Wilks) le homogeneity ea liphapano (teko ea Levene).Hobane sete ea data ha ea ka ea kopana le maikutlo a tlhahlobo ea parametric mme phetoho e ile ea hloleha ho tiisa masala, re entse ANOVA (Kruskal-Wallis) ea litsela tse peli e sa sebetseng ka lintlha tse peli [nako (mekhahlelo e meraro 2, 5, le letsatsi la 8). nako) le fungicide] ho lekola phello ea kalafo ho larval fresh weight, joale lipapiso tsa post hoc nonparametric pairwise li ile tsa etsoa ho sebelisoa tlhahlobo ea Wilcoxon.Re sebelisitse mohlala o tloaelehileng oa linear (GLM) le kabo ea Poisson ho bapisa liphello tsa li-fungicides ho pholoha ho pholletsa le likarolo tse tharo tsa fungicide41,42.Bakeng sa tlhahlobo ea bongata bo fapaneng, palo ea li-amplicon sequence variants (ASVs) e ile ea putlama boemong ba genus.Papiso ea bongata bo fapaneng lipakeng tsa lihlopha tse sebelisang 16S (genus level) le 18S e ngata e amanang le eona e ile ea etsoa ho sebelisoa mohlala o akaretsang oa tlatsetso bakeng sa boemo, sekhahla, le sebopeho (GAMLSS) le kabo ea malapa ea beta zero-inflated (BEZI), e neng e entsoe ka bongata. .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 feela ba lekhetho bo bong le bo bong bo sebelisitsoeng bakeng sa litlhahlobo tse fapaneng.Litlhahlobo tsohle tsa lipalo-palo li entsoe ho sebelisoa R (v. 3.4.3., morero oa CRAN) (Sehlopha sa 2013).
Ho pepesetsoa mancozeb, pyrithiostrobin, le trifloxystrobin ho fokolitse boima ba 'mele haholo ho O. cornifrons (setšoantšo sa 1).Liphello tsena li ne li lula li hlokomeloa bakeng sa litekanyetso tsohle tse tharo tse hlahlobiloeng (setšoantšo sa 1a-c).Cyclostrobin le myclobutanil ha lia ka tsa fokotsa boima ba li-larvae haholo.
Karolelano ea boima bo bocha ba li-stem borer larvae tse lekantsoeng ka nako tse tharo tlas'a mekhoa e mene ea phekolo ea lijo (homogeneous pollen feed + fungicide: control, 0.1X, 0.5X le 1X doses).(a) Tekanyo e tlaase (0.1X): ntlha ea pele (letsatsi la 1): χ2: 30.99, DF = 6;P <0.0001, ntlha ea nako ea bobeli (letsatsi la 5): 22.83, DF = 0.0009;kgetlo la boraro;ntlha (letsatsi la 8): χ2: 28.39, DF = 6;(b) halofo ea tekanyo (0.5X): ntlha ea pele (letsatsi la 1): χ2: 35.67, DF = 6;P <0.0001, ntlha ea nako ea bobeli (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 feletseng (1X): ntlha ea pele (letsatsi la 1) χ2: 20.64, P = 6;P = 0.0326, ntlha ea nako 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 ea nonparametric ea phapang.Mekotla e emela ho bolela ± SE ea ho bapisa ka bobeli (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Ka tekanyo e tlaase ka ho fetisisa (0.1X), boima ba 'mele oa larval bo fokotsehile ka 60% ka trifloxystrobin, 49% le mancozeb, 48% le myclobutanil, le 46% le pyrithistrobin (setšoantšo sa 1a).Ha e pepesehetse halofo ea tekanyo ea tšimo (0.5X), boima ba 'mele oa mancozeb larvae bo fokotsehile ka 86%, pyrithiostrobin ka 52% le trifloxystrobin ka 50% (setšoantšo sa 1b).Tekanyetso e felletseng ea mancozeb (1X) ea mancozeb e fokolitse boima ba 'mele ka 82%, pyrithiostrobin ka 70%, le trifloxystrobin, myclobutanil le sangard ka hoo e ka bang 30% (setšoantšo sa 1c).
Mafu a ne a phahame ka ho fetisisa har'a li-larvae tse feptjoang ka peō e entsoeng ka mancozeb, 'me ea lateloa ke pyrithiostrobin le trifloxystrobin.Ho shoa ho ile ha eketseha ka litekanyetso tse ntseng li eketseha tsa mancozeb le pyritisoline (setšoantšo sa 2; Letlapa la 2).Leha ho le joalo, ho shoa ha corn borer ho ile ha eketseha hanyenyane feela ha likhahla tsa trifloxystrobin li ntse li eketseha;cyprodinil le captan ha lia ka tsa eketsa lefu haholo ha li bapisoa le liphekolo tsa taolo.
Ho shoa ha likokoanyana tsa borer fly ho ile ha bapisoa ka mor'a ho kenngoa ha peo e phofshoana e le 'ngoe e phekoloang ka li-fungicides tse tšeletseng tse fapaneng.Mancozeb le pentopyramide li ne li ameha haholo ha li pepesetsoa molomo ho tšenyane ea poone (GLM: χ = 29.45, DF = 20, P = 0.0059) (mola, moepa = 0.29, P <0.001; moepa = 0.24, P) <0.00).
Ka karolelano, ho pholletsa le liphekolo tsohle, 39.05% ea bakuli e ne e le basali le 60.95% e le banna.Har'a liphekolo tsa taolo, karolo ea basali e ne e le 40% lithutong tsa tekanyo e tlase (0.1X) le halofo ea tekanyo (0.5X), le 30% lithutong tsa tekanyo ea tšimo (1X).Ka tekanyo ea 0.1X, har'a li-larvae tse fepetsoeng ka peo e phofshoana e phekoloang 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 li-larvae tse kholo e ne e le tse tšehali, 44% ea li-larvae tse kholo e le tse tšehali.e tšehali, 41% ea li-larvae tse kholo e ne e le tse tšehali, 'me taolo e ne e le 31% (setšoantšo sa 3a).Ka makhetlo a 0.5, 33% ea liboko tsa batho ba baholo sehlopheng sa mancozeb le pyrithiostrobin e ne e le 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 liborere tse tšehali le tse tona ka mor'a ho pepesehela sebolaea-khuru.(a) Tekanyetso e tlase (0.1X).(b) Halofo ea tekanyo (0.5X).(c) Tekanyo ea tšimo kapa tekanyo e feletseng (1X).
Tlhahlobo ea tatellano ea 16S e bontšitse hore sehlopha sa baktheria se fapane pakeng tsa li-larvae tse feptjoang ka peo e phofshoana ea mancozeb le li-larvae tse feptjoang ka peo e phofo e sa phekoloang (Setšoantšo sa 4a).Lenane la likokoana-hloko tsa li-larvae tse sa phekoloang tse feptjoang ka peō ea limela li ne li le holimo ho feta li-larvae tse feptjoang ka peō e entsoeng ka mancozeb (Setšoantšo sa 4b).Le hoja phapang e bonoang ea leruo pakeng tsa lihlopha e ne e se ea bohlokoa ka lipalo-palo, e ne e le tlase haholo ho feta e ileng ea hlokomeloa bakeng sa li-larvae tse jang peo e phofo e sa phekoloang (Setšoantšo sa 4c).Bongata bo lekanyelitsoeng bo bontšitse hore likokoana-hloko tse feptjoang ke peo e phofshoana ea taolo li ne li fapane ho feta li-larvae tse feptjoang ka li-larvae tse phekoloang ke mancozeb (Setšoantšo sa 5a).Tlhahlobo e hlalosang e ile ea senola ho ba teng ha 28 genera ho laola le lisampole tse tšoaroang ke mancozeb (setšoantšo sa 5b).c Tlhahlobo e sebelisang tatellano ea 18S e senotse liphapang tse kholo (Setšoantšo sa Tlatsetso 2).
Lifaele tsa SAV tse ipapisitseng le tatellano ea 16S li ile tsa bapisoa le borui ba Shannon 'me tsa hlokomela borui boemong ba phylum.(a) Principal coordinate analysis (PCoA) e ipapisitseng le sebopeho sa sechaba sa likokoana-hloko ka har'a peō e feptjoang kapa taolong e sa phekoloang (e putsoa) le larvae e feptjoang ke mancozeb (orange).Ntlha ka 'ngoe ea data e emela sampole e arohaneng.PCoA e ne e baloa ho sebelisoa sebaka sa Bray-Curtis sa kabo ea multivariate t.Li-oval li emela boemo ba 80% ba kholiseho.(b) Boxplot, data e tala ea Shannon ea leruo (lintlha) le c.Leruo le bonahalang.Li-boxplots li bonts'a mabokose a mela e bohareng, interquartile range (IQR), le 1.5 × IQR (n = 3).
Sebopeho sa lihlopha tsa likokoana-hloko tsa li-larvae tse feptjoang ka mancozeb le peo e phofo e sa phekoloang.(a) Bongata bo lekantsoeng ba mofuta oa likokoana-hloko bo baloa ka har'a li-larvae.(b) 'Mapa oa mocheso oa lichaba tsa likokoana-hloko tse khethiloeng.Delftia (karolelano ea liphapang (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 ea 'mapa oa mocheso e kopantsoe ho sebelisoa sebaka sa khokahano le karolelano ea khokahanyo.
Liphetho tsa rona li bonts'a hore ho pepesetsoa ka molomo ho kopana (mancozeb) le systemic (pyrostrobin le trifloxystrobin) fungicides, tse sebelisoang haholo nakong ea lipalesa, li fokolitse boima ba 'mele haholo le ho eketseha ha lefu la poone ea poone.Ntle le moo, mancozeb e fokolitse haholo mefuta-futa le bongata ba microbiome nakong ea prepupal.Myclobutanil, e 'ngoe ea fungicide ea systemic, e fokolitse boima ba 'mele oa larval haholo ka litekanyetso tsohle tse tharo.Phello ena e ile ea bonahala ka nako ea bobeli (letsatsi la 5) le la boraro (letsatsi la 8).Ka lehlakoreng le leng, cyprodinil le captan ha lia ka tsa fokotsa boima ba 'mele kapa ho phela ha li bapisoa le sehlopha sa taolo.Ho ea ka tsebo ea rona, mosebetsi ona ke oa pele oa ho fumana liphello tsa litekanyetso tsa masimo tsa li-fungicides tse fapaneng tse sebelisetsoang ho sireletsa lijalo tsa poone ka ho pepesehela peō ea limela ka ho toba.
Liphekolo tsohle tsa fungicide haholo li fokolitse boima ba 'mele ha li bapisoa le liphekolo tsa taolo.Mancozeb e bile le phello e kholo ho boima ba 'mele oa larval ka phokotso ea karolelano ea 51%, e lateloe ke pyrithiostrobin.Leha ho le joalo, liphuputso tse ling ha lia tlaleha litlamorao tse mpe tsa litekanyetso tsa masimong tsa li-fungicides methating ea larval44.Leha biocides tsa dithiocarbamate li bonts'itsoe li na le chefo e tlase e matla45, ethylene bisdithiocarbamates (EBDCS) joalo ka mancozeb e ka theoleloa ho urea ethylene sulfide.Ka lebaka la liphello tsa eona tsa phetoho liphoofolong tse ling, sehlahisoa sena sa ho senyeha se ka 'na sa ikarabella bakeng sa liphello tse hlokometsoeng46,47.Liphuputso tse fetileng li bontšitse hore sebopeho sa ethylene thiourea se susumetsoa ke lintlha tse kang mocheso o phahameng48, maemo a mongobo49 le bolelele ba polokelo ea lihlahisoa50.Maemo a nepahetseng a polokelo ea li-biocide a ka fokotsa litlamorao tsena.Ho feta moo, European Food Safety Authority e bontšitse ho tšoenyeha ka chefo ea pyrithiopide, e bonts'itsoeng e le carcinogenic tsamaisong ea tšilo ea lijo ea liphoofolo tse ling51.
Tšebeliso ea molomo ea mancozeb, pyrithiostrobin le trifloxystrobin e eketsa ho shoa ha li-larvae tsa poone.Ka lehlakoreng le leng, myclobutanil, ciprocycline le captan li ne li se na phello ho lefu.Liphetho tsena li fapane le tsa Ladurner et al.52, ea bontšitseng hore captan e fokolitse haholo bophelo ba batho ba baholo O. lignaria le Apis mellifera L. (Hymenoptera, Apisidae).Ho feta moo, li-fungicides tse kang captan le boscalid li fumanoe li baka lefu la larval52,53,54 kapa ho fetola boitšoaro ba ho fepa55.Liphetoho tsena, ka lehlakoreng le leng, li ka ama boleng ba phepo e nepahetseng ea peo e phofshoana 'me qetellong e fumana matla a sethaleng sa larval.Lefu le hlokometsoeng sehlopheng sa taolo le ne le lumellana le lithuto tse ling tsa 56,57.
Karolelano ea thobalano e khahlang banna e bonoang mosebetsing oa rona e ka hlalosoa ka lintlha tse kang ho nyallana ho sa lekaneng le maemo a leholimo a mabe nakong ea lipalesa, joalo ka ha ho khothalelitsoe pele bakeng sa O. cornuta ke Vicens le Bosch.Le hoja basali le banna thutong ea rona ba ne ba e-na le matsatsi a mane a ho nyalana (nako eo ka kakaretso e nkoang e lekane bakeng sa ho nyalana ho atlehileng), re ile ra fokotsa matla a khanya ka boomo ho fokotsa khatello ea kelello.Leha ho le joalo, phetoho ena e ka kena-kenana le mokhoa oa ho nyalana ka boomo61.Ho feta moo, linotsi li na le matsatsi a 'maloa a maemo a mabe a leholimo, ho kenyelletsa pula le mocheso o tlase (<5°C), tse ka boela tsa ama katleho ea ho nyalana hampe4,23.
Leha boithuto ba rona bo ne bo tsepamisitse maikutlo ho microbiome eohle ea larval, liphetho tsa rona li fana ka temohisiso mabapi le likamano tse ka bang teng har'a lichaba tsa libaktheria tse ka bang bohlokoa ho phepo ea linotsi le ho pepeseha ha fungicide.Ka mohlala, li-larvae tse fepetsoeng peō e entsoeng ka mancozeb li fokolitse haholo sebopeho sa sechaba sa likokoana-hloko le bongata ha li bapisoa le li-larvae tse feptjoang peo e phofo e sa phekoloang.Ho li-larvae tse neng li ja peō e phofshoana e sa phekoloang, lihlopha tsa baktheria tsa Proteobacteria le Actinobacteria li ne li le matla 'me boholo ba tsona e ne e le aerobic kapa facultatively aerobic.Likokoana-hloko tsa Delft, hangata li amahanngoa le mefuta ea linotsi tse ikemetseng, li tsebahala li na le ts'ebetso ea lithibela-mafu, e bontšang karolo e ka bang teng ea tšireletso khahlanong le likokoana-hloko.Mofuta o mong oa baktheria, Pseudomonas, o ne o e-na le li-larvae tse ngata tse feptjoang peō e phofshoana e sa phekoloang, empa e ile ea fokotseha haholo ka har'a likokoana-hloko tse phekoloang ke mancozeb.Liphetho tsa rona li tšehetsa liphuputso tse fetileng tse khethollang Pseudomonas e le e 'ngoe ea lihlopha tse ngata ka ho fetisisa ho O. bicornis35 le bobi bo bong bo ikemetseng34.Le hoja bopaki ba liteko bakeng sa karolo ea Pseudomonas bophelong bo botle ba O. cornifrons bo e-s'o ithutoe, baktheria ena e bontšitsoe hore e khothalletsa ho qaptjoa ha chefo e sireletsang ho beetle Paederus fuscipes le ho khothalletsa metabolism ea arginine in vitro 35, 65. Lintlha tsena li fana ka maikutlo karolo e ka bang teng ts'ireletsong ea vaerase le baktheria nakong ea nts'etsopele ea li-larvae tsa O. cornifrons.Microbacterium ke leloko le leng le hlwauweng phuputsong ya rona le tlalehwang hore le teng ka dipalo tse hodimo ho dibokwana tsa masole a matsho tlasa maemo a tlala66.Ho li-larvae tsa O. cornifrons, microbacteria e ka kenya letsoho ho leka-lekana le ho tiea ha mala a microbiome tlas'a maemo a khatello ea kelello.Ho feta moo, Rhodococcus e fumaneha ka har'a li-larvae tsa O. cornifrons 'me e tsebahala ka bokhoni ba eona ba ho tlosa chefo67.Mofuta ona o boetse o fumaneha maleng a A. florea, empa ka bongata bo tlase haholo68.Liphetho tsa rona li bonts'a boteng ba mefuta e mengata ea liphatsa tsa lefutso ho pholletsa le mefuta e mengata ea li-microbial taxa tse ka fetolang ts'ebetso ea metabolism ka har'a li-larvae.Leha ho le joalo, kutloisiso e molemo ea mefuta-futa ea ts'ebetso ea O. cornifron e ea hlokahala.
Ka bokhuts'oane, liphetho li bonts'a hore mancozeb, pyrithiostrobin, le trifloxystrobin li fokolitse boima ba 'mele le ho eketseha ha lefu la liboko tsa poone.Leha ho na le ngongoreho e ntseng e hola mabapi le litlamorao tsa li-fungicides ho li-pollinator, ho hlokahala ho utloisisa hamolemo litlamorao tsa mesaletsa ea metabolite ea metsoako ena.Liphetho tsena li ka kenyelletsoa ho likhothaletso bakeng sa mananeo a kopaneng a taolo ea phollina e thusang lihoai ho qoba tšebeliso ea li-fungicides tse itseng pele le nakong ea lipalesa tsa sefate sa litholoana ka ho khetha li-fungicides le ho fetola nako ea ts'ebeliso, kapa ka ho khothaletsa tšebeliso ea mefuta e meng e seng kotsi haholo 36. Tlhahisoleseding ena. e bohlokoa bakeng sa ho hlahisa likhothaletso.mabapi le tšebeliso ea chefo e bolaeang likokonyana, joalo ka ho lokisa mananeo a teng a ho fafatsa le ho fetola nako ea ho fafatsa ha u khetha meriana e bolaeang likokonyana kapa ho khothaletsa tšebeliso ea mefuta e meng e seng kotsi.Ho hlokahala lipatlisiso tse ling mabapi le litlamorao tse mpe tsa li-fungicides mabapi le karo-karolelano ea thobalano, boitšoaro ba ho fepa, gut microbiome, le mekhoa ea limolek'hule e theolelang boima ba 'mele le lefu.
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