Ua tsaug rau koj tuaj xyuas Nature.com. Cov version ntawm browser koj siv tau txwv CSS kev txhawb nqa. Txhawm rau kom tau txais txiaj ntsig zoo tshaj plaws, peb xav kom koj siv qhov tshiab ntawm koj tus browser (lossis lov tes taw Compatibility Hom hauv Internet Explorer). Nyob rau lub sijhawm no, txhawm rau ua kom muaj kev txhawb nqa tsis tu ncua, peb tab tom nthuav tawm lub xaib tsis muaj styling lossis JavaScript.
Cov tshuaj fungicides feem ntau siv thaum tsob ntoo txiv ntoo tawg paj thiab tuaj yeem hem cov kab pollinators. Txawm li cas los xij, me ntsis paub txog yuav ua li cas tsis yog muv pollinators (piv txwv li, ib leeg muv, Osmia cornifrons) teb rau kev sib cuag thiab cov kab mob fungicides feem ntau siv rau cov txiv apples thaum lub paj tawg. Qhov kev paub dhau los no txwv tsis pub cov kev txiav txim siab txiav txim siab txog kev nyab xeeb thiab lub sijhawm ntawm cov tshuaj tua kab mob fungicide. Peb tau soj ntsuam cov teebmeem ntawm ob qhov kev sib cuag fungicides (captan thiab mancozeb) thiab plaub interlayer / phytosystem fungicides (ciprocycline, myclobutanil, pyrostrobin thiab trifloxystrobin). Qhov cuam tshuam rau larval hnyav nce, ciaj sia taus, kev sib deev piv thiab ntau hom kab mob. Kev ntsuam xyuas tau ua los ntawm kev siv lub qhov ncauj bioassay nyob rau hauv uas cov paj ntoos tau kho nyob rau hauv peb koob tshuaj raws li qhov tam sim no pom zoo koob tshuaj rau kev siv (1X), ib nrab koob (0.5X) thiab koob tshuaj tsawg (0.1X). Tag nrho cov koob tshuaj ntawm mancozeb thiab pyritisoline txo qis lub cev hnyav thiab muaj sia nyob. Tom qab ntawd peb tau ua raws li 16S noob los ua tus kab mob larval bacteriome ntawm mancozeb, fungicide lub luag haujlwm rau kev tuag siab tshaj plaws. Peb pom tias cov kab mob sib txawv thiab kev nplua nuj tau txo qis hauv cov larvae noj ntawm mancozeb-kho pollen. Peb qhov kev soj ntsuam kuaj pom tau hais tias kev txau qee yam ntawm cov kab mob fungicides thaum lub sij hawm tawg paj yog qhov tshwj xeeb rau kev noj qab haus huv ntawm O. cornifrons. Cov ntaub ntawv no muaj feem cuam tshuam rau kev txiav txim siab txog kev tswj hwm yav tom ntej hais txog kev siv cov khoom siv tiv thaiv cov txiv hmab txiv ntoo thiab ua lub hauv paus rau cov txheej txheem tswj hwm kev tiv thaiv pollinators.
Tib neeg mason muv Osmia cornifrons (Hymenoptera: Megachilidae) tau qhia rau Tebchaws Meskas los ntawm Nyij Pooj thaum xyoo 1970s thiab thaum ntxov 1980s, thiab cov tsiaj tau ua lub luag haujlwm tseem ceeb hauv kev tswj hwm ecosystems txij thaum ntawd los. Naturalized pejxeem ntawm cov muv no yog ib feem ntawm kwv yees li 50 hom tsiaj qus uas ntxiv rau cov muv uas pollinate almond thiab kua txiv hmab txiv ntoo hauv Tebchaws Meskas 2,3. Mason muv ntsib ntau yam teeb meem, suav nrog thaj chaw tawg, kab mob, thiab tshuaj tua kab3,4. Ntawm cov tshuaj tua kab, fungicides txo lub zog nce, foraging5 thiab lub cev txias 6,7. Txawm hais tias kev tshawb fawb tsis ntev los no qhia tias kev noj qab haus huv ntawm Mason muv yog cuam tshuam ncaj qha los ntawm cov kab mob commensal thiab ectobactic microorganisms, 8,9 vim tias cov kab mob thiab cov kab mob tuaj yeem cuam tshuam rau kev noj zaub mov thiab kev tiv thaiv kab mob, cov teebmeem ntawm cov kab mob fungicide cuam tshuam rau ntau haiv neeg ntawm Mason bees yog nyuam qhuav pib. kawm.
Cov tshuaj fungicides ntawm ntau yam teebmeem (kev sib cuag thiab kev ua haujlwm) yog txau rau hauv orchards ua ntej thiab thaum lub sij hawm flowering los kho cov kab mob xws li kua txob, iab rot, xim av rot thiab powdery mildew10,11. Fungicides suav hais tias tsis muaj teeb meem rau pollinators, yog li lawv raug pom zoo rau cov neeg ua teb thaum lub sij hawm flowering; Kev nthuav tawm thiab kev noj cov kab mob fungicides los ntawm muv yog qhov paub zoo, vim nws yog ib feem ntawm cov txheej txheem sau npe tshuaj tua kab los ntawm US Environmental Protection Agency thiab ntau lwm lub koom haum tswj hwm hauv tebchaws 12,13,14. Txawm li cas los xij, qhov cuam tshuam ntawm fungicides rau cov tsis yog muv tsis tshua paub vim tias lawv tsis tas yuav tsum tau ua raws li kev pom zoo kev lag luam hauv Tebchaws Meskas 15. Tsis tas li ntawd, feem ntau tsis muaj cov txheej txheem kev cai rau kev sim ib leeg bees16,17, thiab tswj cov cheeb tsam uas muab cov muv rau kev sim yog qhov nyuaj18. Kev sim sib txawv ntawm cov muv tswj hwm tau nce zuj zus hauv Tebchaws Europe thiab Tebchaws Asmeskas los kawm txog cov teebmeem ntawm cov tshuaj tua kab ntawm cov muv qus, thiab cov txheej txheem kev cai tsis ntev los no tau tsim rau O. cornifrons19.
Horned muv yog monocytes thiab yog siv nyob rau hauv kev lag luam hauv carp qoob loo raws li ib tug ntxiv los yog hloov rau zib mu muv. Cov muv no tshwm sim thaum lub Peb Hlis thiab Plaub Hlis, nrog rau cov txiv neej precocious tshwm sim peb mus rau plaub hnub ua ntej cov poj niam. Tom qab mating, tus poj niam nquag sau paj ntoos thiab nectar los muab cov noob qes nyob rau hauv lub zes zes kab noj hniav (natural los yog dag) 1,20. Cov qe tso rau ntawm paj ntoos hauv cov hlwb; tus poj niam mam li ua ib phab ntsa av ua ntej npaj lub cell tom ntej. Thawj instar larvae yog nyob rau hauv lub chorion thiab pub rau embryonic kua. Los ntawm lub thib ob mus rau lub thib tsib instar (prepupa), cov larvae pub rau pollen22. Thaum cov paj ntoos tawg tag lawm, cov larvae tsim cocoons, pupate thiab tshwm sim raws li cov neeg laus nyob rau hauv tib brood chamber, feem ntau nyob rau lub caij ntuj sov 20,23. Cov neeg laus tshwm sim tom qab lub caij nplooj ntoos hlav. Cov neeg laus muaj sia nyob yog txuam nrog lub zog nce (qhov hnyav nce) raws li kev noj zaub mov. Yog li, kev noj zaub mov zoo ntawm paj ntoos, nrog rau lwm yam xws li huab cua lossis raug tshuaj tua kab, yog qhov txiav txim ntawm kev ciaj sia thiab kev noj qab haus huv24.
Insecticides thiab fungicides thov ua ntej flowering muaj peev xwm tsiv mus nyob rau hauv cov nroj tsuag vasculature mus rau txawv degrees, los ntawm translaminar (xws li, muaj peev xwm txav los ntawm sab sauv ntawm nplooj mus rau hauv qab, zoo li ib co fungicides) 25 mus rau lub systemic teebmeem. , uas tuaj yeem nkag mus rau hauv cov yas los ntawm cov hauv paus hniav, tuaj yeem nkag mus rau cov kua txiv hmab txiv ntoo ntawm cov paj 26, qhov uas lawv tuaj yeem tua cov neeg laus O. cornifrons27. Qee cov tshuaj tua kab kuj nkag mus rau hauv paj ntoos, cuam tshuam rau kev loj hlob ntawm pob kws larvae thiab ua rau lawv tuag19. Lwm cov kev tshawb fawb tau pom tias qee cov tshuaj fungicides tuaj yeem hloov pauv tus cwj pwm zes ntawm cov hom O. lignaria28. Tsis tas li ntawd, kev soj ntsuam thiab cov kev tshawb fawb hauv teb simulating cov tshuaj tua kab mob tshwm sim (xws li fungicides) tau pom tias cov tshuaj tua kab tsis zoo cuam tshuam rau lub cev 22 morphology 29 thiab kev ciaj sia ntawm zib mu thiab qee cov muv ib leeg. Ntau yam tshuaj fungicidal siv ncaj qha rau qhib paj thaum lub sij hawm paj tuaj yeem kis kab mob paj ntoos sau los ntawm cov neeg laus rau kev loj hlob larval, cov teebmeem uas tseem yuav tau kawm30.
Nws tau paub ntau ntxiv tias kev loj hlob ntawm larval yog cuam tshuam los ntawm paj ntoos thiab cov zej zog microbial ntawm lub plab zom mov. Lub honeybee microbiome cuam tshuam tsis xws li lub cev loj31, metabolic hloov 22 thiab susceptibility rau pathogens32. Cov kev tshawb fawb yav dhau los tau tshuaj xyuas qhov cuam tshuam ntawm theem kev loj hlob, cov as-ham, thiab ib puag ncig ntawm microbiome ntawm cov muv ib leeg. Cov kev tshawb fawb no tau qhia txog qhov sib xws ntawm cov qauv thiab kev nplua nuj ntawm cov kab mob larval thiab paj ntoos microbiomes33, nrog rau cov kab mob ntau tshaj plaws ntawm Pseudomonas thiab Delftia, ntawm cov tsiaj nyeg ib leeg. Txawm li cas los xij, txawm hais tias fungicides tau cuam tshuam nrog cov tswv yim los tiv thaiv muv kev noj qab haus huv, cov teebmeem ntawm fungicides ntawm larval microbiota los ntawm qhov ncauj ncaj qha tseem tsis tau tshawb nrhiav.
Txoj kev tshawb no tau sim qhov tshwm sim ntawm lub ntiaj teb koob tshuaj ntawm 6 feem ntau siv fungicides sau npe siv rau tsob ntoo txiv hmab txiv ntoo hauv Tebchaws Meskas, suav nrog kev sib cuag thiab cov kab mob fungicides tswj qhov ncauj rau pob kws hornworm moth larvae los ntawm cov zaub mov tsis huv. Peb pom tias kev sib cuag thiab cov kab mob fungicides txo cov muv lub cev hnyav nce thiab nce kev tuag, nrog rau cov teebmeem loj tshaj plaws cuam tshuam nrog mancozeb thiab pyrithiopide. Peb mam li muab piv cov microbial ntau haiv neeg ntawm larvae pub rau ntawm mancozeb-kho pollen noj zaub mov nrog cov uas pub rau kev tswj kev noj haus. Peb sib tham txog cov txheej txheem muaj peev xwm hauv qab kev tuag thiab cuam tshuam rau kev sib koom ua ke ntawm kev tswj xyuas kab tsuag thiab pollinator (IPPM)36.
Cov neeg laus O. cornifrons overwintering hauv cocoons tau txais los ntawm Fruit Research Center, Biglerville, PA, thiab khaws cia ntawm −3 txog 2 ° C (± 0.3 ° C). Ua ntej qhov kev sim (600 cocoons nyob rau hauv tag nrho). Thaum lub Tsib Hlis 2022, 100 O. cornifrons cocoons tau hloov txhua hnub rau hauv khob yas (50 cocoons ib khob, DI 5 cm × 15 cm ntev) thiab cov ntaub so tau muab tso rau hauv lub khob los txhawb kev qhib thiab muab lub chewable substrate, txo kev ntxhov siab ntawm pob zeb. qub 37. Muab ob lub khob yas uas muaj cocoons rau hauv ib lub tawb kab (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) nrog 10 ml feeders uas muaj 50% sucrose tov thiab khaws cia rau plaub hnub kom kaw thiab mating. 23 ° C, txheeb ze humidity 60%, photoperiod 10 l (tsawg siv): 14 hnub. 100 mated poj niam thiab txiv neej raug tso tawm txhua tag kis sawv ntxov rau rau hnub (100 ib hnub) rau hauv ob lub zes dag thaum lub caij nplooj ntoos hlav paj (khob zes: dav 33.66 × qhov siab 30.48 × ntev 46.99 cm; Ntxiv daim duab 1). Muab tso rau hauv Pennsylvania State Arboretum, ze cherry (Prunus cerasus 'Eubank' Sweet Cherry Pie™), txiv duaj (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), pear (Pyrus perifolia 'Olympic', Pyrus Perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), coronaria txiv apple ntoo (Malus coronaria) thiab ntau yam txiv apples (Malus coronaria, Malus), cov txiv ntoo hauv tsev 'Co-op 30′ Enterprise™, Malus apple tree 'Co-Op 31′ Winecrisp™, begonia 'Kev ywj pheej', Begonia 'Golden Delicious', Begonia 'Nova Spy'). Txhua xiav yas noog tsev haum rau saum ob lub thawv ntoo. Txhua lub thawv zes muaj 800 khoob kraft ntawv raj (kauv qhib, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) muab tso rau hauv opaque cellophane hlab (0.7 OD pom cov yas ntsaws (T-1X plugs) muab cov chaw zes .
Ob lub zes lub thawv ntsib sab hnub tuaj thiab tau them nrog ntsuab yas vaj fencing (Everbilt qauv # 889250EB12, qhib loj 5 × 5 cm, 0.95 m × 100 m) kom tiv thaiv nas thiab noog nkag thiab muab tso rau ntawm cov av nto ntawm lub zes lub thawv av thawv. Zes lub thawv (Cov duab ntxiv 1a). Pob kws borer qe tau sau txhua hnub los ntawm kev sau 30 raj ntawm zes thiab thauj mus rau qhov chaw kuaj. Siv txiab, ua ib qho kev txiav ntawm qhov kawg ntawm lub raj, ces disassemble lub kauv raj kom nthuav tawm cov brood hlwb. Cov qe thiab lawv cov paj ntoos raug tshem tawm los ntawm kev siv lub spatula nkhaus (Microslide toolkit, BioQuip Products Inc., California). Cov qe tau muab tso rau ntawm daim ntawv lim dej thiab muab tso rau hauv Petri tais rau 2 teev ua ntej siv hauv peb qhov kev sim (Cov duab ntxiv 1b-d).
Hauv chav kuaj, peb ntsuas qhov ncauj toxicity ntawm 6 fungicides siv ua ntej thiab thaum lub sij hawm paj paj ntawm peb qhov concentrations (0.1X, 0.5X, thiab 1X, qhov twg 1X yog lub cim thov rau 100 nkas loos dej / acre. High field dose = concentration. hauv teb). , Tab 1). Txhua qhov concentration tau rov ua dua 16 zaug (n = 16). Ob qho tshuaj tiv thaiv kab mob fungicides (Table S1: mancozeb 2696.14 ppm thiab captan 2875.88 ppm) thiab plaub cov kab mob fungicides (Table S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil. 280.845 ppm) toxicity rau txiv hmab txiv ntoo, zaub thiab ornamental qoob loo. Peb homogenized paj ntoos siv lub grinder, pauv 0.20 g mus rau lub qhov dej (24-zoo Falcon Phaj), thiab ntxiv thiab sib tov 1 μL ntawm fungicide tov los tsim pyramidal paj ntoos nrog 1 hli tob qhov dej rau hauv cov qe tau muab tso rau. Qhov chaw siv lub spatula mini (Daim duab 1c, d). Falcon daim hlau tau khaws cia ntawm chav tsev kub (25 ° C) thiab 70% txheeb ze av noo. Peb piv lawv nrog tswj larvae noj ib homogeneous pollen noj zaub mov kho nrog dej ntshiab. Peb tau sau cov neeg tuag thiab ntsuas qhov hnyav ntawm cov menyuam mos txhua hnub kom txog thaum cov menyuam yaus mus txog lub hnub nyoog prepupal siv qhov kev ntsuas ntsuas (Fisher Scientific, qhov tseeb = 0.0001 g). Thaum kawg, kev sib deev piv tau raug ntsuas los ntawm kev qhib lub cocoon tom qab 2.5 lub hlis.
DNA tau muab rho tawm los ntawm tag nrho O. cornifrons larvae (n = 3 rau ib qho kev kho mob, mancozeb-kho thiab tsis kho paj ntoos) thiab peb tau ua cov tshuaj microbial ntau haiv neeg ntawm cov qauv no, tshwj xeeb tshaj yog vim nyob rau hauv mancozeb qhov kev tuag siab tshaj plaws tau pom nyob rau hauv larvae. txais MnZn. DNA tau nthuav dav, ua kom huv si siv cov khoom siv DNAZymoBIOMICS®-96 MagBead DNA (Zymo Research, Irvine, CA), thiab ua tiav (600 cycles) ntawm Illumina® MiSeq™ siv cov khoom siv v3. Targeted sequencing ntawm cov kab mob 16S ribosomal RNA noob tau ua los ntawm Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) siv primers tsom rau thaj tsam V3-V4 ntawm 16S rRNA noob. Tsis tas li ntawd, 18S sequencing tau ua tiav siv 10% PhiX suav nrog, thiab kev nthuav dav tau ua tiav siv cov primer khub 18S001 thiab NS4.
Ntshuam thiab txheej txheem ua khub reads39 siv QIIME2 pipeline (v2022.11.1). Cov ntawv nyeem no tau muab txiav thiab sib sau ua ke, thiab cov kab ke chimeric raug tshem tawm siv DADA2 plugin hauv QIIME2 (qiime dada2 suab nrov ua ke) 40. Cov chav kawm 16S thiab 18S tau ua tiav siv cov khoom cais tawm plugin Classify-sklearn thiab pre-trained artifact silva-138-99-nb-classifier.
Tag nrho cov ntaub ntawv sim tau raug kuaj xyuas qhov qub (Shapiro-Wilks) thiab homogeneity ntawm qhov sib txawv (Levene's test). Vim hais tias cov ntaub ntawv teev tsis tau raws li qhov kev xav ntawm parametric tsom xam thiab qhov kev hloov pauv tsis ua raws li cov khoom seem, peb tau ua ib qho kev tsis sib xws ntawm ob txoj kev ANOVA (Kruskal-Wallis) nrog ob yam [lub sij hawm (peb-theem 2, 5, thiab 8 hnub). lub sij hawm cov ntsiab lus) thiab fungicide] los soj ntsuam cov txiaj ntsig ntawm kev kho mob ntawm larval tshiab qhov hnyav, tom qab ntawd tom qab hoc nonparametric pairwise kev sib piv tau ua los ntawm Wilcoxon xeem. Peb siv ib tug generalized linear qauv (GLM) nrog ib tug Poisson tis los sib piv cov teebmeem ntawm fungicides rau ciaj sia nyob thoob plaws peb fungicide concentrations41,42. Rau kev txheeb xyuas qhov sib txawv ntawm qhov sib txawv, tus naj npawb ntawm amplicon sequence variants (ASVs) tau poob rau ntawm qib genus. Kev sib piv ntawm qhov sib txawv ntawm qhov sib txawv ntawm cov pab pawg siv 16S (genus theem) thiab 18S txheeb ze kev nplua nuj tau ua los ntawm kev siv tus qauv ntxiv rau txoj haujlwm, nplai, thiab cov duab (GAMLSS) nrog beta xoom-inflated (BEZI) tsev neeg faib khoom, uas tau ua qauv ntawm macro. . Hauv Microbiome R43 (v1.1). 1). Tshem tawm cov kab mob mitochondrial thiab chloroplast ua ntej kev txheeb xyuas qhov sib txawv. Vim tias cov qib taxonomic sib txawv ntawm 18S, tsuas yog qib qis tshaj ntawm txhua tus taxon tau siv rau kev txheeb xyuas qhov sib txawv. Tag nrho cov kev txheeb xyuas txheeb cais tau ua tiav siv R (v. 3.4.3., CRAN project) (Team 2013).
Kev raug mancozeb, pyrithiostrobin, thiab trifloxystrobin ua rau lub cev hnyav nce hauv O. cornifrons (Fig. 1). Cov teebmeem no tau raug soj ntsuam tas li rau tag nrho peb koob tshuaj ntsuam xyuas (Daim duab 1a–c). Cyclostrobin thiab myclobutanil tsis txo qhov hnyav ntawm larvae.
Qhov nruab nrab qhov hnyav ntawm qia borer larvae ntsuas ntawm peb lub sij hawm cov ntsiab lus raws li plaub txoj kev noj haus (homogeneous paj ntoos pub + fungicide: tswj, 0.1X, 0.5X thiab 1X koob tshuaj). (a) Tsawg koob (0.1X): thawj zaug taw tes (hnub 1): χ2: 30.99, DF = 6; P < 0.0001, thib ob point (hnub 5): 22.83, DF = 0.0009; thib peb; point (hnub 8): χ2: 28.39, DF = 6; (b) ib nrab koob tshuaj (0.5X): thawj zaug taw tes (hnub 1): χ2: 35.67, DF = 6; P <0.0001, lub sijhawm thib ob (hnub ib). ): χ2: 15.98, DF = 6; P = 0.0090; thib peb point (hnub 8) χ2: 16.47, DF = 6; (c) Site lossis puv koob tshuaj (1X): thawj zaug point (hnub 1) χ2: 20.64, P = 6; P = 0.0326, thib ob point (hnub 5): χ2: 22.83, DF = 6; P = 0.0009; thib peb point (hnub 8): χ2: 28.39, DF = 6; nonparametric tsom xam ntawm variance. Bars sawv cev txhais tau tias ± SE ntawm kev sib piv sib piv (α = 0.05) (n = 16) * P ≤ 0.05, ** P ≤ 0.001, *** P ≤ 0.0001.
Ntawm qhov koob tshuaj qis tshaj (0.1X), larval lub cev hnyav tau txo los ntawm 60% nrog trifloxystrobin, 49% nrog mancozeb, 48% nrog myclobutanil, thiab 46% nrog pyrithistrobin (Fig. 1a). Thaum raug rau ib nrab ntawm cov koob tshuaj (0.5X), lub cev hnyav ntawm mancozeb larvae raug txo los ntawm 86%, pyrithiostrobin los ntawm 52% thiab trifloxystrobin los ntawm 50% (Fig. 1b). Ib koob tshuaj tag nrho (1X) ntawm mancozeb txo qis qhov hnyav ntawm 82%, pyrithiostrobin los ntawm 70%, thiab trifloxystrobin, myclobutanil thiab sangard los ntawm kwv yees li 30% (Fig. 1c).
Kev tuag yog siab tshaj plaws ntawm cov larvae noj cov paj noob hlis mancozeb, ua raws li pyrithiostrobin thiab trifloxystrobin. Kev tuag tau nce ntxiv nrog kev nce koob tshuaj ntawm mancozeb thiab pyritisoline (Fig. 2; Table 2). Txawm li cas los xij, cov pob kws borer tuag tau nce me ntsis raws li trifloxystrobin ntau ntxiv; cyprodinil thiab captan tsis ua rau muaj kev tuag ntau dua piv nrog kev tswj xyuas.
Kev tuag ntawm borer ya larvae tau muab piv tom qab noj cov paj ntoos ib tus zuj zus kho nrog rau 6 qhov sib txawv fungicides. Mancozeb thiab pentopyramide tau rhiab heev rau qhov ncauj ntawm pob kws maggots (GLM: χ = 29.45, DF = 20, P = 0.0059) (kab, txoj kab nqes = 0.29, P < 0.001; txoj kab nqes = 0.24, P <0.00).
Qhov nruab nrab, thoob plaws txhua qhov kev kho mob, 39.05% ntawm cov neeg mob yog poj niam thiab 60.95% yog txiv neej. Ntawm cov kev tswj hwm kev kho mob, qhov feem pua ntawm cov poj niam yog 40% nyob rau hauv ob qho tib si cov koob tshuaj qis (0.1X) thiab ib nrab koob tshuaj (0.5X) kev tshawb fawb, thiab 30% hauv kev tshawb fawb-dose (1X). Ntawm 0.1X koob tshuaj, ntawm cov paj noob hlis pub noj tshuaj kho nrog mancozeb thiab myclobutanil, 33.33% ntawm cov laus yog poj niam, 22% ntawm cov laus yog poj niam, 44% ntawm cov laus cov menyuam mos yog poj niam, 44% ntawm cov laus cov menyuam yog poj niam. poj niam, 41% ntawm cov laus larvae yog poj niam, thiab tswj tau 31% (Fig. 3a). Ntawm 0.5 npaug ntawm koob tshuaj, 33% ntawm cov neeg laus cab hauv mancozeb thiab pyrithiostrobin pawg yog poj niam, 36% hauv pawg trifloxystrobin, 41% hauv pawg myclobutanil, thiab 46% hauv pawg cyprostrobin. Daim duab no yog 53% hauv pawg. nyob rau hauv pab pawg neeg captan thiab 38% hauv pawg tswj hwm (Fig. 3b). Ntawm 1X koob tshuaj, 30% ntawm pawg mancozeb yog poj niam, 36% ntawm pawg pyrithiostrobin, 44% ntawm pawg trifloxystrobin, 38% ntawm pawg myclobutanil, 50% ntawm pawg tswj hwm yog poj niam - 38.5% (Fig. 3c) .
Feem pua ntawm cov poj niam thiab txiv neej borers tom qab larval theem fungicide raug. (a) Cov koob tshuaj tsawg (0.1X). (b) Ib nrab koob tshuaj (0.5X). (c) Field dose los yog full dose (1X).
16S kev soj ntsuam ib ntus tau pom tias cov kab mob sib txawv ntawm cov kab mob uas tau noj nrog cov paj ntoo uas tau kho mancozeb thiab cov larvae pub nrog cov paj ntoo uas tsis kho (Fig. 4a). Qhov microbial Performance index ntawm cov larvae uas tsis tau kho rau paj paj yog siab dua li cov larvae pub rau ntawm cov paj ntoo uas tsis kho (Fig. 4b). Txawm hais tias qhov pom qhov sib txawv ntawm kev nplua nuj ntawm cov pab pawg tsis yog qhov tseem ceeb, nws tau qis dua qhov pom ntawm cov larvae pub rau cov paj ntoo uas tsis kho (Fig. 4c). Cov txheeb ze muaj txiaj ntsig tau pom tias cov microbiota ntawm cov larvae pub rau kev tswj cov paj ntoos muaj ntau haiv neeg ntau dua li cov larvae pub rau mancozeb-treated larvae (Fig. 5a). Kev txheeb xyuas piav qhia tau pom muaj 28 genera nyob rau hauv kev tswj thiab mancozeb-kho cov qauv (Fig. 5b). c Kev tshuaj xyuas siv 18S ua raws li qhia tau tias tsis muaj qhov sib txawv tseem ceeb (Cov duab ntxiv 2).
SAV profiles raws li 16S ib ntus tau muab piv nrog Shannon nplua nuj thiab pom kev nplua nuj nyob rau theem phylum. (a) Tus Thawj Saib Xyuas Kev Ua Haujlwm (PCoA) raws li tag nrho cov qauv hauv zej zog microbial hauv cov paj ntoo uas tsis tau noj los yog tswj hwm (xiav) thiab mancozeb-fed larvae (txiv kab ntxwv). Txhua cov ntaub ntawv taw qhia sawv cev rau ib qho piv txwv. PCoA tau suav nrog siv Bray-Curtis nrug ntawm qhov sib txawv t tis. Ovals sawv cev rau 80% kev ntseeg siab. (b) Boxplot, raw Shannon wealth data (cov ntsiab lus) thiab c. Observable wealth. Boxplots qhia cov thawv rau kab nruab nrab, qhov sib txawv (IQR), thiab 1.5 × IQR (n = 3).
Kev sib xyaw ntawm cov zej zog microbial ntawm larvae pub rau mancozeb-kho thiab tsis kho paj ntoos. (a) Cov txheeb ze ntau ntawm microbial genera nyeem hauv larvae. (b) Daim ntawv qhia tshav kub ntawm cov zej zog microbial. Delftia (odds ratio (OR) = 0.67, P = 0.0030) thiab Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060); Daim ntawv qhia tshav kub kab yog pawg siv kev sib txheeb ze thiab nruab nrab kev sib txuas.
Peb cov txiaj ntsig tau pom tias qhov ncauj raug rau kev sib cuag (mancozeb) thiab kab mob (pyrostrobin thiab trifloxystrobin) fungicides, dav siv thaum lub sij hawm tawg paj, txo qhov hnyav nce thiab nce kev tuag ntawm pob kws larvae. Tsis tas li ntawd, mancozeb txo qis qhov sib txawv thiab kev nplua nuj ntawm microbiome thaum lub sijhawm prepupal. Myclobutanil, lwm yam kab mob fungicide, txo qis tus menyuam lub cev hnyav nce ntawm tag nrho peb koob. Cov txiaj ntsig no tau tshwm sim nyob rau lub sijhawm thib ob (hnub 5) thiab thib peb (hnub 8) lub sijhawm cov ntsiab lus. Hauv qhov sib piv, cyprodinil thiab captan tsis txo qhov hnyav nce lossis muaj sia nyob piv nrog pawg tswj hwm. Rau peb txoj kev paub, txoj haujlwm no yog thawj zaug los txiav txim siab qhov cuam tshuam ntawm thaj chaw ntawm cov kab mob fungicides sib txawv siv los tiv thaiv cov qoob loo pob kws los ntawm kev raug pollen ncaj qha.
Txhua qhov kev kho mob fungicide ua rau txo lub cev hnyav nce piv rau kev tswj hwm kev kho mob. Mancozeb muaj qhov cuam tshuam loj tshaj plaws ntawm lub cev qhov hnyav nce nrog qhov nruab nrab txo ntawm 51%, ua raws li pyrithiostrobin. Txawm li cas los xij, lwm cov kev tshawb fawb tsis tau qhia txog qhov tsis zoo ntawm kev siv tshuaj tua kab mob fungicides ntawm larval theem 44. Txawm hais tias dithiocarbamate biocides tau pom tias muaj cov tshuaj toxicity tsawg 45, ethylene bisdithiocarbamates (EBDCS) xws li mancozeb tuaj yeem degrade rau urea ethylene sulfide. Muab nws cov teebmeem mutagenic nyob rau hauv lwm yam tsiaj, cov khoom degradation no tej zaum yuav ua rau lub luag hauj lwm rau cov kev pom zoo 46,47. Cov kev tshawb fawb yav dhau los tau pom tias qhov tsim ntawm ethylene thiourea yog cuam tshuam los ntawm cov yam ntxwv xws li kub siab 48, av noo theem 49 thiab qhov ntev ntawm cov khoom cia50. Kev khaws cia zoo rau biocides tuaj yeem txo cov kev mob tshwm sim. Tsis tas li ntawd, European Food Safety Authority tau qhia txog kev txhawj xeeb txog kev lom ntawm pyrithiopide, uas tau pom tias yog carcinogenic rau lub plab zom mov ntawm lwm yam tsiaj51.
Kev tswj qhov ncauj ntawm mancozeb, pyrithiostrobin, thiab trifloxystrobin nce kev tuag ntawm pob kws borer larvae. Hauv qhov sib piv, myclobutanil, ciprocycline thiab captan tsis cuam tshuam rau kev tuag. Cov txiaj ntsig no txawv ntawm cov Ladurner et al.52, uas tau pom tias captan txo qis txoj sia nyob ntawm cov neeg laus O. lignaria thiab Apis mellifera L. (Hymenoptera, Apisidae). Tsis tas li ntawd, cov tshuaj fungicides xws li captan thiab boscalid tau pom tias ua rau cov kab mob tuag taus52,53,54 lossis hloov tus cwj pwm pub mis55. Cov kev hloov no, nyob rau hauv lem, tuaj yeem cuam tshuam cov khoom noj khoom haus zoo ntawm paj ntoos thiab thaum kawg lub zog nce ntawm theem larval. Kev tuag tau pom nyob rau hauv pab pawg tswj tau zoo ib yam nrog lwm cov kev tshawb fawb 56,57.
Cov txiv neej nyiam poj niam txiv neej sib deev pom nyob rau hauv peb txoj hauj lwm tej zaum yuav piav qhia los ntawm tej yam xws li mating tsis txaus thiab huab cua tsis zoo thaum lub sij hawm flowering, raws li yav tas los hais rau O. cornuta los ntawm Vicents thiab Bosch. Txawm hais tias cov poj niam thiab cov txiv neej hauv peb txoj kev tshawb fawb muaj plaub hnub los ua txij nkawm (ib lub sijhawm feem ntau suav tias txaus rau kev sib raug zoo), peb txhob txwm txo qis kev siv zog kom txo qis kev ntxhov siab. Txawm li cas los xij, qhov kev hloov kho no yuav tsis txhob txwm cuam tshuam rau txoj kev sib tw 61. Tsis tas li ntawd, cov muv tau ntsib ntau hnub ntawm huab cua tsis zoo, suav nrog los nag thiab kub qis (<5 ° C), uas tuaj yeem cuam tshuam tsis zoo rau mating kev vam meej4,23.
Txawm hais tias peb txoj kev tshawb fawb tsom mus rau tag nrho cov larval microbiome, peb cov txiaj ntsig tau muab kev nkag siab txog kev sib raug zoo ntawm cov kab mob hauv zej zog uas yuav yog qhov tseem ceeb rau kev noj zaub mov zoo rau muv thiab cov kab mob fungicide. Piv txwv li, larvae pub mancozeb-kho paj ntoos tau txo qis microbial hauv zej zog cov qauv thiab kev nplua nuj piv rau cov larvae pub tsis muaj paj ntoos. Hauv cov menyuam yaus noj cov paj ntoo uas tsis kho, cov kab mob ntawm cov kab mob Proteobacteria thiab Actinobacteria yog cov tseem ceeb thiab feem ntau yog aerobic lossis facultatively aerobic. Cov kab mob Delft, feem ntau cuam tshuam nrog tib hom muv, paub tias muaj cov tshuaj tua kab mob, qhia txog lub luag haujlwm tiv thaiv kab mob. Lwm hom kab mob, Pseudomonas, muaj ntau nyob rau hauv larvae noj tsis kho paj ntoos, tab sis tau txo qis hauv mancozeb-kho larvae. Peb cov txiaj ntsig tau txhawb nqa cov kev tshawb fawb yav dhau los txheeb xyuas Pseudomonas yog ib qho ntawm cov genera tshaj plaws hauv O. bicornis35 thiab lwm yam kab mob tib leeg wasps34. Txawm hais tias kev sim ua pov thawj rau lub luag haujlwm ntawm Pseudomonas hauv kev noj qab haus huv ntawm O. cornifrons tsis tau kawm, cov kab mob no tau pom los txhawb kev sib txuas ntawm kev tiv thaiv co toxins hauv kab Paederus fuscipes thiab txhawb nqa arginine metabolism hauv vitro 35, 65. Cov kev soj ntsuam no qhia. lub luag haujlwm tseem ceeb hauv kev tiv thaiv kab mob thiab kab mob thaum lub sijhawm tsim kho ntawm O. cornifrons larvae. Microbacterium yog lwm hom kab mob uas tau txheeb xyuas hauv peb txoj kev tshawb fawb uas tau tshaj tawm tias muaj cov lej siab hauv cov tub rog dub ya ya nyob rau hauv kev tshaib plab 66. Hauv O. cornifrons larvae, microbacteria tuaj yeem ua rau muaj kev sib npaug thiab kev ua haujlwm ntawm lub plab microbiome nyob rau hauv kev ntxhov siab. Tsis tas li ntawd, Rhodococcus muaj nyob rau hauv O. cornifrons larvae thiab paub txog nws lub peev xwm detoxification67. Cov genus no kuj pom muaj nyob rau hauv lub plab ntawm A. florea, tab sis nyob rau hauv tsawg heev abundance68. Peb cov txiaj ntsig tau pom tias muaj ntau yam kev hloov pauv caj ces hla ntau cov kab mob microbial uas tuaj yeem hloov pauv cov txheej txheem metabolic hauv larvae. Txawm li cas los xij, kev nkag siab zoo dua ntawm kev ua haujlwm ntawm O. cornifrons yog xav tau.
Hauv cov ntsiab lus, cov txiaj ntsig tau qhia tias mancozeb, pyrithiostrobin, thiab trifloxystrobin txo lub cev qhov hnyav thiab nce kev tuag ntawm pob kws borer larvae. Txawm hais tias muaj kev txhawj xeeb loj zuj zus txog qhov cuam tshuam ntawm fungicides ntawm pollinators, yuav tsum tau nkag siab zoo dua qhov cuam tshuam ntawm cov metabolites seem ntawm cov tebchaw. Cov txiaj ntsig no tuaj yeem muab tso rau hauv cov lus pom zoo rau kev tswj hwm pollinator kev sib koom ua ke uas pab cov neeg ua liaj ua teb zam kev siv cov tshuaj fungicides ua ntej thiab thaum lub sij hawm cov txiv hmab txiv ntoo flowering los ntawm kev xaiv cov fungicides thiab sib txawv lub sij hawm ntawm daim ntawv thov, los yog los ntawm kev txhawb nqa kev siv cov kev pab cuam tsawg dua 36. Cov ntaub ntawv no yog ib qho tseem ceeb rau kev tsim cov lus pom zoo. ntawm kev siv tshuaj tua kab, xws li kho cov kev siv tshuaj tsuag uas twb muaj lawm thiab hloov lub sijhawm tshuaj tsuag thaum xaiv cov tshuaj fungicides lossis txhawb kev siv cov kev xaiv tsis zoo. Kev tshawb fawb ntxiv yog xav tau rau cov kev tsis zoo ntawm cov kab mob fungicides ntawm kev sib deev piv, noj tus cwj pwm, plab microbiome, thiab cov txheej txheem molecular hauv qab pob kws borer poob phaus thiab kev tuag.
Cov ntaub ntawv 1, 2 thiab 3 hauv daim duab 1 thiab 2 tau muab tso rau hauv figshare data repository DOI: https://doi.org/10.6084/m9.figshare.24996245 thiab https://doi.org/10.6084/m9. ib.24996233. Cov kab ke txheeb xyuas hauv txoj kev tshawb fawb tam sim no (Fig. 4, 5) muaj nyob rau hauv NCBI SRA repository nyob rau hauv tus lej nkag PRJNA1023565.
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Post lub sij hawm: May-14-2024