Daalụ maka ileta Nature.com.Ụdị ihe nchọgharị ị na-eji nwere nkwado CSS nwere oke.Maka nsonaazụ kacha mma, anyị na-akwado ka ị jiri ụdị ihe nchọgharị gị kachasị ọhụrụ (ma ọ bụ gbanyụọ ọnọdụ ndakọrịta na Internet Explorer).Ka ọ dị ugbu a, iji hụ na nkwado na-aga n'ihu, anyị na-egosipụta saịtị na-enweghị styling ma ọ bụ Javascript.
A na-ejikarị fungicides eme ihe n'oge okooko osisi ma nwee ike iyi ndị na-emetọ ụmụ ahụhụ egwu.Otú ọ dị, ntakịrị ihe a maara banyere otú ndị na-abụghị ndị pollinators (dịka ọmụmaatụ, aṅụ naanị, Osmia cornifrons) na-anabata kọntaktị na usoro fungicides na-ejikarị na apụl n'oge okooko.Oghere ihe ọmụma a na-egbochi mkpebi nchịkwa nke na-ekpebi mkpokọta nchekwa na oge ịgbasa fungicides.Anyị tụlere mmetụta nke abụọ kọntaktị fungicides (captan na mancozeb) na anọ interlayer/phytosystem fungicides (ciprocycline, myclobutanil, pyrostrobin na trifloxystrobin).Mmetụta na oke ibu larval, nlanarị, oke mmekọahụ na ụdị nje dị iche iche.A na-eme nyocha ahụ site na iji bioassay ọnụ na-adịghị ala ala nke a na-agwọ pollen na usoro atọ dabere na usoro akwadoro ugbu a maka iji ubi (1X), ọkara dose (0.5X) na obere dose (0.1X).Usoro onunu ogwu niile nke mancozeb na pyritisoline belatara nke ọma ibu ahụ yana ndụ larval.Anyị wee hazie mkpụrụ ndụ ihe nketa 16S iji mara nje bacteria larval nke mancozeb, ihe fungicides na-akpata ọnwụ kacha elu.Anyị chọpụtara na ụdị nje bacteria dị iche iche na ụbara na-ebelata nke ukwuu na larvae a na-eri na pollen ndị mancozeb na-agwọ.Nsonaazụ ụlọ nyocha anyị na-egosi na ịgbasa ụfọdụ ọgwụ fungicides ndị a n'oge okooko na-emerụ ahụ ike nke O. cornifrons.Ozi a dị mkpa maka mkpebi nchịkwa n'ọdịnihu gbasara iji ngwaahịa nchebe osisi na-adịgide adịgide ma na-eje ozi dị ka ihe ndabere maka usoro nhazi nke iji chebe ndị na-emepụta ihe.
Osmia cornifrons (Hymenoptera: Megachilidae) ewebata na United States site na Japan na njedebe 1970s na mmalite 1980s, na ụdị ahụ ekerewo òkè dị mkpa na pollinator na gburugburu ebe obibi na-achịkwa kemgbe ahụ.Onu ogugu ndi mmadu nke anu a bu akuku nke ihe dika 50 umu anumanu nke na-emeju aṅụ na-emetọ osisi almond na apụl na United States2,3.Aṅụ Mason na-eche ọtụtụ ihe ịma aka ihu, gụnyere nkewa ebe obibi, ọrịa nje, na ọgwụ nje3,4.N'ime ụmụ ahụhụ, fungicides na-ebelata uru ume, nri nri5 na nhazi ahụ6,7.Ọ bụ ezie na nchọpụta na-adịbeghị anya na-egosi na ahụike nke aṅụ Mason na-emetụta kpọmkwem site na commensal na ectobactic microorganisms, 8,9 n'ihi na nje bacteria na fungi nwere ike imetụta nri na-edozi ahụ na nzaghachi mgbochi ọrịa, mmetụta nke fungicides ikpughe na microbial di iche iche nke Mason aṅụ ka na-amalite ịdị. gụrụ akwụkwọ.
Fungicides nke mmetụta dị iche iche (kọntaktị na sistemu) na-efesa n'ubi mkpụrụ osisi tupu na n'oge okooko osisi iji gwọọ ọrịa ndị dị ka scab apụl, ire ilu, ire ere na agba aja aja na mildew powdery10,11.A na-ewere fungicides dị ka ihe adịghị emerụ ahụ nye ndị na-emepụta ihe, ya mere a na-atụ aro ya maka ndị na-elekọta ubi n'oge oge okooko;Mkpughe na ntinye nke ọgwụ ndị a sitere na aṅụ bụ nke a maara nke ọma, n'ihi na ọ bụ akụkụ nke ndebanye aha ọgwụ ahụhụ nke US Environmental Protection Agency na ọtụtụ ụlọ ọrụ na-ahụ maka nchịkwa obodo12,13,14.Otú ọ dị, mmetụta nke fungicides na ndị na-abụghị aṅụ ka amachaghị n'ihi na a chọghị ha n'okpuru nkwekọrịta ikike ịzụ ahịa na United States15.N'ịgbakwụnye, ọ dịghị n'ozuzu ọnweghị ụkpụrụ ọkọlọtọ maka ịnwale otu bees16,17, na idowe obodo ndị na-enye aṅụ maka ule bụ ihe ịma aka18.A na-eme nnwale nke aṅụ dị iche iche na-esiwanye ike na Europe na USA iji nyochaa mmetụta nke ọgwụ na-egbu egbu na anụ ọhịa aṅụ, na e mepụtala ụkpụrụ ọkọlọtọ maka O. cornifrons19 na nso nso a.
Anụ aṅụ nwere mpi bụ monocytes, a na-ejikwa ya eme ihe n'ubi azu kaapu dị ka ihe mgbakwunye ma ọ bụ dochie anya aṅụ mmanụ aṅụ.Aṅụ ndị a na-apụta n'etiti Maachị na Eprel, na ụmụ nwoke na-amụba amụba na-apụta ụbọchị atọ ruo anọ tupu ụmụ nwanyị.Mgbe nwoke na nwanyị na-akpakọrịta, nwanyị na-arụsi ọrụ ike na-anakọta pollen na nectar iji nye usoro mkpụrụ ndụ brood n'ime oghere tubular akwu (nke eke ma ọ bụ artificial)1,20.A na-etinye akwa ndị ahụ na pollen n'ime sel;nwanyị ahụ na-ewu mgbidi ụrọ tupu ya edozi cell ọzọ.A na-emechi larvae nke mbụ na chorion ma na-eri nri na mmiri embrayo.Site na nke abụọ ruo nke ise instar (prepupa), larvae na-eri nri na pollen22.Ozugbo ndị pollen na-ejupụta kpamkpam, larvae na-etolite cocoons, pupate na pụta dị ka ndị okenye n'otu ọnụ ụlọ, na-emekarị na mbubreyo20,23.Ndị okenye na-apụta na mmiri na-esote.A na-ejikọta nlanarị ndị okenye na uru net ume (uru ibu) dabere na nri oriri.Ya mere, ụdị nri pollen na-edozi ahụ, yana ihe ndị ọzọ dị ka ihu igwe ma ọ bụ ikpughe na ọgwụ nje, bụ ihe na-ekpebi nlanarị na ahụike24.
Ọgwụ ahụhụ na fungicides etinyere tupu ifuru na-enwe ike ịkwaga n'ime vasculature osisi ruo ogo dị iche iche, site na translaminar (dịka ọmụmaatụ, nwee ike ịkwaga n'elu elu nke akwụkwọ ahụ gaa n'elu ala, dị ka ụfọdụ fungicides) 25 na-enwe mmetụta nke sistemu., nke nwere ike banye n'ime okpueze sitere na mgbọrọgwụ, nwere ike ịbanye na nectar apple flower26, ebe ha nwere ike igbu okenye O. cornifrons27.Ụfọdụ ọgwụ na-egbu egbu na-abanyekwa n'ime pollen, na-emetụta mmepe nke larvae ọka ma na-egbu ha19.Nnyocha ndị ọzọ egosila na ụfọdụ fungicides nwere ike ịgbanwe nke ọma omume nesting nke ụdị O. lignaria28 metụtara ya.Na mgbakwunye, ụlọ nyocha na ọmụmụ ubi na-egosipụta ọnọdụ mkpughe pesticide (gụnyere fungicides) egosila na ọgwụ nje na-emetụta physiology 22 morphology 29 na-adịghị mma na nlanarị nke aṅụ mmanụ aṅụ na ụfọdụ aṅụ naanị ya.Ọkpụkpọ fungicidal dị iche iche a na-etinye ozugbo na ifuru mepere emepe n'oge okooko nwere ike mebie pollen ndị okenye anakọtara maka mmepe larval, nsonaazụ ya ka ga-amụ30.
A na-amatawanye na mmepe larval na-emetụta pollen na microbial obodo nke usoro nri nri.Microbiome nke honeybee na-emetụta paramita dị ka mass31, mgbanwe metabolic22 na ike nke nje32.Nnyocha ndị gara aga enyochala mmetụta nke ọkwa mmepe, nri nri, na gburugburu ebe obibi na microbiome nke aṅụ naanị ya.Ọmụmụ ihe ndị a kpughere myirịta na nhazi na ụbara nke larval na pollen microbiomes33, yana ụdị nje bacteria Pseudomonas na Delftia na-ahụkarị, n'etiti ụdị aṅụ naanị.Otú ọ dị, ọ bụ ezie na e jikọtara fungicides na atụmatụ iji chebe ahụ ike aṅụ, mmetụta fungicides na microbiota larval site na ikpughe ọnụ kpọmkwem ka anọgideghị na-achọpụta.
Ọmụmụ ihe a nwalere mmetụta nke ọgwụ nje isii a na-ejikarị eme ihe n'ezie bụ nke edebara aha maka iji mkpụrụ osisi dị na United States, gụnyere kọntaktị na usoro fungicides nke a na-enye ọnụ n'ọnụ ọka mpi mpi nla larvae sitere na nri mmetọ.Anyị chọpụtara na kọntaktị na usoro fungicides na-ebelata uru anụ ahụ na-abawanye na ọnwụ, yana mmetụta kachasị njọ metụtara mancozeb na pyrithiopide.Anyị tụlere ụdị dị iche iche nke microbial nke larvae a na-eri na nri pollen nke mancozeb na-agwọ ya na ndị a na-eri nri na-achịkwa.Anyị na-atụle usoro nwere ike ịkpata ọnwụ yana ihe ọ pụtara maka mmemme pest and pollinator management (IPPM) 36 jikọtara ọnụ.
A na-enweta ndị okenye O. cornifrons overwintering na cocoons sitere na Ụlọ Ọrụ Nnyocha Mkpụrụ, Biglerville, PA, ma chekwaa na -3 ruo 2 ° C (± 0.3 ° C).Tupu nnwale (600 cocoons na mkpokọta).Na May 2022, 100 O. cornifrons cocoons e bufere kwa ụbọchị n'ime iko plastik (cocoons 50 kwa iko, DI 5 cm × 15 cm ogologo) na ehichapụkwa n'ime iko ndị ahụ iji kwalite oghere ma nye mkpụrụ nke nwere ike ịta, na-ebelata nrụgide na nkume nkume. aṅụ37 .Debe iko rọba abụọ nwere cocoons n'ime oghere ụmụ ahụhụ (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) yana nri nri 10 ml nwere 50% sucrose ngwọta na chekwaa maka ụbọchị anọ iji hụ na mmechi na mating.23°C, iru iru mmiri 60%, photoperiod 10 l (obere ike): 14 ụbọchị.A tọhapụrụ 100 ụmụ nwanyị na ụmụ nwoke mated kwa ụtụtụ maka ụbọchị isii (100 kwa ụbọchị) n'ime akwụ abụọ arụrụ arụ n'oge okooko osisi apple kacha elu ( akwụkwụ ọnyà: obosara 33.66 × elu 30.48 × ogologo 46.99 cm; Ihe mgbakwunye 1).Edebere na Pennsylvania State Arboretum, nso cherry (Prunus cerasus 'Eubank' Sweet Cherry Pie ™), piich (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), ube (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), osisi apụl coronaria (Malus coronaria) na ọtụtụ ụdị osisi apụl (Malus coronaria, Malus), osisi apụl ụlọ 'Co-op 30' Enterprise ™, Malus apple osisi 'Co- Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy').Ụlọ nnụnụ rọba na-acha anụnụ anụnụ ọ bụla dabara n'elu igbe osisi abụọ.Igbe akwụ nke ọ bụla nwere tubes akwụkwọ kraft efu 800 efu (spiral open, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) etinyere n'ime tubes cellophane opaque (0.7 OD lee Plastic plugs (T-1X plugs) na-enye ebe agbụ agbụ. .
Igbe akwụ abụọ ahụ chere ihu n'ebe ọwụwa anyanwụ ma kpuchie ya na ogige osisi plastik ndụ ndụ (Everbilt nlereanya # 889250EB12, oghere oghere 5 × 5 cm, 0.95 m × 100 m) iji gbochie òké na ohere nnụnụ ma dọba ya n'elu ala n'akụkụ igbe igbe ala. igbe.Igbe akwụkwụ (Nkọwa mgbakwunye 1a).A na-anakọta akwa ndị na-eri ọka kwa ụbọchị site n'ịnakọta tube 30 n' akwụ akwụ ma bufee ya n'ụlọ nyocha.Iji mkpa, mee ka ebibie na njedebe nke tube ahụ, wee kewaa spiral tube iji kpughee mkpụrụ ndụ brood.E wepụrụ àkwá nke ọ bụla na pollen ha site na iji spatula gbagọrọ agbagọ (Microslide Tool kit, BioQuip Products Inc., California).A na-etinye akwa ndị ahụ n'ime akwụkwọ nzacha dị mmiri ma tinye ya na efere Petri maka awa 2 tupu ejiri ya mee ihe na nyocha anyị (Ngbakwunye 1b-d).
N'ime ụlọ nyocha, anyị nyochara nsị ọnụ nke fungicides isii etinyere n'ihu na n'oge ifuru apụl na ntinye atọ (0.1X, 0.5X, na 1X, ebe 1X bụ akara nke etinyere na 100 galọn mmiri / acre. Oke ubi dose = ntinye uche. n'afọ)., Isiokwu 1).A na-atụgharị uche nke ọ bụla ugboro 16 (n = 16).Abụọ kọntaktị fungicides (Table S1: mancozeb 2696.14 ppm na captan 2875.88 ppm) na anọ usoro fungicides (Table S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil 12dinil 75 ppm). mkpụrụ osisi, akwụkwọ nri na ihe ọkụkụ ịchọ mma .Anyị na-ejikọta pollen ahụ site na iji grinder, bufee 0.20 g na olulu mmiri (24-well Falcon Plate), ma gbakwunye ma gwakọta 1 μL nke ngwọta fungicide iji mepụta pollen pyramidal na olulu mmiri 1 mm nke etinyere nsen.Ebe na-eji obere spatula (Ntinye mgbakwunye 1c,d).A na-echekwa efere falcon n'ime ụlọ okpomọkụ (25 Celsius) yana iru mmiri 70%.Anyị jiri ha tụnyere larvae na-achịkwa na-enye nri pollen na-edozi ahụ nke ejiri mmiri dị ọcha emeso ya.Anyị na-edekọ onwu ma tụọ oke ibu kwa ụbọchị ruo mgbe larvae ruru afọ nke prepupal site na iji ntule nyocha (Fisher Scientific, izi ezi = 0.0001 g).N'ikpeazụ, a na-enyocha oke mmekọahụ site na imepe aki oyibo mgbe ọnwa 2.5 gasịrị.
A na-ewepụta DNA site na dum O. cornifrons larvae (n = 3 kwa ọnọdụ ọgwụgwọ, mancozeb-agwọ ọrịa na pollen na-adịghị agwọ ya) na anyị mere nyocha nke microbial diversity na ihe atụ ndị a, karịsịa n'ihi na na mancozeb, a hụrụ ọnwụ kachasị elu na larvae.na-anata MnZn.A kwalitere DNA, sachaa site na iji DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), na usoro (usoro okirikiri 600) na Illumina® MiSeq™ site na iji ngwa v3.Ezubere usoro nke nje nje 16S ribosomal RNA nke nje na-eji ngwa ngwa-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) na-eji primers na-ezubere mpaghara V3-V4 nke mkpụrụ ndụ 16S rRNA.Na mgbakwunye, a na-eme usoro 18S site na iji ntinye 10% PhiX, yana mmụba ka emere site na iji ụzọ mbụ 18S001 na NS4.
Bubata na hazie ọnụ ọgụgụ read39 site na iji pipeline QIIME2 (v2022.11.1).Akpụchapụrụ ma jikọta ọnụ ọgụgụ ndị a, wee wepụ usoro chimeric site na iji ngwa mgbakwunye DADA2 na QIIME2 (qiime dada2 noise pairing)40.Emere ọrụ klaasị 16S na 18S site na iji ihe classifier plugin Classify-sklearn na artifact silva-138-99-nb-classifier a zụrụ nke ọma.
A na-enyocha data nnwale niile maka ịdị mma (Shapiro-Wilks) na ịdị n'otu nke variances (Nnwale Levene).N'ihi na nhazi data ahụ emezughị echiche nke nyocha parametric na mgbanwe ahụ enweghị ike ịhazi ihe ndị fọdụrụnụ, anyị mere ụzọ abụọ ANOVA (Kruskal-Wallis) nke na-abụghị parametric na ihe abụọ [oge (3-phase 2, 5, and 8 day) oge) na fungicides] iji nyochaa mmetụta ọgwụgwọ na-adị na ibu dị ọhụrụ, wee jiri nyocha Wilcoxon mee ntụnyere na-abụghị parametric.Anyị jiri usoro ahịrị n'ozuzu ya (GLM) nwere nkesa Poisson iji tụnyere mmetụta nke fungicides na nlanarị n'ofe mkpokọta fungicides atọ41,42.Maka nyocha ụbara dị iche iche, ọnụọgụ nke amplicon sequence variants (ASVs) dara na ọkwa ọmụmụ.A na-atụnyere ụbara dị iche iche n'etiti otu ndị na-eji 16S (ọkwa genus) na ụbara 18S site na iji usoro mgbakwunye mkpokọta maka ọnọdụ, ọnụ ọgụgụ, na ọdịdị (GAMLSS) na nkesa ezinụlọ beta zero-inflated (BEZI), bụ nke emere na nnukwu nnukwu. .na Microbiome R43 (v1.1).1).Wepu ụdị mitochondrial na chloroplast tupu nyocha nke iche.N'ihi ọkwa taxonomic dị iche iche nke 18S, ọ bụ naanị ọkwa dị ala nke taxon ọ bụla ka ejiri mee nyocha dị iche iche.Emere nyocha ndekọ ọnụ ọgụgụ niile site na iji R (v. 3.4.3., CRAN project) (Team 2013).
Ngosipụta na mancozeb, pyrithiostrobin, na trifloxystrobin budata belata oke ibu ahụ na O. cornifrons (Fig 1).A na-ahụ mmetụta ndị a mgbe niile maka usoro atọ a na-enyocha (Fig 1a-c).Cyclostrobin na myclobutanil ebelataghị ibu nke larvae nke ọma.
Nkezi arọ ọhụrụ nke stem borer larvae tụrụ n'isi oge atọ n'okpuru ọgwụgwọ nri anọ (nri nri pollen + fungicide: njikwa, 0.1X, 0.5X na 1X doses).(a) Ọgwụ dị ala (0.1X): oge mbụ (ụbọchị 1): χ2: 30.99, DF = 6;P <0.0001, oge nke abụọ (ụbọchị 5): 22.83, DF = 0.0009;nke ugboro atọ;isi (ụbọchị 8): χ2: 28.39, DF = 6;(b) ọkara dose (0.5X): oge mbụ (ụbọchị 1): χ2: 35.67, DF = 6;P <0.0001, oge nke abụọ (ụbọchị mbụ).): χ2: 15.98, DF = 6;P = 0.0090;oge nke atọ (ụbọchị 8) χ2: 16.47, DF = 6;(c) Saịtị ma ọ bụ zuru oke (1X): oge mbụ (ụbọchị 1) χ2: 20.64, P = 6;P = 0.0326, oge nke abụọ (ụbọchị 5): χ2: 22.83, DF = 6;P = 0,0009;nkeji oge nke atọ (ụbọchị 8): χ2: 28.39, DF = 6;nonparametric analysis of variance.Ogwe na-anọchi anya ± SE nke ntụnyere ụzọ abụọ (α = 0.05) (n = 16) * P ≤ 0.05, ** P ≤ 0.001, *** P ≤ 0.0001.
N'ihe dị ntakịrị (0.1X), ibu ahụ nke larval belatara site na 60% na trifloxystrobin, 49% na mancozeb, 48% na myclobutanil, na 46% na pyrithistrobin (Fig 1a).Mgbe ekpughere ya na ọkara nke ọgwụ (0.5X), ịdị arọ nke mancozeb larvae belatara site na 86%, pyrithiostrobin site na 52% na trifloxystrobin site na 50% (Fig 1b).Ngwunye ubi zuru oke (1X) nke mancozeb belatara ibu nke oke ahihia site na 82%, pyrithiostrobin site na 70%, na trifloxystrobin, myclobutanil na sangard site na ihe dika 30% (Fig 1c).
Onwu kacha elu n'etiti larvae nyere nri mancozeb pollen, pyrithiostrobin na trifloxystrobin sochiri ya.Ọnwụ na-abawanye site na mmụba nke mancozeb na pyritisoline (Fig 2; Tebụl 2).Otú ọ dị, ọnụ ọgụgụ ndị na-egbu ọka na-amụba amụbawanye ntakịrị ka ọnụọgụ trifloxystrobin na-abawanye;cyprodinil na captan abawanyeghị nke ukwuu ma e jiri ya tụnyere ọgwụgwọ nchịkwa.
A tụlere ọnwụ nke larvae ijiji mgbe ọ risịrị nri pollen n'otu n'otu na ọgwụ fungicides isii dị iche iche.Mancozeb na pentopyramide na-enwe mmetụta nke ọma maka ikpughe ọnụ na ikpuru ọka (GLM: χ = 29.45, DF = 20, P = 0.0059) (akara, mkpọda = 0.29, P <0.001; slope = 0.24, P <0.00)).
Na nkezi, n'ofe ọgwụgwọ niile, 39.05% nke ndị ọrịa bụ nwanyị yana 60.95% bụ nwoke.N'ime usoro ọgwụgwọ ndị a na-achịkwa, ọnụ ọgụgụ ụmụ nwanyị dị 40% na ọmụmụ ihe dị ala (0.1X) na ọkara (0.5X), na 30% na ọmụmụ ihe ubi-dose (1X).Na 0.1X dose, n'etiti larvae na-eri nri pollen na-emeso mancozeb na myclobutanil, 33.33% nke ndị okenye bụ nwanyị, 22% nke ndị okenye bụ nwanyị, 44% nke ndị okenye bụ nwanyị, 44% nke ndị okenye bụ nwanyị.nwanyị, 41% nke ndị okenye larvae bụ ụmụ nwanyị, na njikwa bụ 31% (Fig 3a).Na 0.5 ugboro dose, 33% nke ikpuru ndị okenye na mancozeb na pyrithiostrobin otu bụ nwanyị, 36% na trifloxystrobin otu, 41% na myclobutanil otu na 46% na cyprostrobin otu.Ọnụọgụ a bụ 53% n'ime otu.n'ime otu captan na 38% na otu njikwa (Fig 3b).Na 1X dose, 30% nke mancozeb otu bụ ndị inyom, 36% nke pyrithiostrobin otu, 44% nke trifloxystrobin otu, 38% nke myclobutanil otu, 50% nke akara otu ndị inyom - 38.5% (Fig. 3c). .
Pasent nke nwanyị na nwoke na-agwụ ike mgbe mkpughe fungicides nke larval.(a) Ọgwụ dị ala (0.1X).(b) Ọkara dose (0.5X).(c) Ọgwụ mpaghara ma ọ bụ ọgwụ zuru oke (1X).
Nyocha usoro nke 16S gosiri na otu nje bacteria dị iche n'etiti larvae na-eri nri na mancozeb pollen na larvae na-eri nri na-adịghị edozi (Fig 4a).Ndekọ microbial nke larvae a na-adịghị agwọ ọrịa na-eri nri na pollen dị elu karịa nke larvae a na-eri na pollen a na-agwọ mancozeb (Fig 4b).Ọ bụ ezie na ọdịiche ahụ a hụrụ na ụbara n'etiti otu dị iche iche abụghị ihe ndekọ ọnụ ọgụgụ, ọ dị ntakịrị karịa nke a hụrụ maka nri larvae na pollen na-adịghị edozi (Fig 4c).N'ụba dị iche iche gosiri na microbiota nke larvae a na-eri na pollen njikwa dị iche iche karịa nke larvae na-eri nri na mancozeb na-emeso larvae (Fig 5a).Ntụle nkọwa gosipụtara ọnụnọ nke 28 genera na njikwa na mancozeb-mere samples (Fig. 5b).c Nyocha site na iji usoro 18S ekpughere enweghị nnukwu ọdịiche (Ntụpọ mgbakwunye 2).
Atụnyere profaịlụ SAV dabere na usoro 16S na ịba ụba Shannon wee hụ ọgaranya na ọkwa phylum.(a) Ntụle nhazi nhazi nke isi (PCoA) dabere na nhazi obodo n'ozuzu microbial na nri pollen anaghị agwọta ma ọ bụ njikwa (acha anụnụ anụnụ) na larvae na-enye mancozeb (oroma).Ebe data ọ bụla na-anọchi anya nlele dị iche.A gbakọrọ PCoA site na iji ogologo Bray-Curtis nke nkesa multivariate.Ovals na-anọchi anya ọkwa ntụkwasị obi 80%.(b) Boxplot, raw Shannon akụ data (isi ihe) na c.Akụ na ụba na-ahụ anya.Igbe igbe na-egosi igbe maka ahịrị etiti, oke interquartile (IQR), na 1.5 × IQR (n = 3).
Ihe mejupụtara obodo ụmụ irighiri ihe nke larvae na-eri nri na mancozeb-agwọtaghị na pollen anaghị agwọ ya.(a) A na-agụ n'ụba nke microbial genera na larvae.(b) Maapụ okpomọkụ nke obodo ụmụ nje ndị amapụtara.Delftia (nha nhata (OR) = 0.67, P = 0.0030) na Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060);A na-achịkọta ahịrị maapụ okpomọkụ site na iji njikọ dị anya na nkezi njikọ.
Nsonaazụ anyị na-egosi na ikpughe ọnụ na kọntaktị (mancozeb) na sistemu (pyrostrobin na trifloxystrobin) fungicides, nke a na-etinye n'ọtụtụ ebe n'oge okooko, na-ebelata oke ibu na ụbara ọnwụ nke larvae ọka.Na mgbakwunye, mancozeb belatara nke ukwuu ụdịdị dị iche iche na ụbara microbiome n'oge ogbo prepupal.Myclobutanil, ọgwụ fungicides sistemu ọzọ, na-ebelata oke oke ibu anụ ahụ na usoro atọ niile.Mmetụta a pụtara ìhè na oge nke abụọ (ụbọchị 5) na nke atọ (ụbọchị 8).N'ụzọ dị iche, cyprodinil na captan ebelataghị oke ibu ma ọ bụ nlanarị ma e jiri ya tụnyere otu njikwa.Dị ka anyị maara, ọrụ a bụ nke mbụ iji chọpụta mmetụta nke ọnụ ọgụgụ ubi nke ụdị fungicides dị iche iche na-eji echebe ọka ọka site na ikpughe pollen kpọmkwem.
Ọgwụgwọ fungicides niile belatara oke ibu ahụ ma e jiri ya tụnyere ọgwụgwọ njikwa.Mancozeb nwere mmetụta kachasị ukwuu na oke ibu ahụ larval na mbelata nkezi nke 51%, pyrithiostrobin sochiri ya.Agbanyeghị, ọmụmụ ihe ndị ọzọ akọpụtabeghị mmetụta ọjọọ nke doses nke fungicides dị n'ọhịa na ọkwa larval44.Ọ bụ ezie na egosipụtara na dithiocarbamate biocides nwere obere toxicity45, ethylene bisdithiocarbamates (EBDCS) dị ka mancozeb nwere ike iweda urea ethylene sulfide.Nyere mmetụta mutagenic ya na anụmanụ ndị ọzọ, ngwaahịa a na-emebi emebi nwere ike bụrụ ihe kpatara nsonaazụ a hụrụ46,47.Nnyocha ndị gara aga egosila na nhazi nke ethylene thiourea na-emetụta ihe ndị dị ka okpomọkụ dị elu48, iru mmiri dị elu49 na ogologo nchekwa ngwaahịa50.Ọnọdụ nchekwa nke ọma maka biocides nwere ike ibelata mmetụta ndị a.Na mgbakwunye, European Safety Authority ekwupụtala nchegbu banyere nsi nke pyrithiopide, nke egosiri na ọ bụ carcinogenic na usoro mgbaze nke anụmanụ ndị ọzọ51.
Nlekọta ọnụ nke mancozeb, pyrithiostrobin, na trifloxystrobin na-abawanye ọnwụ nke larvae ọka borer.N'ụzọ dị iche, myclobutanil, ciprocycline na captan enweghị mmetụta na-anwụ anwụ.Nsonaazụ ndị a dị iche na nke Ladurner et al.52, bụ ndị gosipụtara na captan belatara nke ukwuu ndụ nke okenye O. lignaria na Apis mellifera L. (Hymenoptera, Apisidae).Na mgbakwunye, a chọpụtala na fungicides dị ka captan na boscalid na-ebute anwụ anwụ52,53,54 ma ọ bụ gbanwee omume inye nri55.Mgbanwe ndị a, n'aka nke ya, nwere ike imetụta àgwà nri nke pollen na n'ikpeazụ uru ike nke oke ahihia.Ọnwụ anwụ hụrụ n'ime otu njikwa kwekọrọ na ọmụmụ ndị ọzọ 56,57.
Enwere ike ịkọwa oke nwoke na nwanyị na-amasị nwoke na-ahụ n'ọrụ anyị site na ihe ndị dị ka ezughị ezu mating na ọnọdụ ihu igwe na-adịghị mma n'oge okooko, dị ka a tụrụ aro na mbụ maka O. cornuta site na Vicens na Bosch.Ọ bụ ezie na ụmụ nwanyị na ụmụ nwoke na ọmụmụ ihe anyị nwere ụbọchị anọ iji lụọ (oge a na-eche na ọ ga-ezuru maka inwe mmekọahụ na-aga nke ọma), anyị na-ama ụma belata ike ọkụ iji belata nchekasị.Agbanyeghị, mgbanwe a nwere ike itinye aka na usoro mmekọ ahụ na-amaghị ama61.Na mgbakwunye, aṅụ na-enweta ọtụtụ ụbọchị nke ihu igwe ọjọọ, gụnyere mmiri ozuzo na obere okpomọkụ (<5°C), nke nwekwara ike imetụta ihe ịga nke ọma mating na-adịghị mma4,23.
Ọ bụ ezie na ọmụmụ ihe anyị lekwasịrị anya na microbiome dum nke larval, nsonaazụ anyị na-enye nghọta gbasara mmekọrịta dị n'etiti obodo nje ndị nwere ike dị mkpa maka nri aṅụ na ikpughe fungicides.Dịka ọmụmaatụ, ụnwụ pollen a na-agwọ mancozeb belatara nke ukwuu n'usoro obodo na ụbara nke ụmụ irighiri nri ma e jiri ya tụnyere larvae na-enye pollen a na-adịghị edozi ya.N'ime larvae na-eri pollen a na-adịghị agwọ ọrịa, otu nje bacteria Proteobacteria na Actinobacteria bụ ndị na-abụkarị aerobic ma ọ bụ ike ikuku.A maara nje bacteria Delft, nke na-ejikọta ya na ụdị aṅụ naanị ya, nwere ọrụ ọgwụ nje, na-egosi ọrụ nchebe nwere ike imegide nje.Ụdị nje ọzọ, Pseudomonas, dị n'ụba na larvae na-enye pollen a na-adịghị agwọ ya, mana ọ na-ebelata nke ukwuu na larvae ndị mancozeb na-agwọ.Nsonaazụ anyị na-akwado ọmụmụ ihe gara aga na-achọpụta Pseudomonas dị ka otu n'ime mkpụrụ ndụ kachasị na O. bicornis35 na ndị ọzọ naanị wasps34.Ọ bụ ezie na a chọpụtabeghị ihe akaebe nnwale maka ọrụ Pseudomonas na ahụike nke O. cornifrons, egosila na nje bacteria a na-akwalite njikọ nke toxins na-echebe na beetle Paederus fuscipes ma na-akwalite metabolism arginine na vitro 35, 65. Nleba anya ndị a na-atụ aro. enwere ike ịrụ ọrụ na nchekwa nje na nje n'oge mmepe nke O. cornifrons larvae.Microbacterium bụ ụdị ọzọ achọpụtara n'ọmụmụ ihe anyị nke a na-akọ na ọ nọ n'ọnụọgụ dị elu na ndị agha ojii na-efe larvae n'okpuru ọnọdụ agụụ66.Na O. cornifrons larvae, microbacteria nwere ike inye aka na nguzozi na resilience nke microbiome gut n'okpuru ọnọdụ nrụgide.Na mgbakwunye, a na-ahụ Rhodococcus na O. cornifrons larvae ma mara ya maka ikike mkpochapụ ya67.A na-ahụkwa ụdị ụdị a n'ime eriri afọ A. florea, mana n'ọtụtụ dị ala68.Nsonaazụ anyị gosipụtara ọnụnọ nke ọtụtụ ụdị mkpụrụ ndụ ihe nketa gafere ọtụtụ taxa microbial nke nwere ike ịgbanwe usoro metabolic na larvae.Agbanyeghị, ọ dị mkpa ka a ghọtakwuo ụdị ọrụ dị iche iche nke O. cornifrons.
Na nchịkọta, nsonaazụ ya na-egosi na mancozeb, pyrithiostrobin, na trifloxystrobin belatara ibu ahụ yana ụbara ọnwụ nke larvae ọka na-ebu ọka.Ọ bụ ezie na enwere nchegbu na-arịwanye elu banyere mmetụta nke fungicides na pollinators, ọ dị mkpa ịghọta nke ọma mmetụta nke metabolites fọdụrụnụ nke ogige ndị a.Enwere ike itinye nsonaazụ ndị a n'ime ndụmọdụ maka mmemme njikwa pollinator agbakwunyere nke na-enyere ndị ọrụ ugbo aka izere iji ụfọdụ fungicides tupu na n'oge okooko osisi site na ịhọrọ fungicides na mgbanwe oge nke ngwa, ma ọ bụ site n'ịgba ume iji ụzọ ndị ọzọ na-adịghị emerụ ahụ 36. Ozi a. dị mkpa maka ịmepụta ndụmọdụ.na iji ọgwụ nje eme ihe, dị ka ịhazigharị mmemme ịgba mmiri dị ugbu a yana ịgbanwe oge ịgba mgbe ị na-ahọrọ ọgwụ fungicides ma ọ bụ kwalite ojiji nke ọzọ dị ize ndụ.Achọrọ nyocha ọzọ n'ime mmetụta ọjọọ nke fungicides na oke mmekọahụ, omume nri, microbiome gut, na usoro mkpụrụ ndụ ndị na-akpata ọnwụ ọka na-ebelata ibu na ọnwụ.
Edebere data isi mmalite 1, 2 na 3 na Figure 1 na 2 na ebe nchekwa data figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 na https://doi.org/10.6084/m9.figshare.24996233.Usoro nyochara na ọmụmụ ihe ugbu a (Fig 4, 5) dị na ebe nchekwa NCBI SRA n'okpuru nọmba ntinye PRJNA1023565.
Bosch, J. na Kemp, WP Mmepe na nguzobe nke ụdị mmanụ aṅụ dị ka pollinators nke ugbo ugbo: ihe atụ nke genus Osmia.(Hymenoptera: Megachilidae) na osisi mkpụrụ osisi.ehi.Ntomore.akụ.92, 3–16 (2002).
Parker, MG et al.Omume mgbasa ozi na echiche nke ndị ọzọ na-emetọ ihe n'etiti ndị na-akụ apụl na New York na Pennsylvania.imelite.Ọrụ ugbo.usoro nri.35, 1–14 (2020).
Koch I., Lonsdorf EW, Artz DR, Pitts-Singer TL na Ricketts TH Ecology na Economics of almond pollination na-eji aṅụ ala.J. Economics.Ntomore.111, 16–25 (2018).
Lee, E., He, Y., na Park, Y.-L.Mmetụta nke mgbanwe ihu igwe na tragopan phenology: mmetụta maka njikwa ndị mmadụ.Rigo.Gbanwee 150, 305–317 (2018).
Artz, DR na Pitts-Singer, TL Mmetụta nke fungicides na adjuvant sprays na omume nta nke aṅụ abụọ jisiri ike (Osmia lignaria na Megachile rotundata).PloS One 10, e0135688 (2015).
Beauvais, S. et al.Ihe ọkụkụ na-egbu egbu na-egbu egbu (fenbuconazole) na-emetụta akara ngosi ọmụmụ nwoke nke na-ebute mbelata ihe ịga nke ọma n'ime anụ ọhịa aṅụ naanị ya.Ngwa J.ecology.59, 1596–1607 (2022).
Sgolastra F. et al.Neonicotinoid ụmụ ahụhụ na ergosterol biosynthesis na-egbochi ọnwụ fungicides synergistic n'ụdị aṅụ atọ.Njikwa pesti.sayensị.73, 1236–1243 (2017).
Kuhneman JG, Gillung J, Van Dyck MT, Fordyce RF.na Danforth BN Solitary wasp larvae na-agbanwe ụdị nje bacteria dị iche iche nke pollen na-eweta na aṅụ na-eku Osmia cornifrons (Megachilidae).n'ihu.microorganism.13, 1057626 (2023).
Dharampal PS, Danforth BN na Steffan SA Ihe microorganisms ectosymbiotic dị na pollen fermented dị mkpa maka mmepe nke aṅụ naanị dị ka pollen n'onwe ya.ecology.evolushọn.12. e8788 (2022).
Kelderer M, Manici LM, Caputo F na Thalheimer M. Inter-ahiri akuku na osisi apụl iji chịkwaa ọrịa reseeding: a bara uru ọmụmụ dabeere microbial egosi.Ala osisi 357, 381-393 (2012).
Martin PL, Kravchik T., Khodadadi F., Achimovich SG na Peter KA ilu ire ere nke apụl na etiti Atlantic United States: ntule nke causative umu na mmetụta nke mpaghara ihu igwe ọnọdụ na cultivar susceptibility.Phytopathology 111, 966-981 (2021).
Cullen MG, Thompson LJ, Carolan JK, Stout JK.na Stanley DA Fungicides, herbicides na aṅụ: nyochaa nhazi nke nyocha na ụzọ dị ugbu a.PLoS One 14, e0225743 (2019).
Pilling, ED na Jepson, PC Synergistic mmetụta nke EBI fungicides na pyrethroid ụmụ ahụhụ na mmanụ aṅụ (Apis mellifera).pests na sayensị.39, 293–297 (1993).
Mussen, EC, Lopez, JE na Peng, CY Mmetụta nke ahọrọ fungicides na uto na mmepe nke mmanụ aṅụ larvae Apis mellifera L. (Hymenoptera: Apidae).Wenezde.Ntomore.33, 1151-1154 (2004).
Van Dyke, M., Mullen, E., Wickstead, D., na McArt, Ntuzi Mkpebi S. maka iji ọgwụ pesticide mee ihe iji chebe ndị na-eme mkpọtụ na osisi osisi osisi (Mahadum Cornell, 2018).
Iwasaki, JM na Hogendoorn, K. Ngosipụta nke aṅụ na ọgwụ ndị na-abụghị pesticides: nyocha nke ụzọ na nsonaazụ a kọrọ.Ọrụ ugbo.gburugburu ebe obibi.Wenezde.314, 107423 (2021).
Kopit AM, Klinger E, Cox-Foster DL, Ramirez RA.na Pitts-Singer TL Mmetụta nke ụdị ọkọnọ na mkpughe pesticide na mmepe larval nke Osmia lignaria (Hymenoptera: Megachilidae).Wenezde.Ntomore.51, 240–251 (2022).
Ụzọ Kopit AM na Pitts-Sing TL nke ikpughe ọgwụ nje na aṅụ efu efu.Wenezde.Ntomore.47, 499–510 (2018).
Pan, NT et al.Protocol bioassay ingestion ọhụrụ maka inyocha nsí pesticide n'ime ndị okenye aṅụ ndị Japan (Osmia cornifrons).sayensị.Akụkọ 10, 9517 (2020).
Oge nzipu: Mee-14-2024