Mahalo no kou kipa ʻana iā Nature.com.ʻO ka mana o ka polokalamu kele pūnaewele āu e hoʻohana nei he kākoʻo CSS palena ʻole.No nā hualoaʻa maikaʻi loa, paipai mākou e hoʻohana ʻoe i kahi mana hou o kāu polokalamu kele pūnaewele (a i ʻole e hoʻopau i ke ʻano Compatibility Mode ma Internet Explorer).I kēia manawa, e hōʻoia i ke kākoʻo mau ʻana, ke hōʻike nei mākou i ka pūnaewele me ka ʻole styling a JavaScript paha.
Hoʻohana pinepine ʻia nā fungicides i ka wā e pua ai nā hua lāʻau a hiki ke hoʻoweliweli i nā pollinators insect.Eia nō naʻe, ʻaʻole i ʻike nui ʻia e pili ana i ka pane ʻana o nā pollinators non-bee (e laʻa, nā nalo kaʻawale, Osmia cornifrons) i ka hoʻopili ʻana a me nā fungicide ʻōnaehana i hoʻohana mau ʻia ma nā ʻāpala i ka wā e pua ai.Hoʻopili kēia ʻike ʻike i nā hoʻoholo hoʻoponopono e hoʻoholo ana i ka ʻike palekana a me ka manawa o ka hoʻohu ʻana i ka fungicide.Ua loiloi mākou i nā hopena o ʻelua fungicides pili (captan a mancozeb) a me ʻehā interlayer / phytosystem fungicides (ciprocycline, myclobutanil, pyrostrobin a me trifloxystrobin).Nā hopena i ka loaʻa ʻana o ke kaumaha o ka larval, ke ola, ka lakio kāne a me ka ʻokoʻa bacteria.Ua mālama ʻia ka loiloi ma ka hoʻohana ʻana i kahi bioassay waha maʻamau i mālama ʻia ka pollen i ʻekolu mau kau ma muli o ke ʻano i manaʻo ʻia i kēia manawa no ka hoʻohana ʻana i ke kula (1X), ka hapalua (0.5X) a me ka haʻahaʻa haʻahaʻa (0.1X).ʻO nā kumu a pau o ka mancozeb a me ka pyritisoline i hoʻemi nui i ke kaumaha o ke kino a me ke ola ʻana o nā larval.A laila hoʻonohonoho mākou i ka gene 16S e ʻike i ka larval bacteriome o mancozeb, ka fungicide nona ka make kiʻekiʻe loa.Ua ʻike mākou ua hoʻemi nui ʻia ka ʻokoʻa a me ka nui o nā maʻi bacterial i nā larvae i hānai ʻia i ka pollen i mālama ʻia mancozeb.Hōʻike nā hualoaʻa o kā mākou keʻena hoʻokolohua ʻo ka pīpī ʻana i kekahi o kēia mau fungicide i ka wā o ka pua ʻana he mea pōʻino loa ia i ke olakino o O. cornifrons.Pili kēia ʻike no nā hoʻoholo hoʻokele e hiki mai ana e pili ana i ka hoʻohana mau ʻana i nā huahana pale lāʻau huaʻai a lilo i kumu no nā kaʻina hoʻoponopono e pili ana i ka pale ʻana i nā pollinators.
Ua hoʻolauna ʻia ka bee mason bee Osmia cornifrons (Hymenoptera: Megachilidae) i ʻAmelika Hui Pū ʻIa mai Iapana i ka hopena o ka makahiki 1970 a me ka hoʻomaka ʻana o nā makahiki 1980, a ua hoʻokani kēia ʻano i kahi hana pollinator koʻikoʻi i nā kaiaola mālama ʻia mai ia manawa.ʻO nā heluna kanaka maoli o kēia nalo he ʻāpana o kahi o 50 mau ʻano o nā nalo ʻāhiu e hoʻokō i nā pi e pollinate i nā māla ʻalemona a me nā ʻāpala ma ʻAmelika Hui Pū ʻIa2,3.Ke kū nei ka nalo Mason i nā pilikia he nui, e like me ka wehewehe ʻana i ka nohona, nā pathogens, a me nā pesticides3,4.Ma waena o nā insecticides, hoʻemi ka fungicides i ka loaʻa ʻana o ka ikehu, foraging5 a me ke kino kino6,7.ʻOiai ke hōʻike nei nā noiʻi hou e pili pono ana ke olakino o nā pi Mason e nā microorganism commensal a ectobactic, 8,9 no ka mea hiki i ka bacteria a me nā haʻiki ke hoʻoikaika i ka meaʻai a me nā pane kūlohelohe, ʻo nā hopena o ka hoʻolaha ʻana i ka fungicide i ka ʻokoʻa microbial o nā pi Mason ke hoʻomaka nei e lilo. aʻo ʻia.
Hoʻopili ʻia nā fungicides o nā hopena like ʻole (contact and systemic) i loko o nā māla waina ma mua a i ka wā o ka pua ʻana no ka mālama ʻana i nā maʻi e like me ka ʻāpala ʻōpala, ka pala ʻawaʻawa, ka pala ʻeleʻele a me ka pauka10,11.Manaʻo ʻia nā fungicides he mea ʻole i nā pollinators, no laila ua ʻōlelo ʻia lākou i nā māla i ka wā pua;ʻO ka hōʻike ʻana a me ka ʻai ʻana o kēia mau fungicides e nā pi ua ʻike maikaʻi ʻia, ʻoiai he ʻāpana ia o ke kaʻina hoʻopaʻa inoa pesticide e ka US Environmental Protection Agency a me nā ʻoihana hoʻoponopono aupuni ʻē aʻe12,13,14.Eia naʻe, ʻaʻole i ʻike ʻia nā hopena o ka fungicides ma luna o nā pi no ka mea ʻaʻole koi ʻia lākou ma lalo o nā ʻaelike ʻae kūʻai aku ma United States15.Eia hou, ʻaʻohe kuʻina maʻamau no ka hoʻāʻo ʻana i nā nalo hoʻokahi16,17, a paʻakikī ka mālama ʻana i nā kolo e hāʻawi ana i nā pi no ka hoʻāʻo ʻana18.Ke hoʻomau ʻia nei nā hoʻāʻo ʻana o nā pi hoʻokele like ʻole ma ʻEulopa a me ʻAmelika e aʻo i ka hopena o nā pesticides ma luna o nā nalo hihiu, a ua hoʻomohala ʻia nā kuʻuna maʻamau no O. cornifrons19.
He mau monocytes ka nalo liʻiliʻi a hoʻohana ʻia i nā mea kanu carp ma ke ʻano he mea hoʻohui a pani paha no ka meli meli.Puka mai kēia mau nalo ma waena o Malaki a me ʻApelila, me ka puka ʻana mai o nā kane mua i ʻekolu a ʻehā lā ma mua o nā wahine.Ma hope o ka male ʻana, ʻohi ikaika ka wahine i ka pollen a me ka nectar no ka hoʻolako ʻana i nā pūnaewele brood i loko o ka lua pūnana tubular (kūlohelohe a mea hana paha)1,20.Waiho ʻia nā hua ma luna o ka pollen i loko o nā keena;a laila kūkulu ka wahine i ka pā pālolo ma mua o ka hoʻomākaukau ʻana i ke keena aʻe.Hoʻopaʻa ʻia nā larvae instar mua i loko o ka chorion a ʻai i nā wai embryonic.Mai ka lua a hiki i ka lima o ka instar (prepupa), ai na larvae i ka pollen22.I ka pau 'ana o ka pollen, hana 'ia ka larvae i cocoons, pupate a puka mai ma ke 'ano he kanaka makua i loko o ke ke'ena ho'okahi, ma ka hopena o ke kauwela20,23.Puka mai nā mākua i ka pūnāwai aʻe.Hoʻopili ʻia ke ola kanaka makua me ka loaʻa ʻana o ka ikehu ʻupena (paona paona) ma muli o ka ʻai ʻana i ka meaʻai.No laila, ʻo ka maikaʻi o ka meaʻai o ka pollen, a me nā mea ʻē aʻe e like me ke anā a i ʻole ka ʻike ʻana i nā lāʻau pesticides, nā mea e hoʻoholo ai i ke ola a me ke olakino24.
Hiki ke ne'e i loko o ka vasculature o ka mea kanu a me ka fungicides, mai ka translaminar (no ka la'ana, hiki ke ne'e mai ka ili o luna a hiki i ka ili o lalo, e like me kekahi mau fungicides) 25 a hiki i ka hopena maoli., hiki ke komo i ka leialii mai ke a'a mai, hiki ke komo i ka nectar o ka apple pua26, kahi e hiki ai ke pepehi kanaka makua O. cornifrons27.Lilo kekahi mau lāʻau pesticides i loko o ka pollen, e hoʻopilikia ana i ka ulu ʻana o nā ʻōpala maile a make lākou19.Ua hōʻike ʻia nā haʻawina ʻē aʻe e hiki i kekahi mau fungicides ke hoʻololi nui i ke ʻano hana pūnana o nā ʻano pili pili ʻo O. lignaria28.Eia hou, ua hōʻike ʻia nā haʻawina hoʻokolohua a me nā kula e hoʻohālikelike ana i nā hiʻohiʻona ʻike pesticide (me nā fungicides) i ka hopena maikaʻi ʻole o nā pesticides i ka physiology 22 morphology 29 a me ke ola ʻana o nā nalo meli a me kekahi mau nalo mehameha.Hiki ke hoʻohaumia i ka pollen i hōʻiliʻili ʻia e ka poʻe mākua no ka ulu ʻana o ka larval nā ʻano ʻano fungicidal sprays i hoʻopili pololei ʻia i nā pua i ka wā e pua ai.
Ua ʻike nui ʻia ka hoʻomohala ʻana o ka larval e ka pollen a me nā kaiāulu microbial o ka ʻōnaehana digestive.Hoʻopili ka honeybee microbiome i nā ʻāpana e like me ka nui o ke kino31, nā loli metabolic22 a me ka maʻalahi i nā pathogens32.Ua nānā nā haʻawina mua i ka mana o ka pae hoʻomohala, nā meaʻai, a me ke kaiapuni ma luna o ka microbiome o nā nalo kaʻawale.Ua hōʻike ʻia kēia mau haʻawina i nā ʻano like o ke ʻano a me ka nui o ka larval a me ka pollen microbiomes33, a me ka genera bacterial maʻamau ʻo Pseudomonas a me Delftia, ma waena o nā ʻano pi hoʻokahi.Eia nō naʻe, ʻoiai ua pili nā fungicides me nā hoʻolālā e pale ai i ke olakino pi, ʻaʻole i ʻike ʻia nā hopena o ka fungicides ma luna o ka larval microbiota ma o ka hoʻolaha waha.
Ua ho'āʻo kēia haʻawina i nā hopena o nā mea hoʻopaʻa honua maoli o ʻeono i hoʻohana maʻamau ʻia no ka hoʻohana ʻana ma nā hua lāʻau ma ʻAmelika Hui Pū ʻIa, me ka hoʻopili ʻana a me nā fungicides systemic i lawelawe waha ʻia i nā larva hornworm moth mai ka meaʻai i hoʻohaumia ʻia.Ua ʻike mākou ua hoʻemi ʻia ka loaʻa ʻana o ke kaumaha o ke kino pī a me ka hoʻonui ʻana i ka make, me nā hopena koʻikoʻi e pili ana i ka mancozeb a me pyrithiopide.A laila hoʻohālikelike mākou i ka ʻokoʻa microbial o nā larvae i hānai ʻia i ka meaʻai pollen i mālama ʻia mancozeb me nā mea i hānai ʻia i ka meaʻai hoʻomalu.Kūkākūkā mākou i nā ʻano hana e pili ana i ka make a me nā hopena no nā polokalamu hoʻokele pest and pollinator management (IPPM)36.
Ua loaʻa mai ka Hale Noiʻi Huaʻai, Biglerville, PA, ʻo O. cornifrons makua i ka hoʻoilo i ka hoʻoilo, a mālama ʻia ma -3 a 2°C (±0.3°C).Ma mua o ka hoʻokolohua (600 cocoons i ka huina).I ka Mei 2022, ua hoʻoili ʻia he 100 O. cornifrons cocoons i kēlā me kēia lā i loko o nā kīʻaha plastik (50 cocoons no ke kīʻaha, DI 5 cm × 15 cm ka lōʻihi) a waiho ʻia nā holoi holoi i loko o nā kīʻaha e hāpai i ka wehe ʻana a hāʻawi i kahi substrate chewable, e hōʻemi ana i ke kaumaha ma ka pōhaku. nā pi37 .E hoʻokomo i ʻelua kīʻaha plastik i loko o ka hale nalo (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) me nā mea hānai 10 ml me 50% sucrose solution a mālama i nā lā ʻehā e hōʻoia i ka pani ʻana a me ka male ʻana.23°C, haʻahaʻa pili 60%, photoperiod 10 l (kaha haʻahaʻa): 14 lā.Ua hoʻokuʻu ʻia 100 mau wahine male a me nā kāne i kēlā me kēia kakahiaka no nā lā ʻeono (100 i kēlā me kēia lā) i ʻelua pūnana hana i ka wā o ka ulu ʻana o ka ʻāpala kiʻekiʻe.Hoʻonoho ʻia ma Pennsylvania State Arboretum, kokoke i ke cherry (Prunus cerasus 'Eubank' Sweet Cherry Pie ™), peach (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), pear (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), lāʻau ʻāpala coronaria (Malus coronaria) a me nā ʻano ʻano lāʻau ʻāpala he nui (Malus coronaria, Malus), lāʻau ʻāpala hale 'Co-op 30' Enterprise™, lāʻau ʻāpala Malus 'Co- Op 31' Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy').ʻO kēlā me kēia hale manu polū polū ma luna o nā pahu lāʻau ʻelua.Loaʻa i kēlā me kēia pahu pūnana he 800 mau paipu pepa kraft ʻole (spiral open, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) i hoʻokomo ʻia i loko o nā paipu cellophane opaque. .
Huli i ka hikina nā pahu pūnana ʻelua a ua uhi ʻia me nā pā kīhāpai ʻōmaʻomaʻo (Everbilt model #889250EB12, ka nui o ka wehe ʻana he 5 × 5 knm, 0.95 m × 100 m) i mea e pale ai i ke komo ʻana o nā ʻiole a me nā manu a kau ʻia ma ka ʻili lepo e pili ana i ka lepo pahu pūnana. pahu.Pahu pūnana (Hōʻi 1a).Ua hōʻiliʻili ʻia nā hua hua kulina i kēlā me kēia lā ma o ka hōʻiliʻili ʻana i 30 paipu mai nā pūnana a lawe ʻia i ka hale hana.Me ka ʻokiʻoki, e ʻoki i ka hope o ka paipu, a laila e hoʻokaʻawale i ka paipu spiral e hōʻike i nā pūnaewele brood.Wehe ʻia nā hua pākahi a me kā lākou pollen me ka hoʻohana ʻana i ka spatula curved (Microslide tool kit, BioQuip Products Inc., Kaleponi).Hoʻokomo ʻia nā hua manu ma ka pepa kānana wai a waiho ʻia i loko o kahi pā Petri no 2 mau hola ma mua o ka hoʻohana ʻana i kā mākou hoʻokolohua (Supplementary Figure 1b-d).
I loko o ke keʻena hoʻokolohua, ua loiloi mākou i ka ʻawaʻawa waha o ʻeono fungicides i hoʻohana ʻia ma mua a i ka wā o ka ulu ʻana o ka ʻāpala i ʻekolu mau ʻāpana (0.1X, 0.5X, a me 1X, kahi 1X ka māka i hoʻohana ʻia no 100 galani wai/ʻeka. ma ke kula)., Papa 1).Ua hoʻonui hou ʻia kēlā me kēia hoʻopaʻa ʻana i nā manawa 16 (n = 16).ʻElua fungicides pili (Papa S1: mancozeb 2696.14 ppm a me ka captan 2875.88 ppm) a me ʻehā ʻano fungicides systemic (Table S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75.2 ppm , nā mea kanu a me nā mea kanu nani .Hoʻomoʻa mākou i ka pollen me ka hoʻohana ʻana i ka mea wili, hoʻololi i ka 0.20 g i kahi pūnāwai (24-well Falcon Plate), a hoʻohui ʻia a hui pū ʻia me 1 μL o ka hopena fungicide e hana i ka pollen pyramidal me nā luawai hohonu 1 mm kahi i waiho ʻia ai nā hua.E hoʻohana i kahi spatula liʻiliʻi (Hoʻohui Kiʻi 1c,d).Hoʻopaʻa ʻia nā pā Falcon ma ka lumi wela (25 ° C) a me 70% haʻahaʻa haʻahaʻa.Hoʻohālikelike mākou iā lākou me nā larvae control i hānai ʻia i ka meaʻai pollen homogeneous i mālama ʻia me ka wai maʻemaʻe.Ua hoʻopaʻa mākou i ka make a ua ana mākou i ke kaumaha o ka larval i kēlā me kēia lā a hiki i ka hiki ʻana o nā larvae i ka makahiki prepupal me ka hoʻohana ʻana i ke kaulike analytical (Fisher Scientific, pololei = 0.0001 g).ʻO ka hope, ua loiloi ʻia ka lakio kāne ma ka wehe ʻana i ka niu ma hope o 2.5 mahina.
Ua unuhi ʻia ka DNA mai loko mai o O. cornifrons larvae holoʻokoʻa (n = 3 no kēlā me kēia kūlana lapaʻau, mancozeb-treated and untreated pollen) a ua hana mākou i nā ʻano loiloi microbial diversity ma kēia mau laʻana, ʻoi loa no ka mea ma ka mancozeb ʻike ʻia ka make kiʻekiʻe loa i nā larvae.e loaa ana ia MnZn.Ua hoʻonui ʻia ka DNA, hoʻomaʻemaʻe ʻia me ka DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), a hoʻonohonoho ʻia (600 cycles) ma kahi Illumina® MiSeq™ me ka hoʻohana ʻana i ka pahu v3.Hoʻohana ʻia ka hoʻonohonoho ʻana o nā genes 16S ribosomal RNA bacteria me ka hoʻohana ʻana i ka Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) me ka hoʻohana ʻana i nā primers e kuhikuhi ana i ka māhele V3-V4 o ka gene 16S rRNA.Hoʻohui ʻia, ua hana ʻia ke kaʻina hana 18S me ka hoʻohana ʻana i ka 10% PhiX, a ua hoʻokō ʻia ka amplification me ka hoʻohana ʻana i ka pair primer 18S001 a me NS4.
Hoʻokomo a hoʻopaʻa ʻia nā heluhelu39 me ka hoʻohana ʻana i ka paipu QIIME2 (v2022.11.1).Ua ʻoki ʻia kēia mau heluhelu a hoʻohui ʻia, a ua wehe ʻia nā kaʻina chimeric me ka hoʻohana ʻana i ka plugin DADA2 ma QIIME2 (qiime dada2 noise pairing)40.Ua hana ʻia nā hana papa 16S a me 18S me ka hoʻohana ʻana i ka plugin classifier object Classify-sklearn a me ka artifact silva-138-99-nb-classifier.
Ua nānā ʻia nā ʻikepili hoʻokolohua āpau no ka maʻamau (Shapiro-Wilks) a me ka homogeneity o nā ʻano like ʻole (Levene's test).No ka mea ʻaʻole i hoʻokō ka ʻikepili i nā manaʻo o ka nānā ʻana parametric a ʻaʻole i hoʻohālikelike ka hoʻololi ʻana i nā koena, ua hana mākou i kahi ANOVA nonparametric ʻelua ala (Kruskal-Wallis) me nā kumu ʻelua [manawa (ʻekolu manawa 2, 5, a me 8 lā. manawa) a me ka fungicide] no ka loiloi ʻana i ka hopena o ka mālama ʻana i ke kaumaha hou o ka larval, a laila ua hana ʻia nā hoʻohālikelike post hoc nonparametric me ka hoʻohana ʻana i ka hoʻokolohua Wilcoxon.Ua hoʻohana mākou i kahi hiʻohiʻona laina laulā (GLM) me kahi puʻupuʻu Poisson e hoʻohālikelike i nā hopena o nā fungicides i ke ola ʻana ma waena o ʻekolu fungicide concentrations41,42.No ka ʻokoʻa ʻokoʻa abundance analysis, ua hāʻule ka helu o nā amplicon sequence variants (ASVs) i ka pae genus.Hoʻohālikelike ʻia ka nui ʻokoʻa ma waena o nā pūʻulu e hoʻohana ana i ka 16S (genus level) a me 18S pili pili i hana ʻia me ka hoʻohana ʻana i kahi kumu hoʻohui ākea no ke kūlana, ka unahi, a me ke ʻano (GAMLSS) me nā māhele ʻohana beta zero-inflated (BEZI), i hoʻohālike ʻia ma kahi macro. .i ka Microbiome R43 (v1.1).1).Wehe i nā ʻano mitochondrial a me ka chloroplast ma mua o ka nānā ʻana ʻokoʻa.Ma muli o nā pae ʻauhau ʻokoʻa o 18S, ʻo ka pae haʻahaʻa wale nō o kēlā me kēia ʻauhau i hoʻohana ʻia no nā loiloi ʻokoʻa.Ua hana ʻia nā ʻikepili helu a pau me ka hoʻohana ʻana iā R (v. 3.4.3., CRAN project) (Team 2013).
ʻO ka hōʻike ʻana i ka mancozeb, pyrithiostrobin, a me trifloxystrobin i hōʻemi nui i ke kaumaha o ke kino ma O. cornifrons (Fig. 1).Ua ʻike mau ʻia kēia mau hopena no nā ʻekolu a pau i loiloi ʻia (Fig. 1a-c).ʻAʻole i hōʻemi nui ʻo Cyclostrobin a me myclobutanil i ke kaumaha o nā larvae.
ʻAwelika kaumaha hou o nā ʻōniu ʻōpuʻu lāʻau i ana ʻia i ʻekolu mau manawa ma lalo o ʻehā mau lāʻau lapaʻau meaʻai (ʻai pollen homogeneous + fungicide: hoʻomalu, 0.1X, 0.5X a me 1X mau kau.(a) Haʻahaʻa haʻahaʻa (0.1X): ka manawa mua (lā 1): χ2: 30.99, DF = 6;P <0.0001, helu manawa lua (lā 5): 22.83, DF = 0.0009;ke kolu o ka manawa;kiko (lā 8): χ2: 28.39, DF = 6;(b) hapa hapa (0.5X): ka manawa mua (lā 1): χ2: 35.67, DF = 6;P <0.0001, helu manawa ʻelua (lā hoʻokahi).): χ2: 15.98, DF = 6;P = 0.0090;helu ʻekolu (lā 8) χ2: 16.47, DF = 6;(c) Pae a i ʻole ka nui piha (1X): ka manawa mua (lā 1) χ2: 20.64, P = 6;P = 0.0326, helu manawa ʻelua (lā 5): χ2: 22.83, DF = 6;P = 0.0009;ʻekolu manawa helu (lā 8): χ2: 28.39, DF = 6;ka nānā ʻana i nā ʻano like ʻole.Hōʻike nā pahu i ka mean ± SE o nā hoʻohālikelike pālua (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Ma ka haʻahaʻa haʻahaʻa (0.1X), ua hoʻemi ʻia ke kaumaha o ke kino larval e 60% me trifloxystrobin, 49% me mancozeb, 48% me myclobutanil, a me 46% me pyrithistrobin (Fig. 1a).Ke hōʻike ʻia i ka hapalua o ka mahina o ke kahua (0.5X), ua hoʻemi ʻia ke kaumaha o ke kino o nā larvae mancozeb e 86%, pyrithiostrobin e 52% a me trifloxystrobin e 50% (Fig. 1b).ʻO kahi pānaʻi kahua piha (1X) o ka mancozeb i hoʻemi i ke kaumaha larval e 82%, pyrithiostrobin ma 70%, a me trifloxystrobin, myclobutanil a me sangard ma kahi o 30% (Fig. 1c).
ʻOi aku ka nui o ka make ma waena o nā larvae i hānai ʻia mancozeb-treated pollen, a ukali ʻia e pyrithiostrobin a me trifloxystrobin.Hoʻonui ka make me ka hoʻonui ʻana o ka mancozeb a me ka pyritisoline (Fig. 2; Table 2).Eia naʻe, ua piʻi iki ka make ʻana o ka ʻai kulina i ka piʻi ʻana o ka trifloxystrobin;ʻAʻole i hoʻonui nui ka cyprodinil a me ka captan i ka make ma mua o ka mālama ʻana.
Ua hoʻohālikelike ʻia ka make ʻana o nā larvae lele ma hope o ka ʻai ʻana i ka pollen i mālama ʻia me ʻeono fungicides like ʻole.ʻO ka Mancozeb a me ka pentopyramide iʻoi aku ka maʻalahi i kaʻike waha i nā ilo kulina (GLM: χ = 29.45, DF = 20, P = 0.0059) (laina, slope = 0.29, P <0.001; slope = 0.24, P <0.00)).
Ma ka awelika, ma nā lāʻau lapaʻau āpau, 39.05% o nā maʻi he wahine a he 60.95% nā kāne.Ma waena o nā lāʻau hoʻomalu, ʻo ka hapa o nā wahine he 40% ma nā haʻawina haʻahaʻa haʻahaʻa (0.1X) a me ka hapalua (0.5X), a me 30% i nā haʻawina kahua-dose (1X).Ma ka helu 0.1X, ma waena o nā ʻōpala hānai pollen i mālama ʻia me ka mancozeb a me ka myclobutanil, 33.33% o nā pākeke he wahine, 22% o nā pākeke he wahine, 44% o nā ʻōpū makua kāne he wahine, 44% o nā ʻōpū makua he wahine.wahine, he 41% o na larvae makua he wahine, a he 31% ka mana (Fig. 3a).Ma ka 0.5 manawa o ka dose, 33% o nā ilo makua i ka mancozeb a me ka pyrithiostrobin pūʻulu he wahine, 36% i ka trifloxystrobin pūʻulu, 41% i ka myclobutanil hui, a me 46% i ka hui cyprostrobin.He 53% kēia helu ma ka hui.i ka hui captan a me 38% i ka pūʻulu mana (Fig 3b).Ma ka 1X dose, 30% o ka hui mancozeb he mau wahine, 36% o ka hui pyrithiostrobin, 44% o ka hui trifloxystrobin, 38% o ka hui myclobutanil, 50% o ka pūʻulu mana he wahine - 38.5% (Fig 3c) .
Ka pakeneka o na wahine a me na kane borers ma hope o ka larval stage fungicide.(a) Haʻahaʻa haʻahaʻa (0.1X).(b) Ka hapa hapa (0.5X).(c) ʻĀpana kahua a i ʻole ka nui piha (1X).
Ua hōʻike ʻia ka hōʻike ʻana o ke kaʻina hana 16S i ka ʻokoʻa o ka hui bacteria ma waena o nā larvae i hānai ʻia me ka mancozeb-treated pollen a me nā larvae i hānai ʻia me ka pollen i mālama ʻole ʻia (Fig. 4a).ʻOi aku ka kiʻekiʻe o ka helu microbial o nā ʻōpala i mālama ʻole ʻia i hānai ʻia i ka pollen ma mua o ka mea i hānai ʻia i ka pollen i mālama ʻia mancozeb (Fig. 4b).ʻOiai ʻo ka ʻokoʻa i ʻike ʻia i ka waiwai ma waena o nā pūʻulu ʻaʻole i helu nui ʻia, ʻoi aku ka haʻahaʻa ma mua o ka mea i ʻike ʻia no ka ʻai ʻana o nā larvae i ka pollen i mālama ʻole ʻia (Fig. 4c).Ua hōʻike ʻia ka nui o ka microbiota o nā larvae i hānai ʻia i ka pollen control ma mua o ka ʻano o nā larvae i hānai ʻia i nā larvae i mālama ʻia mancozeb (Fig. 5a).Hōʻike ka wehewehe wehewehe ʻana i ka hiki ʻana o nā genera 28 i ka mana a me nā mancozeb-treated samples (Fig. 5b).ʻAʻole i hōʻike ʻia kahi ʻokoʻa koʻikoʻi (Hoʻohui Kiʻi 2).
Ua hoʻohālikelike ʻia nā ʻaoʻao SAV e pili ana i nā kaʻina 16S me ka waiwai ʻo Shannon a nānā i ka waiwai ma ka pae phylum.(a) Ke kālailai hoʻonohonoho kumu (PCoA) e pili ana i ka hoʻolālā kaiaulu microbial holoʻokoʻa i ka hānai pollen a i ʻole ka mana (uliuli) a me nā larvae hānai mancozeb (alani).Hōʻike kēlā me kēia kikoʻikepili i kahi laʻana kaʻawale.Ua helu ʻia ʻo PCoA me ka hoʻohana ʻana i ka mamao Bray-Curtis o ka mahele t multivariate.Hōʻike nā Ovals i ka pae hilinaʻi 80%.(b) Boxplot, ʻike waiwai waiwai Shannon (nā helu) a me c.ʻIke ʻia ka waiwai.Hōʻike nā pahu pahu i nā pahu no ka laina waena, ka laulā interquartile (IQR), a me 1.5 × IQR (n = 3).
ʻO ka hui ʻana o nā kaiāulu microbial o nā larvae i hānai ʻia i ka pollen i mālama ʻia mancozeb a i mālama ʻole ʻia.(a) Heluhelu ʻia ka nui o nā genera microbial i loko o nā larvae.(b) Heat map o nā kaiaulu microbial i ʻike ʻia.Delftia (odds ratio (OR) = 0.67, P = 0.0030) a me Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060);Hoʻopili ʻia nā lālani palapala ʻāina wela me ka hoʻohana ʻana i ka mamao correlation a me ka hoʻopili maʻamau.
Hōʻike kā mākou hopena i ka hoʻolaha waha i ka hoʻopili ʻana (mancozeb) a me ka systemic (pyrostrobin a me trifloxystrobin) fungicides, i hoʻohana nui ʻia i ka wā o ka pua, hoʻemi nui i ka loaʻa kaumaha a hoʻonui i ka make o nā larvae maile.Eia kekahi, ua hoʻemi nui ka mancozeb i ka ʻokoʻa a me ka waiwai o ka microbiome i ka wā prepupal.ʻO Myclobutanil, kekahi mea hoʻomakeʻa ʻōnaehana, hoʻemi nui i ka loaʻa ʻana o ke kaumaha o ke kino larval ma nā ʻano ʻekolu.Ua ʻike ʻia kēia hopena ma ka lua (lā 5) a me ke kolu (lā 8) manawa manawa.I ka hoʻohālikelike ʻana, ʻaʻole i hōʻemi nui ka cyprodinil a me ka captan i ka loaʻa ʻana o ke kaumaha a i ʻole ke ola i hoʻohālikelike ʻia me ka pūʻulu mana.I ko mākou ʻike, ʻo kēia hana ka mea mua e hoʻoholo ai i nā hopena o nā kumukūʻai kula o nā fungicide ʻokoʻa i hoʻohana ʻia no ka pale ʻana i nā hua kulina ma o ka hoʻolaha ʻana i ka pollen.
ʻO nā lāʻau lapaʻau fungicide a pau i hōʻemi nui i ka piʻi ʻana o ke kaumaha o ke kino ke hoʻohālikelike ʻia me nā lāʻau hoʻomalu.ʻO Mancozeb ka hopena nui loa i ka loaʻa ʻana o ke kaumaha o ke kino larval me ka awelika hōʻemi o 51%, a ukali ʻia e pyrithiostrobin.Eia naʻe, ʻaʻole i hōʻike ʻia nā haʻawina ʻē aʻe i nā hopena ʻino o nā pānaʻi kahua o nā fungicides ma nā pae larval44.ʻOiai ua hōʻike ʻia nā biocides dithiocarbamate he mea ʻona haʻahaʻa loa45, hiki i ka ethylene bisdithiocarbamates (EBDCS) e like me mancozeb ke hoʻohaʻahaʻa i ka urea ethylene sulfide.Hāʻawi ʻia i kona hopena mutagenic i nā holoholona ʻē aʻe, hiki i kēia huahana degradation ke kuleana no nā hopena i ʻike ʻia46,47.Ua hōʻike ʻia nā haʻawina mua i ka hoʻokumu ʻia ʻana o ka ethylene thiourea e nā mea e like me ke kiʻekiʻe o ka mahana48, nā pae haʻahaʻa49 a me ka lōʻihi o ka waiho ʻana i nā huahana50.Hiki i nā kūlana mālama kūpono no nā biocides ke hoʻēmi i kēia mau hopena ʻaoʻao.Eia kekahi, ua hōʻike ka European Food Safety Authority i ka hopohopo e pili ana i ka ʻona o ka pyrithiopide, i hōʻike ʻia he carcinogenic i nā ʻōnaehana digestive o nā holoholona ʻē aʻe51.
ʻO ka hoʻohana waha ʻana i ka mancozeb, pyrithiostrobin, a me trifloxystrobin e hoʻonui i ka make o nā larvae corn borer.ʻO ka hoʻohālikelike, ʻaʻohe hopena o ka myclobutanil, ciprocycline a me ka captan i ka make.ʻOkoʻa kēia mau hopena mai ka Ladurner et al.52, nāna i hōʻike i ka hoʻemi nui ʻana o ka captan i ke ola o O. lignaria makua a me Apis mellifera L. (Hymenoptera, Apisidae).Eia kekahi, ua ʻike ʻia nā fungicides e like me captan a me boscalid i mea e make ai nā larval52,53,54 a i ʻole e hoʻololi i ka ʻano hānai55.Hiki i kēia mau hoʻololi ke hoʻololi i ka maikaʻi o ka meaʻai o ka pollen a i ka hopena i ka loaʻa ʻana o ka ikehu o ka pae larval.ʻO ka make i ʻike ʻia ma ka pūʻulu hoʻomalu ua kūlike me nā haʻawina ʻē aʻe 56,57.
Hiki ke wehewehe ʻia ka lakio kāne makemake i ka wahine ma kā mākou hana e nā kumu e like me ka lawa ʻole o ka male ʻana a me ka maikaʻi ʻole o ka wā o ka pua, e like me ka mea i ʻōlelo mua ʻia no O. cornuta e Vicens a me Bosch.ʻOiai ʻo nā wahine a me nā kāne i kā mākou aʻo ʻana he ʻehā mau lā e male ai (kahi manawa i manaʻo ʻia he lawa no ka hoʻopōmaikaʻi ʻana), ua noʻonoʻo mākou e hoʻemi i ka ikaika māmā e hōʻemi i ke kaumaha.Eia naʻe, hiki i kēia hoʻololi ke hoʻopilikia me ka manaʻo ʻole i ke kaʻina hana male61.Eia kekahi, ʻike nā nalo i kekahi mau lā o ka wā ʻino, me ka ua a me nā mahana haʻahaʻa (<5°C), hiki ke hoʻopilikia maikaʻi ʻole i ka kūleʻa male4,23.
ʻOiai kā mākou noiʻi ʻana i ka microbiome larval holoʻokoʻa, hāʻawi kā mākou hopena i ka ʻike i nā pilina pili i waena o nā kaiāulu bacteria e koʻikoʻi i ka meaʻai pī a me ka hoʻolaha ʻana i ka fungicide.No ka laʻana, ua hoʻemi nui ka pollen i hānai ʻia i ka mancozeb i ka nui o ke kaiāulu microbial i hoʻohālikelike ʻia me nā larvae i hānai ʻia i ka pollen i mālama ʻole ʻia.I loko o nā larvae e ʻai ana i ka pollen i mālama ʻole ʻia, ʻo nā pūʻulu bacteria Proteobacteria a me Actinobacteria ka mea nui a ʻoi aku ka nui o ka aerobic a i ʻole facultatively aerobic.ʻO ka bacteria Delft, ka mea maʻamau e pili ana me nā ʻano nalo mehameha, ua ʻike ʻia he hana lāʻau antibiotic, e hōʻike ana i kahi kuleana pale i nā pathogens.ʻO kekahi ʻano bacteria ʻē aʻe, ʻo Pseudomonas, ua nui i loko o nā larvae i hānai ʻia i ka pollen i mālama ʻole ʻia, akā ua hoʻemi nui ʻia i nā larvae i mālama ʻia mancozeb.Kākoʻo kā mākou mau hualoaʻa i nā noiʻi mua e ʻike ana ʻo Pseudomonas kekahi o nā genera nui loa ma O. bicornis35 a me nā nalo mehameha34.ʻOiai ʻaʻole i aʻo ʻia nā hōʻike hoʻokolohua no ka hana o Pseudomonas i ke olakino o O. cornifrons, ua hōʻike ʻia kēia bacterium e hāpai i ka synthesis o nā toxins pale i loko o ka beetle Paederus fuscipes a hoʻoikaika i ka metabolism arginine in vitro 35, 65. Hōʻike kēia mau ʻike. he kuleana koʻikoʻi i ka pale viral a me ka bacteria i ka wā ulu o O. cornifrons larvae.ʻO Microbacterium kekahi ʻano ʻano ʻē aʻe i ʻike ʻia i loko o kā mākou noiʻi ʻana i hōʻike ʻia aia ma nā helu kiʻekiʻe o nā ʻeleʻele lele koa ʻeleʻele ma lalo o nā kūlana pololi66.I loko o O. cornifrons larvae, hiki i ka microbacteria ke kōkua i ke kaulike a me ke kūpaʻa o ka microbiome o ka ʻōpū ma lalo o nā kūlana koʻikoʻi.Eia hou, loaʻa ʻo Rhodococcus i loko o O. cornifrons larvae a ua ʻike ʻia no kona hiki ke hoʻopau ʻana.Loaʻa kēia ʻano ma ka ʻōpū o A. florea, akā haʻahaʻa loa ka nui68.Hōʻike kā mākou mau hopena i ka loaʻa ʻana o nā ʻano hoʻololi genetic ma waena o nā ʻauhau microbial hiki ke hoʻololi i nā kaʻina metabolic i loko o nā larvae.Eia naʻe, pono ka ʻike maikaʻi ʻana i ka ʻokoʻa hana o O. cornifrons.
Ma ka hōʻuluʻulu manaʻo, hōʻike nā hopena i ka mancozeb, pyrithiostrobin, a me trifloxystrobin i hōʻemi i ke kaumaha o ke kino a me ka hoʻonui ʻana i ka make o nā larvae corn borer.ʻOiai ke ulu nei ka hopohopo e pili ana i nā hopena o nā fungicides ma luna o nā pollinators, pono e hoʻomaopopo pono i nā hopena o nā metabolites koena o kēia mau pūhui.Hiki ke hoʻokomo ʻia kēia mau hopena i loko o nā ʻōlelo paipai no nā papahana hoʻokele pollinator i hoʻohui ʻia e kōkua i ka poʻe mahiʻai e pale i ka hoʻohana ʻana i kekahi mau fungicides ma mua a i ka wā o ka ulu ʻana o ka lāʻau hua ma ke koho ʻana i nā fungicides a me ka hoʻololi ʻana i ka manawa o ka noi ʻana, a i ʻole ma ka paipai ʻana i ka hoʻohana ʻana i nā mea ʻoi aku ka pōʻino 36. ʻO kēia ʻike. mea nui no ka hooulu ana i na manao paipai.e pili ana i ka hoʻohana ʻana i ka pesticide, e like me ka hoʻoponopono ʻana i nā polokalamu hoʻoheheʻe e kū nei a me ka hoʻololi ʻana i ka manawa hoʻoheheʻe ʻana i ke koho ʻana i nā fungicide a i ʻole ka paipai ʻana i ka hoʻohana ʻana i nā mea ʻoi aku ka pōʻino.Pono ka noiʻi hou aʻe e pili ana i nā hopena maikaʻi ʻole o nā fungicides i ka lakio wahine, ka hānai ʻana, ka microbiome o ka ʻōpū, a me nā hana molekala i lalo o ka poho kaumaha a me ka make.
Ua waiho ʻia ka ʻikepili kumu 1, 2 a me 3 ma nā Kiʻi 1 a me 2 ma ka waihona ʻikepili figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 a me https://doi.org/10.6084/m9.figshare.24996233.Loaʻa nā kaʻina i hoʻopaʻa ʻia i ka haʻawina o kēia manawa (Fig. 4, 5) i ka waihona NCBI SRA ma lalo o ka helu komo PRJNA1023565.
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Ka manawa hoʻouna: Mei-14-2024