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Sau da yawa ana amfani da fungicides a lokacin furen 'ya'yan itace kuma suna iya barazana ga masu shukar kwari. Duk da haka, ba a san komai ba game da yadda masu shukar da ba sa shukar kudan zuma (misali, ƙudan zuma kaɗai, Osmia cornifrons) ke mayar da martani ga magungunan kashe kwari da aka saba amfani da su a kan apples yayin fure. Wannan gibin ilimi yana iyakance yanke shawara kan ƙa'idoji don tantance yawan lafiya da lokacin fesa maganin kashe kwari. Mun tantance tasirin magungunan kashe kwari guda biyu (captan da mancozeb) da magungunan kashe kwari guda huɗu a cikin Layer/phytosystem (ciprocycline, myclobutanil, pyrostrobin da trifloxystrobin). Tasirin ƙaruwar nauyin tsutsa, rayuwa, rabon jinsi da bambancin ƙwayoyin cuta. An gudanar da kimantawar ta amfani da gwajin bioasay na baki na yau da kullun inda aka yi maganin pollen a allurai uku bisa ga shawarar da aka bayar a yanzu don amfani da filin (1X), rabin allurai (0.5X) da ƙaramin allurai (0.1X). Duk allurai na mancozeb da pyritisoline sun rage nauyin jiki da rayuwar tsutsa sosai. Daga nan muka tsara kwayar halittar 16S don gano ƙwayoyin cuta na tsutsa na mancozeb, maganin kashe ƙwayoyin cuta da ke da alhakin mace-mace mafi girma. Mun gano cewa bambancin ƙwayoyin cuta da yawansu sun ragu sosai a cikin tsutsotsi da aka ciyar da pollen da aka yi wa mancozeb magani. Sakamakon dakin gwaje-gwajenmu ya nuna cewa fesa wasu daga cikin waɗannan magungunan kashe ƙwayoyin cuta yayin fure yana da illa musamman ga lafiyar O. cornifrons. Wannan bayanin ya dace da shawarwarin gudanarwa na gaba game da amfani da samfuran kariya daga bishiyoyi masu ɗorewa kuma yana aiki a matsayin tushen hanyoyin da aka tsara don kare masu yin fure.
An gabatar da ƙudan zuma mai suna Osmia cornifrons (Hymenoptera: Megachilidae) zuwa Amurka daga Japan a ƙarshen shekarun 1970 da farkon shekarun 1980, kuma nau'in ya taka muhimmiyar rawa wajen yin pollinating a cikin yanayin halittu masu sarrafawa tun daga lokacin. Yawan wannan ƙudan zuma da aka haifa a matsayin halitta wani ɓangare ne na kimanin nau'ikan ƙudan zuma 50 na daji waɗanda ke ƙara wa ƙudan zumar da ke yin pollinating a gonakin almond da apple a Amurka2,3. Kudan zumar Mason na fuskantar ƙalubale da yawa, gami da rarrabuwar muhalli, ƙwayoyin cuta, da magungunan kashe ƙwari3,4. Daga cikin magungunan kashe ƙwari, magungunan kashe ƙwari suna rage yawan kuzari, ciyarwa5 da kuma daidaita jiki6,7. Kodayake binciken da aka yi kwanan nan ya nuna cewa lafiyar ƙudan zumar Mason tana da tasiri kai tsaye ta hanyar ƙwayoyin cuta na commensal da ectobactic, 8,9 saboda ƙwayoyin cuta da fungi na iya yin tasiri ga abinci mai gina jiki da martanin garkuwar jiki, tasirin fallasa ƙwayoyin cuta ga bambancin ƙwayoyin cuta na ƙudan zumar Mason ya fara farawa ne kawai.
Ana fesa maganin fungicides masu tasiri daban-daban (wanda aka haɗa da wanda aka haɗa) a gonakin inabi kafin da kuma lokacin fure don magance cututtuka kamar su scab na apple, dattin ruɓewa, launin ruwan kasa da mildew foda10,11. Ana ɗaukar magungunan fungicides marasa lahani ga masu yin fure, don haka ana ba da shawarar su ga masu lambu a lokacin fure; Shafar waɗannan magungunan fungicides ta ƙudan zuma sananne ne, domin yana cikin tsarin yin rijistar magungunan kwari ta Hukumar Kare Muhalli ta Amurka da sauran hukumomin sa ido na ƙasa da yawa12,13,14. Duk da haka, tasirin magungunan fungicides ga waɗanda ba ƙudan zuma ba ba a san su sosai ba saboda ba a buƙatar su a ƙarƙashin yarjejeniyar izinin tallatawa a Amurka15. Bugu da ƙari, gabaɗaya babu ƙa'idodi na yau da kullun don gwada ƙudan zuma ɗaya16,17, kuma kula da wuraren da ke samar da ƙudan zuma don gwaji yana da ƙalubale18. Ana ƙara gudanar da gwaje-gwajen ƙudan zuma daban-daban a Turai da Amurka don nazarin tasirin magungunan kwari akan ƙudan zuma na daji, kuma kwanan nan an ƙirƙiri ƙa'idodi na yau da kullun don O. cornifrons19.
Kudan zuma masu ƙaho suna da ƙwayoyin halitta masu yawa kuma ana amfani da su a fannin amfanin gona na kifi a matsayin kari ko maye gurbin ƙudan zuma. Waɗannan ƙudan zuma suna fitowa tsakanin Maris da Afrilu, inda mazan da suka tsufa ke bayyana kwana uku zuwa huɗu kafin mata. Bayan haɗuwa, macen tana tattara pollen da nectar don samar da jerin ƙwayoyin halittu a cikin ramin gida mai siffar tubular (na halitta ko na wucin gadi)1,20. Ana sanya ƙwai a kan pollen a cikin ƙwayoyin; sai macen ta gina bango na yumbu kafin ta shirya ƙwayar halitta ta gaba. Ƙwayoyin farko na instar suna cikin chorion kuma suna cin ruwan tayi. Daga na biyu zuwa na biyar na instar (prepupa), tsutsotsi suna cin pollen22. Da zarar wadatar pollen ta ƙare gaba ɗaya, tsutsotsi suna yin kokwamba, suna yin jaki kuma suna fitowa a matsayin manya a cikin ɗakin tsuntsaye ɗaya, yawanci a ƙarshen bazara20,23. Manya suna fitowa a bazara mai zuwa. Rayuwar manya tana da alaƙa da ƙaruwar kuzari (ƙarin nauyi) bisa ga abincin da ake ci. Don haka, ingancin abinci mai gina jiki na pollen, da sauran abubuwa kamar yanayi ko fallasa ga magungunan kashe ƙwari, sune abubuwan da ke ƙayyade rayuwa da lafiya24.
Magungunan kashe kwari da maganin kashe kwari da aka yi amfani da su kafin fure suna iya motsawa a cikin jijiyoyin shuka zuwa matakai daban-daban, daga translaminar (misali, suna iya motsawa daga saman ganye zuwa ƙasan saman, kamar wasu magungunan kashe kwari) 25 zuwa tasirin tsarin gaske. , wanda zai iya shiga kambi daga tushen, zai iya shiga nectar na furannin apple26, inda zasu iya kashe manyan O. cornifrons27. Wasu magungunan kashe kwari suma suna shiga cikin pollen, suna shafar ci gaban tsutsotsin masara da haifar da mutuwarsu19. Wasu bincike sun nuna cewa wasu magungunan kashe kwari na iya canza halayen gida na nau'in da ke da alaƙa O. lignaria28. Bugu da ƙari, nazarin dakin gwaje-gwaje da na filin da ke kwaikwayon yanayin fallasa magungunan kashe kwari (gami da magungunan kashe kwari) sun nuna cewa magungunan kashe kwari suna yin mummunan tasiri ga yanayin halittar jiki 22 yanayin halittar zuma da wasu ƙudan zuma guda ɗaya. Feshin kashe kwari daban-daban da aka yi kai tsaye a kan furanni a buɗe yayin fure na iya gurɓata pollen da manya suka tattara don ci gaban tsutsotsi, waɗanda har yanzu ba a yi nazarin tasirinsu ba30.
Ana ƙara fahimtar cewa ci gaban tsutsotsi yana shafar pollen da ƙwayoyin cuta na tsarin narkewar abinci. Kwayoyin halittar zuma suna shafar sigogi kamar girman jiki31, canje-canje na rayuwa22 da kuma sauƙin kamuwa da cututtuka32. Nazarin da aka yi a baya sun binciki tasirin matakin ci gaba, abubuwan gina jiki, da muhalli akan ƙwayoyin cuta na ƙudan zuma guda ɗaya. Waɗannan nazarin sun bayyana kamanceceniya a cikin tsari da yawan ƙwayoyin cuta na tsutsotsi da pollen33, da kuma nau'ikan ƙwayoyin cuta da suka fi yawa a cikin Pseudomonas da Delftia, tsakanin nau'ikan ƙudan zuma guda ɗaya. Duk da haka, kodayake an danganta magungunan fungicides da dabarun kare lafiyar kudan zuma, tasirin magungunan fungicides akan ƙwayoyin cuta na tsutsotsi ta hanyar fallasa kai tsaye ta baki har yanzu ba a bincika su ba.
Wannan binciken ya gwada tasirin allurai shida na maganin kashe kwari da aka saba amfani da su a duniya da aka yi rijista don amfani a kan 'ya'yan itacen a Amurka, gami da maganin kashe kwari da na tsari da aka yi amfani da su ta baki ga tsutsar ƙwaro na masara daga abinci mai gurɓata. Mun gano cewa maganin kashe kwari da na tsari suna rage yawan nauyin jikin kudan zuma da kuma ƙara mace-mace, tare da mafi munin tasirin da ke da alaƙa da mancozeb da pyrithiopide. Sannan muka kwatanta bambancin ƙwayoyin cuta na tsutsotsi da aka ciyar a kan abincin pollen da aka yi wa mancozeb da waɗanda aka ciyar a kan abincin da aka sarrafa. Mun tattauna hanyoyin da za su iya haifar da mace-mace da kuma tasirin shirye-shiryen kula da kwari da pollinators (IPPM)36.
An samo manyan koren O. da ke yin sanyi a cikin koren daga Cibiyar Binciken 'Ya'yan Itace, Biglerville, PA, kuma an adana su a zafin −3 zuwa 2°C (±0.3°C). Kafin gwajin (jimillar koren 600). A watan Mayu na 2022, an tura koren cornifrons 100 O. kowace rana zuwa kofunan filastik (koren 50 a kowace kofi, tsawon DI 5 cm × 15 cm) kuma an sanya goge a cikin kofunan don haɓaka buɗewa da samar da wani abu mai taunawa, wanda ke rage damuwa ga ƙudan zuma masu duwatsu37. Sanya kofuna biyu na filastik da ke ɗauke da koren a cikin kejin kwari (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) tare da masu ciyarwa 10 ml waɗanda ke ɗauke da maganin sucrose 50% kuma a adana na tsawon kwanaki huɗu don tabbatar da rufewa da haɗuwa. 23°C, ɗanɗanon dangi 60%, photoperiod 10 l (ƙarancin ƙarfi): kwanaki 14. Ana sakin mata da maza 100 da suka haɗu kowace safiya na tsawon kwanaki shida (100 a kowace rana) a cikin gidaje biyu na wucin gadi yayin da apple ke fure (gidan tarko: faɗi 33.66 × tsayi 30.48 × tsayi 46.99 cm; Ƙarin Hoto na 1). An sanya shi a Pennsylvania State Arboretum, kusa da ceri (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'), coronaria apple tree (Malus coronaria) da nau'ikan bishiyoyin apple da yawa (Malus coronaria, Malus), itacen apple na gida 'Co-op 30′ Enterprise™, Malus apple tree 'Co-Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy'). Kowace gidan tsuntsaye mai launin shuɗi ya dace da akwatunan katako guda biyu. Kowace akwatin gida tana ɗauke da bututun takarda kraft marasa komai guda 800 (wanda aka buɗe a karkace, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) da aka saka a cikin bututun cellophane marasa haske (0.7 OD duba matosai na filastik (matosai na T-1X) suna samar da wuraren zama.
Akwatunan gida biyu suna fuskantar gabas kuma an rufe su da shingen lambun kore na filastik (Everbilt model #889250EB12, girman buɗewa 5 × 5 cm, 0.95 m × 100 m) don hana beraye da tsuntsaye shiga kuma an sanya su a saman ƙasa kusa da akwatunan ƙasa na akwatin gida. Akwatin gida (Hoto na Ƙarin 1a). Ana tattara ƙwai na masara kowace rana ta hanyar tattara bututu 30 daga gida sannan a kai su dakin gwaje-gwaje. Ta amfani da almakashi, a yanke a ƙarshen bututun, sannan a wargaza bututun mai karkace don fallasa ƙwayoyin tsuntsaye. An cire ƙwai ɗaya da pollen ɗinsu ta amfani da spatula mai lanƙwasa (Kit ɗin Kayan Aiki na Microslide, BioQuip Products Inc., California). An saka ƙwai a kan takardar tace mai ɗanɗano kuma an sanya su a cikin kwano na Petri na tsawon awanni 2 kafin a yi amfani da su a gwaje-gwajenmu (Hoto na Ƙarin 1b-d).
A dakin gwaje-gwaje, mun kimanta gubar baki na magungunan kashe kwari guda shida da aka yi amfani da su kafin da kuma lokacin furen apple a yawan ruwa guda uku (0.1X, 0.5X, da 1X, inda 1X shine alamar da aka yi amfani da ita a kowace galan 100 na ruwa/eka. Babban adadin filin = yawan ruwa a cikin filin). , Tebur 1). An maimaita kowace yawan ruwa sau 16 (n = 16). Magungunan kashe kwari guda biyu (Tebur S1: mancozeb 2696.14 ppm da captan 2875.88 ppm) da magungunan kashe kwari guda huɗu (Tebur S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75 .12 ppm; cyprodinil 280.845 ppm) guba ga 'ya'yan itatuwa, kayan lambu da amfanin gona na ado. Mun haɗa ƙwayar furen ta amfani da injin niƙa, muka mayar da 0.20 g zuwa rijiya (Falcon Plate mai rijiyoyi 24), sannan muka ƙara kuma muka haɗa 1 μL na maganin kashe ƙwayoyin cuta don samar da ƙwayar fure mai siffar pyramidal tare da rijiyoyin zurfin mm 1 inda aka sanya ƙwai. A sanya ta amfani da ƙaramin spatula (Hoto na Ƙarin 1c,d). An adana faranti na Falcon a zafin ɗaki (25°C) da kuma ɗanɗano 70%. Mun kwatanta su da tsutsotsi masu sarrafawa waɗanda aka ciyar da su da abinci iri ɗaya na furen da aka shafa da ruwa mai tsarki. Mun rubuta mace-mace kuma muka auna nauyin tsutsotsi kowace rana har sai tsutsotsin ya kai shekarun shiryawa ta amfani da ma'aunin nazari (Fisher Scientific, daidaito = 0.0001 g). A ƙarshe, an tantance rabon jinsi ta hanyar buɗe kwakwar bayan watanni 2.5.
An cire DNA daga dukkan tsutsotsin O. cornifrons (n = 3 a kowace yanayin magani, pollen da aka yi wa mancozeb da kuma pollen da ba a yi wa magani ba) kuma mun yi nazarin bambancin ƙwayoyin cuta a kan waɗannan samfuran, musamman saboda a cikin mancozeb an ga mafi girman mace-mace a cikin tsutsotsi. Ana samun MnZn. An ƙara girman DNA, an tsarkake ta amfani da kayan DNA na DNAZymoBIOMICS®-96 MagBead (Zymo Research, Irvine, CA), kuma an yi jerin (zagaye 600) akan kayan Illumina® MiSeq™ ta amfani da kayan v3. An yi jerin kwayoyin halittar RNA ribosomal 16S na ƙwayoyin cuta ta amfani da Kayan Shirya Laburare na Quick-16S™ (Zymo Research, Irvine, CA) ta amfani da kayan share fage da ke kai hari ga yankin V3-V4 na kwayar halittar 16S rRNA. Bugu da ƙari, an yi jerin 18S ta amfani da 10% na haɗa PhiX, kuma an yi ƙarin girma ta amfani da kayan share fage na primer 18S001 da NS4.
Shigo da aiwatar da reads39 ta amfani da bututun QIIME2 (v2022.11.1). An gyara waɗannan karatun kuma an haɗa su, kuma an cire jerin chimeric ta amfani da plugin ɗin DADA2 a cikin QIIME2 (qiime dada2 noise pairing)40. An yi ayyukan aji na 16S da 18S ta amfani da plugin ɗin classifier Classify-sklearn da kayan tarihi silva-138-99-nb-classifier da aka riga aka horar.
An duba dukkan bayanan gwaji don ganin daidaito (Shapiro-Wilks) da kuma daidaiton bambance-bambancen (gwajin Levene). Saboda saitin bayanai bai cika zato na nazarin parametric ba kuma canjin ya kasa daidaita sauran, mun yi ANOVA mai hanyoyi biyu mara parametric (Kruskal-Wallis) tare da abubuwa biyu [lokaci (maki uku na 2, 5, da 8 na lokacin kwana) da kuma maganin kashe ƙwayoyin cuta] don kimanta tasirin magani akan sabon nauyin tsutsa, sannan aka yi kwatancen ba tare da parametric ba bayan hoc ta amfani da gwajin Wilcoxon. Mun yi amfani da samfurin layi na gabaɗaya (GLM) tare da rarraba Poisson don kwatanta tasirin maganin kashe ƙwayoyin cuta akan rayuwa a cikin yawan maganin kashe ƙwayoyin cuta guda uku41,42. Don nazarin yawan bambancin, an ruguza adadin nau'ikan amplicon (ASVs) a matakin nau'in. An yi kwatancen bambancin yawan ƙungiyoyi tsakanin ƙungiyoyi masu amfani da 16S (matakin genus) da yawan 18S ta amfani da samfurin ƙari na gabaɗaya don matsayi, sikelin, da siffa (GAMLSS) tare da rarrabawar iyali ta beta sifili (BEZI), waɗanda aka yi wa samfurin a kan macro. a cikin Microbiome R43 (v1.1). 1). Cire nau'in mitochondrial da chloroplast kafin nazarin bambance-bambance. Saboda matakan rarrabuwa daban-daban na 18S, an yi amfani da mafi ƙarancin matakin kowane taxon kawai don nazarin bambance-bambance. An yi duk nazarin ƙididdiga ta amfani da R (v. 3.4.3., aikin CRAN) (Ƙungiya 2013).
Fuskantar mancozeb, pyrithiostrobin, da trifloxystrobin sun rage yawan nauyin jiki a cikin O. cornifrons (Hoto na 1). An lura da waɗannan tasirin akai-akai a duk allurai uku da aka kimanta (Hoto na 1a–c). Cyclostrobin da myclobutanil ba su rage nauyin tsutsotsi sosai ba.
Matsakaicin nauyin sabbin tsutsotsi masu rarrafe a lokacin da aka auna a maki uku a ƙarƙashin maganin abinci guda huɗu (abincin pollen iri ɗaya + maganin kashe ƙwari: sarrafawa, allurai 0.1X, 0.5X da 1X). (a) Ƙaramin allurai (0.1X): lokacin farko (rana ta 1): χ2: 30.99, DF = 6; P < 0.0001, lokacin karo na biyu (rana ta 5): 22.83, DF = 0.0009; lokaci na uku; lokaci (rana ta 8): χ2: 28.39, DF = 6; (b) rabin allurai (0.5X): lokacin farko (rana ta 1): χ2: 35.67, DF = 6; P < 0.0001, lokacin karo na biyu (rana ta farko). ): χ2: 15.98, DF = 6; P = 0.0090; Lokaci na uku (rana ta 8) χ2: 16.47, DF = 6; (c) Wurin da aka yi amfani da shi ko cikakken magani (1X): lokaci na farko (rana ta 1) χ2: 20.64, P = 6; P = 0.0326, lokaci na biyu (rana ta 5): χ2: 22.83, DF = 6; P = 0.0009; lokaci na uku (rana ta 8): χ2: 28.39, DF = 6; nazarin bambance-bambance marasa daidaituwa. Sanduna suna wakiltar matsakaicin ± SE na kwatancen biyu (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
A mafi ƙarancin allurar (0.1X), nauyin tsutsa ya ragu da kashi 60% tare da trifloxystrobin, kashi 49% tare da mancozeb, kashi 48% tare da myclobutanil, da kashi 46% tare da pyrithistrobin (Hoto na 1a). Lokacin da aka fallasa shi ga rabin allurar da aka yi amfani da ita a filin (0.5X), nauyin tsutsa mancozeb ya ragu da kashi 86%, pyrithiostrobin da kashi 52% da trifloxystrobin da kashi 50% (Hoto na 1b). Cikakken allurar mancozeb (1X) ya rage nauyin tsutsa da kashi 82%, pyrithiostrobin da kashi 70%, da trifloxystrobin, myclobutanil da sangard da kusan kashi 30% (Hoto na 1c).
Mutuwar ta fi yawa a tsakanin furannin da aka yi wa mancozeb magani, sai kuma pyrithiostrobin da trifloxystrobin. Mutuwar ta ƙaru tare da ƙaruwar allurai na mancozeb da pyritisoline (Hoto na 2; Tebur na 2). Duk da haka, mutuwar masu fama da cutar masara ta ƙaru kaɗan ne kawai yayin da yawan trifloxystrobin ya ƙaru; cyprodinil da captan ba su ƙara mace-mace sosai ba idan aka kwatanta da magungunan da aka yi amfani da su wajen magance cutar.
An kwatanta mutuwar tsutsotsin ƙudaje masu ƙura bayan an sha ƙudan zuma daban-daban da aka yi wa magani da maganin kashe ƙwayoyin cuta guda shida. Mancozeb da pentopyramide sun fi saurin kamuwa da tsutsotsin masara (GLM: χ = 29.45, DF = 20, P = 0.0059) (layi, gangare = 0.29, P < 0.001; gangare = 0.24, P <0.00)).
A matsakaici, a duk hanyoyin magani, kashi 39.05% na marasa lafiya mata ne kuma kashi 60.95% maza ne. Daga cikin hanyoyin magani, kashi 40% na mata ne a cikin binciken da aka yi a ƙananan allurai (0.1X) da rabin allurai (0.5X), da kuma kashi 30% a cikin binciken da aka yi a fili (1X). A cikin allurai 0.1X, tsakanin tsutsotsin da aka ciyar da pollen da aka yi wa magani da mancozeb da myclobutanil, kashi 33.33% na manya mata ne, kashi 22% na manya mata ne, kashi 44% na tsutsotsin manya mata ne, kashi 44% na tsutsotsin manya mata ne. Mata, kashi 41% na tsutsotsin manya mata ne, kuma sarrafawa 31% (Hoto na 3a). A kashi 0.5 cikin ɗari na maganin, kashi 33 cikin ɗari na tsutsotsi manya a cikin ƙungiyar mancozeb da pyrithiostrobin mata ne, kashi 36 cikin ɗari a cikin ƙungiyar trifloxystrobin, kashi 41 cikin ɗari a cikin ƙungiyar myclobutanil, da kuma kashi 46 cikin ɗari a cikin ƙungiyar cyprostrobin. Wannan adadi ya kasance kashi 53 cikin ɗari a cikin ƙungiyar captan da kuma kashi 38 cikin ɗari a cikin ƙungiyar sarrafawa (Hoto na 3b). A kashi 1 cikin ɗari na maganin, kashi 30 cikin ɗari na ƙungiyar mancozeb mata ne, kashi 36 cikin ɗari na ƙungiyar pyrithiostrobin, kashi 44 cikin ɗari na ƙungiyar trifloxystrobin, kashi 38 cikin ɗari na ƙungiyar myclobutanil, kashi 50 cikin ɗari na ƙungiyar sarrafawa mata ne - kashi 38.5 cikin ɗari (Hoto na 3c).
Kashi na mata da maza masu kamuwa da cutar bayan kamuwa da cutar fungal. (a) Ƙaramin allurai (0.1X). (b) Rabin allurai (0.5X). (c) Yawan allurai a fili ko cikakken allurai (1X).
Binciken jerin kwayoyin cuta na 16S ya nuna cewa rukunin ƙwayoyin cuta ya bambanta tsakanin tsutsotsi da aka ciyar da pollen da aka yi wa mancozeb da kuma tsutsotsi da aka ciyar da pollen da ba a yi wa magani ba (Hoto na 4a). Ma'aunin ƙwayoyin cuta na tsutsotsi da ba a yi wa magani ba da aka ciyar da pollen ya fi na tsutsotsi da aka ciyar da pollen da aka yi wa mancozeb (Hoto na 4b). Duk da cewa bambancin da aka lura a cikin wadata tsakanin ƙungiyoyi bai kasance mai mahimmanci a kididdiga ba, ya yi ƙasa da wanda aka lura da shi ga tsutsotsi da ke ciyar da pollen da ba a yi wa magani ba (Hoto na 4c). Yawan da aka samu ya nuna cewa ƙwayoyin cuta na tsutsotsi da aka ciyar da pollen da ba a yi wa magani ya fi bambancin da na tsutsotsi da aka ciyar da pollen da ba a yi wa magani ba (Hoto na 5a). Binciken kwatancen ya nuna kasancewar nau'ikan halittu 28 a cikin sarrafawa da samfuran da aka yi wa mancozeb (Hoto na 5b). c Binciken da aka yi amfani da jerin 18S bai nuna wani bambanci mai mahimmanci ba (Hoto na Ƙarin 2).
An kwatanta bayanan SAV bisa ga jerin 16S da wadatar Shannon kuma an lura da wadata a matakin phylum. (a) Babban nazarin daidaitawa (PCoA) bisa ga tsarin al'umma na ƙwayoyin cuta gabaɗaya a cikin tsutsotsi da aka ciyar da pollen ko sarrafawa (shuɗi) da mancozeb (lemu). Kowane ma'aunin bayanai yana wakiltar samfuri daban. An ƙididdige PCoA ta amfani da nisan Bray-Curtis na rarraba t mai yawa. Ovals suna wakiltar matakin amincewa na 80%. (b) Boxplot, bayanan arzikin Shannon marasa amfani (maki) da c. Arzikin da ake iya gani. Akwatunan zane suna nuna akwatuna don layin tsakiya, kewayon interquartile (IQR), da 1.5 × IQR (n = 3).
Haɗuwar al'ummomin ƙwayoyin cuta na tsutsotsi da aka ciyar da su a kan pollen da aka yi wa mancozeb magani da kuma wanda ba a yi masa magani ba. (a) Yawan adadin ƙwayoyin cuta a cikin tsutsotsi. (b) Taswirar zafi na al'ummomin ƙwayoyin cuta da aka gano. Delftia (rabowar rashin daidaito (OR) = 0.67, P = 0.0030) da Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060); Layukan taswirar zafi an haɗa su ta amfani da nisan haɗin kai da matsakaicin haɗin kai.
Sakamakon bincikenmu ya nuna cewa shan taba ta baki ga magungunan kashe kwari (mancozeb) da na rigakafi (pyrostrobin da trifloxystrobin), waɗanda ake amfani da su sosai a lokacin fure, sun rage yawan kiba sosai da kuma ƙaruwar mace-macen tsutsotsin masara. Bugu da ƙari, mancozeb ya rage bambancin da wadatar ƙwayoyin cuta a lokacin shirye-shiryen. Myclobutanil, wani maganin kashe kwari na tsari, ya rage yawan kiba a jiki sosai a dukkan allurai uku. Wannan tasirin ya bayyana a lokacin na biyu (rana ta 5) da na uku (rana ta 8). Sabanin haka, cyprodinil da captan ba su rage yawan kiba ko rayuwa sosai ba idan aka kwatanta da ƙungiyar da ke kula da su. A saninmu, wannan aikin shine na farko da ya tantance tasirin yawan ƙwayoyin cuta daban-daban da ake amfani da su don kare amfanin gonakin masara ta hanyar fallasa pollen kai tsaye.
Duk magungunan kashe ƙwayoyin cuta sun rage yawan nauyin jiki sosai idan aka kwatanta da magungunan da ake amfani da su wajen magance cututtuka. Mancozeb ya fi tasiri kan karuwar nauyin jiki tare da matsakaicin raguwar kashi 51%, sai kuma pyrithiostrobin. Duk da haka, wasu bincike ba su bayar da rahoton mummunan tasirin da alluran maganin kashe ƙwayoyin cuta na gona ke yi a matakan tsutsotsi ba. Duk da cewa an nuna cewa dithiocarbamate biocides yana da ƙarancin guba mai tsanani45, ethylene bisdithiocarbamates (EBDCS) kamar mancozeb na iya raguwa zuwa urea ethylene sulfide. Ganin tasirinsa na maye gurbi a wasu dabbobi, wannan samfurin na iya zama sanadin tasirin da aka lura46,47. Nazarin da aka yi a baya ya nuna cewa samuwar ethylene thiourea yana da tasiri ta hanyar abubuwa kamar yawan zafin jiki48, matakan danshi49 da tsawon ajiyar samfur50. Yanayin ajiya mai kyau don biocides na iya rage waɗannan illolin. Bugu da ƙari, Hukumar Tsaron Abinci ta Turai ta nuna damuwa game da gubar pyrithiopide, wanda aka nuna yana haifar da cutar kansa ga tsarin narkewar abinci na wasu dabbobi51.
Shan mancozeb, pyrithiostrobin, da trifloxystrobin ta baki yana ƙara mace-macen tsutsotsin masara. Sabanin haka, myclobutanil, ciprocycline da captan ba su da wani tasiri a kan mace-mace. Waɗannan sakamakon sun bambanta da na Ladurner et al.52, waɗanda suka nuna cewa captan ya rage rayuwar manya O. lignaria da Apis mellifera L. (Hymenoptera, Apisidae). Bugu da ƙari, an gano cewa magungunan fungi kamar captan da boscalid suna haifar da mutuwar tsutsotsi52,53,54 ko canza halayen ciyarwa55. Waɗannan canje-canjen, bi da bi, na iya shafar ingancin abinci mai gina jiki na pollen da kuma ƙarshe samun kuzarin matakin tsutsotsi. Mutuwar da aka lura a cikin ƙungiyar kulawa ta yi daidai da sauran bincike 56,57.
Ana iya bayyana bambancin jinsi da maza ke fifita a aikinmu ta hanyar abubuwan da suka haɗa da rashin isasshen haɗuwa da mummunan yanayi yayin fure, kamar yadda Vicens da Bosch suka ba da shawara a baya ga O. cornuta. Duk da cewa mata da maza a cikin bincikenmu suna da kwanaki huɗu don haɗuwa (lokacin da aka ɗauka ya isa ga nasarar haɗuwa), mun rage ƙarfin haske da gangan don rage damuwa. Duk da haka, wannan gyare-gyare na iya tsoma baki cikin tsarin haɗuwa ba tare da gangan ba61. Bugu da ƙari, ƙudan zuma suna fuskantar kwanaki da yawa na mummunan yanayi, gami da ruwan sama da ƙarancin zafi (<5°C), wanda kuma zai iya yin mummunan tasiri ga nasarar haɗuwa4,23.
Duk da cewa bincikenmu ya mayar da hankali kan dukkan ƙwayoyin cuta na tsutsotsi, sakamakonmu ya ba da haske game da alaƙar da ke tsakanin al'ummomin ƙwayoyin cuta waɗanda za su iya zama mahimmanci ga abinci mai gina jiki na kudan zuma da kuma fallasa ƙwayoyin cuta. Misali, ƙudan zuma da aka yi wa mancozeb da tsutsotsi ya rage tsarin al'umma da yawan ƙwayoyin cuta idan aka kwatanta da ƙudan zuma da aka ba wa pollen da ba a yi wa magani ba. A cikin tsutsotsi da ke cin pollen da ba a yi wa magani ba, ƙungiyoyin ƙwayoyin cuta na Proteobacteria da Actinobacteria sun fi yawa kuma galibi suna da aerobic ko aerobic. Kwayoyin cuta na Delft, galibi suna da alaƙa da nau'in ƙudan zuma ɗaya, an san suna da aikin maganin rigakafi, wanda ke nuna yuwuwar rawar kariya daga ƙwayoyin cuta. Wani nau'in ƙwayoyin cuta, Pseudomonas, yana da yawa a cikin tsutsotsi da aka ba wa pollen da ba a yi wa magani ba, amma an rage shi sosai a cikin tsutsotsi da aka yi wa magani da mancozeb. Sakamakonmu yana goyan bayan binciken da ya gabata wanda ya gano Pseudomonas a matsayin ɗaya daga cikin mafi yawan nau'ikan ƙwayoyin cuta a cikin O. bicornis35 da sauran kudan zuma ɗaya34. Duk da cewa ba a yi nazarin shaidar gwaji kan rawar da Pseudomonas ke takawa a lafiyar O. cornifrons ba, an nuna cewa wannan ƙwayar cuta tana haɓaka haɗakar gubar kariya a cikin ƙwaro Paederus fuscipes kuma tana haɓaka metabolism na arginine a cikin vitro 35, 65. Waɗannan abubuwan da aka lura sun nuna yuwuwar rawa a cikin kariyar ƙwayoyin cuta da ƙwayoyin cuta a lokacin haɓaka tsutsotsin O. cornifrons. Microbacterium wani nau'in halitta ne da aka gano a cikin bincikenmu wanda aka ruwaito yana cikin adadi mai yawa a cikin tsutsotsin fararen fata a ƙarƙashin yanayin yunwa66. A cikin tsutsotsin O. cornifrons, ƙwayoyin cuta na iya ba da gudummawa ga daidaito da juriya na ƙwayoyin cuta na hanji a ƙarƙashin yanayin damuwa. Bugu da ƙari, ana samun Rhodococcus a cikin tsutsotsin O. cornifrons kuma an san shi da iyawarsa ta kawar da gubobi67. Wannan nau'in kuma ana samunsa a cikin hanjin A. florea, amma yana da ƙarancin yawa68. Sakamakonmu yana nuna kasancewar bambance-bambancen kwayoyin halitta da yawa a cikin nau'ikan ƙwayoyin cuta da yawa waɗanda zasu iya canza hanyoyin rayuwa a cikin tsutsotsi. Duk da haka, ana buƙatar fahimtar bambancin aiki na O. cornifrons.
A taƙaice, sakamakon ya nuna cewa mancozeb, pyrithiostrobin, da trifloxystrobin sun rage yawan nauyin jiki da kuma ƙaruwar mace-macen tsutsotsin masara. Duk da cewa akwai damuwa game da tasirin fungicides akan masu yin pollinators, akwai buƙatar fahimtar tasirin sauran metabolites na waɗannan mahadi. Ana iya haɗa waɗannan sakamakon cikin shawarwari don shirye-shiryen kula da pollinators waɗanda ke taimaka wa manoma su guji amfani da wasu fungicides kafin da kuma lokacin fure bishiyoyin 'ya'yan itace ta hanyar zaɓar fungicides da canza lokacin amfani, ko ta hanyar ƙarfafa amfani da madadin da ba shi da illa 36. Wannan bayanin yana da mahimmanci don haɓaka shawarwari. kan amfani da magungunan kashe kwari, kamar daidaita shirye-shiryen feshi da ake da su da canza lokacin feshi lokacin zaɓar fungicides ko haɓaka amfani da madadin da ba shi da haɗari. Ana buƙatar ƙarin bincike kan mummunan tasirin fungicides akan rabon jinsi, halayen ciyarwa, ƙwayoyin cuta na hanji, da hanyoyin kwayoyin halitta waɗanda ke haifar da asarar nauyi da mace-mace na masara.
An ajiye bayanan tushe na 1, 2 da 3 a cikin Figures 1 da 2 a cikin ma'ajiyar bayanai ta figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 da https://doi.org/10.6084/m9. figshare.24996233. Jerin da aka yi nazari a cikin binciken na yanzu (Hotuna na 4, 5) suna samuwa a cikin ma'ajiyar NCBI SRA a ƙarƙashin lambar shiga PRJNA1023565.
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