Zikomo pochezera Nature.com. Mtundu wa msakatuli womwe mukugwiritsa ntchito uli ndi chithandizo chochepa cha CSS. Kuti mupeze zotsatira zabwino, tikukulimbikitsani kuti mugwiritse ntchito mtundu watsopano wa msakatuli wanu (kapena letsani Compatibility Mode mu Internet Explorer). Pakadali pano, kuti tiwonetsetse kuti chithandizo chikupitilira, tikuwonetsa tsamba lino popanda kukongoletsa kapena JavaScript.
Mankhwala ophera bowa nthawi zambiri amagwiritsidwa ntchito popanga maluwa a zipatso za mitengo ndipo amatha kuopseza tizilombo toyambitsa bowa. Komabe, sizikudziwika bwino momwe tizilombo tomwe sitili njuchi (monga njuchi zodzipatula, Osmia cornifrons) timayankhira tizilombo toyambitsa bowa tomwe timagwiritsidwa ntchito pa maapulo nthawi ya maluwa. Kusiyana kwa chidziwitso kumeneku kumalepheretsa zisankho zoyendetsera zomwe zimatsimikiza kuchuluka kwa bowa komanso nthawi yopopera bowa. Tinayang'ana zotsatira za mankhwala awiri ophera bowa (captan ndi mancozeb) ndi mankhwala anayi ophera bowa (ciprocycline, myclobutanil, pyrostrobin ndi trifloxystrobin). Zotsatira pa kulemera kwa mphutsi, kupulumuka, chiŵerengero cha amuna ndi akazi komanso kusiyanasiyana kwa mabakiteriya. Kuwunikaku kunachitika pogwiritsa ntchito njira yodziwira matenda yomwe mungu unachiritsidwa m'njira zitatu kutengera mlingo womwe ukulimbikitsidwa pano kuti ugwiritsidwe ntchito m'munda (1X), theka la mlingo (0.5X) ndi mlingo wochepa (0.1X). Mlingo wonse wa mancozeb ndi pyritisoline unachepetsa kwambiri kulemera kwa thupi ndi kupulumuka kwa mphutsi. Kenako tinasanthula jini ya 16S kuti tidziwe bacteriome ya mphutsi ya mancozeb, yomwe ndi fungicide yomwe imayambitsa imfa zambiri. Tinapeza kuti kusiyanasiyana kwa mabakiteriya ndi kuchuluka kwawo kunachepa kwambiri mu mphutsi zomwe zimadyedwa ndi mungu wothiridwa ndi mancozeb. Zotsatira zathu za labotale zikusonyeza kuti kupopera ena mwa fungicides awa panthawi yophukira maluwa ndikoopsa kwambiri pa thanzi la O. cornifrons. Izi ndizofunikira pazisankho zamtsogolo zokhudzana ndi kugwiritsa ntchito zinthu zoteteza mitengo ya zipatso mokhazikika ndipo zimagwiritsidwa ntchito ngati maziko a njira zowongolera zomwe cholinga chake ndi kuteteza oyambitsa mungu.
Njuchi ya mason yokha ya Osmia cornifrons (Hymenoptera: Megachilidae) inabweretsedwa ku United States kuchokera ku Japan kumapeto kwa zaka za m'ma 1970 ndi kumayambiriro kwa zaka za m'ma 1980, ndipo mtundu uwu wakhala ndi gawo lofunika kwambiri pakukula kwa mungu m'malo otetezedwa kuyambira nthawi imeneyo. Mitundu ya njuchi iyi ndi mbali ya mitundu pafupifupi 50 ya njuchi zakuthengo zomwe zimathandizira njuchi zomwe zimakula mungu m'minda ya amondi ndi maapulo ku United States2,3. Njuchi za Mason zimakumana ndi mavuto ambiri, kuphatikizapo kugawikana kwa malo okhala, tizilombo toyambitsa matenda, ndi mankhwala ophera tizilombo3,4. Pakati pa mankhwala ophera tizilombo, mankhwala ophera tizilombo amachepetsa mphamvu, kufunafuna chakudya5 ndi kukonza thupi6,7. Ngakhale kafukufuku waposachedwapa akusonyeza kuti thanzi la njuchi za Mason limakhudzidwa mwachindunji ndi tizilombo toyambitsa matenda komanso tizilombo toyambitsa matenda, 8,9 chifukwa mabakiteriya ndi bowa amatha kukhudza zakudya ndi chitetezo cha mthupi, zotsatira za kufalikira kwa fungicide pa mitundu yosiyanasiyana ya tizilombo toyambitsa matenda ya njuchi za Mason zikungoyamba kumene kuphunziridwa.
Mankhwala ophera bowa omwe ali ndi zotsatira zosiyanasiyana (zokhudzana ndi thupi lonse) amathiridwa m'minda ya zipatso asanayambe komanso panthawi ya maluwa kuti achiritse matenda monga nkhanambo ya apulo, kuola kowawa, kuola kofiirira ndi powdery mildew10,11. Mankhwala ophera bowa amaonedwa kuti ndi osavulaza kwa obereketsa mungu, kotero amalimbikitsidwa kwa alimi panthawi ya maluwa; Kuwonekera ndi kumeza kwa mankhwalawa ndi njuchi kumadziwika bwino, chifukwa ndi gawo la njira yolembetsera mankhwala ophera tizilombo ndi US Environmental Protection Agency ndi mabungwe ena ambiri olamulira dziko lonse12,13,14. Komabe, zotsatira za mankhwala ophera bowa pa anthu omwe si njuchi sizidziwika kwenikweni chifukwa sizikufunika malinga ndi mgwirizano wololeza malonda ku United States15. Kuphatikiza apo, nthawi zambiri palibe njira zoyezera njuchi imodzi16,17, ndipo kusunga njuchi zomwe zimayesa njuchi n'kovuta18. Mayeso a njuchi zosiyanasiyana zoyang'aniridwa akuchulukirachulukira ku Europe ndi USA kuti aphunzire za zotsatira za mankhwala ophera tizilombo pa njuchi zakuthengo, ndipo njira zoyezera zapangidwa posachedwapa za O. cornifrons19.
Njuchi zokhala ndi nyanga ndi ma monocyte ndipo zimagwiritsidwa ntchito m'mafakitale a carp ngati chowonjezera kapena cholowa m'malo mwa njuchi za uchi. Njuchi izi zimatuluka pakati pa Marichi ndi Epulo, ndipo zamphongo zokhwima zimatuluka masiku atatu kapena anayi akazi asanakwatirane. Pambuyo poberekana, yaikazi imasonkhanitsa mungu ndi timadzi tokoma kuti ipereke maselo angapo a ana mkati mwa chisa cha chubu (chachilengedwe kapena chopangidwa)1,20. Mazira amaikidwa pa mungu mkati mwa maselo; yaikazi imamanga khoma ladothi isanakonzekere selo lotsatira. Mphutsi yoyamba ya instar imatsekedwa mu chorion ndikudyetsa madzi a m'mimba. Kuyambira instar yachiwiri mpaka yachisanu (prepupa), mphutsi zimadya mungu22. Mungu ukatha kwathunthu, mphutsi zimapanga ma cocoons, pupate ndipo zimatuluka ngati zazikulu m'chipinda chomwecho cha ana, nthawi zambiri kumapeto kwa chilimwe20,23. Akuluakulu amatuluka masika otsatira. Kupulumuka kwa akuluakulu kumalumikizidwa ndi kuwonjezeka kwa mphamvu (kulemera) kutengera kudya. Chifukwa chake, ubwino wa mungu, komanso zinthu zina monga nyengo kapena kukhudzana ndi mankhwala ophera tizilombo, ndizomwe zimapangitsa kuti munthu akhale ndi moyo komanso thanzi labwino24.
Mankhwala ophera tizilombo ndi ma fungicides omwe amagwiritsidwa ntchito asanayambe maluwa amatha kuyenda mkati mwa mitsempha ya zomera mpaka madigiri osiyanasiyana, kuyambira pa translaminar (monga, kutha kuyenda kuchokera pamwamba pa masamba kupita pansi, monga ma fungicides ena) 25 mpaka zotsatira zenizeni za thupi. , zomwe zimatha kulowa m'mutu kuchokera ku mizu, zimatha kulowa mu timadzi ta maluwa a apulo 26, komwe zimatha kupha O. cornifrons akuluakulu 27. Mankhwala ena ophera tizilombo amalowanso mu mungu, zomwe zimakhudza kukula kwa mphutsi za chimanga ndikuzipha 19. Kafukufuku wina wasonyeza kuti ma fungicides ena amatha kusintha kwambiri momwe mitundu yofananira ya O. lignaria imakhalira zisa. Kuphatikiza apo, maphunziro a labotale ndi amunda omwe amatsanzira zochitika zodziwika ndi mankhwala ophera tizilombo (kuphatikizapo ma fungicides) awonetsa kuti mankhwala ophera tizilombo amakhudza kwambiri kapangidwe ka thupi 22 29 ndi kupulumuka kwa njuchi za uchi ndi njuchi zina zokha. Ma spray osiyanasiyana ophera tizilombo omwe amagwiritsidwa ntchito mwachindunji ku maluwa otseguka panthawi ya maluwa amatha kuipitsa mungu womwe umasonkhanitsidwa ndi akuluakulu kuti apange mphutsi, zomwe zotsatira zake sizikuphunziridwa 30.
Zikudziwika bwino kuti kukula kwa mphutsi kumakhudzidwa ndi mungu ndi mabakiteriya m'magawo a m'mimba. Tizilombo ta njuchi timeneti timakhudza zinthu monga kulemera kwa thupi, kusintha kwa kagayidwe kachakudya, komanso kukhudzidwa ndi matenda opatsirana. Kafukufuku wakale adafufuza momwe siteji ya chitukuko, zakudya, ndi malo ozungulira njuchi zodziyimira pawokha zimakhudzira tizilombo toyambitsa matenda. Kafukufukuyu adawonetsa kufanana kwa kapangidwe ndi kuchuluka kwa mphutsi ndi mphutsi zodziyimira pawokha, komanso mtundu wa mabakiteriya ofala kwambiri a Pseudomonas ndi Delftia, pakati pa mitundu ya njuchi zodziyimira pawokha. Komabe, ngakhale kuti mankhwala ophera fungicide akhala akugwirizana ndi njira zotetezera thanzi la njuchi, zotsatira za mankhwala ophera fungicide pa tizilombo toyambitsa matenda kudzera mukamwa mwachindunji sizikufufuzidwa.
Kafukufukuyu adayesa zotsatira za mankhwala asanu ndi limodzi ophera tizilombo toyambitsa matenda omwe amagwiritsidwa ntchito kwambiri omwe adalembetsedwa kuti agwiritsidwe ntchito pa zipatso za mitengo ku United States, kuphatikizapo mankhwala ophera tizilombo toyambitsa matenda omwe amaperekedwa pakamwa ku mphutsi za corn hornworm moth kuchokera ku chakudya chodetsedwa. Tapeza kuti mankhwala ophera tizilombo toyambitsa matenda omwe amakhudzana ndi tizilombo toyambitsa matenda amachepetsa kulemera kwa thupi la njuchi komanso kufa kwambiri, zomwe zimakhala ndi zotsatirapo zoopsa kwambiri zokhudzana ndi mancozeb ndi pyrithiopide. Kenako tinayerekeza kusiyanasiyana kwa tizilombo toyambitsa matenda komwe kumadyedwa pa zakudya za mungu zomwe zimadyedwa ndi mancozeb ndi zomwe zimadyedwa pa zakudya zowongolera. Tikukambirana njira zomwe zingayambitse imfa komanso zotsatira zake pa mapulogalamu ophatikizana a tizilombo toyambitsa matenda ndi mungu (IPPM).
Ma cocoons akuluakulu a O. cornifrons omwe amamera nthawi yozizira m'makokoon adapezeka ku Fruit Research Center, Biglerville, PA, ndipo amasungidwa pa −3 mpaka 2°C (± 0.3°C). Asanayesere (makokoon 600 onse). Mu Meyi 2022, ma cocoons 100 a O. cornifrons ankasamutsidwa tsiku lililonse m'makapu apulasitiki (makokoon 50 pa chikho, DI 5 cm × 15 cm kutalika) ndipo zopukutira zidayikidwa mkati mwa makapu kuti zitseguke bwino ndikupereka substrate yotafuna, kuchepetsa kupsinjika kwa njuchi zamwala37. Ikani makapu awiri apulasitiki okhala ndi makokoon mu khola la tizilombo (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) ndi zodyetsa 10 ml zokhala ndi yankho la 50% la sucrose ndikusunga kwa masiku anayi kuti zitsimikizire kutsekedwa ndi kukwerana. 23°C, chinyezi chaching'ono 60%, nthawi yowonera 10 l (mphamvu yotsika): masiku 14. Akazi ndi amuna 100 okwatiwa ankatulutsidwa m'mawa uliwonse kwa masiku asanu ndi limodzi (100 patsiku) m'zisa ziwiri zopanga panthawi ya maluwa a apulo (chisa cha msampha: m'lifupi 33.66 × kutalika 30.48 × kutalika 46.99 cm; Chithunzi Chowonjezera 1). Anayikidwa ku Pennsylvania State Arboretum, pafupi ndi chitumbuwa (Prunus cerasus 'Eubank' Sweet Cherry Pie™), pichesi (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), peyala (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), mtengo wa apulo wa coronaria (Malus coronaria) ndi mitundu yambiri ya mitengo ya apulo (Malus coronaria, Malus), mtengo wa apulo wapakhomo 'Co-op 30′ Enterprise™, mtengo wa apulo wa Malus 'Co-Op 31′ Winecrisp™, begonia 'Freedom', Begonia 'Golden Delicious', Begonia 'Nova Spy'). Nyumba iliyonse ya mbalame ya pulasitiki yabuluu imayikidwa pamwamba pa mabokosi awiri amatabwa. Bokosi lililonse la chisa linali ndi machubu 800 opanda kanthu a mapepala a kraft (otseguka mozungulira, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) omwe amaikidwa m'machubu a cellophane opaque (0.7 OD onani mapulagi apulasitiki (mapulagi a T-1X) amapereka malo oti azikamera.
Mabokosi onse awiri okhala ndi zisa anayang'ana kum'mawa ndipo anaphimbidwa ndi mpanda wobiriwira wa pulasitiki (Everbilt model #889250EB12, kukula kotseguka 5 × 5 cm, 0.95 m × 100 m) kuti apewe kulowa kwa makoswe ndi mbalame ndipo anaikidwa pamwamba pa nthaka pafupi ndi mabokosi a nthaka a bokosi la zisa. Bokosi la zisa (Chithunzi Chowonjezera 1a). Mazira a borer a chimanga ankasonkhanitsidwa tsiku lililonse posonkhanitsa machubu 30 kuchokera ku zisa ndikuwanyamula kupita nawo ku labotale. Pogwiritsa ntchito lumo, dulani kumapeto kwa chubucho, kenako masulani chubu chozungulira kuti muwone maselo a ana. Mazira amodzi ndi mungu wawo anachotsedwa pogwiritsa ntchito spatula yokhota (Microslide tool kit, BioQuip Products Inc., California). Mazirawo anaikidwa pa pepala lonyowa losefera ndikuyikidwa mu mbale ya Petri kwa maola awiri asanayambe kugwiritsidwa ntchito mu zoyeserera zathu (Chithunzi Chowonjezera 1b-d).
Mu labotale, tinayesa poizoni wa mankhwala asanu ndi limodzi ophera fungicides omwe anagwiritsidwa ntchito asanayambe komanso panthawi ya maluwa a apulo pamlingo wachitatu (0.1X, 0.5X, ndi 1X, pomwe 1X ndiye chizindikiro chomwe chinagwiritsidwa ntchito pa magaloni 100 a madzi/ekala. Mlingo wambiri wamunda = kuchuluka kwa nthaka m'munda). , Gome 1). Kuchuluka kulikonse kunabwerezedwa nthawi 16 (n = 16). Mankhwala awiri ophera fungicides (Gome S1: mancozeb 2696.14 ppm ndi captan 2875.88 ppm) ndi mankhwala anayi ophera fungicides (Gome S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil azole 75 .12 ppm; cyprodinil 280.845 ppm) kuopsa kwa zipatso, ndiwo zamasamba ndi mbewu zokongoletsera. Tinapanga mungu pogwiritsa ntchito chopukusira, tinasamutsa 0.20 g ku chitsime (24-well Falcon Plate), ndipo tinawonjezera ndikusakaniza 1 μL ya yankho la fungicide kuti tipange mungu wa pyramidal ndi zitsime zakuya 1 mm momwe mazira anayikidwa. Ikani pogwiritsa ntchito spatula yaying'ono (Chithunzi Chowonjezera 1c, d). Ma Falcon plates anasungidwa kutentha kwa chipinda (25°C) ndi chinyezi cha 70%. Tinawayerekeza ndi mphutsi zoyang'anira zomwe zimadyetsedwa chakudya cha mungu chofanana chomwe chimathandizidwa ndi madzi oyera. Tinalemba imfa ndipo tinayesa kulemera kwa mphutsi tsiku lililonse mpaka mphutsi zitafika msinkhu wokhwima pogwiritsa ntchito kulinganiza kosanthula (Fisher Scientific, accuracy = 0.0001 g). Pomaliza, chiŵerengero cha amuna ndi akazi chinayesedwa potsegula chikwanje patatha miyezi 2.5.
DNA idatengedwa kuchokera ku mphutsi zonse za O. cornifrons (n = 3 pa vuto lililonse la chithandizo, mungu wochiritsidwa ndi mancozeb komanso wosachiritsidwa) ndipo tidachita kafukufuku wosiyanasiyana wa tizilombo toyambitsa matenda pa zitsanzo izi, makamaka chifukwa mu mancozeb imfa yayikulu idawonedwa mu mphutsi. Kulandira MnZn. DNA idakulitsidwa, kuyeretsedwa pogwiritsa ntchito DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), ndikutsatiridwa (ma cycles 600) pa Illumina® MiSeq™ pogwiritsa ntchito v3 kit. Kutsata kwa majini a ribosomal RNA a bakiteriya a 16S kunachitika pogwiritsa ntchito Quick-16S™ NGS Library Prep Kit (Zymo Research, Irvine, CA) pogwiritsa ntchito ma primer omwe akuyang'ana dera la V3-V4 la jini la 16S rRNA. Kuphatikiza apo, kutsata kwa 18S kunachitika pogwiritsa ntchito 10% PhiX inclusion, ndipo kukulitsa kunachitika pogwiritsa ntchito primer pair 18S001 ndi NS4.
Kulowetsa ndi kukonza ma reads39 pogwiritsa ntchito payipi ya QIIME2 (v2022.11.1). Ma reads awa adadulidwa ndikuphatikizidwa, ndipo ma chimeric sequences adachotsedwa pogwiritsa ntchito DADA2 plugin mu QIIME2 (qiime dada2 noise pairing)40. Ma assignments a kalasi ya 16S ndi 18S adachitika pogwiritsa ntchito plugin ya object classifier Classify-sklearn ndi pre-trained artifact silva-138-99-nb-classifier.
Deta yonse yoyesera idayang'aniridwa ngati ili bwino (Shapiro-Wilks) komanso kufanana kwa kusiyana (mayeso a Levene). Chifukwa detayo sinakwaniritse malingaliro a kusanthula kwa parametric ndipo kusinthaku sikunathe kulinganiza zotsalira, tidachita nonparametric two-way ANOVA (Kruskal-Wallis) yokhala ndi zinthu ziwiri [nthawi (magawo atatu a nthawi ya 2, 5, ndi masiku 8) ndi fungicide] kuti tiwone momwe chithandizochi chimakhudzira kulemera kwa mphutsi zatsopano, kenako kufananiza kwa post hoc nonparametric pairwise kunachitika pogwiritsa ntchito mayeso a Wilcoxon. Tidagwiritsa ntchito generalized linear model (GLM) yokhala ndi Poisson distribution kuti tiyerekeze zotsatira za fungicide pa kupulumuka pa kuchuluka kwa fungicide katatu41,42. Pa kusanthula kosiyanasiyana kwa kuchuluka, chiwerengero cha amplicon sequence variants (ASVs) chidagwa pamlingo wa genus. Kuyerekeza kuchuluka kosiyana pakati pa magulu pogwiritsa ntchito 16S (genus level) ndi 18S relative abundance kunachitika pogwiritsa ntchito generalized additive model for position, scale, and shape (GAMLSS) ndi beta zero-inflated (BEZI) family distributions, zomwe zinapangidwa pa macro . mu Microbiome R43 (v1.1). 1). Chotsani mitundu ya mitochondrial ndi chloroplast musanayese kusanthula kosiyana. Chifukwa cha milingo yosiyanasiyana ya taxonomic ya 18S, mulingo wotsika kwambiri wa taxon iliyonse ndi womwe unagwiritsidwa ntchito pakuwunika kosiyana. Kusanthula konse kwa ziwerengero kunachitika pogwiritsa ntchito R (v. 3.4.3., CRAN project) (Team 2013).
Kukumana ndi mancozeb, pyrithiostrobin, ndi trifloxystrobin kunachepetsa kwambiri kulemera kwa thupi mu O. cornifrons (Chithunzi 1). Zotsatirazi zinawonedwa nthawi zonse pa milingo yonse itatu yomwe inayesedwa (Chithunzi 1a–c). Cyclostrobin ndi myclobutanil sizinachepetse kwambiri kulemera kwa mphutsi.
Kulemera kwapakati kwa mphutsi zatsopano za stem borer kumayesedwa katatu pa nthawi yopatsidwa mankhwala anayi (chakudya chofanana cha mungu + fungicide: kulamulira, 0.1X, 0.5X ndi 1X mlingo). (a) Mlingo wochepa (0.1X): nthawi yoyamba (tsiku 1): χ2: 30.99, DF = 6; P < 0.0001, nthawi yachiwiri (tsiku 5): 22.83, DF = 0.0009; nthawi yachitatu; nthawi (tsiku 8): χ2: 28.39, DF = 6; (b) theka la mlingo (0.5X): nthawi yoyamba (tsiku 1): χ2: 35.67, DF = 6; P < 0.0001, nthawi yachiwiri (tsiku loyamba). ): χ2: 15.98, DF = 6; P = 0.0090; nthawi yachitatu (tsiku 8) χ2: 16.47, DF = 6; (c) Malo kapena mlingo wonse (1X): nthawi yoyamba (tsiku 1) χ2: 20.64, P = 6; P = 0.0326, nthawi yachiwiri (tsiku 5): χ2: 22.83, DF = 6; P = 0.0009; nthawi yachitatu (tsiku 8): χ2: 28.39, DF = 6; kusanthula kosagwirizana ndi magawo a kusiyana. Mipiringidzo ikuyimira avereji ± SE ya kufananiza kwa awiriawiri (α = 0.05) (n = 16) *P ≤ 0.05, **P ≤ 0.001, ***P ≤ 0.0001.
Pa mlingo wotsika kwambiri (0.1X), kulemera kwa thupi la mphutsi kunachepetsedwa ndi 60% ndi trifloxystrobin, 49% ndi mancozeb, 48% ndi myclobutanil, ndi 46% ndi pyrithistrobin (Chithunzi 1a). Atapatsidwa theka la mlingo wa m'munda (0.5X), kulemera kwa thupi la mphutsi za mancozeb kunachepetsedwa ndi 86%, pyrithiostrobin ndi 52% ndi trifloxystrobin ndi 50% (Chithunzi 1b). Mlingo wonse wa m'munda (1X) wa mancozeb unachepetsa kulemera kwa mphutsi ndi 82%, pyrithiostrobin ndi 70%, ndi trifloxystrobin, myclobutanil ndi sangard ndi pafupifupi 30% (Chithunzi 1c).
Chiwerengero cha imfa chinali chachikulu kwambiri pakati pa mphutsi zomwe zimadyetsedwa mungu wothiridwa ndi mancozeb, kutsatiridwa ndi pyrithiostrobin ndi trifloxystrobin. Chiwerengero cha imfa chinawonjezeka ndi kuchuluka kwa mancozeb ndi pyritisoline (Chithunzi 2; Gome 2). Komabe, imfa ya anthu oyambitsa matenda a chimanga inawonjezeka pang'ono pamene kuchuluka kwa trifloxystrobin kunawonjezeka; cyprodinil ndi captan sizinawonjezere kwambiri imfa poyerekeza ndi mankhwala oletsa.
Kufa kwa mphutsi za borer kunayerekezeredwa atadya mungu wothiridwa ndi mankhwala asanu ndi limodzi osiyanasiyana a fungicides. Mancozeb ndi pentopyramide zinali zosavuta kukhudzidwa ndi mphutsi za chimanga pakamwa (GLM: χ = 29.45, DF = 20, P = 0.0059) (mzere, slope = 0.29, P < 0.001; slope = 0.24, P <0.00)).
Pa avareji, pa chithandizo chonse, 39.05% ya odwala anali akazi ndipo 60.95% anali amuna. Pakati pa chithandizo chowongolera, chiwerengero cha akazi chinali 40% m'maphunziro onse a mlingo wochepa (0.1X) ndi theka (0.5X), ndi 30% m'maphunziro a mlingo wa field-dose (1X). Pa mlingo wa 0.1X, pakati pa mphutsi zodyetsedwa mungu zomwe zinapatsidwa mancozeb ndi myclobutanil, 33.33% ya akuluakulu anali akazi, 22% ya akuluakulu anali akazi, 44% ya mphutsi zazikulu zinali akazi, 44% ya mphutsi zazikulu zinali akazi. Akazi, 41% ya mphutsi zazikulu zinali akazi, ndipo owongolera anali 31% (Chithunzi 3a). Pa nthawi ya mlingo wa 0.5, 33% ya nyongolotsi zazikulu mu gulu la mancozeb ndi pyrithiostrobin zinali zachikazi, 36% mu gulu la trifloxystrobin, 41% mu gulu la myclobutanil, ndi 46% mu gulu la cyprostrobin. Chiwerengerochi chinali 53% mu gulu la captan ndi 38% mu gulu lolamulira (Chithunzi 3b). Pa mlingo wa 1X, 30% ya gulu la mancozeb anali akazi, 36% ya gulu la pyrithiostrobin, 44% ya gulu la trifloxystrobin, 38% ya gulu la myclobutanil, 50% ya gulu lolamulira anali akazi - 38.5% (Chithunzi 3c).
Chiwerengero cha ziweto zazikazi ndi zazimuna zobereka pambuyo poti zakhudzidwa ndi fungus. (a) Mlingo wochepa (0.1X). (b) Mlingo wa theka (0.5X). (c) Mlingo wonse kapena wathunthu (1X).
Kusanthula kwa 16S sequence kunasonyeza kuti gulu la mabakiteriya limasiyana pakati pa mphutsi zomwe zimadyetsedwa ndi mungu wothiridwa ndi mancozeb ndi mphutsi zomwe zimadyetsedwa ndi mungu wosathiridwa (Chithunzi 4a). Chiwerengero cha tizilombo toyambitsa matenda chomwe chimadyetsedwa ndi mungu wothiridwa ndi mancozeb chinali chokwera kuposa cha mphutsi zomwe zimadyetsedwa ndi mungu wothiridwa ndi mancozeb (Chithunzi 4b). Ngakhale kusiyana komwe kunawonedwa pakati pa magulu sikunali kofunikira kwambiri, kunali kotsika kwambiri kuposa komwe kunawonedwa kwa mphutsi zomwe zimadya mungu wosathiridwa (Chithunzi 4c). Kuchulukana kunasonyeza kuti microbiota ya mphutsi zomwe zimadyetsedwa ndi mungu wothiridwa ndi mancozeb zinali zosiyana kwambiri kuposa za mphutsi zomwe zimadyetsedwa ndi mphutsi zomwe zimadyetsedwa ndi mancozeb (Chithunzi 5a). Kusanthula kofotokozera kunawonetsa kukhalapo kwa mitundu 28 mu zitsanzo zoyang'anira ndi zoyesedwa ndi mancozeb (Chithunzi 5b). c Kusanthula pogwiritsa ntchito 18S sequencing sikunawonetse kusiyana kwakukulu (Chithunzi Chowonjezera 2).
Ma profiles a SAV ozikidwa pa 16S sequences adayerekezeredwa ndi Shannon richness ndipo adawona richness pa phylum level. (a) Principal coordinate analysis (PCoA) kuzikidwa pa kapangidwe ka tizilombo toyambitsa matenda tomwe sitinalandire mungu kapena control (buluu) ndi mancozeb-fed mphutsi (lalanje). Deta iliyonse imayimira chitsanzo chosiyana. PCoA idawerengedwa pogwiritsa ntchito mtunda wa Bray-Curtis wa multivariate t distribution. Ovals amaimira 80% confidence level. (b) Boxplot, raw Shannon wealth data (points) ndi c. Observable wealth. Boxplots ikuwonetsa mabokosi a median line, interquartile range (IQR), ndi 1.5 × IQR (n = 3).
Kapangidwe ka magulu a tizilombo toyambitsa matenda a mphutsi zomwe zimadyedwa ndi mungu wothiridwa ndi mancozeb komanso wosachiritsidwa. (a) Kuchuluka kwa tizilombo toyambitsa matenda kumawerengedwa mu mphutsi. (b) Mapu a kutentha a magulu a tizilombo toyambitsa matenda omwe adziwika. Delftia (chiŵerengero cha mwayi (OR) = 0.67, P = 0.0030) ndi Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060); Mizere ya mapu a kutentha imayikidwa m'magulu pogwiritsa ntchito mtunda wolumikizana ndi kulumikizana kwapakati.
Zotsatira zathu zikusonyeza kuti kumwa mankhwala ophera tizilombo toyambitsa matenda (mancozeb) ndi mankhwala ophera tizilombo toyambitsa matenda (pyrostrobin ndi trifloxystrobin), omwe amagwiritsidwa ntchito kwambiri nthawi ya maluwa, kunachepetsa kwambiri kulemera kwa tizilombo toyambitsa matenda komanso kufa kwa mphutsi za chimanga. Kuphatikiza apo, mancozeb inachepetsa kwambiri kusiyanasiyana ndi kuchuluka kwa tizilombo toyambitsa matenda panthawi yokonzekera. Myclobutanil, mankhwala ena ophera tizilombo toyambitsa matenda, inachepetsa kwambiri kulemera kwa mphutsi pa mlingo wonse wa milingo itatu. Izi zinaonekera kwambiri pa nthawi yachiwiri (tsiku la 5) ndi lachitatu (tsiku la 8). Mosiyana ndi zimenezi, cyprodinil ndi captan sizinachepetse kwambiri kulemera kapena kupulumuka poyerekeza ndi gulu lolamulira. Monga momwe tikudziwira, ntchito iyi ndi yoyamba kudziwa zotsatira za kuchuluka kwa fungicides m'munda zomwe zimagwiritsidwa ntchito kuteteza mbewu za chimanga kudzera mu mungu mwachindunji.
Mankhwala onse a fungicide achepetsa kwambiri kulemera kwa thupi poyerekeza ndi mankhwala oletsa. Mancozeb inali ndi zotsatirapo zabwino kwambiri pa kulemera kwa thupi la mphutsi ndi kuchepa kwapakati pa 51%, kutsatiridwa ndi pyrithiostrobin. Komabe, maphunziro ena sananene za zotsatirapo zoyipa za milingo ya fungicide m'munda pa magawo a mphutsi44. Ngakhale kuti ma dithiocarbamate biocides awonetsedwa kuti ali ndi poizoni wochepa45, ethylene bisdithiocarbamates (EBDCS) monga mancozeb imatha kuwonongeka kukhala urea ethylene sulfide. Popeza zotsatira zake zosinthika mwa nyama zina, mankhwalawa akhoza kukhala omwe amachititsa zotsatira zomwe zawonedwa46,47. Kafukufuku wakale wasonyeza kuti kupangika kwa ethylene thiourea kumakhudzidwa ndi zinthu monga kutentha kwakukulu48, kuchuluka kwa chinyezi49 ndi kutalika kwa kusungidwa kwa mankhwala50. Malo oyenera osungira ma biocides amatha kuchepetsa zotsatirazi. Kuphatikiza apo, European Food Safety Authority yawonetsa nkhawa za poizoni wa pyrithiopide, yomwe yawonetsedwa kuti imayambitsa khansa m'maselo am'mimba a nyama zina51.
Kumwa mancozeb, pyrithiostrobin, ndi trifloxystrobin kumawonjezera kufa kwa mphutsi za chimanga. Mosiyana ndi zimenezi, myclobutanil, ciprocycline ndi captan sizinakhudze imfa. Zotsatirazi zimasiyana ndi za Ladurner et al.52, omwe adawonetsa kuti captan inachepetsa kwambiri kupulumuka kwa akuluakulu O. lignaria ndi Apis mellifera L. (Hymenoptera, Apisidae). Kuphatikiza apo, mankhwala ophera fungicides monga captan ndi boscalid apezeka kuti amayambitsa kufa kwa mphutsi52,53,54 kapena kusintha khalidwe la kudya55. Kusintha kumeneku, kungakhudze ubwino wa zakudya za mungu ndipo pamapeto pake mphamvu ya mphutsiyo imapezeka. Imfa yomwe idawonedwa m'gulu lolamulira idagwirizana ndi maphunziro ena 56,57.
Chiŵerengero cha amuna ndi akazi chomwe chimakonda amuna chomwe chawonedwa mu ntchito yathu chikhoza kufotokozedwa ndi zinthu monga kusakwanira kukwerana ndi nyengo yoipa panthawi ya maluwa, monga momwe Vicens ndi Bosch adanenera kale za O. cornuta. Ngakhale akazi ndi amuna mu kafukufuku wathu anali ndi masiku anayi kuti akwerena (nthawi yomwe nthawi zambiri imaganiziridwa kuti ndi yokwanira kukwerana bwino), tinachepetsa dala mphamvu ya kuwala kuti tichepetse kupsinjika. Komabe, kusinthaku kungasokoneze njira yokwerana mosadziwa61. Kuphatikiza apo, njuchi zimakumana ndi masiku angapo a nyengo yoipa, kuphatikizapo mvula ndi kutentha kochepa (<5°C), zomwe zingasokonezenso kupambana kwa kukwerana4,23.
Ngakhale kuti kafukufuku wathu unayang'ana kwambiri pa tizilombo toyambitsa matenda tomwe timayambitsa mphutsi, zotsatira zathu zikuwonetsa ubale womwe ungakhalepo pakati pa magulu a mabakiteriya womwe ungakhale wofunikira kwambiri pa zakudya za njuchi komanso kufalikira kwa fungicide. Mwachitsanzo, mungu wodyetsedwa ndi mphutsi wothandizidwa ndi mancozeb unachepetsa kwambiri kapangidwe ka tizilombo toyambitsa matenda komanso kuchuluka kwa tizilombo toyambitsa matenda poyerekeza ndi mungu wodyetsedwa ndi mphutsi wosachiritsidwa. Mu mphutsi zomwe zimadya mungu wosachiritsidwa, magulu a mabakiteriya a Proteobacteria ndi Actinobacteria anali amphamvu ndipo anali othamanga kwambiri kapena othamanga kwambiri. Mabakiteriya a Delft, omwe nthawi zambiri amagwirizanitsidwa ndi mitundu ya njuchi zokha, amadziwika kuti ali ndi mphamvu zopha mabakiteriya, zomwe zikusonyeza kuti angathe kuteteza matenda opatsirana. Mtundu wina wa mabakiteriya, Pseudomonas, unali wochuluka mu mungu wosachiritsidwa, koma unachepa kwambiri mu mphutsi zomwe zimadwala mancozeb. Zotsatira zathu zikuthandizira maphunziro am'mbuyomu omwe adazindikira Pseudomonas ngati imodzi mwa mitundu yochuluka kwambiri mu O. bicornis35 ndi mavu ena okhaokha34. Ngakhale umboni woyesera wa ntchito ya Pseudomonas pa thanzi la O. cornifrons sunafufuzidwe, bacterium iyi yawonetsedwa kuti ikulimbikitsa kupanga poizoni woteteza mu mbewa ya Paederus fuscipes ndikulimbikitsa kagayidwe ka arginine mu vitro 35, 65. Zomwe zapezekazi zikusonyeza kuti ikhoza kukhala ndi gawo pa chitetezo cha mavairasi ndi mabakiteriya panthawi yakukula kwa mphutsi za O. cornifrons. Microbacterium ndi mtundu wina womwe wapezeka mu kafukufuku wathu womwe wanenedwa kuti ulipo kwambiri mu mphutsi zakuda za ntchentche zomwe zimakhala ndi njala66. Mu mphutsi za O. cornifrons, microbacteria ingathandize kuti microbiome ya m'matumbo ikhale yolimba komanso yolimba pamene ili ndi mavuto. Kuphatikiza apo, Rhodococcus imapezeka mu mphutsi za O. cornifrons ndipo imadziwika ndi mphamvu zake zochotsa poizoni67. Mtundu uwu umapezekanso m'matumbo a A. florea, koma uli ndi kuchuluka kochepa kwambiri68. Zotsatira zathu zikuwonetsa kukhalapo kwa mitundu yosiyanasiyana ya majini m'mitundu yosiyanasiyana ya tizilombo toyambitsa matenda yomwe ingasinthe njira zamagetsi mu mphutsi. Komabe, kumvetsetsa bwino kusiyanasiyana kwa ntchito za O. cornifrons ndikofunikira.
Mwachidule, zotsatira zake zikusonyeza kuti mancozeb, pyrithiostrobin, ndi trifloxystrobin zinachepetsa kulemera kwa thupi ndi kufa kwa mphutsi za chimanga. Ngakhale kuti pali nkhawa yowonjezereka yokhudza zotsatira za fungicides pa zonyamula mungu, pali kufunika komvetsetsa bwino zotsatira za metabolites zotsalira za mankhwala awa. Zotsatirazi zitha kuphatikizidwa mu malangizo a mapulogalamu oyendetsera zonyamula mungu omwe amathandiza alimi kupewa kugwiritsa ntchito fungicides zina asanayambe komanso panthawi ya maluwa a mitengo ya zipatso posankha fungicides ndikusinthasintha nthawi yogwiritsira ntchito, kapena polimbikitsa kugwiritsa ntchito njira zina zosavulaza 36. Chidziwitsochi ndi chofunikira popanga malangizo pakugwiritsa ntchito mankhwala ophera tizilombo, monga kusintha mapulogalamu omwe alipo kale opopera ndikusintha nthawi yopopera posankha fungicides kapena kulimbikitsa kugwiritsa ntchito njira zina zosavulaza. Kafukufuku wowonjezereka akufunika pa zotsatira zoyipa za fungicides pa chiŵerengero cha amuna ndi akazi, khalidwe la kudya, tizilombo toyambitsa matenda m'matumbo, ndi njira zama molekyulu zomwe zimayambitsa kuchepa ndi kufa kwa fungicides.
Deta yochokera 1, 2 ndi 3 mu Zithunzi 1 ndi 2 yasungidwa mu malo osungira deta a figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 ndi https://doi.org/10.6084/m9. figshare.24996233. Ma sequences omwe adasanthulidwa mu kafukufuku wapano (Zithunzi 4, 5) akupezeka mu malo osungira a NCBI SRA omwe ali ndi nambala yovomerezeka ya PRJNA1023565.
Bosch, J. ndi Kemp, WP Kupanga ndi kukhazikitsa mitundu ya njuchi za uchi monga zonyamula mungu ku mbewu zaulimi: chitsanzo cha mtundu wa Osmia. (Hymenoptera: Megachilidae) ndi mitengo ya zipatso. ng'ombe. Ntomore. resource. 92, 3–16 (2002).
Parker, MG et al. Machitidwe obereketsa ndi malingaliro a alimi a apulo ku New York ndi Pennsylvania pankhani yobereketsa ndi kubereketsa mitundu ina ya apulo. zosintha. Ulimi. machitidwe azakudya. 35, 1–14 (2020).
Koch I., Lonsdorf EW, Artz DR, Pitts-Singer TL ndi Ricketts TH Zachilengedwe ndi zachuma za kupukutira kwa amondi pogwiritsa ntchito njuchi zakomweko. J. Economics. Ntomore. 111, 16–25 (2018).
Lee, E., He, Y., ndi Park, Y.-L. Zotsatira za kusintha kwa nyengo pa phenology ya tragopan: zotsatira zake pa kasamalidwe ka anthu. Climb. Change 150, 305–317 (2018).
Artz, DR ndi Pitts-Singer, TL Zotsatira za mankhwala ophera tizilombo toyambitsa matenda ndi mankhwala othandizira pa njuchi ziwiri zomwe zimasamalidwa zokha (Osmia lignaria ndi Megachile rotundata). PloS One 10, e0135688 (2015).
Beauvais, S. ndi ena. Fungicide yochokera ku mbewu yopanda poizoni (fenbuconazole) imasokoneza zizindikiro za kubereka kwa amuna zomwe zimapangitsa kuti njuchi zakuthengo zokha zisamapambane. J. Apps. ecology. 59, 1596–1607 (2022).
Sgolastra F. ndi ena. Mankhwala ophera tizilombo a Neonicotinoid ndi ergosterol biosynthesis amaletsa kufa kwa fungicide m'mitundu itatu ya njuchi. Kulamulira tizilombo. sayansi. 73, 1236–1243 (2017).
Kuhneman JG, Gillung J, Van Dyck MT, Fordyce RF. ndi Danforth BN Mphutsi za mavu zokha zimasintha kusiyanasiyana kwa mabakiteriya omwe amaperekedwa ndi mungu kwa njuchi zoberekera tsinde la Osmia cornifrons (Megachilidae). front. microorganism. 13, 1057626 (2023).
Dharampal PS, Danforth BN ndi Steffan SA Tizilombo toyambitsa matenda tomwe timapezeka mu mungu wovunda ndi ofunikira kwambiri pakukula kwa njuchi zokhala zokha monga momwe munguwo umakhalira. ecology. evolution. 12. e8788 (2022).
Kelderer M, Manici LM, Caputo F ndi Thalheimer M. Kubzala pakati pa mizere m'minda ya zipatso za maapulo kuti muchepetse matenda obzalanso mbewu: kafukufuku wogwira ntchito bwino wozikidwa pa zizindikiro za tizilombo toyambitsa matenda. Plant Soil 357, 381–393 (2012).
Martin PL, Kravchik T., Khodadadi F., Achimovich SG ndi Peter KA Kuwola kowawa kwa maapulo pakati pa Atlantic United States: kuwunika mitundu ya tizilombo toyambitsa matenda ndi momwe nyengo ya m'madera osiyanasiyana imakhudzira komanso momwe mbewu zimavutikira. Phytopathology 111, 966–981 (2021).
Cullen MG, Thompson LJ, Carolan JK, Stout JK. ndi Stanley DA Mankhwala ophera bowa, mankhwala ophera udzu ndi njuchi: kuwunikanso mwadongosolo kafukufuku ndi njira zomwe zilipo. PLoS One 14, e0225743 (2019).
Pilling, ED ndi Jepson, PC Zotsatira za EBI fungicides ndi pyrethroid insecticides pa njuchi za uchi (Apis mellifera) zimawononga sayansi. 39, 293–297 (1993).
Mussen, EC, Lopez, JE ndi Peng, CY Zotsatira za mankhwala osankhidwa a fungicides pa kukula ndi chitukuko cha mphutsi za njuchi za uchi Apis mellifera L. (Hymenoptera: Apidae). Lachitatu. Ntomore. 33, 1151-1154 (2004).
Van Dyke, M., Mullen, E., Wickstead, D., ndi McArt, S. Buku Lotsogolera Zosankha Zogwiritsira Ntchito Mankhwala Ophera Tizilombo Poteteza Oyambitsa Mungu M'minda ya Mitengo (Cornell University, 2018).
Iwasaki, JM ndi Hogendoorn, K. Kukumana ndi njuchi ku mankhwala osapha tizilombo: kuwunikanso njira ndi zotsatira zomwe zanenedwa. Ulimi. zachilengedwe. Lachitatu. 314, 107423 (2021).
Kopit AM, Klinger E, Cox-Foster DL, Ramirez RA. ndi Pitts-Singer TL Zotsatira za mtundu wa chakudya ndi kufalikira kwa mankhwala ophera tizilombo pakukula kwa mphutsi za Osmia lignaria (Hymenoptera: Megachilidae). Lachitatu. Ntomore. 51, 240–251 (2022).
Kopit AM ndi Pitts-Singer TL Njira zopewera tizilombo toyambitsa matenda ku njuchi zopanda kanthu. Lachitatu. Ntomore. 47, 499–510 (2018).
Pan, NT et al. Njira yatsopano yoyesera momwe mungamwe mankhwala ophera tizilombo toyambitsa matenda m'njuchi zazikulu zaku Japan (Osmia cornifrons). sayansi. Malipoti 10, 9517 (2020).
Nthawi yotumizira: Meyi-14-2024