Ukuze ngempumelelolawula omiyanekanye nokunciphisa izinga lezifo abazithwalayo, izindlela zamasu, ezisimeme nezingasizungezile imvelo esikhundleni sezibulala-zinambuzane zamakhemikhali.Sihlole ukudla kwembewu okuvela ku-Brassicaceae ethile (umndeni we-Brassica) njengomthombo wama-isothiocyanate atholakala ezitshalweni akhiqizwe yi-enzymatic hydrolysis ye-biologically inactive glucosinolates ukuze isetshenziswe ekulawuleni i-Egypt Aedes (L., 1762).Isidlo sembewu esinezinhlamvu ezinhlanu (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 kanye ne-Thlaspi arvense - izinhlobo ezintathu eziyinhloko zokuchithwa kwe-thermal kanye ne-enzymatic imikhiqizo Ukunquma ubuthi (LC50) be-allyl isothiocyanate, i-benzyl isothiocyanate kanye ne-4-hydroxybenzylisothiocyanate ku-Aedes aegypti izibungu ekuchayekeni kwamahora angu-24 = 0.04 g/120 ml dH2O).Amanani we-LC50 wesinaphi, lwesinaphi esimhlophe nomsila wehhashi.ukudla kwembewu kwakuyi-0.05, 0.08 kanye ne-0.05 ngokulandelanayo uma kuqhathaniswa ne-allyl isothiocyanate (LC50 = 19.35 ppm) kanye ne-4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) yayinobuthi kakhulu kuzibungu nge-20 ml nge-20.Le miphumela ihambisana nokukhiqizwa kwembewu ye-alfalfa.Ukusebenza okuphezulu kwe-benzyl esters kuhambisana namanani abaliwe e-LC50.Ukusebenzisa imbewu yembewu kunganikeza indlela ephumelelayo yokulawula omiyane.ukuphumelela kwembewu ye-cruciferous powder kanye nezingxenye zayo zamakhemikhali eziyinhloko ngokumelene nezibungu zomiyane futhi kubonisa ukuthi izinhlanganisela zemvelo ku-cruciferous seed powder zingasebenza kanjani njenge-larvicide ethembisayo engokwemvelo yokulawula umiyane.
Izifo ezithwalwa yizilwane ezibangelwa omiyane be-Aedes zihlala ziyinkinga enkulu yezempilo yomphakathi emhlabeni wonke.Izehlakalo zezifo ezithwalwa omiyane zisabalala endaweni1,2,3 futhi ziphinde ziqhamuke, okuholela ekuqubukeni kwezifo ezinzima4,5,6,7.Ukusabalala kwezifo phakathi kwabantu nezilwane (isb., i-chikungunya, udenga, i-Rift Valley fever, i-yellow fever kanye negciwane leZika) akukaze kwenzeke.I-Dengue fever iyodwa ibeka cishe abantu abayizigidi eziyizinkulungwane ezingu-3.6 engcupheni yokutheleleka ezindaweni ezishisayo, kanti izifo ezibalelwa ezigidini ezingu-390 zenzeka minyaka yonke, okuholela ekufeni kwabantu abayizi-6,100–24,300 ngonyaka8.Ukuvela kabusha kanye nokuqubuka kwegciwane leZika eNingizimu Melika sekudonse ukunaka emhlabeni wonke ngenxa yokulimala kwengqondo okubangela ezinganeni ezizalwa ngabesifazane abanaleli gciwane2.U-Kremer et al 3 babikezela ukuthi uhla lwezindawo zomiyane i-Aedes luzoqhubeka nokukhula nokuthi ngo-2050, ingxenye yabantu bomhlaba izoba sengozini yokutheleleka ngama-arbovirus athwalwa omiyane.
Ngaphandle kwemithi yokugomela udenga ne-yellow fever esanda kwakhiwa, imithi yokugomela izifo eziningi ezithwalwa omiyane ayikasungulwa9,10,11.Imithi yokugoma isatholakala ngenani elilinganiselwe futhi isetshenziswa kuphela ezivivinyweni zokwelashwa.Ukulawulwa kwezifo zomiyane kusetshenziswa izibulala-zinambuzane zokwenziwa kube yisu elibalulekile lokulawula ukusabalala kwezifo ezithwalwa omiyane12,13.Nakuba izibulala-zinambuzane zokwenziwa zisebenza ngempumelelo ekubulaleni omiyane, ukusetshenziswa okuqhubekayo kwezibulala-zinambuzane zokwenziwa kuthinta kabi izinto eziphilayo ezingahlosiwe futhi kungcolisa imvelo14,15,16.Okwethusayo nakakhulu umkhuba wokwanda kokumelana nomiyane ezinambuzaneni zamakhemikhali17,18,19.Lezi zinkinga ezihlobene nezibulala-zinambuzane ziye zasheshisa ukufuna ezinye izindlela eziphumelelayo nezingasizungezile imvelo zokulawula amagciwane.
Izitshalo ezihlukahlukene ziye zathuthukiswa njengemithombo ye-phytopesticides yokulawula izinambuzane20,21.Izitshalo ngokuvamile azivumelani nemvelo ngoba ziyakwazi ukubola futhi zinobuthi obuncane noma obunganakwa ezilwaneni ezingahlosiwe njengezilwane ezincelisayo, izinhlanzi kanye nezilwane ezihlala emanzini20,22.Amalungiselelo e-Herbal ayaziwa ngokukhiqiza izinhlobonhlobo ze-bioactive compounds nezinqubo ezahlukene zokwenza ukuze kulawulwe ngokuphumelelayo izigaba zokuphila ezihlukahlukene zomiyane23,24,25,26.Izinhlanganisela ezithathwe ezitshalweni ezifana namafutha abalulekile nezinye izithako zezitshalo ezisebenzayo ziye zathola ukunakwa futhi zavula indlela yamathuluzi amasha okulawula amagciwane omiyane.Amafutha abalulekile, ama-monoterpenes kanye nama-sesquiterpenes asebenza njengezinto ezixoshayo, izinqamuleli zokudla kanye nama-ovicides27,28,29,30,31,32,33.Amafutha amaningi emifino abangela ukufa kwezibungu zomiyane, iziphungumangathi kanye nabantu abadala34,35,36, okuthinta izinzwa, ukuphefumula, i-endocrine nezinye izinhlelo ezibalulekile zezinambuzane37.
Ucwaningo lwakamuva lunikeze ukuqonda mayelana nokusetshenziswa okungenzeka kwezitshalo zesinaphi kanye nembewu yazo njengomthombo wezinhlanganisela ze-bioactive.Impuphu yembewu yesinaphi ihlolwe njenge-biofumigant38,39,40,41 futhi yasetshenziswa njengesichibiyelo senhlabathi yokucindezelwa kokhula42,43,44 kanye nokulawula amagciwane ezitshalo atholakala emhlabathini45,46,47,48,49,50, ukondleka kwezitshalo.i-nematodes 41,51, 52, 53, 54 kanye nezinambuzane 55, 56, 57, 58, 59, 60. Umsebenzi wokubulala isikhunta walezi zimpushana zembewu kuthiwa ubangelwa izinhlanganisela ezivikela izitshalo ezibizwa ngokuthi ama-isothiocyanates38,42,60.Ezitshalweni, lezi zinhlanganisela zokuzivikela zigcinwa kumaseli ezitshalo ngendlela yama-non-bioactive glucosinolates.Kodwa-ke, lapho izitshalo zonakaliswa ukudla kwezinambuzane noma ukutheleleka kwe-pathogen, i-glucosinolates i-hydrolyzed nge-myrosinase ibe yi-bioactive isothiocyanates55,61.I-Isothiocyanates yizinhlanganisela eziguquguqukayo ezaziwa ngokuba nomsebenzi obanzi we-antimicrobial kanye ne-insecticidal, futhi isakhiwo sawo, umsebenzi wezinto eziphilayo kanye nokuqukethwe kuyahluka kakhulu phakathi kwezinhlobo ze-Brassicaceae42,59,62,63.
Nakuba ama-isothiocyanate atholakala esidlweni sembewu yesinaphi kwaziwa ukuthi anomsebenzi wokubulala izinambuzane, idatha yomsebenzi webhayoloji emelene nama-arthropod vectors abalulekile ngokwezokwelapha ayantuleka.Ucwaningo lwethu luhlole umsebenzi we-larvicidal wezimpushana ezine zembewu encibilikile ngokumelene nomiyane be-Aedes.Izibungu ze-Aedes aegypti.Inhloso yocwaningo kwakuwukuhlola ukusetshenziswa kwazo okungase kube khona njengama-biopesticides angayingeni imvelo ekulawuleni omiyane.Izingxenye ezintathu zamakhemikhali ezinkulu zesidlo sembewu, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC), ne-4-hydroxybenzylisothiocyanate (4-HBITC) nazo zahlolwa ukuze kuhlolwe umsebenzi webhayoloji walezi zingxenye zamakhemikhali ezibungwini zomiyane.Lona umbiko wokuqala wokuhlola ukusebenza kahle kwezimpushana zembewu yeklabishi ezine kanye nezingxenye zazo zamakhemikhali eziyinhloko ngokumelene nezibungu zomiyane.
Amakholoni aselabhorethri e-Aedes aegypti (uhlobo lwe-Rockefeller) ayenakekelwa ku-26°C, 70% womswakama ohlobene (RH) kanye no-10:14 h (L:D photoperiod).Ezinsikazi eziganile zahlaliswa emakhejini epulasitiki (ubude obuyi-11 cm nobubanzi obungu-9.5 cm) futhi zondliwa ngohlelo lokuphakela ngebhodlela kusetshenziswa igazi lenkomo elicijile (HemoStat Laboratories Inc., Dixon, CA, USA).Ukuphakelwa kwegazi kwenziwa njengokujwayelekile kusetshenziswa i-membrane ye-multi-glass feeder (Chemglass, Life Sciences LLC, Vineland, NJ, USA) exhunywe kushubhu lokugeza lamanzi elizungezayo (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) elinezinga lokushisa ukulawula 37 °C.Yelula ifilimu yeParafilm M ngaphansi kwegumbi lokuphakelayo lengilazi ngalinye (indawo engu-154 mm2).Isiphakeli ngasinye sabe sesibekwa phezu kwegridi engaphezulu emboze ikheji elinensikazi ekhwelayo.Cishe u-350–400 μl wegazi lenkomo lengezwe kufaneli yokuphakela ingilazi kusetshenziswa i-Pasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) kanye nezikelemu ezindala zavunyelwa ukuba zidonse okungenani ihora elilodwa.Abesifazane abakhulelwe babe sebenikezwa isixazululo se-sucrose esingu-10% futhi bavunyelwa ukuba bazalele amaqanda ephepheni lokuhlunga elimanzi elifakwe ezinkombeni ze-soufflé ezicacile (usayizi ongu-1.25 fl oz, i-Dart Container Corp., i-Mason, i-MI, e-USA).ikheji ngamanzi.Faka iphepha lokuhlunga elinamaqanda esikhwameni esivaliwe (SC Johnsons, Racine, WI) bese ugcine ku-26°C.Amaqanda achanyuselwa futhi cishe izibungu ezingu-200–250 zakhuliswa ngamathileyi epulasitiki aqukethe ingxube ye-rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) kanye nempushana yesibindi (MP Biomedicals, LLC, Solon, OH, USA).kanye nefillet (TetraMin, Tetra GMPH, Meer, Germany) ngesilinganiso esingu-2:1:1.Izibungu zasemuva ze-instar yesithathu zakamuva zisetshenziswe kuma-bioassays ethu.
Imbewu yezitshalo esetshenziswe kulolu cwaningo itholwe emithonjeni yezohwebo nekahulumeni elandelayo: i-Brassica juncea (brown mustard-Pacific Gold) kanye ne-Brassica juncea (white mustard-Ida Gold) evela ku-Pacific Northwest Farmers' Cooperative, e-Washington State, USA;(Garden Cress) evela ku-Kelly Seed and Hardware Co., Peoria, IL, USA kanye ne-Thlaspi arvense (Field Pennycress-Elisabeth) evela ku-USDA-ARS, Peoria, IL, USA;Ayikho imbewu esetshenziswe ocwaningweni eye yaphathwa ngezibulala-zinambuzane.Zonke izinto zembewu zacutshungulwa futhi zasetshenziswa kulolu cwaningo ngokuhambisana nemithetho yendawo kanye nekazwelonke futhi ngokuhambisana nayo yonke imithetho yendawo efanele kanye nekazwelonke.Lolu cwaningo aluzange luhlole izinhlobo zezitshalo ze-transgenic.
I-Brassica juncea (PG), i-Alfalfa (Ls), i-White mustard (IG), imbewu ye-Thlaspi arvense (DFP) igaywe yaba impushana kusetshenziswa isigayo se-Retsch ZM200 ultracentrifugal (Retsch, Haan, Germany) esifakelwe inenetha elingu-0.75 mm kanye ne-Stainless i-rotor yensimbi, amazinyo ayi-12, i-10,000 rpm (Ithebula 1).Impushana yembewu egayiwe idluliselwe esiqwini sephepha bese icoliswa nge-hexane kumshini we-Soxhlet amahora angu-24.Isampula encane yensimu yesinaphi encishisiwe iphathwe ukushisa ku-100 °C ihora elingu-1 ukuze i-denature myrosinase futhi ivimbele i-hydrolysis yama-glucosinolate ukuze kwakheke ama-isothiocyanate asebenzayo.Impushana yembewu yehhashi ephethwe ukushisa (DFP-HT) isetshenziswe njengokulawula okungalungile ngokukhipha i-myrosinase.
Okuqukethwe kwe-Glucosinolate kokudla kwembewu encishiwe kwanqunywa ngokuphindwe kathathu kusetshenziswa i-high-performance liquid chromatography (HPLC) ngokuvumelana nephrothokholi eshicilelwe ngaphambilini engu-64.Kafushane, u-3 mL we-methanol wengezwe kusampula engu-250 mg yempushane yembewu encishiwe.Isampula ngayinye yayifakwe emanzini okugeza ngamanzi imizuzu engu-30 futhi ishiywe ebumnyameni ku-23 ° C amahora angu-16.I-aliquot engu-1 mL yesendlalelo sezinto eziphilayo yabe isihlungwa ngesihlungi esingu-0.45 μm sayisampler esizenzakalelayo.Isebenza ngohlelo lwe-Shimadzu HPLC (amaphampu amabili e-LC 20AD; i-SIL 20A autosampler; i-DGU 20As degasser; umtshina we-SPD-20A UV-VIS ozogadwa ku-237 nm; kanye nemojuli yebhasi yokuxhumana ye-CBM-20A), okuqukethwe kwe-glucosinolate kokudla kwembewu kwanqunywa. ngokuphindwe kathathu .usebenzisa inguqulo yesofthiwe ye-Shimadzu LC Solution 1.25 (Shimadzu Corporation, Columbia, MD, USA).Ikholomu bekuyikholomu yesigaba sokuhlehla se-C18 Inertsil (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA).Izimo zesigaba sokuqala seselula zazisethwe ku-12% methanol/88% 0.01 M tetrabutylammonium hydroxide emanzini (TBAH; Sigma-Aldrich, St. Louis, MO, USA) ngesilinganiso sokugeleza esingu-1 mL/min.Ngemuva komjovo we-15 μl wesampula, izimo zokuqala zagcinwa imizuzu engu-20, bese isilinganiso se-solvent salungiswa saba yi-methanol engu-100%, nesikhathi sokuhlaziya isampula esiphelele semizuzu engama-65.Ijika elijwayelekile (elisekelwe ku-nM/mAb) lakhiqizwa ukuhlanjululwa okulandelanayo kwezindinganiso ezisanda kulungiswa ze-sinapine, i-glucosinolate kanye ne-myrosin (Sigma-Aldrich, St. Louis, MO, USA) ukuze kulinganiswe okuqukethwe kwesibabule kokudla kwembewu ekhululiwe.ama-glucosinolates.Ukugxila kwe-Glucosinolate kumasampuli kuhlolwe ku-Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) kusetshenziswa inguqulo ye-OpenLAB CDS ChemStation (C.01.07 SR2 [255]) efakwe ikholomu efanayo futhi kusetshenziswa indlela echazwe ngaphambilini.Ukugxila kwe-Glucosinolate kwanqunywa;iqhathaniswe phakathi kwezinhlelo ze-HPLC.
I-Allyl isothiocyanate (94%, stable) ne-benzyl isothiocyanate (98%) ithengwe kwa-Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA).I-4-Hydroxybenzylisothiocyanate ithengwe kwa-ChemCruz (Santa Cruz Biotechnology, CA, USA).Lapho i-enzymatically i-hydrolyzed nge-myrosinase, i-glucosinolates, i-glucosinolates, ne-glucosinolates yakha i-allyl isothiocyanate, i-benzyl isothiocyanate, ne-4-hydroxybenzylisothiocyanate, ngokulandelana.
Ukuhlolwa kwe-laboratory kwenziwa ngokwendlela kaMuturi et al.32 nezinguqulo.Kusetshenziswe ukudla okuhlanu kwembewu okunamafutha aphansi ocwaningweni: i-DFP, i-DFP-HT, i-IG, i-PG ne-Ls.Izibungu ezingamashumi amabili zafakwa ku-400 mL we-beaker enezindlela ezintathu ezilahlwayo (VWR International, LLC, Radnor, PA, USA) equkethe 120 mL amanzi enziwe nge-deionized (dH2O).Ukugxila kwembewu eyisikhombisa kwahlolelwa ubuthi bezibungu zomiyane: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 kanye no-0.12 g seed meal/120 ml dH2O ye-DFP seed meal , DFP-HT, IG kanye ne-PG.Ukuhlolwa kokuqala kwe-bioassay kubonisa ukuthi ufulawa wembewu ye-Ls osuthiwe unobuthi kakhulu kunamanye amafulawa embewu amane ahloliwe.Ngakho-ke, silungise ukugxiliswa kokwelashwa okuyisikhombisa kwesidlo sembewu ye-Ls kuya ekugxilweni okulandelayo: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, kanye no-0.075 g/120 mL dH2O.
Iqembu lokulawula elingalashwa (dH20, elingenaso isithako sembewu yokudla) lafakwa ukuze kuhlolwe ukufa kwezinambuzane okuvamile ngaphansi kwezimo zokuhlola.Ama-bioassay ezinto ezinobuthi ekudleni ngakunye kwembewu ahlanganisa amabhekha amathathu emithambeka amathathu aphindaphindayo (izibungu ezingama-20 zenkanyezi yesithathu ebholeni ngalinye), ngesamba sezitsha eziyi-108.Iziqukathi ezilashiwe zazigcinwa ekamelweni lokushisa (20-21 ° C) futhi ukufa kwezibungu kwabhalwa phakathi namahora angu-24 kanye nama-72 wokuchayeka okuqhubekayo ekugxilweni kokwelashwa.Uma umzimba womiyane nama-appendages kunganyakazi lapho kubhobozwa noma kuthintwa nge-spatula encane yensimbi engagqwali, izibungu zomiyane zibhekwa njengezifile.Izibungu ezifile zivame ukuhlala zinganyakazi endaweni engemuva noma ye-ventral ngaphansi kwesitsha noma phezu kwamanzi.Ukuhlolwa kwaphindwa kathathu ngezinsuku ezihlukene kusetshenziswa amaqembu ahlukene ezibungu, ingqikithi yezibungu ezingu-180 ezivezwe ekugxilweni ngakunye kokwelashwa.
Ubuthi be-AITC, BITC, kanye ne-4-HBITC ezibungwini zomiyane buhlolwe kusetshenziswa inqubo efanayo ye-bioassay kodwa ngokwelashwa okuhlukile.Lungiselela izixazululo zesitoko ezingu-100,000 ppm zekhemikhali ngayinye ngokwengeza u-100 µL wekhemikhali ku-900 µL we-ethanol ephelele kushubhu ye-centrifuge engu-2-mL futhi unyakazise imizuzwana engu-30 ukuze uxube kahle.Ukugxila kokwelashwa kwanqunywa ngokusekelwe ekuhlolweni kwethu kokuqala kwe-bioassays, okuthole i-BITC inobuthi kakhulu kune-AITC kanye ne-4-HBITC.Ukunquma ubuthi, ukugxiliswa okungu-5 kwe-BITC (1, 3, 6, 9 kanye no-12 ppm), ukugxilisa ingqondo okungu-7 kwe-AITC (5, 10, 15, 20, 25, 30 kanye no-35 ppm) nokugxilisa okungu-6 kwe-4-HBITC (15). , 15, 20, 25, 30 kanye 35 ppm).30, 45, 60, 75 kanye 90 ppm).Ukwelashwa kokulawula kwajovwa ngo-108 μL we-ethanol ephelele, okulingana nevolumu enkulu yokwelashwa ngamakhemikhali.Ama-Bioassay aphindaphindiwe njengenhla, okuveza ingqikithi yezibungu eziyi-180 ekugxilweni kokwelashwa ngakunye.Ukufa kwezibungu kuye kwarekhodwa ekugxiliseni ngakunye kwe-AITC, BITC, ne-4-HBITC ngemva kwamahora angu-24 wokuchayeka okuqhubekayo.
Ukuhlaziywa kwe-probit kwedatha yokufa ehlobene nomthamo engama-65 kwenziwa kusetshenziswa isofthiwe ye-Polo (i-Polo Plus, i-LeOra Software, inguqulo 1.0) ukuze kubalwe ukuhlushwa okubulalayo okungu-50% (LC50), ukuhlushwa okubulalayo okungu-90% (LC90), ukuthambeka, i-coefficient yomthamo obulalayo, kanye nama-95 % ukugxila okubulalayo.ngokusekelwe ezinkathini zokuzethemba zezilinganiso zethamo elibulalayo lokugxilisa okuguqulelwe kwilogi kanye namajika omthamo wokufa.Idatha yokufa isekelwe kudatha eyimpinda ehlanganisiwe yezibungu ezingu-180 ezivezwe ekugxilweni ngakunye kokwelashwa.Ukuhlaziywa okunokwenzeka kwenziwa ngokuhlukene ngokudla kwembewu ngayinye kanye nengxenye ngayinye yamakhemikhali.Ngokusekelwe esikhathini sokuzethemba esingu-95% sesilinganiso sethamo elibulalayo, ubuthi besidlo sembewu kanye nezakhi zamakhemikhali ezibungwini zomiyane kubhekwa njengehluke kakhulu, ngakho isikhathi sokuzethemba esiqukethe inani elingu-1 sasingehluke kakhulu, P = 0.0566.
Imiphumela ye-HPLC yokunqunywa kwama-glucosinolates amakhulu kufulawa wembewu oncibilikisiwe i-DFP, IG, PG kanye no-Ls ibalwe kuThebula 1. Ama-glucosinolate amakhulu kufulawa wembewu ahloliwe ayehluka ngaphandle kwe-DFP ne-PG, womabili ayequkethe i-myrosinase glucosinolates.Okuqukethwe kwe-myrosinin ku-PG kwakuphakeme kune-DFP, 33.3 ± 1.5 kanye no-26.5 ± 0.9 mg/g, ngokulandelanayo.I-Ls imbewu powder iqukethe i-36.6 ± 1.2 mg/g glucoglycone, kuyilapho i-IG imbewu powder iqukethe i-38.0 ± 0.5 mg/g ye-sinapine.
Izibungu ze-Ae.Omiyane be-Aedes aegypti babulawa lapho belashwa ngesidlo sembewu enciphile, nakuba ukusebenza kahle kokwelashwa kwakuhlukahluka kuye ngezinhlobo zezitshalo.I-DFP-NT kuphela eyayingenabo ubuthi ezibungwini zomiyane ngemva kwamahora angama-24 no-72 wokuchayeka (Ithebula 2).Ubuthi be-powder yembewu esebenzayo bukhuphuke ngokugxila okwandayo (Fig. 1A, B).Ubuthi besidlo sembewu kuzibungu zomiyane buhluka kakhulu ngokususelwa ku-95% CI wesilinganiso sethamo elibulalayo lamanani we-LC50 ekuhlolweni kwamahora angama-24 namahora angama-72 (Ithebula 3).Ngemuva kwamahora angama-24, umphumela onobuthi wokudla kwembewu ye-Ls wawumkhulu kunezinye izindlela zokwelapha zokudla kwembewu, nomsebenzi ophakeme kakhulu kanye nobuthi obukhulu ezibungwini (LC50 = 0.04 g/120 ml dH2O).Izibungu zazingazweli kangako ku-DFP ngamahora angu-24 uma kuqhathaniswa ne-IG, Ls kanye ne-PG yokwelashwa kwempushana yembewu, ngamavelu e-LC50 angu-0.115, 0.04 kanye no-0.08 g/120 ml dH2O ngokulandelanayo, ayephezulu ngokwezibalo kunenani le-LC50.0.211 g/120 ml dH2O (Ithebula 3).Amanani e-LC90 e-DFP, IG, PG kanye ne-Ls abengu-0.376, 0.275, 0.137 kanye no-0.074 g/120 ml dH2O, ngokulandelanayo (Ithebula 2).Ukuhlushwa okuphezulu kakhulu kwe-DPP kwaba ngu-0.12 g/120 ml dH2O.Ngemva kwamahora angu-24 okuhlolwa, isilinganiso sokufa kwezibungu sasingu-12% kuphela, kuyilapho isilinganiso sokufa kwezibungu ze-IG kanye ne-PG sifinyelele ku-51% kanye nama-82% ngokulandelana.Ngemva kwamahora angu-24 okuhlola, isilinganiso sokufa kwezibungu sokuhlushwa okuphezulu kakhulu kokwelashwa kwembewu ye-Ls (0.075 g/120 ml dH2O) kwaba ngu-99% (Fig. 1A).
Amajika okufa alinganiselwa kusukela empendulweni yomthamo (Probit) ye-Ae.Izibungu zaseGibhithe (izibungu ze-3rd instar) ekugxilweni kwembewu yembewu amahora angu-24 (A) namahora angu-72 (B) ngemva kokwelashwa.Umugqa wamachashazi umele i-LC50 yokwelashwa kwembewu yembewu.I-DFP Thlaspi arvense, i-DFP-HT Ukushisa kungasebenzi i-Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Ekuhlolweni kwamahora angama-72, amanani e-LC50 e-DFP, IG kanye ne-PG seed meal ayengu-0.111, 0.085 kanye no-0.051 g/120 ml dH2O, ngokulandelana.Cishe zonke izibungu ezichayeke ku-Ls seed meal zafa ngemva kwamahora angu-72 wokuchayeka, ngakho idatha yokufa yayingahambisani nokuhlaziywa kwe-Probit.Uma kuqhathaniswa nokunye ukudla kwembewu, izibungu zazingazweli kangako ekwelashweni kwembewu ye-DFP futhi zinamanani aphezulu e-LC50 ngokwezibalo (Amathebula 2 no-3).Ngemuva kwamahora angama-72, amanani e-LC50 okwelashwa kwembewu ye-DFP, IG kanye ne-PG alinganiselwa ukuthi abe ngu-0.111, 0.085 kanye no-0.05 g/120 ml dH2O, ngokulandelana.Ngemuva kwamahora angama-72 okuhlolwa, amanani e-LC90 we-DFP, IG kanye ne-PG imbewu powders abengu-0.215, 0.254 kanye no-0.138 g/120 ml dH2O, ngokulandelana.Ngemva kwamahora angu-72 okuhlolwa, isilinganiso sokufa kwezibungu zemithi yokwelashwa kwembewu ye-DFP, IG kanye ne-PG ekugxiliseni okukhulu kuka-0.12 g/120 ml dH2O kwaba ngu-58%, 66% kanye nama-96%, ngokulandelana (Fig. 1B).Ngemva kokuhlolwa kwamahora angu-72, ukudla kwembewu ye-PG kwatholakala kunobuthi kakhulu kunokudla kwembewu ye-IG ne-DFP.
Ama-synthetic isothiocyanate, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC) kanye ne-4-hydroxybenzylisothiocyanate (4-HBITC) zingabulala ngempumelelo izibungu zomiyane.Emahoreni angu-24 ngemva kokwelashwa, i-BITC yayinobuthi kakhulu ezibungwini ezinenani le-LC50 lika-5.29 ppm uma kuqhathaniswa no-19.35 ppm we-AITC kanye no-55.41 ppm we-4-HBITC (Ithebula 4).Uma kuqhathaniswa ne-AITC ne-BITC, i-4-HBITC inobuthi obuphansi kanye nenani eliphakeme le-LC50.Kunomehluko omkhulu kubuthi bezibungu zomiyane be-isothiocyanate ezimbili ezinkulu (Ls kanye ne-PG) ekudleni kwembewu okunamandla kakhulu.Ubuthi obususelwe esilinganisweni somthamo obulalayo wamanani e-LC50 phakathi kwe-AITC, BITC, ne-4-HBITC bubonise umehluko wezibalo kangangokuthi u-95% CI wesilinganiso sethamo elibulalayo le-LC50 alizange lifake inani elingu-1 (P = 0.05, Ithebula 4).Ukugxila okuphezulu kakhulu kwakho kokubili i-BITC ne-AITC kulinganiselwa ukuthi kuzobulala u-100% wezibungu ezihloliwe (Umfanekiso 2).
Amajika okufa alinganiselwa kusukela empendulweni yomthamo (Probit) ye-Ae.Amahora angu-24 ngemva kokwelashwa, izibungu zaseGibhithe (izibungu ze-3rd instar) zafinyelela ekugxilweni kwe-synthetic isothiocyanate.Umugqa wamachashazi umele i-LC50 yokwelashwa kwe-isothiocyanate.I-Benzyl isothiocyanate BITC, i-allyl isothiocyanate AITC ne-4-HBITC.
Ukusetshenziswa kwama-biopesticides ezitshalo njengama-ejenti okulawula ama-mosquito vector sekuyisikhathi eside kwacwaningwa.Izitshalo eziningi zikhiqiza amakhemikhali emvelo anomsebenzi wokubulala izinambuzane37.Izinhlanganisela zabo ze-bioactive zinikeza enye indlela ekhangayo yezibulala-zinambuzane zokwenziwa ezinamandla amakhulu okulawula izinambuzane, kuhlanganise nomiyane.
Izitshalo zesinaphi zitshalwa njengesitshalo sembewu yazo, zisetshenziswa njengesinongo kanye nomthombo wamafutha.Lapho amafutha e-mustard ekhishwa embewini noma lapho kukhishwa isinaphi ukuze kusetshenziswe njenge-biofuel, 69 umkhiqizo ophumayo uwukudla kwembewu enamafutha.Lokhu kudla kwembewu kugcina izingxenye zako eziningi zemvelo ze-biochemical kanye nama-enzyme e-hydrolytic.Ubuthi balesi sidlo sembewu bubangelwa ukukhiqizwa kwe-isothiocyanates55,60,61.Ama-Isothiocyanates akhiwa yi-hydrolysis ye-glucosinolates yi-enzyme myrosinase ngesikhathi sokugijiswa kwembewu yembewu38,55,70 futhi aziwa ngokuba ne-fungicidal, bactericidal, nematicidal kanye ne-insecticidal imiphumela, kanye nezinye izakhiwo ezihlanganisa imiphumela yezinzwa zamakhemikhali kanye nezakhiwo ze-chemotherapeutic61,62, 70.Ucwaningo oluningana lubonise ukuthi izitshalo zesinaphi kanye nesidlo sembewu kusebenza ngempumelelo njengemfucuza ngokumelene nenhlabathi kanye nezinambuzane zokudla ezigciniwe57,59,71,72.Kulolu cwaningo, sihlole ubuthi bempuphu yezinhlamvu ezine kanye nemikhiqizo yayo emithathu ye-bioactive AITC, BITC, kanye ne-4-HBITC kuya kwezibungu zomiyane i-Aedes.I-Aedes yaseGibhithe.Ukwengeza ukudla kwembewu ngokuqondile emanzini aqukethe izibungu zomiyane kulindeleke ukuthi kusebenze izinqubo ze-enzymatic ezikhiqiza ama-isothiocyanate anobuthi ezibungwini zomiyane.Le biotransformation yaboniswa ngokwengxenye umsebenzi wokubulala izibungu wesidlo sembewu kanye nokulahlekelwa umsebenzi wokubulala izinambuzane lapho impuphu encane yembewu yesinaphi ilashwa ngaphambi kokusetshenziswa.Ukwelashwa kokushisa kulindeleke ukuthi kucekele phansi ama-enzyme e-hydrolytic enza ama-glucosinolates asebenze, ngaleyo ndlela kuvimbele ukwakheka kwama-isothiocyanates e-bioactive.Lolu wucwaningo lokuqala lokuqinisekisa izici zokubulala izinambuzane zempushana yembewu yeklabishi ngokumelene nomiyane endaweni ehlala emanzini.
Phakathi kwezimpushana zembewu ezihloliwe, i-watercress seed powder (Ls) yayinobuthi kakhulu, okubangela ukufa okuphezulu kwe-Aedes albopictus.Izibungu ze-Aedes aegypti zacutshungulwa ngokuqhubekayo amahora angama-24.Izimpushana ezintathu ezisele zembewu (PG, IG kanye ne-DFP) zazinomsebenzi omncane futhi zisabangela ukufa okuphawulekayo ngemva kwamahora angu-72 wokwelashwa okuqhubekayo.Ukudla kwembewu ye-Ls kuphela okuqukethe amanani abalulekile ama-glucosinolate, kuyilapho i-PG ne-DFP iqukethe i-myrosinase futhi i-IG iqukethe i-glucosinolate njenge-glucosinolate enkulu (Ithebula 1).I-Glucotropaeolin i-hydrolyzed ibe yi-BITC futhi i-sinalbine i-hydrolyzed ibe yi-4-HBITC61,62.Imiphumela yethu ye-bioassay ibonisa ukuthi kokubili ukudla kwembewu ye-Ls kanye ne-synthetic BITC kuyingozi kakhulu ezibungwini zomiyane.Ingxenye eyinhloko ye-PG ne-DFP sembewu yokudla i-myrosinase glucosinolate, eyenziwa nge-hydrolyzed ibe yi-AITC.I-AITC iyasebenza ekubulaleni izibungu zomiyane ngenani le-LC50 lika-19.35 ppm.Uma kuqhathaniswa ne-AITC ne-BITC, i-4-HBITC isothiocyanate iwubuthi obuncane ezibungwini.Nakuba i-AITC inobuthi obuncane kune-BITC, amanani abo e-LC50 aphansi kunamafutha amaningi abalulekile ahlolwe izibungu zomiyane32,73,74,75.
I-powder yethu yembewu ye-cruciferous ukuze isetshenziswe ngokumelene nezibungu zomiyane iqukethe i-glucosinolate eyodwa enkulu, ibalwa ngaphezu kwe-98-99% yenani lama-glucosinolates njengoba kunqunywe yi-HPLC.Kutholwe amanani amanye ama-glucosinolate, kodwa amazinga awo ayengaphansi kuka-0.3% wengqikithi yama-glucosinolate.I-Watercress (L. sativum) imbewu ye-powder iqukethe i-glucosinolates yesibili (i-sinigrin), kodwa ingxenye yabo ingu-1% wesamba se-glucosinolates, futhi okuqukethwe kwabo kusengabalulekile (mayelana ne-0.4 mg / g yembewu yempushana).Nakuba i-PG ne-DFP iqukethe i-glucosinolate eyinhloko efanayo (myrosin), umsebenzi wokuqeda izibungu zokudla kwazo wembewu wehluka kakhulu ngenxa yamanani azo e-LC50.Iyahlukahluka ngobuthi kuya ku-powdery mildew.Ukuvela kwezibungu ze-Aedes aegypti kungase kube ngenxa yomehluko emsebenzini we-myrosinase noma ukuzinza phakathi kokuphakelayo kwembewu okubili.Umsebenzi we-Myrosinase udlala indima ebalulekile ku-bioavailability yemikhiqizo ye-hydrolysis efana ne-isothiocyanates ezitshalweni ze-Brassicaceae76.Imibiko yangaphambilini ka-Pocock et al.77 kanye no-Wilkinson et al.78 ibonise ukuthi izinguquko emsebenzini we-myrosinase nokuzinza kungase futhi kuhlotshaniswe nezici zofuzo nezemvelo.
Okuqukethwe kwe-isothiocyanate ye-bioactive okulindelekile kubalwe ngokusekelwe kumanani e-LC50 okudla kwembewu ngayinye emahoreni angama-24 nangama-72 (Ithebula 5) ukuze kuqhathaniswe nokusetshenziswa kwamakhemikhali okuhambisanayo.Ngemuva kwamahora angama-24, ama-isothiocyanates esidlweni sembewu ayenobuthi kakhulu kunezinhlanganisela ezimsulwa.Amanani e-LC50 abalwe ngokusekelwe ezingxenyeni zesigidi ngasinye (ppm) zokwelashwa kwembewu ye-isothiocyanate ayengaphansi kwamanani e-LC50 kuzinhlelo zokusebenza ze-BITC, AITC, kanye ne-4-HBITC.Sibone izibungu zidla ama-pellets okudla kwembewu (Umfanekiso 3A).Ngakho-ke, izibungu zingase zithole ukuchayeka kakhudlwana kuma-isothiocyanate anobuthi ngokudla amapellets embewu.Lokhu kubonakale kakhulu ekwelashweni kwembewu ye-IG kanye ne-PG ekuchayekeni kwamahora angama-24, lapho ukugxila kwe-LC50 bekuku-75% kanye no-72% okuphansi kunokwelashwa kwe-AITC okumsulwa kanye ne-4-HBITC, ngokulandelana.Ukwelashwa kwe-Ls ne-DFP bekunobuthi kakhulu kune-isothiocyanate emsulwa, enamanani e-LC50 angama-24% nama-41% aphansi, ngokulandelana.Izibungu ekwelapheni kokulawula zikhukhumeze ngempumelelo (Fig. 3B), kuyilapho izibungu eziningi ekwelapheni ukudla kwembewu azizange ziqhume futhi ukuthuthukiswa kwezibungu kwabambezeleka kakhulu (Fig. 3B, D).Ku-Spodopteralitura, ama-isothiocyanate ahlotshaniswa nokuncipha kokukhula kanye nokulibaziseka kokuthuthuka79.
Izibungu ze-Ae.Omiyane be-Aedes aegypti babelokhu bechayeka ku-Brassica imbewu powder amahora angu-24-72.(A) Izibungu ezifile ezinezinhlayiya zembewu ezingxenyeni zomlomo (eziyindilinga);(B) Ukwelashwa kokulawula (i-dH20 ngaphandle kwesidlo sembewu eyengeziwe) kubonisa ukuthi izibungu zikhula ngokujwayelekile futhi ziqala ukuzala ngemva kwamahora angu-72 (C, D) Izibungu eziphathwa ngokudla kwembewu;ukudla kwembewu kubonise umehluko ekuthuthukisweni futhi akuzange kuphuke.
Asizange sifunde indlela yemiphumela enobuthi ye-isothiocyanates ezibungwini zomiyane.Kodwa-ke, izifundo zangaphambilini ezintuthwaneni ezibomvu zomlilo (i-Solenopsis invicta) zibonise ukuthi ukuvinjelwa kwe-glutathione S-transferase (GST) ne-esterase (EST) kuyindlela eyinhloko ye-isothiocyanate bioactivity, futhi i-AITC, ngisho nasemsebenzini ophansi, ingavimbela umsebenzi we-GST. .izintuthwane zomlilo ezingenisiwe ezibomvu ezindaweni eziphansi.Umthamo ngu-0.5 µg/ml80.Ngokuphambene, i-AITC ivimbela i-acetylcholinesterase kuma-weevil ommbila omdala (Sitophilus zeamais)81.Izifundo ezifanayo kufanele zenziwe ukuze kucaciswe indlela yokusebenza kwe-isothiocyanate ezibungwini zomiyane.
Sisebenzisa ukwelashwa kwe-DFP engasebenzi ukushisa ukuze sisekele isiphakamiso sokuthi i-hydrolysis ye-plant glucosinolates ukwakha i-isothiocyanates esebenzayo isebenza njengendlela yokulawula izibungu zomiyane ngesidlo sembewu yesinaphi.Isidlo sembewu ye-DFP-HT sasingenabo ubuthi ngamanani okufaka isicelo ahloliwe.Lafarga et al.82 ibike ukuthi ama-glucosinolate azwela ukuwohloka emazingeni okushisa aphezulu.Ukwelashwa okushisa nakho kulindeleke ukuthi kuguqule i-enzyme ye-myrosinase esidlweni sembewu futhi kuvimbele i-hydrolysis yama-glucosinolates akhe ama-isothiocyanates asebenzayo.Lokhu kuphinde kwaqinisekiswa ngu-Okunade et al.75 ibonise ukuthi i-myrosinase izwela izinga lokushisa, okubonisa ukuthi umsebenzi we-myrosinase wawungasebenzi ngokuphelele lapho lwesinaphi, lwesinaphi esimnyama, kanye nezinhlamvu ze-bloodroot zichayeka emazingeni okushisa angaphezu kuka-80°.C. Lezi zindlela zingase zibangele ukulahlekelwa umsebenzi wokubulala izinambuzane wesidlo sembewu ye-DFP esifakwe ngokushisa.
Ngakho, imbewu yesinaphi kanye nama-isothiocyanate ayo amathathu amakhulu anobuthi ezibungwini zomiyane.Uma kubhekwa lo mehluko phakathi kwesidlo sembewu nokwelashwa ngamakhemikhali, ukusetshenziswa kwembewu yembewu kungase kube indlela ephumelelayo yokulawula umiyane.Kunesidingo sokuhlonza ukwakheka okufanele kanye nezinhlelo zokulethwa ezisebenzayo ukuze kuthuthukiswe ukusebenza kahle kanye nokuzinza kokusetshenziswa kwezimpushana zembewu.Imiphumela yethu ikhombisa ukuthi kungenzeka ukusetshenziswa kwembewu yesinaphi esikhundleni sokusebenzisa izibulala-zinambuzane zokwenziwa.Lobu buchwepheshe bungase bube yithuluzi elisha lokulawula ama-vectors omiyane.Ngenxa yokuthi izibungu zomiyane zikhula kahle ezindaweni ezihlala emanzini futhi ama-glucosinolate embewu yokudla aguqulwa nge-enzymatic abe ama-isothiocyanate asebenzayo lapho kufakwe amanzi, ukusetshenziswa kwembewu yesinaphi emanzini agcwele omiyane kunikeza amandla amakhulu okulawula.Nakuba umsebenzi we-larvicidal we-isothiocyanates uhluka (BITC > AITC > 4-HBITC), ucwaningo olwengeziwe luyadingeka ukuze kutholakale ukuthi ukuhlanganisa ukudla kwembewu nama-glucosinolates amaningi kwandisa ubuthi yini.Lolu ucwaningo lokuqala ukukhombisa imiphumela ebulala izinambuzane yokudla kwembewu ye-cruciferous enciphile kanye nama-isothiocyanate amathathu e-bioactive komiyane.Imiphumela yalolu cwaningo igqamisa isisekelo esisha ngokubonisa ukuthi impuphu yembewu yeklabishi enamafutha, i-byproduct kawoyela ekhishwe ezinhlamvini, ingase isebenze njenge-ejenti ethembisayo yokubulala izibungu ekulawuleni omiyane.Lolu lwazi lungasiza ekuqhubekiseni phambili ukutholakala kwama-ejenti okulawula isitshalo kanye nokuthuthuka kwawo njengezibulala-zinambuzane ezishibhile, ezisebenzisekayo, nezingangeni imvelo.
Amasethi edatha akhiqizelwe lolu cwaningo kanye nokuhlaziya okuwumphumela kuyatholakala kumbhali ohambisanayo ngesicelo esiphusile.Ekupheleni kocwaningo, zonke izinto ezisetshenziswa ocwaningweni (izinambuzane nempuphu yembewu) zacekelwa phansi.
Isikhathi sokuthumela: Jul-29-2024