Ukuze ngempumelelolawula omiyanekanye nokunciphisa ukwanda kwezifo abazithwalayo, kudingeka ezinye izindlela ezihlakaniphile, eziqhubekayo nezinobungani nemvelo esikhundleni sezibulala-zinambuzane zamakhemikhali. Sihlole ukudla kwembewu okuvela ku-Brassicaceae ethile (umndeni we-Brassica) njengomthombo wama-isothiocyanates atholakala ezitshalweni akhiqizwa yi-enzyme hydrolysis yama-glucosinolates angasebenzi ngokwezinto eziphilayo ukuze asetshenziswe ekulawuleni ama-Egyptian Aedes (L., 1762). Ukudla kwembewu okunamafutha amahlanu (iBrassica juncea (L) Czern., 1859, iLepidium sativum L., 1753, iSinapis alba L., 1753, iThlaspi arvense L., 1753 kanye neThlaspi arvense – izinhlobo ezintathu eziyinhloko zokungasebenzi kahle kokushisa kanye nokuwohloka kwe-enzymatic Imikhiqizo yamakhemikhali Ukunquma ubuthi (LC50) be-allyl isothiocyanate, i-benzyl isothiocyanate kanye ne-4-hydroxybenzylisothiocyanate kuma-larvae e-Aedes aegypti ekuvezweni kwamahora angama-24 = 0.04 g/120 ml dH2O). Amanani e-LC50 e-mustard, i-white mustard kanye ne-horsetail. Ukudla kwembewu kwakungu-0.05, 0.08 kanye no-0.05 ngokulandelana uma kuqhathaniswa ne-allyl isothiocyanate (LC50 = 19.35 ppm) kanye no-4. -I-Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) yayinobuthi kakhulu ezibungwini emahoreni angama-24 ngemva kokwelashwa kune-0.1 g/120 ml dH2O ngokulandelana. Le miphumela ihambisana nokukhiqizwa kokudla kwembewu ye-alfalfa. Ukusebenza kahle okuphezulu kwama-benzyl esters kuhambisana namanani e-LC50 abaliwe. Ukusebenzisa ukudla kwembewu kunganikeza indlela ephumelelayo yokulawula omiyane. Ukusebenza kahle kwempuphu yembewu ye-cruciferous kanye nezingxenye zayo eziyinhloko zamakhemikhali ngokumelene nezibungu zomiyane futhi kukhombisa ukuthi amakhemikhali emvelo kumpuphu yembewu ye-cruciferous angasebenza kanjani njengesibulala-zinambuzane esithembisayo esinobungani nemvelo sokulawula omiyane.
Izifo ezithwalwa yizinambuzane ezibangelwa omiyane base-Aedes zisalokhu ziyinkinga enkulu yezempilo yomphakathi emhlabeni jikelele. Ukwanda kwezifo ezithwalwa omiyane kusakazeka ngokwendawo1,2,3 futhi kuphinde kuvele, okuholela ekuqubukeni kwesifo esibi4,5,6,7. Ukusabalala kwezifo phakathi kwabantu nezilwane (isb. i-chikungunya, i-dengue, i-Rift Valley fever, i-yellow fever kanye ne-Zika virus) akukaze kwenzeke. I-dengue fever iyodwa ibeka abantu ababalelwa ezigidini ezingu-3.6 engozini yokutheleleka ezindaweni ezishisayo, kanti kulinganiselwa ukuthi kutheleleke abantu abayizigidi ezingu-390 minyaka yonke, okuholela ekufeni kwabantu abangu-6,100–24,300 ngonyaka8. Ukuvela kabusha kanye nokuqubuka kwegciwane le-Zika eNingizimu Melika kudonsele ukunaka emhlabeni wonke ngenxa yomonakalo wobuchopho obuwubangela ezinganeni ezizalwa ngabesifazane abanegciwane2. UKremer nabanye 3 babikezela ukuthi ububanzi bezindawo zomiyane base-Aedes buzoqhubeka nokukhula nokuthi ngo-2050, ingxenye yabantu emhlabeni izoba sengozini yokutheleleka ngama-arbovirus athwalwa omiyane.
Ngaphandle kwemithi yokugomela esanda kukhiqizwa yokulwa nodenga kanye nomkhuhlane ophuzi, imithi yokugomela izifo eziningi ezithwalwa omiyane ayikakathuthukiswa9,10,11. Imijovo isatholakala ngobuningi obulinganiselwe futhi isetshenziswa kuphela ezivivinyweni zemitholampilo. Ukulawulwa kwemithi yomiyane esebenzisa imithi yokubulala izinambuzane yokwenziwa kube isu elibalulekile lokulawula ukusabalala kwezifo ezithwalwa omiyane12,13. Nakuba imithi yokubulala izinambuzane yokwenziwa isebenza kahle ekubulaleni omiyane, ukusetshenziswa okuqhubekayo kwemithi yokubulala izinambuzane yokwenziwa kuthinta kabi izinto eziphilayo ezingahlosiwe futhi kungcolisa imvelo14,15,16. Okwethusa nakakhulu umkhuba wokwanda kokumelana nomiyane nemithi yokubulala izinambuzane yamakhemikhali17,18,19. Lezi zinkinga ezihlobene nemithi yokubulala izinambuzane ziye zasheshisa ukusesha ezinye izindlela ezisebenzayo nezinobungani nemvelo zokulawula imithi yokubulala izinambuzane.
Izitshalo ezahlukahlukene ziye zathuthukiswa njengemithombo yemithi yokubulala izinambuzane yokulawula izinambuzane20,21. Izinto zezitshalo ngokuvamile zinobungani nemvelo ngoba ziyabola futhi zinobuthi obuphansi noma obuncane ezintweni eziphilayo ezingahlosiwe njengezilwane ezincelisayo, izinhlanzi kanye nezilwane eziphila emanzini nasemanzini20,22. Amalungiselelo ezitshalo ayaziwa ngokukhiqiza izinhlobo ezahlukene zamakhemikhali asebenzayo anezindlela ezahlukene zokusebenza ukulawula ngempumelelo izigaba zokuphila ezahlukene zomiyane23,24,25,26. Amakhemikhali atholakala ezitshalweni afana namafutha abalulekile nezinye izithako zezitshalo ezisebenzayo athole ukunakwa futhi avula indlela yamathuluzi amasha okulawula izinambuzane ezithwala omiyane. Amafutha abalulekile, ama-monoterpenes kanye nama-sesquiterpenes asebenza njengemithi yokuxosha, okuvimbela ukudla kanye nama-ovicides27,28,29,30,31,32,33. Amafutha amaningi emifino abangela ukufa kwezibungu zomiyane, ama-pupae kanye nabantu abadala34,35,36, okuthinta imizwa, ukuphefumula, i-endocrine kanye nezinye izinhlelo ezibalulekile zezinambuzane37.
Izifundo zakamuva zinikeze ukuqonda ngokusetshenziswa okungenzeka kwezitshalo zesinaphi kanye nembewu yazo njengomthombo wamakhemikhali asebenzayo. Ukudla kwembewu yesinaphi kuhlolwe njenge-biofumigant38,39,40,41 futhi kusetshenziswe njengesilungiso senhlabathi sokuqeda ukhula42,43,44 kanye nokulawulwa kwamagciwane ezitshalo athwalwa umhlabathi45,46,47,48,49,50, ukudla okunomsoco kwezitshalo. ama-nematode 41,51, 52, 53, 54 kanye nezinambuzane 55, 56, 57, 58, 59, 60. Umsebenzi wokubulala isikhunta walawa maphepha embewu ubangelwa amakhemikhali avikela izitshalo abizwa ngokuthi ama-isothiocyanates38,42,60. Ezitshalweni, la makhemikhali avikelayo agcinwa kumaseli ezitshalo ngendlela yama-glucosinolates angewona asebenzayo. Kodwa-ke, lapho izitshalo zonakaliswa ukudla kwezinambuzane noma ukutheleleka ngamagciwane, ama-glucosinolates afakwa i-hydrolyzed yi-myrosinase abe yi-isothiocyanates55,61. Ama-Isothiocyanates ayizinhlanganisela eziguquguqukayo ezaziwa ngokuba nomsebenzi wokulwa namagciwane kanye nokubulala izinambuzane obanzi, futhi isakhiwo sazo, umsebenzi wezinto eziphilayo kanye nokuqukethwe kwazo kuyahlukahluka kakhulu phakathi kwezinhlobo ze-Brassicaceae42,59,62,63.
Nakuba ama-isothiocyanate atholakala ekudleni kwembewu yesinaphi aziwa ukuthi anomsebenzi wokubulala izinambuzane, idatha ngomsebenzi wezinto eziphilayo ngokumelene nezidalwa ezithwala izinambuzane ezibalulekile kwezokwelapha ayitholakali. Ucwaningo lwethu luhlole umsebenzi wokubulala izinambuzane wempuphu yembewu ezine ezikhishwe amafutha ngokumelene nomiyane base-Aedes. Izibungu zase-Aedes aegypti. Inhloso yalolu cwaningo kwakuwukuhlola ukusetshenziswa kwazo njengemithi yokubulala izinambuzane enobungani nemvelo yokulawula omiyane. Izingxenye ezintathu ezinkulu zamakhemikhali zokudla kwembewu, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC), kanye ne-4-hydroxybenzyllisothiocyanate (4-HBITC) nazo zahlolwa ukuze kuhlolwe umsebenzi wezinto eziphilayo walezi zingxenye zamakhemikhali ezibunguni zomiyane. Lona umbiko wokuqala wokuhlola ukusebenza kahle kwezimpuphu zembewu yeklabishi ezine kanye nezingxenye zazo zamakhemikhali eziyinhloko ngokumelene nezibungu zomiyane.
Amakholoni elabhorethri e-Aedes aegypti (uhlobo lweRockefeller) agcinwa ku-26°C, umswakama ongu-70% (RH) kanye no-10:14 h (isikhathi sokukhanya kwelanga). Izinsikazi ezizalwe zagcinwa ezindlini zepulasitiki (ukuphakama okungu-11 cm nobubanzi obungu-9.5 cm) futhi zondliwa ngesistimu yokondla ngebhodlela kusetshenziswa igazi lezinkomo elicitshiwe (iHemoStat Laboratories Inc., iDixon, CA, USA). Ukondla ngegazi kwenziwa njengokujwayelekile kusetshenziswa i-membrane multi-glass feeder (iChemglass, Life Sciences LLC, iVineland, NJ, USA) exhunywe kuthubhu lokugeza amanzi elijikelezayo (i-HAAKE S7, iThermo-Scientific, iWaltham, MA, USA) ngokulawula izinga lokushisa okungu-37°C. Yelula ifilimu yeParafilm M phansi kwegumbi ngalinye lokondla ngengilazi (indawo engu-154 mm2). Okondla ngakunye kwabekwa kugridi ephezulu emboza ikheji eliqukethe insikazi ezalwe. Cishe ama-350–400 μl egazi lezinkomo afakwe ku-funnel yokuphakela ingilazi kusetshenziswa i-Pasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) kanti izibungu ezindala zavunyelwa ukuphuma okungenani ihora elilodwa. Abesifazane abakhulelwe babe sebenikwa isisombululo se-sucrose esingu-10% futhi bavunyelwe ukubeka amaqanda ephepheni lokuhlunga elimanzi elifakwe ezinkomishini ze-soufflé ezicacile (usayizi we-1.25 fl oz, Dart Container Corp., Mason, MI, USA). Faka amanzi ekhejini. Beka iphepha lokuhlunga elinamaqanda esikhwameni esivaliwe (SC Johnsons, Racine, WI) bese ugcina ku-26°C. Amaqanda achanyuselwa futhi cishe izibungu ezingu-200–250 zakhuliswa emathileyi epulasitiki aqukethe ingxube ye-rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) kanye nempuphu yesibindi (MP Biomedicals, LLC, Solon, OH, USA). kanye ne-fish fillet (TetraMin, Tetra GMPH, Meer, Germany) ngesilinganiso esingu-2:1:1. Izibungu ze-instar zesikhathi sokugcina sesithathu zisetshenziswe ekuhlolweni kwethu kwe-bioassays.
Izinto zembewu yezitshalo ezisetshenziswe kulolu cwaningo zitholakale emithonjeni elandelayo yezentengiselwano kanye nekahulumeni: i-Brassica juncea (brown mustard-Pacific Gold) kanye ne-Brassica juncea (white mustard-Ida Gold) evela ku-Pacific Northwest Farmers' Cooperative, eWashington State, e-USA; (Garden Cress) evela ku-Kelly Seed and Hardware Co., ePeoria, IL, e-USA kanye ne-Thlaspi arvense (Field Pennycress-Elisabeth) evela ku-USDA-ARS, ePeoria, IL, e-USA; Ayikho imbewu esetshenziswe kulolu cwaningo ephathwe ngezibulala-zinambuzane. Zonke izinto zembewu zacutshungulwa futhi zasetshenziswa kulolu cwaningo ngokuhambisana nemithetho yendawo neyezwe kanye nokuhambisana nayo yonke imithetho efanele yesifundazwe neyezwe. Lolu cwaningo aluzange luhlole izinhlobo zezitshalo eziguqulwe izakhi zofuzo.
Imbewu ye-Brassica juncea (PG), i-Alfalfa (Ls), i-White mustard (IG), i-Thlaspi arvense (DFP) yagaywa yaba yimpuphu ecolekile kusetshenziswa umshini wokugaya we-Retsch ZM200 ultracentrifugal (Retsch, Haan, Germany) ohlonyiswe nge-mesh engu-0.75 mm kanye ne-rotor yensimbi engagqwali, amazinyo angu-12, i-10,000 rpm (Ithebula 1). Impuphu yembewu egayiwe yadluliselwa ku-thimble yephepha futhi yasuswa amafutha nge-hexane kudivayisi ye-Soxhlet amahora angu-24. Isampula encane ye-mustard yasensimini ekhishwe amafutha yaphathwa ngokushisa ku-100 °C ihora eli-1 ukuze isuse i-myrosinase futhi ivimbele i-hydrolysis yama-glucosinolates ukuze kwakheke ama-isothiocyanates asebenzayo ngokwebhayoloji. Impuphu yembewu ye-horsetail ephathwe ngokushisa (DFP-HT) yasetshenziswa njengokulawula okungekuhle ngokususa i-myrosinase.
Okuqukethwe kwe-glucosinolate kokudla kwembewu okunamafutha kunqunywe kathathu kusetshenziswa i-high-performance liquid chromatography (HPLC) ngokusho kwephrothokholi eshicilelwe ngaphambilini 64. Kafushane, i-3 mL ye-methanol yanezelwa kusampula engu-250 mg yempuphu yembewu enamafutha. Isampula ngayinye yafakwa ku-sonic emanzini okugeza imizuzu engama-30 futhi yashiywa ebumnyameni ku-23°C amahora ayi-16. I-aliquot engu-1 mL yesendlalelo sezinto eziphilayo yabe isihlungwa ngesihlungi esingu-0.45 μm ibe yi-autosampler. Isebenza ohlelweni lwe-Shimadzu HPLC (amaphampu amabili e-LC 20AD; i-SIL 20A autosampler; i-DGU 20As degasser; i-SPD-20A UV-VIS detector yokuqapha ku-237 nm; kanye ne-CBM-20A communication bus module), okuqukethwe kwe-glucosinolate kokudla kwembewu kunqunywe kathathu. kusetshenziswa isofthiwe ye-Shimadzu LC Solution version 1.25 (Shimadzu Corporation, Columbia, MD, USA). Ikholomu yayiyikholomu yesigaba sokubuyela emuva se-C18 Inertsil (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA). Izimo zesigaba sokuqala sokuhamba zabekwa ku-12% methanol/88% 0.01 M tetrabutylammonium hydroxide emanzini (TBAH; Sigma-Aldrich, St. Louis, MO, USA) ngesilinganiso sokugeleza esingu-1 mL/min. Ngemva kokujova isampula engu-15 μl, izimo zokuqala zagcinwa imizuzu engu-20, bese isilinganiso se-solvent salungiswa saba yi-100% methanol, ngesikhathi sokuhlaziywa kwesampula esiphelele semizuzu engu-65. Ijika elijwayelekile (elisekelwe ku-nM/mAb) lakhiqizwa ngokuncishiswa okulandelanayo kwamazinga e-sinapine asanda kulungiswa, i-glucosinolate kanye ne-myrosin (Sigma-Aldrich, St. Louis, MO, USA) ukuze kulinganiswe okuqukethwe kwe-sulfur kokudla kwembewu okunamafutha. Ukuhlushwa kwe-Glucosinolate kumasampula 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. Ukuhlushwa kwe-Glucosinolate kunqunywe; kuqhathaniswe phakathi kwezinhlelo ze-HPLC.
I-Allyl isothiocyanate (94%, eqinile) kanye ne-benzyl isothiocyanate (98%) zithengwe kwa-Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). I-4-Hydroxybenzylisothiocyanate ithengwe kwa-ChemCruz (Santa Cruz Biotechnology, CA, USA). Uma i-enzyme hydrolyzed yi-myrosinase, ama-glucosinolates, ama-glucosinolates, kanye nama-glucosinolates akha i-allyl isothiocyanate, i-benzyl isothiocyanate, kanye ne-4-hydroxybenzylisothiocyanate, ngokulandelana.
Ukuhlolwa kwe-bioassay kwelebhu kwenziwa ngokwendlela kaMuturi et al. 32 kanye nokuguqulwa. Kusetshenziswe ukudla kwembewu okuhlanu okunamafutha aphansi ocwaningweni: i-DFP, i-DFP-HT, i-IG, i-PG kanye ne-L. Izibungu ezingamashumi amabili zafakwa ebhekheni elingama-400 mL elilahlwayo elinezindlela ezintathu (i-VWR International, LLC, i-Radnor, i-PA, i-USA) eliqukethe amanzi angenawo i-120 mL (i-dH2O). Ukugxila kokudla kwembewu okuyisikhombisa kwahlolwa ubuthi bezibungu zomiyane: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 kanye no-0.12 g kokudla kwembewu/120 ml dH2O kokudla kwembewu ye-DFP, i-DFP-HT, i-IG kanye ne-PG. Ukuhlolwa kokuqala kwe-bioassays kubonisa ukuthi ufulawa wembewu ye-Ls osuliwe unobuthi kakhulu kunezinye ufulawa wembewu ezine ezihlolwe. Ngakho-ke, silungise amazinga okwelashwa ayisikhombisa okudla kwembewu ye-Ls abe amazinga alandelayo: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, kanye no-0.075 g/120 mL dH2O.
Iqembu lokulawula elingalashwanga (i-dH20, akukho isengezo sokudla kwembewu) lafakwa ukuze kuhlolwe ukufa okuvamile kwezinambuzane ngaphansi kwezimo zokuhlolwa. Ukuhlolwa kwe-biological okunobuthi kokudla ngakunye kwembewu kwakuhlanganisa ama-beakers amathathu aphindaphindwayo amathathu (ama-larvae angu-20 asezingeni lesithathu e-beaker ngayinye), ngama-vial angu-108. Izitsha ezilashiwe zagcinwa ekushiseni kwegumbi (20-21°C) futhi ukufa kwama-larva kwaqoshwa phakathi namahora angu-24 no-72 okuchayeka okuqhubekayo ekugxilweni kokwelashwa. Uma umzimba womiyane kanye nezitho zomzimba zinganyakazi lapho zibhobozwa noma zithintwa nge-spatula encane yensimbi engagqwali, ama-larvae omiyane abhekwa njengafile. Ama-larvae afile avame ukuhlala enganyakazi endaweni yangemuva noma yangaphakathi phansi kwesitsha noma phezu kwamanzi. Ukuhlolwa kwaphindwa kathathu ezinsukwini ezahlukene kusetshenziswa amaqembu ahlukene ama-larvae, ngama-larvae angu-180 avulelwe ekugxilweni ngakunye kokwelashwa.
Ubuthi be-AITC, i-BITC, kanye ne-4-HBITC ezibungwini zomiyane buhlolwe kusetshenziswa inqubo efanayo yokuhlolwa kwe-bioassay kodwa ngezindlela ezahlukene zokwelapha. Lungisa izixazululo zesitoko eziyi-100,000 ppm zekhemikhali ngayinye ngokungeza i-100 µL yekhemikhali ku-900 µL ye-ethanol ephelele epayipini le-centrifuge elingu-2-mL bese unyakazisa imizuzwana engama-30 ukuze kuhlanganiswe kahle. Ubuningi bokwelashwa bunqunywe ngokusekelwe ku-bioassays yethu yokuqala, ethole ukuthi i-BITC inobuthi kakhulu kune-AITC kanye ne-4-HBITC. Ukuze kutholakale ubuthi, amazinga ama-5 e-BITC (1, 3, 6, 9 kanye ne-12 ppm), amazinga ayi-7 e-AITC (5, 10, 15, 20, 25, 30 kanye ne-35 ppm) kanye namazinga ama-6 e-4-HBITC (15, 15, 20, 25, 30 kanye ne-35 ppm). 30, 45, 60, 75 kanye no-90 ppm). Ukwelashwa kokulawula kwajovwa nge-108 μL ye-ethanol ephelele, okulingana nomthamo omkhulu wokwelashwa ngamakhemikhali. Ukuhlolwa kwe-bioassays kwaphindwa njengoba kushiwo ngenhla, kwadalula inani lama-larvae angu-180 ngokuhlushwa ngakunye kokwelashwa. Ukufa kwama-larvae kwaqoshwa ngokuhlushwa ngakunye kwe-AITC, BITC, kanye ne-4-HBITC ngemva kwamahora angu-24 okuvezwa okuqhubekayo.
Ukuhlaziywa kwe-probit kwedatha yokufa ehlobene nomthamo engu-65 kwenziwa kusetshenziswa isofthiwe yePolo (Polo Plus, LeOra Software, inguqulo 1.0) ukuze kubalwe ukuhlushwa okubulalayo okungu-50% (LC50), ukuhlushwa okubulalayo okungu-90% (LC90), ukuthambeka, i-letal dose coefficient, kanye nokuhlushwa okubulalayo okungu-95%. ngokusekelwe ezikhawulweni zokuzethemba zezilinganiso zomthamo obulalayo zokuhlushwa okuguquliwe kwelogi kanye nama-dose-death curves. Idatha yokufa isekelwe kudatha ephindaphindiwe ehlanganisiwe yezibungu ezingu-180 ezivezwe ekuhlushweni ngakunye kokwelashwa. Ukuhlaziywa okungenzeka kwenziwe ngokwehlukana kokudla ngakunye kwembewu kanye nengxenye ngayinye yamakhemikhali. Ngokusekelwe esikhawulweni sokuzethemba esingu-95% sesilinganiso somthamo obulalayo, ubuthi bokudla kwembewu kanye nezinto zamakhemikhali ezibunguni zomiyane kwakubhekwa njengokuhlukile kakhulu, ngakho-ke isikhawu sokuzethemba esiqukethe inani elingu-1 sasingahlukile kakhulu, P = 0.0566.
Imiphumela ye-HPLC yokunqunywa kwama-glucosinolates amakhulu kufulawa wembewu osunamafutha i-DFP, IG, PG kanye ne-Ls ibhalwe kuThebula 1. Ama-glucosinolates amakhulu kufulawa wembewu ovivinyiwe ahluka ngaphandle kwe-DFP kanye ne-PG, zombili ezaziqukethe ama-glucosinolates e-myrosinase. Okuqukethwe kwe-myrosinin ku-PG kwakuphakeme kune-DFP, okungu-33.3 ± 1.5 kanye no-26.5 ± 0.9 mg/g, ngokulandelana. Impuphu yembewu ye-Ls yayiqukethe i-gluglycone engu-36.6 ± 1.2 mg/g, kanti impuphu yembewu ye-IG yayiqukethe i-sinapine engu-38.0 ± 0.5 mg/g.
Izibungu zomiyane i-Ae. Aedes aegypti zabulawa lapho zelashwa ngesidlo sembewu esinamafutha, yize ukusebenza kahle kokwelashwa kwahlukahluka kuye ngohlobo lwesitshalo. I-DFP-NT kuphela eyayingeyona ubuthi ezibungwini zomiyane ngemva kwamahora angu-24 nangu-72 okuchayeka (Ithebula 2). Ubuthi bempuphu yembewu esebenzayo banda ngokuhlushwa okwandayo (Isithombe 1A, B). Ubuthi besidlo sembewu ezibungwini zomiyane bahlukahluka kakhulu ngokusekelwe ku-95% CI yesilinganiso somthamo obulalayo samanani e-LC50 ekuhlolweni kwamahora angu-24 namahora angu-72 (Ithebula 3). Ngemuva kwamahora angu-24, umphumela onobuthi wesidlo sembewu i-Ls wawumkhulu kunezinye izindlela zokwelapha zesidlo sembewu, ngomsebenzi ophezulu kakhulu kanye nobuthi obukhulu ezibungwini (LC50 = 0.04 g/120 ml dH2O). Ama-larvae ayengazweli kakhulu ku-DFP emahoreni angama-24 uma kuqhathaniswa nokwelashwa ngempuphu yembewu ye-IG, Ls kanye ne-PG, ngamanani e-LC50 angu-0.115, 0.04 kanye no-0.08 g/120 ml dH2O ngokulandelana, ayephakeme ngokwezibalo kunenani le-LC50. 0.211 g/120 ml dH2O (Ithebula 3). Amanani e-LC90 e-DFP, IG, PG kanye ne-Ls ayengu-0.376, 0.275, 0.137 kanye no-0.074 g/120 ml dH2O, ngokulandelana (Ithebula 2). Ukuhlushwa okuphezulu kakhulu kwe-DPP kwakungu-0.12 g/120 ml dH2O. Ngemuva kwamahora angama-24 okuhlolwa, isilinganiso sokufa kwama-larvae sasingu-12% kuphela, kanti isilinganiso sokufa kwama-larvae e-IG kanye ne-PG sifinyelele ku-51% kanye no-82%, ngokulandelana. Ngemva kwamahora angu-24 okuhlolwa, isilinganiso sokufa kwama-larval ngenxa yokuhlushwa okuphezulu kakhulu kokwelashwa kokudla kwembewu ye-Ls (0.075 g/120 ml dH2O) kwaba ngu-99% (Isithombe 1A).
Ama-curve okufa alinganiswe kusukela empendulweni yomthamo (Probit) we-Ae. Egyptian larvae (izibungu ze-instar yesithathu) kuya ekugxilweni kokudla kwembewu emahoreni angama-24 (A) kanye namahora angama-72 (B) ngemuva kokwelashwa. Umugqa onamachashazi umele i-LC50 yokwelashwa kokudla kwembewu. I-DFP Thlaspi arvense, i-DFP-HT Heat inactivated Thlaspi arvense, i-IG Sinapsis alba (Ida Gold), i-PG Brassica juncea (Pacific Gold), i-Ls Lepidium sativum.
Ekuhlolweni kwamahora angu-72, amanani e-LC50 okudla kwembewu ye-DFP, IG kanye ne-PG ayengu-0.111, 0.085 kanye no-0.051 g/120 ml dH2O, ngokulandelana. Cishe zonke izibungu ezivezwe ekudleni kwembewu ye-L zafa ngemva kwamahora angu-72 okuvezwe, ngakho idatha yokufa yayingahambisani nokuhlaziywa kwe-Probit. Uma kuqhathaniswa nokunye ukudla kwembewu, izibungu zazingezwani kakhulu nokwelashwa kokudla kwembewu ye-DFP futhi zazinamanani aphezulu e-LC50 ngokwezibalo (Amathebula 2 no-3). Ngemuva kwamahora angu-72, amanani e-LC50 okwelashwa kokudla kwembewu ye-DFP, IG kanye ne-PG alinganiselwa ku-0.111, 0.085 kanye no-0.05 g/120 ml dH2O, ngokulandelana. Ngemuva kwamahora angu-72 okuhlolwa, amanani e-LC90 ompushana wembewu ye-DFP, IG kanye ne-PG ayengu-0.215, 0.254 kanye no-0.138 g/120 ml dH2O, ngokulandelana. Ngemva kwamahora angu-72 okuhlolwa, isilinganiso sokufa kwezibungu kokwelashwa kokudla kwembewu ye-DFP, IG kanye ne-PG ekuhlungeni okuphezulu okungu-0.12 g/120 ml dH2O kwaba ngu-58%, 66% kanye no-96%, ngokulandelana (Isithombe 1B). Ngemva kokuhlolwa kwamahora angu-72, ukudla kwembewu ye-PG kwatholakala ukuthi kunobuthi kakhulu kunokudla kwembewu ye-IG kanye ne-DFP.
Ama-isothiocyanate okwenziwa, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC) kanye ne-4-hydroxybenzylisothiocyanate (4-HBITC) angabulala ngempumelelo izibungu zomiyane. Ngemva kwamahora angama-24 ukwelashwa, i-BITC yayinobuthi kakhulu ezibunguni ngenani le-LC50 elingu-5.29 ppm uma kuqhathaniswa ne-19.35 ppm ye-AITC kanye ne-55.41 ppm ye-4-HBITC (Ithebula 4). Uma kuqhathaniswa ne-AITC kanye ne-BITC, i-4-HBITC inobuthi obuphansi kanye nenani le-LC50 eliphakeme. Kunomehluko omkhulu ebuthi bezibungu zomiyane ze-isothiocyanate ezimbili ezinkulu (Ls kanye ne-PG) ekudleni kwembewu okunamandla kakhulu. Ubuthi obusekelwe esilinganisweni somthamo obulalayo wamanani e-LC50 phakathi kwe-AITC, i-BITC, kanye ne-4-HBITC kubonise umehluko wezibalo kangangokuthi i-95% CI yesilinganiso somthamo obulalayo we-LC50 ayizange ifake inani elingu-1 (P = 0.05, Ithebula 4). Amazinga aphezulu kakhulu e-BITC kanye ne-AITC alinganiselwa ukuthi abulale u-100% wezibungu ezihlolwe (Isithombe 2).
Ama-curve okufa alinganiswe kusukela empendulweni yomthamo (Probit) we-Ae. Ngemva kwamahora angu-24 ekwelashweni, izibungu zaseGibhithe (izibungu ze-instar yesithathu) zifinyelele amazinga e-isothiocyanate okwenziwa. Umugqa onamachashazi umele i-LC50 yokwelashwa kwe-isothiocyanate. I-Benzyl isothiocyanate BITC, i-allyl isothiocyanate AITC kanye ne-4-HBITC.
Ukusetshenziswa kwemithi yokubulala izinambuzane yezitshalo njengezinto zokulawula izinambuzane ezithwala omiyane sekuyisikhathi eside kucwaningwa. Izitshalo eziningi zikhiqiza amakhemikhali emvelo anomsebenzi wokubulala izinambuzane37. Amakhemikhali azo asebenzayo ahlinzeka ngenye indlela ekhangayo kunemithi yokubulala izinambuzane yokwenziwa enamandla amakhulu okulawula izinambuzane, okuhlanganisa omiyane.
Izitshalo zesinaphi zikhuliswa njengesitshalo sembewu yazo, ezisetshenziswa njengesinongo kanye nomthombo wamafutha. Lapho amafutha esinaphi ekhishwa embewini noma lapho kukhishwa isinaphi ukuze sisetshenziswe njengophethiloli we-biofuel, umkhiqizo ophumayo uwukudla kwembewu okukhishwe amafutha. Lokhu kudla kwembewu kugcina izingxenye zayo eziningi zemvelo ze-biochemical kanye nama-enzyme e-hydrolytic. Ubuthi balokhu kudla kwembewu bubangelwa ukukhiqizwa kwe-isothiocyanates55,60,61. Ama-Isothiocyanates akhiwa yi-hydrolysis yama-glucosinolates yi-enzyme i-myrosinase ngesikhathi sokumanzisa ukudla kwembewu38,55,70 futhi ayaziwa ukuthi anemiphumela yokubulala isikhunta, ukubulala amagciwane, ukubulala izinambuzane kanye nokubulala izinambuzane, kanye nezinye izakhiwo ezifaka imiphumela yokuzwa kwamakhemikhali kanye nezakhiwo zokwelapha ngamakhemikhali61,62,70. Izifundo eziningana zibonise ukuthi izitshalo zesinaphi kanye nokudla kwembewu kusebenza kahle njengezifutho ngokumelene nezinambuzane ezidla inhlabathi kanye nezilwane ezigciniwe57,59,71,72. Kulolu cwaningo, sihlole ubuthi bokudla okunembewu ezine kanye nemikhiqizo yako emithathu esebenzayo ye-AITC, BITC, kanye ne-4-HBITC kuma-Aedes mosquito larvae. I-Aedes aegypti. Ukwengeza ukudla okunembewu ngqo emanzini aqukethe ama-mosquito larvae kulindeleke ukuthi kusebenze izinqubo ze-enzyme ezikhiqiza ama-isothiocyanate anobuthi kuma-mosquito larvae. Lokhu kuguqulwa kwe-biotransformation kuboniswe ngokwengxenye ngomsebenzi wokubulala ama-larvae obonwe wokudla okunembewu kanye nokulahlekelwa umsebenzi wokubulala izinambuzane lapho ukudla okunembewu yesinaphi encane kwelashwa ngokushisa ngaphambi kokusetshenziswa. Ukwelashwa ngokushisa kulindeleke ukuthi kubhubhise ama-enzyme e-hydrolytic avuselela ama-glucosinolates, ngaleyo ndlela kuvimbele ukwakheka kwama-isothiocyanate asebenzayo. Lolu ucwaningo lokuqala lokuqinisekisa izakhiwo zokubulala izinambuzane zempuphu yembewu yeklabishi ngokumelene nomiyane endaweni yasemanzini.
Phakathi kwempushana yembewu ehlolwe, impushana yembewu ye-watercress (Ls) yayiyiyona enobuthi kakhulu, yabangela ukufa okuphezulu kwe-Aedes albopictus. Izibungu ze-Aedes aegypti zacutshungulwa njalo amahora angama-24. Izimpushana ezintathu zembewu ezisele (PG, IG kanye ne-DFP) zazinomsebenzi ohamba kancane futhi zaqhubeka nokubulala okukhulu ngemva kwamahora angama-72 okwelashwa okuqhubekayo. Ukudla kwembewu ye-Ls kuphela okwakunenani elikhulu lama-glucosinolates, kanti i-PG kanye ne-DFP kwakune-myrosinase kanti i-IG yayine-glucosinolate njenge-glucosinolate enkulu (Ithebula 1). I-Glucotropaeolin i-hydrolyzed ibe yi-BITC kanti i-sinalbine i-hydrolyzed ibe yi-4-HBITC61,62. Imiphumela yethu ye-bioassay ikhombisa ukuthi kokubili ukudla kwembewu ye-Ls kanye ne-BITC yokwenziwa kuyingozi kakhulu ezibunguni zomiyane. Ingxenye eyinhloko yokudla kwembewu ye-PG kanye ne-DFP yi-myrosinase glucosinolate, e-hydrolyzed ibe yi-AITC. I-AITC iyasebenza ekubulaleni izibungu zomiyane ngenani le-LC50 elingu-19.35 ppm. Uma kuqhathaniswa ne-AITC ne-BITC, i-isothiocyanate ye-4-HBITC ayinabo ubuthi obuncane kakhulu ezibungwini. Nakuba i-AITC ingenabo ubuthi obuncane kune-BITC, amanani ayo e-LC50 aphansi kunamafutha amaningi abalulekile ahlolwe ezibungwini zomiyane32,73,74,75.
Impuphu yethu yembewu e-cruciferous yokusetshenziswa ngokumelene nezibungu zomiyane iqukethe i-glucosinolate eyodwa enkulu, equkethe ngaphezu kwama-98-99% ewonke ama-glucosinolates njengoba kunqunywe yi-HPLC. Inani elincane lamanye ama-glucosinolates atholakale, kodwa amazinga awo ayengaphansi kuka-0.3% we-glucosinolates iyonke. Impuphu yembewu ye-Watercress (L. sativum) iqukethe ama-glucosinolates esibili (sinigrin), kodwa isilinganiso sawo siyi-1% yempuphu yembewu iyonke, futhi okuqukethwe kwawo kusabalulekile (cishe i-0.4 mg/g yempuphu yembewu). Nakuba i-PG ne-DFP ziqukethe i-glucosinolate (myrosin) efanayo eyinhloko, umsebenzi wokubulala izibungu ekudleni kwazo kwembewu uhluka kakhulu ngenxa yamanani azo e-LC50. Kuyahlukahluka ngobuthi ku-powdery mildew. Ukuvela kwezibungu ze-Aedes aegypti kungase kube ngenxa yomehluko emsebenzini we-myrosinase noma ukuzinza phakathi kokudla kwembewu okubili. Umsebenzi we-Myrosinase udlala indima ebalulekile ekutholakaleni kwemikhiqizo ye-hydrolysis njenge-isothiocyanates ezitshalweni ze-Brassicaceae76. Imibiko yangaphambilini kaPocock et al.77 noWilkinson et al.78 ikhombisile ukuthi izinguquko emsebenzini we-myrosinase kanye nokuzinza kungahlotshaniswa nezici zofuzo kanye nezemvelo.
Okuqukethwe kwe-isothiocyanate esebenzayo okulindelwe kubalwe ngokusekelwe kumanani e-LC50 okudla ngakunye kwembewu emahoreni angama-24 nangama-72 (Ithebula 5) ukuze kuqhathaniswe nokusetshenziswa kwamakhemikhali okuhambisanayo. Ngemva kwamahora angama-24, ama-isothiocyanate ekudleni kwembewu ayenobuthi kakhulu kunamakhemikhali ahlanzekile. Amanani e-LC50 abalwe ngokusekelwe ezingxenyeni ngesigidi (ppm) sokwelashwa kwembewu ye-isothiocyanate ayephansi kunamanani e-LC50 okusetshenziswa kwe-BITC, AITC, kanye ne-4-HBITC. Sibone izibungu zidla ama-pellets okudla kwembewu (Isithombe 3A). Ngenxa yalokho, izibungu zingase zithole ukuchayeka okugxile kakhulu kuma-isothiocyanate anobuthi ngokudla ama-pellets okudla kwembewu. Lokhu kwabonakala kakhulu ekwelashweni kokudla kwembewu kwe-IG kanye ne-PG emahoreni angama-24 okuvezwa, lapho ukugcwala kwe-LC50 kwakuphansi ngo-75% no-72% kunokwelashwa kwe-AITC ehlanzekile kanye ne-4-HBITC, ngokulandelana. Ukwelashwa kwe-Ls kanye ne-DFP kwakunobuthi kakhulu kune-isothiocyanate emsulwa, ngamanani e-LC50 angu-24% kanye no-41% aphansi, ngokulandelana. Ama-larvae ekwelashweni kokulawula aphumile ngempumelelo (Isithombe 3B), kuyilapho iningi lama-larvae ekwelashweni kokudla kwembewu azange aphumile futhi ukukhula kwama-larval kwabambezeleka kakhulu (Isithombe 3B,D). Ku-Spodopteralitura, ama-isothiocyanates ahlotshaniswa nokulibaziseka kokukhula kanye nokubambezeleka kokukhula79.
Izibungu ze-Ae. Omiyane be-Aedes aegypti babehlala bevezwa yimpuphu yembewu ye-Brassica amahora angama-24-72. (A) Izibungu ezifile ezinezinhlayiya zokudla kwembewu emlonyeni (ezijikeleziwe); (B) Ukwelashwa kokulawula (dH20 ngaphandle kokudla kwembewu okwengeziwe) kubonisa ukuthi izibungu zikhula ngendlela evamile futhi ziqala ukuphuphuzela ngemva kwamahora angama-72 (C, D) Izibungu ziphathwe ngokuphuphuzela imbewu; ukudla kwembewu kubonise umehluko ekukhuleni futhi akuzange kuphuphuzele.
Asikayifundi indlela esebenza ngayo imiphumela enobuthi ye-isothiocyanates ezibungwini zomiyane. Kodwa-ke, izifundo zangaphambilini ezintuthwaneni ezibomvu zomlilo (i-Solenopsis invicta) zikhombisile ukuthi ukuvinjelwa kwe-glutathione S-transferase (GST) kanye ne-esterase (EST) kuyindlela eyinhloko yokusebenza kwe-isothiocyanate, kanti i-AITC, ngisho noma ingasebenzi kahle, ingavimbela nomsebenzi we-GST. Izintuthwane ezibomvu ezingenisiwe ezingeni eliphansi. Umthamo ungu-0.5 µg/ml80. Ngokuphambene nalokho, i-AITC ivimbela i-acetylcholinesterase ezibungwini zommbila zabantu abadala (i-Sitophilus zeamais)81. Izifundo ezifanayo kumele zenziwe ukuze kucaciswe indlela esebenza ngayo i-isothiocyanate ezibungwini zomiyane.
Sisebenzisa ukwelashwa kwe-DFP okungasebenzi ngokushisa ukusekela isiphakamiso sokuthi i-hydrolysis yama-glucosinolates ezitshalo ukuze kwakheke ama-isothiocyanates asebenzayo isebenza njengendlela yokulawula izibungu zomiyane ngokudla kwembewu yesinaphi. Ukudla kwembewu ye-DFP-HT bekungenobuthi ngamanani okusetshenziswa ahlolwe. ULafarga et al. 82 babike ukuthi ama-glucosinolates azwela ukuwohloka emazingeni okushisa aphezulu. Ukwelashwa ngokushisa kulindeleke ukuthi kunciphise i-enzyme ye-myrosinase ekudleni kwembewu futhi kuvimbele i-hydrolysis yama-glucosinolates ukuze kwakheke ama-isothiocyanates asebenzayo. Lokhu kwaqinisekiswa futhi ngu-Okunade et al. 75 babonise ukuthi i-myrosinase izwela ekushiseni, okubonisa ukuthi umsebenzi we-myrosinase awuzange usebenze ngokuphelele lapho imbewu yesinaphi, isinaphi esimnyama, kanye nezimpande zegazi zivezwa emazingeni okushisa angaphezu kwama-80°. C. Lezi zindlela zingase ziholele ekulahlekelweni komsebenzi wokubulala izinambuzane wokudla kwembewu ye-DFP ephathwe ngokushisa.
Ngakho-ke, ukudla kwembewu yesinaphi kanye nama-isothiocyanate ayo amathathu amakhulu kuyingozi ezibungwini zomiyane. Njengoba kunikezwe lo mehluko phakathi kokudla kwembewu kanye nokwelashwa ngamakhemikhali, ukusetshenziswa kokudla kwembewu kungaba yindlela ephumelelayo yokulawula omiyane. Kunesidingo sokubona amafomula afanele kanye nezinhlelo zokulethwa ezisebenzayo ukuthuthukisa ukusebenza kahle kanye nokuqina kokusetshenziswa kwezimpushana zembewu. Imiphumela yethu ikhombisa ukusetshenziswa okungenzeka kokudla kwembewu yesinaphi njengendlela ehlukile kunezibulala-zinambuzane zokwenziwa. Lobu buchwepheshe bungaba ithuluzi elisha lokulawula izinambuzane. Ngenxa yokuthi izibungu zomiyane ziyachuma ezindaweni zasemanzini futhi ama-glucosinolate okudla kwembewu aguqulwa ngama-enzyme abe ama-isothiocyanate asebenzayo lapho emanzi, ukusetshenziswa kokudla kwembewu yesinaphi emanzini agcwele omiyane kunikeza amandla amakhulu okulawula. Nakuba umsebenzi wokubulala izibungu wama-isothiocyanate uyahlukahluka (BITC > AITC > 4-HBITC), kudingeka ucwaningo olwengeziwe ukuze kunqunywe ukuthi ukuhlanganisa ukudla kwembewu nama-glucosinolates amaningi kukhulisa ubuthi ngokusebenzisana. Lolu ucwaningo lokuqala olubonisa imiphumela yokubulala izinambuzane yokudla kwembewu yesinaphi esinamafutha kanye nama-isothiocyanate amathathu asebenzayo komiyane. Imiphumela yalolu cwaningo ivula indlela entsha ngokukhombisa ukuthi ukudla kwembewu yeklabishi okunamafutha, okuwumkhiqizo ophuma kumafutha akhishwe embewini, kungase kusebenze njenge-ejenti yokubulala izibungu ethembisayo yokulawula omiyane. Lolu lwazi lungasiza ekutholeni ama-ejenti okulawula izitshalo kanye nentuthuko yawo njengemithi yokubulala izibungu eshibhile, esebenzayo, futhi enobungani nemvelo.
Amasethi edatha akhiqizwe kulolu cwaningo kanye nokuhlaziywa okutholiwe kuyatholakala kumbhali ohambisanayo uma kucelwe ngokufanele. Ekupheleni kocwaningo, zonke izinto ezisetshenziswe ocwaningweni (izinambuzane kanye nokudla kwembewu) zabhujiswa.
Isikhathi sokuthunyelwe: Julayi-29-2024