Ukuze ngokufanelekileyolawula iingcongconikunye nokunciphisa ukwanda kwezifo abaziphetheyo, kufuneka ezinye iindlela ezinobuchule, ezizinzileyo nezinobuhlobo nokusingqongileyo endaweni yezibulali-zinambuzane zeekhemikhali. Sivavanye ukutya kwembewu evela kwiBrassicaceae ethile (usapho lweBrassica) njengomthombo we-isothiocyanates ezivela kwizityalo eziveliswa yi-enzymatic hydrolysis ye-glycosinolates engasebenziyo ngokwebhayoloji ukuze isetyenziswe kulawulo lwe-Egyptian Aedes (L., 1762). Isidlo sembewu esihluthwe amanqatha ezintlanu (iBrassica juncea (L) Czern., 1859, iLepidium sativum L., 1753, iSinapis alba L., 1753, iThlaspi arvense L., 1753 kunye neThlaspi arvense – iintlobo ezintathu eziphambili zokungavuseleli ubushushu kunye nokubola kwe-enzymatic Iimveliso zeekhemikhali Ukufumanisa ubuthi (LC50) be-allyl isothiocyanate, i-benzyl isothiocyanate kunye ne-4-hydroxybenzylisothiocyanate kwi-Aedes aegypti larvae xa isasazwa iiyure ezingama-24 = 0.04 g/120 ml dH2O). Amaxabiso e-LC50 kwi-mustard, i-white mustard kunye ne-horsetail. Isidlo sembewu sasiyi-0.05, 0.08 kunye ne-0.05 ngokulandelelana xa kuthelekiswa ne-allyl isothiocyanate (LC50 = 19.35 ppm) kunye ne-4. -I-Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) yayinobuthi ngakumbi kwi-larvicide emva kweeyure ezingama-24 emva konyango kune-0.1 g/120 ml dH2O ngokulandelelana. Ezi ziphumo zihambelana nokuveliswa kwesidlo sembewu ye-alfalfa. Ukusebenza okuphezulu kwe-benzyl esters kuhambelana namaxabiso e-LC50 abaliweyo. Ukusebenzisa isidlo sembewu kunokubonelela ngendlela esebenzayo yokulawula iingcongconi. ukusebenza kwe-cruciferous seed powder kunye neekhemikhali zayo eziphambili ngokuchasene ne-mosquito larvicide kwaye kubonisa indlela iikhompawundi zendalo kwi-cruciferous seed powder ezinokusebenza ngayo njenge-larvicide enobuhlobo nokusingqongileyo yokulawula iingcongconi.
Izifo ezithwalwa ziintsholongwane ezibangelwa ziingcongconi ze-Aedes ziseyingxaki enkulu kwimpilo yoluntu kwihlabathi liphela. Ukuxhaphaka kwezifo ezithwalwa ziingcongconi kusasazeka ngokwendawo1,2,3 kwaye kuphinde kuvele, nto leyo ekhokelela ekuqhambukeni kwesifo esibi4,5,6,7. Ukusasazeka kwezifo phakathi kwabantu nezilwanyana (umz., i-chikungunya, i-dengue, i-Rift Valley fever, i-yellow fever kunye ne-Zika virus) akukaze kwenzeke ngaphambili. I-dengue fever yodwa ibeka abantu abamalunga ne-3.6 yeebhiliyoni emngciphekweni wokosuleleka kwiindawo ezishushu, apho kuqikelelwa ukuba zizigidi ezingama-390 zosulelo ezenzeka minyaka le, nto leyo ebangela ukufa kwabantu abayi-6,100–24,300 ngonyaka8. Ukuvela kwakhona nokuqhambuka kwe-Zika virus eMzantsi Melika kuye kwatsala ingqalelo kwihlabathi liphela ngenxa yomonakalo wengqondo ewubangelayo kubantwana abazelwe ngabafazi abasulelekileyo2. UKremer et al 3 baxela kwangaphambili ukuba uluhlu lweendawo zeengcongconi ze-Aedes luya kuqhubeka lukhula kwaye ngo-2050, isiqingatha sabemi behlabathi siya kuba semngciphekweni wokosuleleka zii-arbovirus ezithwalwa ziingcongconi.
Ngaphandle kwemithi yokugonya esandul’ ukuveliswa yokulwa ne-dengue kunye ne-yellow fever, imithi yokugonya uninzi lwezifo ezithwalwa yingcongconi ayikaphuhliswa9,10,11. Imithi yokugonya isafumaneka ngobuninzi obuncinci kwaye isetyenziswa kuphela kwizilingo zeklinikhi. Ulawulo lwezilwanyana ezithwala iingcongconi kusetyenziswa imithi yokugonya yokwenziwa ibe licebo eliphambili lokulawula ukusasazeka kwezifo ezithwalwa yingcongconi12,13. Nangona imithi yokugonya yokwenziwa isebenza kakuhle ekubulaleni iingcongconi, ukusetyenziswa okuqhubekayo kwezibulali-zinambuzane zokwenziwa kuchaphazela kakubi izinto eziphilayo ezingajoliswanga kwaye kungcolisa okusingqongileyo14,15,16. Okuxhalabisa ngakumbi yindlela yokwanda kokuxhathisa iingcongconi kwimithi yokugonya yokwenziwa yikhemikhali17,18,19. Ezi ngxaki ezinxulumene nemithi yokugonya ziye zakhawulezisa ukukhangela ezinye iindlela ezisebenzayo nezinobuhlobo nokusingqongileyo zokulawula izinambuzane zezifo.
Izityalo ezahlukeneyo ziye zaphuhliswa njengemithombo ye-phytopesticides yokulawula izinambuzane20,21. Izinto zezityalo zihlala zinobuhlobo nokusingqongileyo kuba ziyabola kwaye zinobuthi obuphantsi okanye obungenamsebenzi kwizinto eziphilayo ezingajoliswanga njengezilwanyana ezanyisayo, iintlanzi kunye nezilwanyana ezihlala emanzini20,22. Amalungiselelo ezityalo ayaziwa ngokuvelisa iintlobo ngeentlobo zeekhompawundi ezisebenzayo eziphilayo kunye neendlela ezahlukeneyo zokusebenza ukulawula ngempumelelo amanqanaba obomi ahlukeneyo eengcongconi23,24,25,26. Iikhompawundi ezivela kwizityalo ezifana neeoyile ezibalulekileyo kunye nezinye izithako zezityalo ezisebenzayo ziye zafumana ingqalelo kwaye zavula indlela yezixhobo ezintsha zokulawula iivectors zeengcongconi. Iioyile ezibalulekileyo, ii-monoterpenes kunye nee-sesquiterpenes zisebenza njengezigxothi, izithinteli zokutya kunye nee-ovicides27,28,29,30,31,32,33. Iioyile ezininzi zemifuno zibangela ukufa kweengcongconi, iipupae kunye nabantu abadala34,35,36, ezichaphazela imithambo-luvo, ukuphefumla, i-endocrine kunye nezinye iinkqubo ezibalulekileyo zezinambuzane37.
Izifundo zakutshanje zibonelele ngengqiqo malunga nokusetyenziswa okunokwenzeka kwezityalo zemastadi kunye nembewu yazo njengomthombo weekhompawundi ezisebenzayo. Umgubo wembewu yemustard uvavanyiwe njenge-biofumigant38,39,40,41 kwaye usetyenziswa njengotshintsho lomhlaba wokunciphisa ukhula42,43,44 kunye nolawulo lweentsholongwane zezityalo ezithwalwa ngumhlaba45,46,47,48,49,50, izondlo zezityalo. ii-nematodes 41,51, 52, 53, 54 kunye nezinambuzane 55, 56, 57, 58, 59, 60. Umsebenzi wokubulala iifungus wale mibhobho yembewu ubangelwa ziikhompawundi ezikhusela izityalo ezibizwa ngokuba yi-isothiocyanates38,42,60. Kwizityalo, ezi khompawundi zikhuselayo zigcinwa kwiiseli zezityalo ngendlela ye-glucosinolates ezingezizo ezisebenzayo. Nangona kunjalo, xa izityalo zonakaliswe kukutya kwezinambuzane okanye usulelo lwe-pathogen, ii-glucosinolates ziyanyibilikiswa yi-myrosinase zibe yi-isothiocyanates ezisebenzayo55,61. Ii-Isothiocyanates ziikhompawundi eziguquguqukayo ezaziwa ngokuba nomsebenzi wokulwa neentsholongwane kunye nokubulala izinambuzane obanzi, kwaye ulwakhiwo lwazo, umsebenzi webhayoloji kunye nomxholo wazo zahlukile kakhulu phakathi kweentlobo zeBrassicaceae42,59,62,63.
Nangona ii-isothiocyanates ezithathwe kwisidlo sembewu ye-mustard zaziwa ngokuba zinokubulala izinambuzane, idatha malunga nomsebenzi webhayoloji ngokuchasene ne-arthropod vectors ebalulekileyo kwezonyango ayikho. Uphononongo lwethu luhlolisise umsebenzi wokubulala i-larvicidal weempuphu ezine zembewu ezicociweyo nxamnye neengcongconi ze-Aedes. Ii-Larvae ze-Aedes aegypti. Injongo yolu phononongo yayikukuvavanya ukusetyenziswa kwazo njenge-biopesticides enobuhlobo nokusingqongileyo yokulawula iingcongconi. Iinxalenye ezintathu eziphambili zeekhemikhali zesidlo sembewu, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC), kunye ne-4-hydroxybenzylisothiocyanate (4-HBITC) nazo zavavanywa ukuvavanya umsebenzi webhayoloji wezi nxalenye zeekhemikhali kwi-mother larvae. Le yingxelo yokuqala yokuvavanya ukusebenza kakuhle kweempuphu ezine zembewu yekhaphetshu kunye neenxalenye zazo eziphambili zeekhemikhali ngokuchasene neengcongconi.
Iikholoni zelabhoratri ze-Aedes aegypti (uhlobo lweRockefeller) zigcinwe kuma-26°C, umswakama ongama-70% (RH) kunye ne-10:14 h (L:D photoperiod). Iimazi ezizalelweyo zigcinwe kwiikheyiji zeplastiki (ubude obuyi-11 cm kunye nobubanzi obuyi-9.5 cm) kwaye zondliwa ngenkqubo yokondla ngebhotile kusetyenziswa igazi leenkomo eliqinisiweyo (HemoStat Laboratories Inc., Dixon, CA, USA). Ukondla ngegazi kwenziwa njengesiqhelo kusetyenziswa i-membrane multi-glass feeder (Chemglass, Life Sciences LLC, Vineland, NJ, USA) eqhagamshelwe kwityhubhu yokuhlambela yamanzi ejikelezayo (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) enolawulo lobushushu obuyi-37°C. Yolula ifilimu yeParafilm M ezantsi kwegumbi ngalinye lokondla ngeglasi (indawo eyi-154 mm2). I-feeder nganye yabekwa kwigridi ephezulu egubungela ikheyiji equlethe imazi ezalelweyo. Malunga ne-350–400 μl yegazi leenkomo longezwe kwi-funnel yeglasi kusetyenziswa i-Pasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) kwaye iimpethu ezindala zavunyelwa ukuba zikhuphe amanzi ubuncinane iyure enye. Iimazi ezikhulelweyo zanikwa isisombululo se-10% se-sucrose zaza zavunyelwa ukuba zibeke amaqanda kwiphepha lesihluzo elimanzi elifakwe kwiikomityi ze-soufflé ezicacileyo (1.25 fl oz ubukhulu, Dart Container Corp., Mason, MI, USA). Faka amanzi kwikheyiji. Beka iphepha lesihluzo elinamaqanda kwibhegi evaliweyo (SC Johnsons, Racine, WI) kwaye ugcine kuma-26°C. Amaqanda aqanduselwa kwaye malunga ne-200–250 larvae zakhuliswa kwiitreyi zeplastiki eziqulethe umxube we-rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) kunye nomgubo wesibindi (MP Biomedicals, LLC, Solon, OH, USA). kunye ne-fish fillet (TetraMin, Tetra GMPH, Meer, Germany) ngomlinganiselo we-2:1:1. Iimbungu ze-instar zexesha lokugqibela zisetyenzisiwe kwi-bioassays yethu.
Izinto zembewu yezityalo ezisetyenzisiweyo kolu phononongo zifunyenwe kwimithombo elandelayo yezorhwebo nekarhulumente: iBrassica juncea (brown mustard-Pacific Gold) kunye neBrassica juncea (white mustard-Ida Gold) ezivela kwiPacific Northwest Farmers' Cooperative, eWashington State, e-USA; (Garden Cress) ezivela kwiKelly Seed and Hardware Co., ePeoria, IL, e-USA kunye neThlaspi arvense (Field Pennycress-Elisabeth) ezivela kwi-USDA-ARS, ePeoria, IL, e-USA; Akukho nanye imbewu esetyenzisiweyo kolu phononongo eyanyangwa ngezibulali-zinambuzane. Zonke izinto zembewu zacutshungulwa zaza zasetyenziswa kolu phononongo ngokuhambelana nemigaqo yengingqi neyelizwe kwaye ngokuhambelana nayo yonke imithetho yengingqi neyelizwe. Olu phononongo aluzange luhlolisise iintlobo zezityalo eziguqulweyo.
Imbewu yeBrassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) zicoliwe zaba ngumgubo ocolekileyo kusetyenziswa iRetsch ZM200 ultracentrifugal mill (Retsch, Haan, Germany) exhotyiswe nge-0.75 mm mesh kunye ne-Stainless steel rotor, amazinyo ali-12, i-10,000 rpm (Itheyibhile 1). Umgubo wembewu ecoliweyo udluliselwe kwi-thimble yephepha kwaye wasuswa amafutha nge-hexane kwisixhobo seSoxhlet iiyure ezingama-24. Isampulu encinci ye-mustard yasentsimini ecoliweyo yanyangwa ngobushushu kwi-100 °C kangangeyure e-1 ukuze isuse i-myrosinase kwaye ithintele i-hydrolysis ye-glucosinolates ukuze yenze i-isothiocyanates esebenzayo ngokwebhayoloji. Umgubo wembewu ye-horsetail ecociweyo ngobushushu (DFP-HT) wasetyenziswa njengolawulo olubi ngokususa i-myrosinase.
Umxholo we-glucosinolate womgubo wembewu enamafutha uchongiwe kathathu kusetyenziswa i-high-performance liquid chromatography (HPLC) ngokweprotokholi epapashwe ngaphambili 64. Ngamafutshane, i-3 mL ye-methanol yongezwa kwisampulu ye-250 mg yomgubo wembewu enamafutha. Isampulu nganye yafakwa kwi-sonic bath emanzini imizuzu engama-30 yaze yashiywa ebumnyameni kwi-23°C kangangeeyure ezili-16. I-1 mL aliquot yomaleko wendalo yahluzwa ngesihluzo se-0.45 μm kwi-autosampler. Isebenza kwinkqubo yeShimadzu HPLC (iipompo ezimbini ze-LC 20AD; i-SIL 20A autosampler; i-DGU 20As degasser; i-SPD-20A UV-VIS detector yokujonga kwi-237 nm; kunye ne-CBM-20A communication bus module), umxholo we-glucosinolate womgubo wembewu uchongiwe kathathu. kusetyenziswa isoftware yeShimadzu LC Solution version 1.25 (Shimadzu Corporation, Columbia, MD, USA). Ikholamu yayiyikholamu yesigaba sokubuyela umva ye-C18 Inertsil (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA). Iimeko zokuqala zesigaba sokuhamba zazimiselwe kwi-12% methanol/88% 0.01 M tetrabutylammonium hydroxide emanzini (TBAH; Sigma-Aldrich, St. Louis, MO, USA) ngesantya sokuhamba se-1 mL/min. Emva kokufakwa kwe-15 μl yesampuli, iimeko zokuqala zagcinwa imizuzu engama-20, kwaye emva koko umlinganiselo we-solvent walungiswa kwi-100% methanol, kunye nexesha elipheleleyo lohlalutyo lwesampuli lemizuzu engama-65. I-standard curve (esekelwe kwi-nM/mAb) yenziwe ngokuxutywa kwe-serial kwe-sinapine esandul’ ukulungiswa, i-glucosinolate kunye ne-myrosin standards (Sigma-Aldrich, St. Louis, MO, USA) ukuqikelela umxholo wesulfure womgubo wembewu enamafutha. Uxinzelelo lweGlucosinolate kwiisampuli luvavanywe kwi-Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) kusetyenziswa inguqulelo ye-OpenLAB CDS ChemStation (C.01.07 SR2 [255]) exhotyiswe ngekholamu efanayo kwaye kusetyenziswa indlela echazwe ngaphambili. Uxinzelelo lweGlucosinolate lufunyenwe; lunokuthelekiseka phakathi kweenkqubo ze-HPLC.
I-Allyl isothiocyanate (94%, izinzile) kunye ne-benzyl isothiocyanate (98%) zithengwe kwiFisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). I-4-Hydroxybenzylisothiocyanate ithengwe kwiChemCruz (Santa Cruz Biotechnology, CA, USA). Xa i-enzymatically hydrolyzed yi-myrosinase, i-glucosinolates, i-glucosinolates, kunye ne-glucosinolates zenza i-allyl isothiocyanate, i-benzyl isothiocyanate, kunye ne-4-hydroxybenzylisothiocyanate, ngokwahlukeneyo.
Uvavanyo lwebhayoloji lwenziwe ngokwendlela kaMuturi et al. 32 kunye notshintsho. Ukutya kwembewu okuhlanu okunamafutha aphantsi kusetyenziswe kolu phando: iDFP, iDFP-HT, iIG, iPG kunye neL. Iimbungu ezingamashumi amabini zafakwa kwi-400 mL disposable three-way beaker (VWR International, LLC, Radnor, PA, USA) equlethe amanzi acocekileyo angama-120 mL (dH2O). Uxinaniso lwembewu ezisixhenxe lwavavanywa ukuze kujongwe ubuthi beembungu zeengcongconi: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 kunye ne-0.12 g yembewu yokutya/120 ml dH2O yembewu yokutya kwembewu yeDFP, iDFP-HT, iIG kunye nePG. Uvavanyo lwebhayoloji lokuqala lubonisa ukuba umgubo wembewu yeLs ocociweyo unetyhefu ngakumbi kuneminye imipuphu emine yembewu evavanyiweyo. Ngoko ke, sihlengahlengise amanqanaba asixhenxe onyango lomgubo wembewu ye-Ls kula manqanaba alandelayo: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, kunye ne-0.075 g/120 mL dH2O.
Iqela lolawulo olunganyangwanga (i-dH20, akukho songezelelo sesidlo sembewu) lifakiwe ukuvavanya ukufa okuqhelekileyo kwezinambuzane phantsi kweemeko zovavanyo. Uvavanyo lwebhayoloji olunobungozi kwisidlo ngasinye sembewu luquka iibhika ezintathu eziphindaphindwayo ezintathu (ii-larvae ezingama-20 ezikwinqanaba lesithathu kwibhika nganye), kwiibhotile ezili-108 zizonke. Izikhongozeli ezinyangiweyo zigcinwe kubushushu begumbi (20-21°C) kwaye ukufa kwee-larvae kwarekhodwa ngexesha leeyure ezingama-24 kunye nama-72 zokuchatshazelwa rhoqo kumanqanaba onyango. Ukuba umzimba wengcongconi kunye nezitho azishukumi xa zihlatywa okanye zichukunyiswa nge-spatula encinci yentsimbi engagqwaliyo, ii-larvae zengcongconi zithathwa njengezifileyo. Ii-larvae ezifileyo zihlala zingashukumi kwindawo engasemva okanye ezantsi kwesikhongozeli okanye phezu kwamanzi. Olu vavanyo luphindwe kathathu kwiintsuku ezahlukeneyo kusetyenziswa amaqela ahlukeneyo ee-larvae, kwii-larvae ezili-180 ezichatshazelwe kumanqanaba onyango ngalunye.
Ubuthi be-AITC, i-BITC, kunye ne-4-HBITC kwi-mosquito larvae buhlolwe kusetyenziswa inkqubo efanayo ye-bioassay kodwa ngonyango olwahlukileyo. Lungisa izisombululo zesitokhwe se-100,000 ppm kwikhemikhali nganye ngokongeza i-100 µL yekhemikhali kwi-900 µL ye-ethanol epheleleyo kwityhubhu ye-centrifuge ye-2-mL kwaye ushukumise imizuzwana engama-30 ukuze ixutywe kakuhle. Ubungakanani bonyango bumiselwe ngokusekelwe kuvavanyo lwethu lokuqala lwe-bioassays, olufumanise ukuba i-BITC inetyhefu kakhulu kune-AITC kunye ne-4-HBITC. Ukufumanisa ubuthi, ubuninzi be-BITC obu-5 (1, 3, 6, 9 kunye ne-12 ppm), ubuninzi be-AITC obu-7 (5, 10, 15, 20, 25, 30 kunye ne-35 ppm) kunye nobunzima be-6 be-4-HBITC (15, 15, 20, 25, 30 kunye ne-35 ppm). 30, 45, 60, 75 kunye ne-90 ppm). Unyango lolawulo lufakwe i-108 μL ye-ethanol epheleleyo, elingana nomthamo ophezulu wonyango lweekhemikhali. Uvavanyo lwe-bioassays luphindwe njengoko kungasentla, luveza inani elipheleleyo le-larvae ezili-180 ngoxinzelelo ngalunye lonyango. Ukufa kwe-larvae kubhalwe phantsi kuxinzelelo ngalunye lwe-AITC, i-BITC, kunye ne-4-HBITC emva kweeyure ezingama-24 zokuvezwa rhoqo.
Uhlalutyo lwe-probit lwedatha yokufa enxulumene nedosi engama-65 lwenziwe kusetyenziswa isoftware yePolo (Polo Plus, LeOra Software, inguqulelo 1.0) ukubala uxinaniso olubulalayo oluyi-50% (LC50), uxinaniso olubulalayo oluyi-90% (LC90), i-slope, i-letal dose coefficient, kunye noxinaniso olubulalayo oluyi-95%. ngokusekelwe kwizithuba zokuzithemba zemilinganiselo yedosi ebulalayo yoxinaniso oluguqulweyo lwelog kunye ne-dose-death curves. Idatha yokufa isekelwe kwidatha edibeneyo ephindaphindwayo yee-larvae ezili-180 ezivezwe kuxinaniso ngalunye lonyango. Uhlalutyo olunokwenzeka lwenziwe ngokwahlukeneyo kwisidlo ngasinye sembewu kunye nenxalenye nganye yekhemikhali. Ngokusekelwe kwisithuba sokuzithemba se-95% somlinganiselo wedosi ebulalayo, ubuthi bokutya kwembewu kunye nezinto ezibangela iikhemikhali kwi-mosquito lathathwa njengezahluke kakhulu, ngoko ke isithuba sokuzithemba esinexabiso le-1 sasingahlukanga kakhulu, P = 0.0566.
Iziphumo ze-HPLC zokumisela ii-glucosinolates eziphambili kwimibhobho yembewu enamafutha i-DFP, i-IG, i-PG kunye ne-L zidweliswe kwiTheyibhile 1. Ii-glucosinolates eziphambili kwimibhobho yembewu evavanyiweyo zahlukile ngaphandle kwe-DFP kunye ne-PG, zombini ezazine-myrosinase glucosinolates. Umxholo we-myrosinin kwi-PG wawuphezulu kune-DFP, 33.3 ± 1.5 kunye ne-26.5 ± 0.9 mg/g, ngokwahlukeneyo. Umgubo wembewu ye-Ls wawune-36.6 ± 1.2 mg/g ye-gluglycone, ngelixa umgubo wembewu ye-IG wawune-38.0 ± 0.5 mg/g ye-sinapine.
Iimbungu ze-Ae. Aedes aegypti mosquitoes zabulawa xa zinyangwa ngesidlo sembewu esicociweyo, nangona ukusebenza konyango kwahluka ngokuxhomekeke kuhlobo lwesityalo. Yi-DFP-NT kuphela eyayingenabungozi kwimbungu yezinambuzane emva kweeyure ezingama-24 kunye nama-72 zokuchaphazeleka (Itheyibhile 2). Ubuthi bomgubo wembewu esebenzayo banda ngokukhula koxinzelelo (Umzobo 1A, B). Ubuthi bomgubo wembewu kwimbungu yezinambuzane bahluka kakhulu ngokusekelwe kwi-95% CI yomlinganiselo wedosi ebulalayo yamaxabiso e-LC50 kwiiyure ezingama-24 kunye novavanyo lweeyure ezingama-72 (Itheyibhile 3). Emva kweeyure ezingama-24, isiphumo esinobuthi somgubo wembewu ye-Ls sasingaphezulu kunezinye iindlela zonyango lomgubo wembewu, kunye nomsebenzi ophezulu kunye nobuthi obukhulu kwimbungu (LC50 = 0.04 g/120 ml dH2O). Iimbungu bezingenangxaki kangako kwi-DFP kwiiyure ezingama-24 xa kuthelekiswa nonyango lwe-IG, Ls kunye ne-PG seed powder, kunye namaxabiso e-LC50 angama-0.115, 0.04 kunye ne-0.08 g/120 ml dH2O ngokwahlukeneyo, awayephezulu ngokwezibalo kunexabiso le-LC50. 0.211 g/120 ml dH2O (Itheyibhile 3). Amaxabiso e-LC90 e-DFP, IG, PG kunye ne-Ls ayengama-0.376, 0.275, 0.137 kunye ne-0.074 g/120 ml dH2O, ngokwahlukeneyo (Itheyibhile 2). Uxinzelelo oluphezulu lwe-DPP yayiyi-0.12 g/120 ml dH2O. Emva kweeyure ezingama-24 zovavanyo, umyinge wokufa kweembungu wawuyi-12% kuphela, ngelixa umyinge wokufa kweembungu ze-IG kunye ne-PG ufikelele kwi-51% kunye ne-82%, ngokwahlukeneyo. Emva kweeyure ezingama-24 zovavanyo, umndilili wokufa kwee-larval kwingxinano ephezulu yonyango lwembewu ye-Ls (0.075 g/120 ml dH2O) yayiyi-99% (Umzobo 1A).
Iigrafu zokufa ziqikelelwe ukusuka kwimpendulo yedosi (Probit) ye-Ae. Egyptian larvae (3rd instar larvae) ukuya kuxinaniso lomgubo wembewu kwiiyure ezingama-24 (A) kunye neeyure ezingama-72 (B) emva konyango. Umgca onamachaphaza umele i-LC50 yonyango lomgubo wembewu. 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.
Kuvavanyo lweeyure ezingama-72, amaxabiso e-LC50 e-DFP, IG kunye ne-PG seed meal ayeyi-0.111, 0.085 kunye ne-0.051 g/120 ml dH2O, ngokwahlukeneyo. Phantse zonke ii-larvae ezivezwe kwi-Ls seed meal zafa emva kweeyure ezingama-72 zokuvezwa, ngoko ke idatha yokufa yayingangqinelani nohlalutyo lweProbit. Xa kuthelekiswa nezinye ii-larvae, ii-larvae zazingenangxaki kangako nonyango lwe-DFP seed meal kwaye zazinamaxabiso aphezulu e-LC50 ngokwezibalo (Iitheyibhile 2 kunye ne-3). Emva kweeyure ezingama-72, amaxabiso e-LC50 e-DFP, IG kunye ne-PG seed meal aqikelelwa ukuba yi-0.111, 0.085 kunye ne-0.05 g/120 ml dH2O, ngokwahlukeneyo. Emva kweeyure ezingama-72 zovavanyo, amaxabiso e-LC90 e-DFP, IG kunye ne-PG seed powders ayeyi-0.215, 0.254 kunye ne-0.138 g/120 ml dH2O, ngokwahlukeneyo. Emva kweeyure ezingama-72 zovavanyo, umndilili wokufa kwee-larval kwi-DFP, IG kunye ne-PG seed meal kuxinzelelo oluphezulu lwe-0.12 g/120 ml dH2O yayiyi-58%, 66% kunye ne-96%, ngokulandelanayo (Umzobo 1B). Emva kovavanyo lweeyure ezingama-72, isidlo sembewu se-PG sifunyenwe sinetyhefu ngaphezu kwesidlo sembewu se-IG kunye ne-DFP.
Ii-isothiocyanates ezenziweyo, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC) kunye ne-4-hydroxybenzylisothiocyanate (4-HBITC) zinokubulala ngempumelelo iimbungu zengcongconi. Kwiiyure ezingama-24 emva konyango, i-BITC yayinobungozi ngakumbi kwiimbungu ngexabiso le-LC50 le-5.29 ppm xa kuthelekiswa ne-19.35 ppm ye-AITC kunye ne-55.41 ppm ye-4-HBITC (Itheyibhile 4). Xa kuthelekiswa ne-AITC kunye ne-BITC, i-4-HBITC inetyhefu ephantsi kwaye ixabiso le-LC50 liphezulu. Kukho umahluko omkhulu kwityhefu yeembungu zengcongconi ze-isothiocyanates ezimbini eziphambili (ii-L kunye ne-PG) kwisidlo sembewu esinamandla kakhulu. Ubuthi obusekelwe kumlinganiselo wedosi ebulalayo yamaxabiso e-LC50 phakathi kwe-AITC, i-BITC, kunye ne-4-HBITC bubonise umahluko wezibalo kangangokuba i-95% CI yomlinganiselo wedosi ebulalayo ye-LC50 ayizange ibandakanye ixabiso le-1 (P = 0.05, Itheyibhile 4). Amanqanaba aphezulu e-BITC kunye ne-AITC aqikelelwa ukuba abulale i-100% yeembungu ezivavanyiweyo (Umfanekiso 2).
Iigrafu zokufa ziqikelelwe kwimpendulo yedosi (Probit) ye-Ae. Kwiiyure ezingama-24 emva konyango, ii-larvae zase-Egypt (ii-larvae ze-instar yesithathu) zifikelele kumanqanaba e-isothiocyanate adibeneyo. Umgca onamachaphaza umele i-LC50 yonyango lwe-isothiocyanate. I-Benzyl isothiocyanate BITC, i-allyl isothiocyanate AITC kunye ne-4-HBITC.
Ukusetyenziswa kwezityalo ezibulala iintsholongwane njengeearhente zokulawula iingcongconi kuye kwafundwa kudala. Izityalo ezininzi zivelisa iikhemikhali zendalo ezinomsebenzi wokubulala izinambuzane37. Iikhompawundi zazo ezisebenzayo zibonelela ngenye indlela ekhangayo kunezibulala-zinambuzane ezenziwe ngokwenziwa ezinamandla amakhulu okulawula izinambuzane, kuquka neengcongconi.
Izityalo ze-mustard zikhuliswa njengesityalo sembewu yazo, zisetyenziswa njengesiqholo kunye nomthombo weoyile. Xa ioyile ye-mustard ikhutshwa kwimbewu okanye xa i-mustard ikhutshwa ukuze isetyenziswe njenge-biofuel, 69 imveliso ephumayo yimbewu ecociweyo. Esi sidlo sembewu sigcina uninzi lwezinto zaso zendalo ze-biochemical kunye nee-enzymes ze-hydrolytic. Ubuthi besi sidlo sembewu bubangelwa kukuveliswa kwe-isothiocyanates55,60,61. Ii-Isothiocyanates zenziwa yi-hydrolysis ye-glucosinolates yi-enzyme myrosinase ngexesha lokufuthwa kwembewu38,55,70 kwaye zaziwa ngokuba nemiphumo yokubulala iifungi, iintsholongwane, ii-nematic kunye nezinambuzane, kunye nezinye iimpawu kubandakanya iziphumo zemvakalelo yeekhemikhali kunye neempawu zonyango lwe-chemotherapeutic61,62,70. Izifundo ezininzi zibonise ukuba izityalo ze-mustard kunye nembewu yembewu zisebenza ngokufanelekileyo njengezifunxi-mafutha nxamnye nezinambuzane zomhlaba kunye nokutya okugciniweyo57,59,71,72. Kolu phononongo, sivavanye ubuthi bomgubo wembewu ezine kunye neemveliso zawo ezintathu ezisebenzayo kwi-AITC, BITC, kunye ne-4-HBITC kwi-Aedes mosquito larvae. I-Aedes aegypti. Ukongeza umgubo wembewu ngqo emanzini ane-mosquito larvae kulindeleke ukuba kusebenze iinkqubo ze-enzyme ezivelisa i-isothiocyanates ezinobungozi kwi-mosquito larvae. Olu tshintsho lwe-biotransformation lubonakaliswe ngokuyinxenye ngumsebenzi wokubulala ii-larvicate womgubo wembewu kunye nokulahleka komsebenzi wokubulala ii-nambuzane xa umgubo wembewu ye-mustard omfutshane unyangwa ngobushushu ngaphambi kokusetyenziswa. Unyango lobushushu kulindeleke ukuba lutshabalalise ii-enzymes ze-hydrolytic ezivuselela i-glucosinolates, ngaloo ndlela zithintele ukwakheka kwe-isothiocyanates ezisebenzayo kwi-nambuzane. Olu luphononongo lokuqala lokuqinisekisa iipropati zokubulala ii-nambuzane zomgubo wembewu yekhaphetshu kwi-nambuzane kwindawo yasemanzini.
Phakathi kweempuphu zembewu ezivavanyiweyo, umgubo wembewu ye-watercress (Ls) yayiyeyona inobungozi kakhulu, nto leyo eyabangela ukufa okuphezulu kwe-Aedes albopictus. Iimbungu ze-Aedes aegypti zacutshungulwa rhoqo iiyure ezingama-24. Iimpungu zembewu ezintathu eziseleyo (PG, IG kunye ne-DFP) zazisebenza kancinci kwaye zabangela ukufa okukhulu emva kweeyure ezingama-72 zonyango oluqhubekayo. Kuphela yi-Ls seed meal eyayine-glucosinolates ezininzi, ngelixa i-PG kunye ne-DFP zazine-myrosinase kwaye i-IG yayine-glucosinolate njenge-glucosinolate ephambili (Itheyibhile 1). I-Glucotropaeolin i-hydrolyzed kwi-BITC kwaye i-sinalbine i-hydrolyzed kwi-4-HBITC61,62. Iziphumo zethu ze-bioassay zibonisa ukuba zombini i-Ls seed meal kunye ne-synthetic BITC ziyingozi kakhulu kwiimbungu ze-mosquito. Icandelo eliphambili le-PG kunye ne-DFP seed meal yi-myrosinase glucosinolate, e-hydrolyzed kwi-AITC. I-AITC iyasebenza ekubulaleni iimbungu ze-mosquito ngexabiso le-LC50 le-19.35 ppm. Xa kuthelekiswa ne-AITC kunye ne-BITC, i-4-HBITC isothiocyanate yeyona inobungozi obuncinci kwi-larvae. Nangona i-AITC inobungozi obuncinci kune-BITC, amaxabiso ayo e-LC50 aphantsi kunee-oyile ezininzi ezibalulekileyo ezivavanyiweyo kwi-larvae ye-mosquito32,73,74,75.
Umgubo wethu wembewu eqholiweyo osetyenziselwa ukulwa neengcongconi uneglucosinolate enye enkulu, ebalelwa ngaphezulu kwe-98-99% yeeglucosinolates zizonke njengoko kumiselwe yi-HPLC. Izixa ezimbalwa zezinye iiglucosinolates zifunyenwe, kodwa amanqanaba azo ayengaphantsi kwe-0.3% yeeglucosinolates zizonke. Umgubo wembewu yeWatercress (L. sativum) uqulethe iiglucosinolates zesibini (sinigrin), kodwa umlinganiselo wazo yi-1% yeeglucosinolates zizonke, kwaye umxholo wazo awukabalulekanga (malunga ne-0.4 mg/g umgubo wembewu). Nangona i-PG kunye ne-DFP ziqulethe i-glucosinolate (myrosin) efanayo ephambili, umsebenzi wokubulala ii-larvicidal wokutya kwazo kwembewu wahlukile kakhulu ngenxa yamaxabiso azo e-LC50. Ubungozi buyahluka kwi-powdery mildew. Ukuvela kwee-Aedes aegypti larvae kunokubangelwa kukungafani komsebenzi we-myrosinase okanye ukuzinza phakathi kokutya kwembewu ezimbini. Umsebenzi we-Myrosinase udlala indima ebalulekileyo ekufumanekeni kwezinto eziphilayo kwiimveliso ze-hydrolysis ezifana ne-isothiocyanates kwizityalo zaseBrassicaceae76. Iingxelo zangaphambili zikaPocock et al.77 kunye noWilkinson et al.78 zibonise ukuba utshintsho kumsebenzi we-myrosinase kunye nokuzinza kunokunxulunyaniswa nezinto zemfuza kunye nokusingqongileyo.
Umxholo we-isothiocyanate osebenzayo kwi-bioactive ubalwe ngokusekelwe kumaxabiso e-LC50 yesidlo ngasinye sembewu kwiiyure ezingama-24 kunye nama-72 (Itheyibhile 5) ukuze kuthelekiswe nokusetyenziswa kweekhemikhali ezifanelekileyo. Emva kweeyure ezingama-24, ii-isothiocyanates kwisidlo sembewu zazinobuthi ngaphezu kweekhompawundi ezicocekileyo. Amaxabiso e-LC50 abalwe ngokusekelwe kwiindawo ngesigidi (ppm) zonyango lwembewu ye-isothiocyanate ayephantsi kunamaxabiso e-LC50 kwizicelo ze-BITC, AITC, kunye ne-4-HBITC. Sibone ii-larvae zisitya ii-pellets zembewu (Umfanekiso 3A). Ngenxa yoko, ii-larvae zinokufumana ukuvezwa okunzulu kwi-isothiocyanates ezinobuthi ngokutya ii-pellets zembewu. Oku kwabonakala kakhulu kunyango lwembewu ye-IG kunye ne-PG kwi-24-hour exposure, apho ukugxila kwe-LC50 kwakungama-75% kunye nama-72% ngaphantsi kunonyango lwe-AITC kunye ne-4-HBITC ecocekileyo, ngokwahlukeneyo. Unyango lwe-Ls kunye ne-DFP belunetyhefu ngakumbi kune-isothiocyanate ecocekileyo, kunye namaxabiso e-LC50 angama-24% kunye nama-41% aphantsi, ngokulandelelana. Iimbungu kunyango lolawulo ziphumelele ukufuya (Umzobo 3B), ngelixa uninzi lweembungu kunyango lomgubo wembewu azizange zifuyane kwaye uphuhliso lweembungu lwalibaziseka kakhulu (Umzobo 3B,D). Kwi-Spodopteralitura, ii-isothiocyanates zinxulunyaniswa nokulibaziseka kokukhula kunye nokulibaziseka kophuhliso79.
Iimbungu ze-Ae. Iingcongconi ze-Aedes aegypti bezisoloko zichatshazelwa yimpuphu yembewu yeBrassica kangangeeyure ezingama-24–72. (A) Iimbungu ezifileyo ezineenxalenye zomgubo wembewu emlonyeni (zijikelezwe); (B) Unyango lolawulo (dH20 ngaphandle kokutya kwembewu okongeziweyo) lubonisa ukuba iimbungu zikhula ngokuqhelekileyo kwaye ziqala ukuphuhla emva kweeyure ezingama-72 (C, D) Iimbungu ziphathwe ngomgubo wembewu; isidlo sembewu sibonise umahluko ekuphuhleni kwaye asizange siphuhlane.
Asikayifundi indlela ezisebenza ngayo ii-isothiocyanates kwimibungu yeengcongconi. Nangona kunjalo, izifundo zangaphambili kwiimbovane ezibomvu zomlilo (iSolenopsis invicta) zibonise ukuba ukuthintela i-glutathione S-transferase (GST) kunye ne-esterase (EST) yeyona ndlela iphambili yokusebenza kwe-isothiocyanate, kwaye i-AITC, nokuba ingasebenzi kakhulu, inokuthintela umsebenzi we-GST. I-AITC, nokuba iyasebenza kancinci, inokuthintela umsebenzi we-GST. I-AITC ithintela i-acetylcholinesterase kwi-corn weevils yabantu abadala (iSitophilus zeamais)81. Izifundo ezifanayo kufuneka zenziwe ukuze kucaciswe indlela esebenza ngayo i-isothiocyanate kwimibungu yeengcongconi.
Sisebenzisa unyango lwe-DFP olungasebenzisi ubushushu ukuxhasa isindululo sokuba i-hydrolysis ye-glucosinolates yezityalo yenze i-isothiocyanates esebenzayo isebenza njengendlela yokulawula i-larval yeengcongconi ngomgubo wembewu ye-mustard. Umgubo wembewu ye-DFP-HT wawungenabungozi kumazinga okusetyenziswa avavanyiweyo. ULafarga et al. 82 baxele ukuba ii-glucosinolates ziyachaphazeleka kukuwohloka kubushushu obuphezulu. Unyango lobushushu lukwalindeleke ukuba lutshintshe i-enzyme ye-myrosinase kwisidlo sembewu kwaye luthintele i-hydrolysis ye-glucosinolates ukuba yenze i-isothiocyanates esebenzayo. Oku kwaqinisekiswa ngu-Okunade et al. 75 babonise ukuba i-myrosinase inobuthathaka kubushushu, nto leyo ebonisa ukuba umsebenzi we-myrosinase awuzange usebenze ngokupheleleyo xa imbewu ye-mustard, i-black mustard, kunye ne-bloodroot ivezwe kubushushu obungaphezulu kwama-80°. C. Ezi ndlela zinokubangela ukulahleka komsebenzi wokubulala izinambuzane womgubo wembewu ye-DFP ephathwe ngobushushu.
Ngoko ke, isidlo sembewu yemustard kunye nee-isothiocyanates zayo ezintathu eziphambili ziyingozi kwiingcongconi. Ngenxa yolu mahluko phakathi kokutya kwembewu kunye nonyango lweekhemikhali, ukusetyenziswa kokutya kwembewu kunokuba yindlela esebenzayo yokulawula iingcongconi. Kukho imfuneko yokuchonga iindlela ezifanelekileyo kunye neenkqubo zokuhambisa ezisebenzayo ukuphucula ukusebenza kakuhle kunye nokuzinza kokusetyenziswa kwempuphu yembewu. Iziphumo zethu zibonisa ukusetyenziswa okunokwenzeka kokutya kwembewu yemustard njengendlela eyahlukileyo kwi-synthetic pesticides. Le teknoloji inokuba sisixhobo esitsha sokulawula i-vectors yengcongconi. Ngenxa yokuba iingcongconi zikhula kakuhle kwiindawo zasemanzini kwaye i-seed meal glucosinolates ziguqulwa nge-enzyme zibe yi-isothiocyanates esebenzayo xa zimanzi, ukusetyenziswa kokutya kwembewu yemustard emanzini azaliswe yingcongconi kunika amandla okulawula abalulekileyo. Nangona umsebenzi wokubulala ii-larvicidal we-isothiocyanates wahlukile (BITC > AITC > 4-HBITC), uphando olongezelelweyo luyadingeka ukumisela ukuba ukudibanisa isidlo sembewu kunye ne-glucosinolates ezininzi kuyandisa na ityhefu. Olu luphononongo lokuqala olubonisa iziphumo zokubulala izinambuzane zesidlo sembewu ye-cruciferous esinamafutha kunye ne-isothiocyanates ezintathu ezisebenzayo kwiingcongconi. Iziphumo zolu phononongo zivule indlela entsha ngokubonisa ukuba umgubo wembewu yekhaphetshu ocoliweyo, imveliso ephuma kwioyile ethathwe kwimbewu, unokusebenza njengearhente yokubulala iintsholongwane ethembisayo ekulawuleni iingcongconi. Olu lwazi lunokunceda ekufumaneni ngakumbi iiarhente zokulawula izityalo kunye nophuhliso lwazo njengee-biopesticides ezingabizi, ezisebenzayo, nezinobuhlobo nokusingqongileyo.
Iiseti zedatha eziveliswe kolu phononongo kunye nohlalutyo oluvela kuzo ziyafumaneka kumbhali ohambelanayo xa kucelwe ngokufanelekileyo. Ekupheleni kophando, zonke izixhobo ezisetyenzisiweyo kolu phando (izinambuzane kunye nomgubo wembewu) zatshatyalaliswa.
Ixesha leposi: Julayi-29-2024