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Umsebenzi webhayoloji wembewu yekhaphetshu ngumgubo kunye neekhompawundi zayo njengesibulala-mbumbulu esichasene neengcongconi

Ukuze ngempumeleloukulawula iingcongconikunye nokunciphisa izehlo zezifo abaziphetheyo, iindlela ezinobuchule, ezizinzileyo nezinobungani nokusingqongileyo endaweni yezibulali-zinambuzane zemichiza ziyafuneka.Sivavanye ukutya kwembewu kwi-Brassicaceae ethile (intsapho yaseBrassica) njengomthombo we-isothiocyanates ephuma kwisityalo eveliswa yi-enzymatic hydrolysis ye-biologically inactive glucosinolates ukuze isetyenziswe kulawulo lwe-Aedes yaseYiputa (L., 1762).Ukutya kwembewu okune-five-defatted seed (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 kunye neThlaspi arvense - iintlobo ezintathu eziphambili ze-thermal inactivation kunye ne-en iimveliso Ukumisela ubuthi (LC50) ye-allyl isothiocyanate, i-benzyl isothiocyanate kunye ne-4-hydroxybenzylisothiocyanate kwi-Aedes aegypti larvae kwi-24-yure exposure = 0.04 g / 120 ml dH2O).Ixabiso le-LC50 lemostade, i-mustard emhlophe kunye nomsila wehashe.ukutya kwembewu yi-0.05, i-0.08 kunye ne-0.05 ngokulandelanayo xa kuthelekiswa ne-allyl isothiocyanate (LC50 = 19.35 ppm) kunye ne-4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) yayinetyhefu ngakumbi kwi-larvae nge-20 ml emva kweeyure ze-20.Ezi ziphumo zihambelana nokuveliswa komgubo wembewu ye-alfalfa.Ukusebenza okuphezulu kwe-benzyl esters kuhambelana nexabiso le-LC50 elibaliweyo.Ukusebenzisa ukutya kwembewu kunokubonelela ngendlela esebenzayo yokulawula iingcongconi.ukusebenza komgubo wembewu ye-cruciferous kunye namacandelo ayo aphambili ekhemikhali ngokuchasene nemibungu yeengcongconi kwaye ibonisa indlela iikhompawundi zendalo kwi-cruciferous seed powder zingasebenza njenge-larvicide ethembisayo yokusingqongileyo yokulawula iingcongconi.
Izifo ezithwalwa ziintsholongwane ezibangelwa ziingcongconi ze-Aedes zihlala ziyingxaki enkulu yempilo yoluntu yehlabathi.Iziganeko zezifo ezibangelwa yingcongconi zisasazeka ngokwelizwe1,2,3 kwaye ziphinde zivele, okukhokelela ekuqhambukeni kwezifo ezinzima4,5,6,7.Ukusasazeka kwezifo phakathi kwabantu kunye nezilwanyana (umzekelo, i-chikungunya, i-dengue, i-Rift Valley fever, i-yellow fever kunye ne-Zika virus) yinto engazange ibonwe ngaphambili.I-Dengue fever iyodwa ibeka malunga ne-3.6 yezigidigidi zabantu emngciphekweni wosulelo kwiindawo ezitshisayo, kunye nosulelo oluqikelelwa kwi-390 yezigidi ezenzeka ngonyaka, okukhokelela ekufeni kwe-6,100-24,300 ngonyaka8.Ukuvela kwakhona kunye nokuqhambuka kwentsholongwane yeZika eMzantsi Melika kutsale umdla kwihlabathi jikelele ngenxa yomonakalo wobuchopho owenzayo kubantwana abazelwe ngabasetyhini abosulelekileyo2.U-Kremer et al 3 baqikelela ukuba uluhlu lweengcongconi ze-Aedes luza kuqhubeka lusanda kwaye ngo-2050, isiqingatha sabemi behlabathi siya kuba semngciphekweni wokosuleleka zii-arboviruses ezithwalwa ziingcongconi.
Ngaphandle kwezitofu zokugonya ezichasene nedengue kunye ne-yellow fever, izitofu zokugonya uninzi lwezifo ezithwalwa ziingcongconi azikaphuhliswa9,10,11.Izitofu zokugonya zisafumaneka ngokwezixa ezimbalwa kwaye zisetyenziswa kuphela kulingo lwezonyango.Ukulawulwa kwee-mosquito vectors usebenzisa i-synthetic insecticide ibe yinkqubo ephambili yokulawula ukusasazeka kwezifo ezibangelwa yi-mosquito12,13.Nangona i-pesticides yokwenziwa iyasebenza ekubulaleni iimiyane, ukuqhubeka nokusetyenziswa kwe-pesticides yokwenziwa kuchaphazela kakubi izinto eziphilayo ezingajoliswanga kwaye kungcolise indalo14,15,16.Okumangalisa ngakumbi yindlela yokunyuka kokuchasana kweengcongconi kwi-insecticide yekhemikhali17,18,19.Ezi ngxaki zinxulunyaniswa nezibulali-zinambuzane ziye zakhawulezisa ukufuna ezinye iindlela ezisebenzayo nezilungele ukusingqongileyo zokulawula izifo ezithwala izifo.
Izityalo ezahlukeneyo ziye zaphuhliswa njengemithombo ye-phytopesticides yokulawula izinambuzane20,21.Izinto zezityalo ziqhelene nokusingqongileyo kuba ziyakwazi ukubola kwaye zinetyhefu ephantsi okanye engabalulekanga kwizinto ezingajoliswanga njengezilwanyana ezanyisayo, iintlanzi kunye nezilwanyana ezihlala emanzini20,22.Amalungiselelo e-Herbal ayaziwa ukuba avelise iindidi ze-bioactive compounds kunye neendlela ezahlukeneyo zesenzo sokulawula ngokufanelekileyo izigaba zobomi bommiyane23,24,25,26.Iikhompawundi eziphuma kwizityalo ezifana neeoli eziyimfuneko kunye nezinye izithako zezityalo ezisebenzayo ziye zafumana ingqwalasela kwaye zavula indlela yezixhobo ezintsha zokulawula ii-mosquito vectors.I-oyile ebalulekileyo, i-monoterpenes kunye ne-sesquiterpenes zisebenza njenge-repellents, izithinteli zokutya kunye ne-ovicides27,28,29,30,31,32,33.Ioli eninzi yemifuno ibangela ukufa kweembungu zeengcongconi, i-pupae kunye nabantu abadala34,35,36, echaphazela i-nervous, breathing, endocrine kunye nezinye iinkqubo ezibalulekileyo zezinambuzane37.
Uphononongo lwakutsha nje lunike ingqiqo malunga nokusetyenziswa okunokwenzeka kwezityalo zemostade kunye nembewu yazo njengomthombo weekhompawundi ze-bioactive.Ukutya kwembewu yemostade kuye kwavavanywa njenge-biofumigant38,39,40,41 kwaye isetyenziswe njengesilungiso somhlaba ukucudisa ukhula42,43,44 kunye nolawulo lwezifo zezityalo ezifumaneka emhlabeni45,46,47,48,49,50, isondlo sezityalo.i-nematodes 41, 51, 52, 53, 54 kunye nezinambuzane 55, 56, 57, 58, 59, 60. Umsebenzi we-fungicidal wale miqulu yembewu ibangelwa kwizityalo ezikhuselayo ezibizwa ngokuba yi-isothiocyanates38,42,60.Kwizityalo, ezi zixhobo zokukhusela zigcinwa kwiiseli zezityalo ngendlela ye-non-bioactive glucosinolates.Nangona kunjalo, xa izityalo zonakaliswe ngokutya kwezinambuzane okanye ukusuleleka kwe-pathogen, i-glucosinolates i-hydrolyzed yi-myrosinase ibe yi-bioactive isothiocyanates55,61.I-Isothiocyanates ziyi-compounds eziguquguqukayo ezaziwa ngokuba ne-wide-spectrum antimicrobial and insecticidal activity, kunye nesakhiwo sabo, umsebenzi we-biological kunye nomxholo uyahluka ngokubanzi phakathi kwe-Brassicaceae iintlobo42,59,62,63.
Nangona i-isothiocyanates ephuma kumgubo wembewu yemostade isaziwa njengomsebenzi wokubulala izinambuzane, akukho datha yebhayoloji echasene ne-arthropod vectors ebalulekileyo kwezonyango.Uphononongo lwethu luvavanye umsebenzi we-larvicidal we-powder ezine zembewu ehluthiweyo ngokuchasene neengcongconi ze-Aedes.Imibungu yeAedes aegypti.Injongo yolu phononongo yayikukuvavanya ukusetyenziswa kwazo okunokubakho njengezibulali-zinambuzane ezisingqongileyo zokulawula iingcongconi.Amacandelo amathathu eekhemikhali ezinkulu zesidlo sembewu, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC), kunye ne-4-hydroxybenzylisothiocyanate (4-HBITC) nazo zavavanywa ukuvavanya umsebenzi webhayoloji wale nxalenye yemichiza kwimibungu yeengcongconi.Le yingxelo yokuqala yokuvavanya ukusebenza komgubo wembewu yekhaphetshu ezine kunye namalungu azo aphambili ekhemikhali ngokuchasene nemibungu yeengcongconi.
Iikholoni zeLebhu ze-Aedes aegypti (i-Rockefeller strain) zigcinwe kwi-26 ° C, i-70% ye-humidity ehambelanayo (RH) kunye ne-10: 14 h (L: D photoperiod).Iimazi ezitshatileyo zazihlaliswa kwiikheji zeplastiki (ubude be-11 cm kunye nobubanzi obuyi-9.5 cm) kwaye zondliwa ngenkqubo yokutyisa ibhotile kusetyenziswa igazi lenkomo elicinyiweyo (HemoStat Laboratories Inc., Dixon, CA, USA).Ukutya ngegazi kwakuqhutyelwa njengesiqhelo kusetyenziswa i-membrane ye-multi-glass feeder (i-Chemglass, i-Life Sciences LLC, i-Vineland, i-NJ, i-USA) exhunywe kwi-tube yokuhlamba amanzi ajikelezayo (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) kunye nobushushu. ukulawula 37 °C.Yolula ifilim yeParafilm M kumzantsi wegumbi leglasi nganye yokutya (indawo eyi-154 mm2).Isondlo ngasinye sasibekwe kwigridi ephezulu egquma ikheji enemazi yokukhwelana.Malunga ne-350–400 μl yegazi lenkomo yongezwa kwifaneli yeglasi yokutya kusetyenziswa iPasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) kunye neentshulube ezikhulileyo ziye zavunyelwa ukuba zikhuphe iyure enye ubuncinci.Abasetyhini abakhulelweyo banikwa isisombululo se-sucrose ye-10% kwaye bavunyelwa ukuba babeke amaqanda kwiphepha lokucoca elifumileyo elifakwe kwiikomityi ze-soufflé ezicacileyo (1.25 fl oz size, Dart Container Corp., Mason, MI, USA).ikheji ngamanzi.Beka iphepha lokucoca elinamaqanda engxoweni etywiniweyo (SC Johnsons, Racine, WI) kwaye uyigcine ku-26°C.Amaqanda aqanduselwa kwaye malunga ne-200-250 imibungu yaphakanyiswa kwiitreyi zeplastiki eziqulethe umxube we-rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) kunye nomgubo wesibindi (MP Biomedicals, LLC, Solon, OH, I-USA).kunye ne-fish fillet (TetraMin, Tetra GMPH, Meer, Germany) kumlinganiselo we-2: 1: 1.Imibungu yasemva kwexesha yesithathu isetyenziswe kwi-bioassays yethu.
Imbewu yezityalo esetyenzisiweyo kolu phononongo ifunyenwe kule mithombo ilandelayo yorhwebo kunye norhulumente: i-Brassica juncea (iGolide yemostade emdaka-ePasifiki) kunye ne-Brassica juncea (igolide yemostade emhlophe ye-Ida) evela kwi-Pacific Northwest Farmers' Cooperative, eWashington State, eMelika;(Garden Cress) evela kuKelly Seed kunye ne-Hardware Co., Peoria, IL, USA kunye neThlaspi arvense (iField Pennycress-Elisabeth) esuka kwi-USDA-ARS, Peoria, IL, USA;Akukho nanye kwiimbewu ezisetyenzisiweyo kuphononongo eyaye yanyangwa ngamayeza okubulala izitshabalalisi.Zonke izinto zembewu zicutshungulwe kwaye zisetyenziswe kwesi sifundo ngokuhambelana nemimiselo yendawo kunye nelizwe kunye nokuthotyelwa kwayo yonke imimiselo yendawo yendawo kunye nelizwe.Olu phononongo alukhange luhlolisise iintlobo zezityalo eziguqukayo.
Brassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) imbewu yacolwa yaba ngumgubo ocolekileyo kusetyenziswa iRetsch ZM200 ultracentrifugal mill (Retsch, Haan, Germany) exhotyiswe ngomnatha oyi-0.75 mm kunye neStainless i-rotor yensimbi, amazinyo angama-12, i-10,000 rpm (Itheyibhile 1).Imbewu yomhlaba engumgubo idluliselwe kwi-thimble yephepha kwaye yathanjiswa nge-hexane kwisixhobo se-Soxhlet i-24 h.I-subsample ye-mustard yentsimi ehluthiweyo yayiphathwe ukushisa kwi-100 ° C kwi-1 h ukuze i-denature myrosinase kunye nokukhusela i-hydrolysis ye-glucosinolates ukwenza i-isothiocyanates esebenzayo ye-biologically.I-powder yembewu ye-horsetail ephathwa ngobushushu (i-DFP-HT) isetyenziswe njengolawulo olubi ngokukhupha i-myrosinase.
Umxholo we-Glucosinolate we-defatted seed meal unqunywe kwi-triplicate usebenzisa i-high-performance liquid chromatography (HPLC) ngokwe-protocol epapashwe ngaphambili ye-64.Ngokufutshane, i-3 mL ye-methanol yongezwa kwisampulu ye-250 mg yomgubo wembewu ehluthiweyo.Isampuli nganye yayifakwe kwi-sonicated kwindawo yokuhlambela amanzi imizuzu engama-30 kwaye ishiywe ebumnyameni kwi-23 ° C kwiiyure ze-16.I-aliquot ye-1 mL yomaleko ophilayo yahluzwa nge-0.45 μm yokucoca kwi-autosampler.Ukusebenza kwinkqubo ye-Shimadzu HPLC (iipompo ezimbini ze-LC 20AD; i-SIL 20A autosampler; i-DGU 20As degasser; i-SPD-20A UV-VIS detector yokubeka iliso kwi-237 nm; kunye ne-CBM-20A imodyuli yebhasi yonxibelelwano), umxholo we-glucosinolate wesidlo sembewu unqunywe. kumacala amathathu .usebenzisa i-Shimadzu LC Solution software version 1.25 (Shimadzu Corporation, Columbia, MD, USA).Ikholomu yayiyikholomu yesigaba se-C18 Inertsil reverse (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA).Iimeko zesigaba sokuqala seselula zibekwe kwi-12% methanol / 88% 0.01 M tetrabutylammonium hydroxide emanzini (TBAH; Sigma-Aldrich, St. Louis, MO, USA) kunye nesantya sokuhamba kwe-1 mL / min.Emva kwesitofu se-15 μl yesampuli, iimeko zokuqala zigcinwe imizuzu engama-20, kwaye emva koko umlinganiselo we-solvent ulungelelaniswe kwi-100% ye-methanol, kunye nexesha elipheleleyo lokuhlalutya isampuli yemizuzu engama-65.I-curve esemgangathweni (i-nM / mAb esekelwe) yenziwe yi-serial dilutions ye-sinapine esanda kulungiswa, i-glucosinolate kunye nemigangatho ye-myrosin (i-Sigma-Aldrich, i-St.iiglucosinolates.Ugxininiso lwe-Glucosinolate kwiisampuli zavavanywa kwi-Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) usebenzisa i-OpenLAB CDS ChemStation version (C.01.07 SR2 [255]) exhotywe ngekholomu efanayo kunye nokusebenzisa indlela echazwe ngaphambili.Ukugxininiswa kwe-Glucosinolate kwamiselwa;inokuthelekiseka phakathi kweenkqubo ze-HPLC.
I-Allyl isothiocyanate (i-94%, i-stable) kunye ne-benzyl isothiocyanate (98%) yathengwa kwi-Fisher Scientific (i-Thermo Fisher Scientific, Waltham, MA, USA).I-4-Hydroxybenzylisothiocyanate yathengwa kwi-ChemCruz (i-Santa Cruz Biotechnology, CA, USA).Xa i-enzymatic hydrolyzed yi-myrosinase, i-glucosinolates, i-glucosinolates, kunye ne-glucosinolates yenza i-allyl isothiocyanate, i-benzyl isothiocyanate, kunye ne-4-hydroxybenzylisothiocyanate, ngokulandelanayo.
Ii-bioassays zeLabhoratri zenziwa ngokwendlela ye-Muuri et al.32 kunye nohlengahlengiso.Ukutya kwembewu okuhlanu okunamafutha aphantsi kwasetyenziswa kuphononongo: DFP, DFP-HT, IG, PG kunye neLs.Imibungu engamashumi amabini yafakwa kwi-400 mL yebeaker eneendlela ezintathu elahlwayo (VWR International, LLC, Radnor, PA, USA) equlethe i-120 mL yamanzi adiyoniyo (dH2O).Ukutya kwembewu ezisixhenxe kwavavanywa ubutyhefu bengcongconi: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 kunye ne-0.12 g seed meal/120 ml dH2O yeDFP seed meal , DFP-HT, IG kunye nePG.Uvavanyo lokuqala lwebhayoloji lubonisa ukuba umgubo wembewu ye-Ls othanjisweyo unetyhefu ngakumbi kuneminye imihluma yembewu evavanyiweyo.Ngoko ke, silungelelanise izigxina ezisixhenxe zonyango lwe-Ls seed meal kule miba ilandelayo: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, kunye ne-0.075 g / 120 mL dH2O.
Iqela lolawulo elinganyangwanga (dH20, akukho songezelelo sembewu yokutya) libandakanyiwe ukuvavanya ukufa kwezinambuzane eziqhelekileyo phantsi kweemeko zovavanyo.Uvavanyo lwebhayoloji yeToxicological kwisidlo sembewu nganye siquka iibhekile ezintathu eziphindaphindayo ezithambekele ezintathu (imibungu engama-20 yasemva kwesithathu kwibeaker nganye), kwiibhotile ezili-108 zizonke.Izikhongozeli eziphathwayo zigcinwe kwindawo yokushisa (i-20-21 ° C) kwaye ukufa kombungu kwabhalwa ngexesha le-24 kunye neeyure ze-72 zokuvezwa ngokuqhubekayo kugxininiso lonyango.Ukuba umzimba wengcongconi kunye ne-appendages ayishukumi xa ihlatywa okanye ichukunyiswe nge-spatula yensimbi ebhityileyo, i-larvae yengcongconi ithathwa njengefile.Imibungu efileyo idla ngokuhlala ingashukumi kwindawo ehlala irholi okanye i-ventral ezantsi kwesikhongozeli okanye kumphezulu wamanzi.Uvavanyo luphindwe kathathu ngeentsuku ezahlukeneyo kusetyenziswa amaqela ahlukeneyo emibungu, kwi-180 iyonke imibungu evezwe kugxininiso lonyango ngalunye.
Ubutyhefu be-AITC, BITC, kunye ne-4-HBITC kwimibungu yeengcongconi yavavanywa kusetyenziswa inkqubo efanayo yovavanyo lwebhayoloji kodwa ngonyango olwahlukileyo.Lungiselela izisombululo zesitokhwe se-100,000 ppm kwikhemikhali nganye ngokudibanisa i-100 µL yekhemikhali kwi-900 µL ye-ethanol epheleleyo kwi-2-mL ye-centrifuge ityhubhu kwaye ungcangcazele imizuzwana engama-30 ukuze uxube ngokucokisekileyo.Ugxininiso lonyango luye lwamiselwa ngokusekelwe kuvavanyo lwethu lokuqala lwe-bioassays, olufumene i-BITC inetyhefu kakhulu kune-AITC kunye ne-4-HBITC.Ukumisela ubutyhefu, ugxininiso lwe-5 lwe-BITC (1, 3, 6, 9 kunye ne-12 ppm), i-7 yoxinzelelo lwe-AITC (5, 10, 15, 20, 25, 30 kunye ne-35 ppm) kunye ne-6 yoxinzelelo lwe-4-HBITC (15). , 15, 20, 25, 30 kunye ne-35 ppm).30, 45, 60, 75 kunye ne-90 ppm).Unyango lolawulo lwatofwa nge-108 μL ye-ethanol epheleleyo, elingana nomthamo omkhulu wonyango lwekhemikhali.Uvavanyo lwe-Bioassays lwaphindwa njengangentla apha, luveza umbungu opheleleyo we-180 ngoxinaniso lonyango.Ukufa kombungu kushicilelwe kuxinaniso ngalunye lwe-AITC, BITC, kunye ne-4-HBITC emva kwe-24 h yokuvezwa okuqhubekayo.
Uhlalutyo lweprobit yedatha yokufa enxulumene nedosi ye-65 yenziwa kusetyenziswa isoftware yePolo (i-Polo Plus, iLeOra Software, inguqulo 1.0) ukubala i-50% yoxinaniso olubulalayo (LC50), i-90% yoxinaniso olubulalayo (LC90), i-slope, i-coefficient yedosi ebulalayo, kunye ne-95 % ugxininiso olubulalayo.ngokusekwe kumaxesha okuzithemba kwimilinganiselo yedosi ebulalayo yoxinaniso oluguqulweyo lwelogi kunye neegophe zokufa kwedosi.Idatha yokufa isekelwe kwidatha ephindwe kabini ye-larvae ye-180 evezwe kugxininiso lonyango ngalunye.Uhlalutyo olunokwenzeka lwenziwa ngokwahlukileyo kwisidlo ngasinye sembewu kunye necandelo ngalinye lekhemikhali.Ngokusekelwe kwi-95% yokuzithemba kwexesha lomlinganiselo wedosi ebulalayo, ubutyhefu besidlo sembewu kunye neekhemikhali kwimibungu yeengcongconi yayithathwa njengento eyahlukileyo kakhulu, ngoko ke ixesha lokuzithemba eliqulethe ixabiso le-1 alizange lihluke kakhulu, P = 0.0566.
Iziphumo ze-HPLC zokumisela i-glucosinolates ezinkulu kwi-defatted seed flours DFP, IG, PG kunye ne-Ls zidweliswe kwiThebhile 1. I-glucosinolates enkulu kwimbewu yembewu evavanyiweyo yahluka ngaphandle kwe-DFP kunye ne-PG, zombini iqulethe i-myrosinase glucosinolates.Umxholo we-myrosinin kwi-PG wawuphezulu kune-DFP, i-33.3 ± 1.5 kunye ne-26.5 ± 0.9 mg / g, ngokulandelanayo.I-Ls powder powder iqulethe i-36.6 ± 1.2 mg / g glucoglycone, kanti i-IG yembewu ye powder iqulethe i-38.0 ± 0.5 mg / g i-sinapine.
Imibungu ye-Ae.Iingcongconi ze-Aedes aegypti zabulawa xa zinyangwa ngesidlo sembewu encitshisiweyo, nangona ukusebenza konyango kwahluka ngokuxhomekeke kwiintlobo zezityalo.Kuphela i-DFP-NT yayingenayo ityhefu kwimibungu yeengcongconi emva kwe-24 kunye ne-72 h yokuvezwa (Itheyibhile 2).Ubuthi be-powder esebenzayo yembewu bunyuke ngokunyuka koxinzelelo (Umfanekiso 1A, B).Ubuthi bokutya kwembewu kwimibungu yeengcongconi yahluka kakhulu ngokusekwe kwi-95% CI yomlinganiselo wedosi ebulalayo yexabiso le-LC50 kwiiyure ezingama-24 kunye novavanyo lweeyure ezingama-72 (Itheyibhile 3).Emva kweeyure ezingama-24, isiphumo esinetyhefu se-Ls seed meal sasisikhulu kunezinye iindlela zonyango zembewu, kunye nomsebenzi ophezulu kunye ne-toxicity ephezulu kwi-larvae (LC50 = 0.04 g / 120 ml dH2O).I-Larvae yayingaphantsi kwe-DFP kwiiyure ze-24 xa kuthelekiswa ne-IG, i-Ls kunye ne-PG yonyango lwembewu yembewu, kunye nexabiso le-LC50 ye-0.115, 0.04 kunye ne-0.08 g / 120 ml dH2O ngokulandelelanayo, eyayiphezulu ngokwezibalo kunexabiso le-LC50.0.211 g / 120 ml dH2O (Itheyibhile 3).Amanani e-LC90 ye-DFP, IG, PG kunye ne-Ls yayiyi-0.376, 0.275, 0.137 kunye ne-0.074 g / 120 ml dH2O, ngokulandelanayo (Itheyibhile 2).Uxinzelelo oluphezulu lwe-DPP lwaluyi-0.12 g / 120 ml dH2O.Emva kweeyure ezingama-24 zovavanyo, i-avareji yokufa kwemibungu yayiyi-12% kuphela, ngelixa i-avareji yokufa kwe-IG kunye ne-PG larvae ifikelele kwi-51% kunye ne-82%, ngokulandelanayo.Emva kweeyure ezingama-24 zovavanyo, umyinge wokufa kombungu weyona ndawo iphezulu yonyango lwembewu ye-Ls yonyango (0.075 g/120 ml dH2O) yayingama-99% (umzobo 1A).
Iijika zokufa ziqikelelwe kwimpendulo yethamo (iProbit) ye-Ae.Iimbungu zaseYiputa (i-3rd instar larvae) kwi-concentration yembewu yembewu iiyure ezingama-24 (A) kunye neeyure ze-72 (B) emva kwonyango.Umgca onamachaphaza umele i-LC50 yonyango lokutya kwembewu.I-DFP Thlaspi arvense, i-DFP-HT Ubushushu obungasetyenziswanga i-Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Kuvavanyo lweeyure ezingama-72, amaxabiso e-LC50 e-DFP, i-IG kunye ne-PG yembewu yokutya yayiyi-0.111, 0.085 kunye ne-0.051 g/120 ml dH2O, ngokulandelanayo.Phantse yonke imibungu evezwe kwisidlo sembewu ye-Ls yafa emva kwe-72 h yokuvezwa, ngoko ke idatha yokufa yayingahambelani nohlalutyo lweProbit.Xa kuthelekiswa nolunye uhlobo lwesidlo sembewu, imibungu yayingakhathali kangako kunyango lwembewu ye-DFP kwaye yayinamaxabiso aphezulu e-LC50 (Itheyibhile 2 kunye ne-3).Emva kweeyure ezingama-72, amaxabiso e-LC50 e-DFP, i-IG kunye ne-PG yonyango lwembewu yokutya aqikelelwa ukuba yi-0.111, 0.085 kunye ne-0.05 g/120 ml dH2O, ngokulandelelanayo.Emva kweeyure ze-72 zovavanyo, ixabiso le-LC90 ye-DFP, i-IG kunye ne-PG yembewu yomgubo yayiyi-0.215, 0.254 kunye ne-0.138 g / 120 ml dH2O, ngokulandelanayo.Emva kweeyure ze-72 zovavanyo, umyinge wokufa kombungu we-DFP, i-IG kunye ne-PG yonyango lwembewu yokutya kwi-concentration ephezulu ye-0.12 g / 120 ml dH2O yayingu-58%, 66% kunye ne-96%, ngokulandelanayo (Fig. 1B).Emva kovavanyo lweeyure ezingama-72, ukutya kwembewu ye-PG kwafunyaniswa kunetyhefu ngakumbi kunokutya kwembewu ye-IG kunye ne-DFP.
I-synthetic isothiocyanates, i-allyl isothiocyanate (AITC), i-benzyl isothiocyanate (BITC) kunye ne-4-hydroxybenzylisothiocyanate (4-HBITC) inokubulala ngokufanelekileyo imibungu yeengcongconi.Kwiiyure ezingama-24 emva konyango, i-BITC yayinetyhefu ngakumbi kwimibungu enexabiso 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 ine-toxicity ephantsi kunye nexabiso eliphezulu le-LC50.Kukho iiyantlukwano ezibalulekileyo kwityhefu yengcongconi yombungu we-isothiocyanates ezimbini ezinkulu (Ls kunye ne-PG) kwesona sidlo sinamandla sembewu.Ubuthi obusekwe kumlinganiselo wethamo elibulalayo lamaxabiso eLC50 phakathi kwe-AITC, BITC, kunye ne-4-HBITC ibonise umahluko wezibalo kangangokuba i-95% CI ye-LC50 yedosi ebulalayo ayiquki ixabiso le-1 (P = 0.05, Itheyibhile 4).Ugxininiso oluphezulu lwe-BITC kunye ne-AITC kuqikelelwa ukubulala i-100% yombungu ovavanyiweyo (Umfanekiso 2).
Iijika zokufa ziqikelelwe kwimpendulo yethamo (iProbit) ye-Ae.Iiyure ezingama-24 emva kokonyango, i-larvae yaseYiputa (i-3rd instar larvae) yafikelela kwi-synthetic isothiocyanate concentrations.Umgca onamachaphaza umele i-LC50 yonyango lwe-isothiocyanate.I-Benzyl isothiocyanate BITC, i-allyl isothiocyanate AITC kunye ne-4-HBITC.
Ukusetyenziswa kwezibulali-zinambuzane zezityalo njengee-arhente zokulawula iingcongconi kudala kuphononongwa.Izityalo ezininzi zivelisa imichiza yendalo enomsebenzi wokubulala izinambuzane37.Iikhompawundi zabo ze-bioactive zibonelela ngenye indlela enomtsalane kwizitshabalalisi zokwenziwa ezinamandla okulawula izinambuzane ezitshabalalisayo, kuquka iingcongconi.
Izityalo zemostade zikhuliswa njengesityalo sembewu yazo, zisetyenziswa njengesinongo kunye nomthombo weoli.Xa ioli yemostade itsalwa kwiimbewu okanye xa isardadi itsalwa ukuze isetyenziswe njenge-biofuel, i-69 imveliso ephuma kwimveliso yi-defatted seed meal.Esi sidlo sembewu sigcina ezininzi zendalo ze-biochemical components kunye ne-hydrolytic enzymes.Ityhefu yesi sidlo sembewu ibangelwa ukuveliswa kwe-isothiocyanates55,60,61.I-Isothiocyanates yenziwa yi-hydrolysis ye-glucosinolates nge-enzyme myrosinase ngexesha lokuxutywa kwembewu yokutya38,55,70 kwaye iyaziwa ngokuba ne-fungicidal, bactericidal, nemacidal kunye ne-insecticidal effects, kunye nezinye iipropathi ezibandakanya imiphumo ye-chemical sensory kunye ne-chemotherapeutic properties61,62, 70.Uphononongo oluninzi lubonise ukuba izityalo zemostade kunye nembewu yembewu zisebenza ngokufanelekileyo njengezinto ezitshabalalisa umhlaba kunye nezitshabalalisi zokutya ezigciniweyo57,59,71,72.Kolu phononongo, sivavanye ubutyhefu besidlo sembewu ezine kunye neemveliso zayo ezintathu ze-bioactive AITC, BITC, kunye ne-4-HBITC kwi-Aedes mosquito larvae.Aedes yaseYiputa.Ukongeza ukutya kwembewu ngokuthe ngqo kumanzi aqulethe imibungu yeengcongconi kulindeleke ukuba kusebenze iinkqubo ze-enzymatic ezivelisa i-isothiocyanates ezinetyhefu kwimibungu yeengcongconi.Olu tshintsho lwebhayoloji lubonakaliswe ngokuyinxenye ngumsebenzi wokutshabalalisa imibungu kwisidlo sembewu kunye nokulahleka komsebenzi wokubulala izitshabalalisi xa umgubo wembewu yemostade encinci unyangwa bubushushu phambi kokusetyenziswa.Unyango lokushisa kulindeleke ukuba lutshabalalise i-enzymes ye-hydrolytic eyenza i-glucosinolates isebenze, ngaloo ndlela ikhusela ukubunjwa kwe-isothiocyanates ye-bioactive.Olu luphononongo lokuqala lokuqinisekisa iimpawu zokubulala izinambuzane zembewu yekhaphetshu engumgubo kwiingcongconi kwindawo ehlala emanzini.
Phakathi kweepowder zembewu ezivavanyiweyo, i-watercress seed powder (Ls) yayiyityhefu kakhulu, ebangela ukufa okuphezulu kwe-Aedes albopictus.Imibungu yeAedes aegypti yayisenziwa ngokuqhubekayo iiyure ezingama-24.I-powders yembewu emithathu eseleyo (PG, IG kunye ne-DFP) yayinomsebenzi ocothayo kwaye isabangela ukufa okuphawulekayo emva kweeyure ze-72 zonyango oluqhubekayo.Kuphela kwesidlo sembewu ye-Ls esinezixa ezibalulekileyo ze-glucosinolates, ngelixa i-PG kunye ne-DFP iqulethe i-myrosinase kunye ne-IG iqulethe i-glucosinolate njengeyona glucosinolate enkulu (Itheyibhile 1).I-Glucotropaeolin i-hydrolyzed kwi-BITC kunye ne-sinalbine i-hydrolyzed kwi-4-HBITC61,62.Iziphumo zethu ze-bioassay zibonisa ukuba zombini ukutya kwembewu ye-Ls kunye ne-synthetic BITC ziyityhefu kakhulu kwimibungu yeengcongconi.Inxalenye ephambili ye-PG kunye ne-DFP yembewu yokutya yi-myrosinase glucosinolate, e-hydrolyzed kwi-AITC.I-AITC iyasebenza ekubulaleni imibungu yeengcongconi ngexabiso le-LC50 le-19.35 ppm.Xa kuthelekiswa ne-AITC kunye ne-BITC, i-4-HBITC isothiocyanate yeyona tyhefu incinci kwimibungu.Nangona i-AITC inetyhefu encinci kune-BITC, ixabiso labo le-LC50 lingaphantsi kuneoli eninzi ebalulekileyo evavanyiweyo kwi-larvae yengcongconi32,73,74,75.
Imbewu yethu ye-cruciferous powder yokusetyenziswa ngokuchasene nezibungu zeengcongconi iqulethe enye i-glucosinolate enkulu, ibalwa ngaphezu kwe-98-99% ye-glucosinolates iyonke njengoko inqunywe yi-HPLC.Ukulandelela iimali zezinye i-glucosinolates zifunyenwe, kodwa amanqanaba abo ayengaphantsi kwe-0.3% ye-glucosinolates iyonke.I-Watercress (i-L. sativum) i-powder yembewu iqulethe i-glucosinolates yesibini (i-sinigrin), kodwa umlinganiselo wabo ngu-1% we-glucosinolates iyonke, kwaye umxholo wabo usengabalulekanga (malunga ne-0.4 mg / g powder powder).Nangona i-PG kunye ne-DFP ziqulethe eyona glucosinolate (myrosin), umsebenzi we-larvicidal wesidlo sabo sembewu wahluka kakhulu ngenxa yexabiso labo le-LC50.Iyahluka kwityhefu kwi powdery mildew.Ukuvela kwemibungu ye-Aedes aegypti kusenokuba ngenxa yomahluko kumsebenzi we-myrosinase okanye uzinzo phakathi kokutya kwembewu ezimbini.Umsebenzi we-Myrosinase udlala indima ebalulekileyo kwi-bioavailability yeemveliso ze-hydrolysis ezifana ne-isothiocyanates kwizityalo ze-Brassicaceae76.Iingxelo zangaphambili zikaPocock et al.77 kunye noWilkinson et al.78 baye babonisa ukuba utshintsho kumsebenzi we-myrosinase kunye nokuzinza kunokudibaniswa nezinto zofuzo kunye nokusingqongileyo.
Umxholo we-isothiocyanate olindelweyo we-bioactive ubalwe ngokusekelwe kumaxabiso e-LC50 okutya kwembewu nganye kwiiyure ezingama-24 kunye neeyure ezingama-72 (Itheyibhile 5) xa kuthelekiswa nezicelo zeekhemikhali ezihambelanayo.Emva kweeyure ze-24, i-isothiocyanates kwisidlo sembewu yayinetyhefu ngaphezu kweekhompawundi ezicocekileyo.Amaxabiso e-LC50 abalwe ngokusekwe kumalungu kwisigidi (ppm) zonyango lwembewu ye-isothiocyanate ayengaphantsi kumaxabiso e-LC50 ye-BITC, AITC, kunye ne-4-HBITC yezicelo.Siye saqwalasela imibungu isitya iipellets zokutya zembewu (Umfanekiso 3A).Ngenxa yoko, imibungu inokufumana utyhileko oluthe kratya kwi-isothiocyanates enetyhefu ngokuginya iipellets zembewu.Oku kwabonakala kakhulu kwi-IG kunye ne-PG yonyango lokutya kwembewu kwi-24-h exposure, apho i-LC50 igxininiso yayiyi-75% kunye ne-72% ephantsi kune-AITC ecocekileyo kunye ne-4-HBITC yonyango, ngokulandelanayo.Unyango lwe-Ls kunye ne-DFP lwalunetyhefu ngakumbi kune-isothiocyanate esulungekileyo, enexabiso le-LC50 24% kunye ne-41% esezantsi, ngokulandelanayo.Imibungu kunyango lolawulo pupated ngempumelelo (Fig. 3B), ngelixa uninzi imibungu kunyango imbewu ukutya akazange pupate kunye nophuhliso umbungu kwabambezeleka kakhulu (Fig. 3B, D).Kwi-Spodopteralitura, i-isothiocyanates inxulunyaniswa nokuncipha kokukhula kunye nokulibaziseka kophuhliso79.
Imibungu ye-Ae.Iimiyane ze-Aedes aegypti zazihlala zibonakaliswe kwi-Brassica imbewu powder kwiiyure ze-24-72.(A) Imibungu efileyo enamasuntswana omgubo wembewu kumalungu omlomo (ebiyelwe);(B) Ulawulo lonyango (dH20 ngaphandle kwesidlo sembewu eyongezelelweyo) lubonisa ukuba imibungu ikhula ngokuqhelekileyo kwaye iqala ukuhluma emva kweeyure ezingama-72 (C, D) Imibungu enyangwa ngokutya kwembewu;isidlo sembewu sibonise umahluko ekuphuhliseni kwaye asizange sikhule.
Asizange sifunde indlela yemiphumo eyityhefu ye-isothiocyanates kwimibungu yeengcongconi.Nangona kunjalo, izifundo zangaphambili kwiintuthwane ezibomvu zomlilo (i-Solenopsis invicta) zibonise ukuba inhibition ye-glutathione S-transferase (GST) kunye ne-esterase (EST) yindlela ephambili ye-isothiocyanate bioactivity, kunye ne-AITC, nakwimisebenzi ephantsi, inokuthintela umsebenzi we-GST. .iimbovane zomlilo ezivela kumazwe angaphandle ezibomvu kwiindawo eziphantsi.Ithamo yi-0.5 µg/ml80.Ngokwahlukileyo, i-AITC inqanda i-acetylcholinesterase kwi-weevils yabantu abadala (Sitophilus zeamais)81.Izifundo ezifanayo kufuneka zenziwe ukucacisa indlela yokusebenza kwe-isothiocyanate kwimibungu yeengcongconi.
Sisebenzisa unyango lwe-DFP olungasebenzi ubushushu ukuxhasa isindululo sokuba i-hydrolysis ye-plant glucosinolates ukwenza i-isothiocyanates esebenzayo isebenza njengendlela yokulawula imibungu yeengcongconi ngesidlo sembewu yemostade.Ukutya kwembewu ye-DFP-HT kwakungekho tyhefu kumazinga okufaka isicelo avavanyiweyo.Lafarga et al.I-82 ibike ukuba i-glucosinolates ivakalelwa ukuthotywa kwamaqondo aphezulu.Unyango olushushu lukwalindeleke ukuba lukhuphe i-enzyme ye-myrosinase kwisidlo sembewu kwaye ithintele i-hydrolysis ye-glucosinolates ukwenza i-isothiocyanates esebenzayo.Oku kwaqinisekiswa nguOkunade et al.I-75 ibonise ukuba i-myrosinase inobunzulu bobushushu, ebonisa ukuba umsebenzi we-myrosinase wawungasebenzi ngokupheleleyo xa i-mustard, i-mustard emnyama, kunye nembewu ye-bloodroot yavezwa kubushushu obungaphezulu kwe-80 °.C. Ezi ndlela zinokubangela ukulahlekelwa ngumsebenzi wokubulala izinambuzane kwisidlo sembewu ye-DFP ephethwe bubushushu.
Ke, umgubo wembewu yemostade kunye ne-isothiocyanates zayo ezinkulu ezintathu ziyityhefu kwimibungu yeengcongconi.Ukunika lo mahluko phakathi kwesidlo sembewu kunye nonyango lwemichiza, ukusetyenziswa kwesidlo sembewu kunokuba yindlela esebenzayo yokulawula iingcongconi.Kukho imfuneko yokuchonga iifomyula ezifanelekileyo kunye neenkqubo zokuhanjiswa ezisebenzayo zokuphucula ukusebenza kunye nokuzinza kokusetyenziswa kwe powders yembewu.Iziphumo zethu zibonisa ukusetyenziswa okunokubakho kwembewu yemostade njengenye indlela yokubulala izitshabalalisi zokwenziwa.Obu buchwepheshe bunokuba sisixhobo esitsha sokulawula iintsholongwane zeengcongconi.Ngenxa yokuba imibungu yeengcongconi ikhula kakuhle kwindawo ehlala emanzini kwaye imbewu yokutya iglucosinolates iguqulwa nge-enzymatically ibe yi-isothiocyanates esebenzayo xa i-hydration, ukusetyenziswa komgubo wembewu yemostade emanzini azaliswe ziingcongconi kunika amandla amakhulu okulawula.Nangona umsebenzi we-larvicidal we-isothiocyanates uhluka (BITC> AITC> 4-HBITC), uphando olongezelelweyo luyafuneka ukuze kuqinisekiswe ukuba ukudibanisa ukutya kwembewu kunye ne-glucosinolates ezininzi nge-synergistically kwandisa ubuthi.Olu luphononongo lokuqala lokubonisa iziphumo ze-insecticidal ze-defatted cruciferous seed meal kunye ne-isothiocyanates ezintathu ze-bioactive kwiingcongconi.Iziphumo zolu phononongo zaphula umhlaba omtsha ngokubonisa ukuba umgubo wembewu yekhaphetshu encitshisiweyo, imveliso ye-oyile etsalwa kwiimbewu, unokusebenza njenge-ejenti ethembisayo ye-larvicidal kulawulo lweengcongconi.Olu lwazi lunokunceda ngakumbi ukufunyaniswa kwee-arhente zolawulo lwebhayoloji yezityalo kunye nophuhliso lwazo njengezibulali-zinambuzane ezishibhileyo, ezisebenzisekayo, nezingenabungozi kokusingqongileyo.
Iiseti zedatha ezenzelwe olu phononongo kunye nohlalutyo olubangelwayo luyafumaneka kumbhali ohambelanayo ngesicelo esifanelekileyo.Ekupheleni kophononongo, zonke izinto ezisetyenzisiweyo kuphononongo (izinambuzane kunye nesidlo sembewu) zatshatyalaliswa.


Ixesha lokuposa: Jul-29-2024