Kev ua haujlwm ntawm cov hmoov noob zaub qhwv thiab nws cov tshuaj ua ib qho tshuaj tua kab uas zoo rau ib puag ncig

Ua kom zootswj cov yoov tshaj cumthiab txo qhov muaj mob uas lawv nqa, kev xaiv cov tshuaj tua kab uas zoo, ruaj khov thiab zoo rau ib puag ncig yog qhov xav tau. Peb tau soj ntsuam cov noob zaub mov los ntawm qee yam Brassicaceae (tsev neeg Brassica) ua qhov chaw ntawm cov isothiocyanates los ntawm cov nroj tsuag uas tsim los ntawm enzymatic hydrolysis ntawm biologically inactive glucosinolates rau kev siv hauv kev tswj hwm ntawm Egyptian Aedes (L., 1762). Pluas noob tsib-defatted (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 thiab Thlaspi arvense - peb hom tseem ceeb ntawm thermal inactivation thiab enzymatic degradation Cov khoom siv tshuaj lom neeg Txhawm rau txiav txim siab txog kev lom (LC50) ntawm allyl isothiocyanate, benzyl isothiocyanate thiab 4-hydroxybenzylisothiocyanate rau Aedes aegypti larvae ntawm 24-teev raug = 0.04 g / 120 ml dH2O). LC50 tus nqi rau mustard, mustard dawb thiab horsetail. Cov noob hmoov yog 0.05, 0.08 thiab 0.05 feem piv rau allyl isothiocyanate (LC50 = 19.35 ppm) thiab 4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) muaj tshuaj lom ntau dua rau cov kab me me ntawm 24 teev tom qab kev kho mob dua li 0.1 g / 120 ml dH2O feem. Cov txiaj ntsig no yog sib xws nrog kev tsim cov noob hmoov alfalfa. Kev ua haujlwm zoo dua ntawm benzyl esters sib raug rau cov nqi LC50 xam. Kev siv noob hmoov tuaj yeem muab txoj hauv kev zoo ntawm kev tswj yoov tshaj cum. qhov ua tau zoo ntawm cov hmoov noob cruciferous thiab nws cov tshuaj lom neeg tseem ceeb tiv thaiv cov kab me me thiab qhia tau tias cov tshuaj ntuj hauv cov hmoov noob cruciferous tuaj yeem ua haujlwm ua cov kab me me zoo rau ib puag ncig rau kev tswj yoov tshaj cum.
Cov kab mob uas kis los ntawm cov yoov tshaj cum Aedes tseem yog ib qho teeb meem loj rau kev noj qab haus huv thoob ntiaj teb. Qhov tshwm sim ntawm cov kab mob uas kis los ntawm yoov tshaj cum kis mus rau ntau qhov chaw1,2,3 thiab rov tshwm sim dua, ua rau muaj kev sib kis ntawm cov kab mob hnyav4,5,6,7. Kev sib kis ntawm cov kab mob ntawm tib neeg thiab tsiaj txhu (piv txwv li, chikungunya, dengue, Rift Valley fever, yellow fever thiab Zika virus) tsis tau muaj dua li. Dengue fever ib leeg ua rau kwv yees li 3.6 billion tus neeg muaj feem yuav kis tus kab mob hauv thaj chaw sov, nrog rau kwv yees li 390 lab tus kab mob tshwm sim txhua xyoo, ua rau muaj 6,100–24,300 tus neeg tuag txhua xyoo8. Qhov rov tshwm sim thiab kev sib kis ntawm tus kab mob Zika hauv South America tau nyiam thoob ntiaj teb vim yog kev puas tsuaj rau lub hlwb uas nws ua rau cov menyuam yaus yug los ntawm cov poj niam muaj kab mob2. Kremer et al 3 kwv yees tias qhov ntau ntawm cov yoov tshaj cum Aedes yuav txuas ntxiv nthuav dav thiab tias los ntawm xyoo 2050, ib nrab ntawm cov pej xeem hauv ntiaj teb yuav muaj feem yuav kis tus kab mob los ntawm cov yoov tshaj cum arboviruses.
Tsuas yog cov tshuaj tiv thaiv kab mob dengue thiab mob ua npaws daj uas nyuam qhuav tsim tawm xwb, cov tshuaj tiv thaiv kab mob feem ntau uas kis los ntawm yoov tshaj cum tseem tsis tau tsim tawm9,10,11. Cov tshuaj tiv thaiv kab mob tseem muaj tsawg thiab tsuas yog siv rau hauv kev sim tshuaj xwb. Kev tswj cov kab mob uas kis los ntawm yoov tshaj cum siv cov tshuaj tua kab yog ib qho tseem ceeb los tswj kev kis tus kab mob uas kis los ntawm yoov tshaj cum12,13. Txawm hais tias cov tshuaj tua kab ua tau zoo hauv kev tua yoov tshaj cum, kev siv cov tshuaj tua kab ua txuas ntxiv mus cuam tshuam tsis zoo rau cov kab mob uas tsis yog lub hom phiaj thiab ua rau ib puag ncig puas tsuaj14,15,16. Qhov txaus ntshai tshaj plaws yog qhov sib txawv ntawm kev tiv thaiv yoov tshaj cum rau cov tshuaj tua kab17,18,19. Cov teeb meem no cuam tshuam nrog cov tshuaj tua kab tau ua rau kev tshawb nrhiav lwm txoj hauv kev zoo thiab zoo rau ib puag ncig los tswj cov kab mob.
Muaj ntau hom nroj tsuag tau tsim los ua cov tshuaj tua kab tsuag rau kev tswj kab tsuag20,21. Cov tshuaj nroj tsuag feem ntau yog cov phooj ywg ib puag ncig vim tias lawv lwj tau thiab muaj kev lom tsawg lossis tsis muaj kev phom sij rau cov kab mob uas tsis yog lub hom phiaj xws li tsiaj txhu, ntses thiab amphibians20,22. Cov tshuaj ntsuab tau paub tias tsim ntau yam tshuaj lom neeg nrog ntau yam txheej txheem ua haujlwm kom tswj tau cov kab mob sib txawv ntawm lub neej23,24,25,26. Cov tshuaj los ntawm cov nroj tsuag xws li cov roj tseem ceeb thiab lwm yam khoom xyaw ntawm cov nroj tsuag tau txais kev saib xyuas thiab tau tsim txoj hauv kev rau cov cuab yeej tshiab los tswj cov kab mob yoov tshaj cum. Cov roj tseem ceeb, monoterpenes thiab sesquiterpenes ua haujlwm ua cov tshuaj tua kab, cov tshuaj tiv thaiv kev noj zaub mov thiab cov tshuaj tua kab tsuag27,28,29,30,31,32,33. Ntau cov roj zaub ua rau cov kab mob yoov tshaj cum tuag, cov menyuam dev thiab cov neeg laus34,35,36, cuam tshuam rau lub paj hlwb, kev ua pa, endocrine thiab lwm yam kab mob tseem ceeb37.
Cov kev tshawb fawb tsis ntev los no tau muab kev nkag siab txog kev siv cov nroj tsuag mustard thiab lawv cov noob ua ib qho chaw ntawm cov tshuaj lom neeg. Cov hmoov noob mustard tau raug sim ua biofumigant38,39,40,41 thiab siv ua kev hloov pauv av rau kev tswj cov nroj tsuag42,43,44 thiab tswj cov kab mob cog hauv av45,46,47,48,49,50, kev noj zaub mov ntawm cov nroj tsuag. nematodes 41,51, 52, 53, 54 thiab kab tsuag 55, 56, 57, 58, 59, 60. Cov haujlwm fungicidal ntawm cov hmoov noob no yog vim cov tshuaj tiv thaiv cov nroj tsuag hu ua isothiocyanates38,42,60. Hauv cov nroj tsuag, cov tshuaj tiv thaiv no khaws cia rau hauv cov hlwb ntawm cov nroj tsuag hauv daim ntawv ntawm cov glucosinolates uas tsis yog bioactive. Txawm li cas los xij, thaum cov nroj tsuag raug puas tsuaj los ntawm kev noj kab lossis kev kis kab mob, glucosinolates raug hydrolyzed los ntawm myrosinase rau hauv bioactive isothiocyanates55,61. Isothiocyanates yog cov tshuaj sib xyaw ua ke uas paub tias muaj cov tshuaj tua kab mob thiab tua kab dav dav, thiab lawv cov qauv, kev ua haujlwm ntawm lub cev thiab cov ntsiab lus sib txawv ntawm cov hom Brassicaceae42,59,62,63.
Txawm hais tias cov isothiocyanates uas tau los ntawm cov noob mustard hmoov paub tias muaj kev ua haujlwm tua kab, cov ntaub ntawv ntawm kev ua haujlwm ntawm cov kab mob arthropod tseem ceeb rau kev kho mob tseem tsis muaj. Peb txoj kev tshawb fawb tau tshuaj xyuas qhov kev ua haujlwm tua kab mob ntawm plaub cov hmoov noob uas tau tshem tawm cov rog tawm tsam Aedes yoov tshaj cum. Cov kab ntawm Aedes aegypti. Lub hom phiaj ntawm txoj kev tshawb fawb yog los ntsuam xyuas lawv qhov kev siv tau ua cov tshuaj tua kab mob zoo rau ib puag ncig rau kev tswj kab. Peb cov khoom siv tshuaj lom neeg tseem ceeb ntawm cov noob hmoov, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC), thiab 4-hydroxybenzylisothiocyanate (4-HBITC) kuj tau raug sim los sim qhov kev ua haujlwm ntawm cov tshuaj lom neeg no rau cov kab mob yoov tshaj cum. Qhov no yog thawj daim ntawv tshaj tawm los ntsuam xyuas qhov ua tau zoo ntawm plaub cov hmoov noob zaub qhwv thiab lawv cov khoom siv tshuaj lom neeg tseem ceeb tawm tsam cov kab mob yoov tshaj cum.
Cov kab Aedes aegypti (Rockefeller strain) hauv chav kuaj tau khaws cia rau ntawm 26°C, 70% qhov av noo (RH) thiab 10:14 teev (L:D photoperiod). Cov poj niam uas tau sib yuav tau muab tso rau hauv cov tawb yas (qhov siab 11 cm thiab txoj kab uas hla 9.5 cm) thiab pub los ntawm lub raj mis siv cov ntshav nyuj citrated (HemoStat Laboratories Inc., Dixon, CA, USA). Kev pub ntshav tau ua raws li niaj zaus siv lub membrane multi-glass feeder (Chemglass, Life Sciences LLC, Vineland, NJ, USA) txuas nrog lub raj dej da dej (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) nrog kev tswj qhov kub thiab txias 37°C. Ncab ib daim zaj duab xis ntawm Parafilm M rau hauv qab ntawm txhua lub tais pub iav (cheeb tsam 154 mm2). Txhua lub tais pub tau muab tso rau ntawm daim phiaj saum toj kawg nkaus uas npog lub tawb uas muaj poj niam sib yuav. Kwv yees li ntawm 350–400 μl ntawm cov ntshav nyuj tau ntxiv rau hauv lub raj mis iav siv lub raj mis Pasteur (Fisherbrand, Fisher Scientific, Waltham, MA, USA) thiab cov kab laus tau cia kom ntws tsawg kawg ib teev. Cov poj niam cev xeeb tub tau muab 10% sucrose kua thiab cia lawv tso qe rau ntawm daim ntawv lim dej uas muaj dej noo uas tau muab tso rau hauv lub tawb nrog dej. Muab daim ntawv lim uas muaj qe tso rau hauv lub hnab kaw (SC Johnsons, Racine, WI) thiab khaws cia rau ntawm 26°C. Cov qe tau tawm thiab kwv yees li 200–250 tus menyuam kab tau yug hauv cov tais yas uas muaj cov hmoov luav (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) thiab hmoov siab (MP Biomedicals, LLC, Solon, OH, USA) thiab ntses fillet (TetraMin, Tetra GMPH, Meer, Lub Tebchaws Yelemees) hauv qhov sib piv ntawm 2:1:1. Cov kab menyuam hauv qib thib peb kawg tau siv rau hauv peb cov kev kuaj mob bioassays.
Cov noob cog uas siv rau hauv txoj kev tshawb no tau txais los ntawm cov chaw lag luam thiab tsoomfwv hauv qab no: Brassica juncea (xim av mustard-Pacific Gold) thiab Brassica juncea (dawb mustard-Ida Gold) los ntawm Pacific Northwest Farmers' Cooperative, Washington State, Tebchaws Meskas; (Garden Cress) los ntawm Kelly Seed and Hardware Co., Peoria, IL, Tebchaws Meskas thiab Thlaspi arvense (Field Pennycress-Elisabeth) los ntawm USDA-ARS, Peoria, IL, Tebchaws Meskas; Tsis muaj cov noob siv hauv txoj kev tshawb no tau kho nrog tshuaj tua kab. Tag nrho cov noob khoom tau ua tiav thiab siv hauv txoj kev tshawb no raws li cov cai hauv zos thiab lub tebchaws thiab ua raws li txhua txoj cai hauv xeev thiab lub tebchaws. Txoj kev tshawb no tsis tau tshuaj xyuas cov nroj tsuag transgenic.
Cov noob Brassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) raug zom ua hmoov zoo siv lub tshuab Retsch ZM200 ultracentrifugal mill (Retsch, Haan, Lub Tebchaws Yelemees) uas muaj lub mesh 0.75 hli thiab Stainless hlau rotor, 12 hniav, 10,000 rpm (Rooj 1). Cov hmoov noob av raug xa mus rau lub thimble ntawv thiab defatted nrog hexane hauv Soxhlet apparatus rau 24 teev. Ib qho subsample ntawm defatted teb mustard raug cua sov kho ntawm 100 ° C rau 1 teev kom denature myrosinase thiab tiv thaiv hydrolysis ntawm glucosinolates los tsim biologically active isothiocyanates. Cov hmoov noob horsetail kho cua sov (DFP-HT) tau siv ua qhov kev tswj tsis zoo los ntawm denaturing myrosinase.
Cov ntsiab lus glucosinolate ntawm cov noob hmoov defatted tau txiav txim siab ua peb daim ntawv siv cov kua chromatography ua haujlwm siab (HPLC) raws li cov txheej txheem luam tawm ua ntej 64. Hauv luv luv, 3 mL ntawm methanol tau ntxiv rau 250 mg qauv ntawm cov hmoov noob defatted. Txhua tus qauv tau sonicated hauv dej da dej rau 30 feeb thiab tso rau hauv qhov tsaus ntuj ntawm 23 ° C rau 16 teev. Ib qho 1 mL aliquot ntawm cov txheej organic ces lim los ntawm lub lim dej 0.45 μm rau hauv lub autosampler. Khiav ntawm lub kaw lus Shimadzu HPLC (ob lub twj tso kua mis LC 20AD; SIL 20A autosampler; DGU 20As degasser; SPD-20A UV-VIS ntes rau kev saib xyuas ntawm 237 nm; thiab CBM-20A kev sib txuas lus tsheb npav module), cov ntsiab lus glucosinolate ntawm cov noob hmoov tau txiav txim siab ua peb daim ntawv. siv Shimadzu LC Solution software version 1.25 (Shimadzu Corporation, Columbia, MD, USA). Lub kem yog C18 Inertsil rov qab theem kem (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA). Cov xwm txheej theem txav tau teeb tsa ntawm 12% methanol / 88% 0.01 M tetrabutylammonium hydroxide hauv dej (TBAH; Sigma-Aldrich, St. Louis, MO, USA) nrog tus nqi ntws ntawm 1 mL / min. Tom qab txhaj tshuaj ntawm 15 μl ntawm cov qauv, cov xwm txheej pib tau khaws cia rau 20 feeb, thiab tom qab ntawd qhov piv ntawm cov kuab tshuaj tau kho rau 100% methanol, nrog rau lub sijhawm tshuaj xyuas cov qauv tag nrho ntawm 65 feeb. Ib qho nkhaus txheem (nM / mAb raws li) tau tsim los ntawm kev sib xyaw ua ke ntawm cov qauv tshiab npaj sinapine, glucosinolate thiab myrosin (Sigma-Aldrich, St. Louis, MO, USA) los kwv yees cov ntsiab lus sulfur ntawm cov noob hmoov defatted. glucosinolates. Cov glucosinolate concentration hauv cov qauv tau sim rau ntawm Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) siv OpenLAB CDS ChemStation version (C.01.07 SR2 [255]) nruab nrog tib kem thiab siv txoj kev piav qhia ua ntej. Cov glucosinolate concentration tau txiav txim siab; yuav sib piv tau ntawm HPLC systems.
Allyl isothiocyanate (94%, ruaj khov) thiab benzyl isothiocyanate (98%) tau yuav los ntawm Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). 4-Hydroxybenzylisothiocyanate tau yuav los ntawm ChemCruz (Santa Cruz Biotechnology, CA, USA). Thaum enzymatically hydrolyzed los ntawm myrosinase, glucosinolates, glucosinolates, thiab glucosinolates tsim allyl isothiocyanate, benzyl isothiocyanate, thiab 4-hydroxybenzylisothiocyanate, raws li.
Cov kev sim tshuaj ntsuam xyuas hauv chav kuaj tau ua raws li txoj kev ntawm Muturi et al. 32 nrog kev hloov kho. Tsib lub noob pub uas tsis muaj roj ntau tau siv rau hauv kev tshawb fawb no: DFP, DFP-HT, IG, PG thiab Ls. Nees nkaum tus menyuam tau muab tso rau hauv lub khob peb-txoj kev 400 mL pov tseg (VWR International, LLC, Radnor, PA, USA) uas muaj 120 mL dej deionized (dH2O). Xya qhov concentration ntawm cov noob tau kuaj rau qhov lom ntawm cov menyuam yoov tshaj cum: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 thiab 0.12 g noob hmoov / 120 ml dH2O rau DFP noob hmoov, DFP-HT, IG thiab PG. Cov kev sim tshuaj ntsuam xyuas thawj zaug qhia tias cov hmoov noob Ls uas tau defatted muaj tshuaj lom ntau dua li plaub lwm cov hmoov noob uas tau sim. Yog li ntawd, peb tau kho xya qhov kev kho mob ntawm Ls noob hmoov rau cov kev sib xyaw ua ke hauv qab no: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, thiab 0.075 g/120 mL dH2O.
Ib pawg tswj tsis tau kho (dH20, tsis muaj noob hmoov ntxiv) tau suav nrog los ntsuas qhov kev tuag ntawm cov kab hauv qab cov xwm txheej sim. Kev tshuaj xyuas tshuaj lom rau txhua pluas noj noob suav nrog peb lub khob peb-qaum (20 tus menyuam kab hauv qib thib peb lig ib lub khob), rau tag nrho 108 lub raj mis. Cov thawv kho tau khaws cia rau hauv chav sov (20-21 ° C) thiab kev tuag ntawm cov menyuam kab tau sau tseg thaum 24 thiab 72 teev ntawm kev siv cov tshuaj kho tas mus li. Yog tias lub cev thiab cov ceg ntawm tus yoov tshaj cum tsis txav thaum raug chob lossis kov nrog lub spatula hlau tsis xeb nyias, cov menyuam kab raug suav tias tuag lawm. Cov menyuam kab tuag feem ntau nyob twj ywm tsis txav hauv qhov chaw dorsal lossis ventral ntawm qab ntawm lub thawv lossis ntawm qhov dej. Qhov kev sim tau rov ua dua peb zaug rau hnub sib txawv siv cov pab pawg sib txawv ntawm cov menyuam kab, rau tag nrho 180 tus menyuam kab uas raug siv txhua qhov kev kho.
Qhov tshuaj lom ntawm AITC, BITC, thiab 4-HBITC rau cov kab laug sab tau raug soj ntsuam siv tib txoj kev bioassay tab sis nrog kev kho mob sib txawv. Npaj 100,000 ppm cov tshuaj rau txhua yam tshuaj los ntawm kev ntxiv 100 µL ntawm cov tshuaj rau 900 µL ntawm ethanol hauv lub raj centrifuge 2-mL thiab co rau 30 vib nas this kom sib tov kom huv si. Kev kho mob concentration tau txiav txim siab raws li peb cov bioassays ua ntej, uas pom BITC muaj tshuaj lom ntau dua li AITC thiab 4-HBITC. Txhawm rau txiav txim siab txog kev lom, 5 concentration ntawm BITC (1, 3, 6, 9 thiab 12 ppm), 7 concentration ntawm AITC (5, 10, 15, 20, 25, 30 thiab 35 ppm) thiab 6 concentration ntawm 4-HBITC (15, 15, 20, 25, 30 thiab 35 ppm). 30, 45, 60, 75 thiab 90 ppm). Kev kho mob tswj tau txhaj nrog 108 μL ntawm ethanol kiag li, uas yog sib npaug rau qhov ntim siab tshaj plaws ntawm kev kho mob tshuaj. Bioassays tau rov ua dua li saum toj no, qhia txog tag nrho 180 tus kab menyuam rau ib qho kev kho mob concentration. Kev tuag ntawm cov kab menyuam tau sau tseg rau txhua qhov concentration ntawm AITC, BITC, thiab 4-HBITC tom qab 24 teev ntawm kev raug tshuaj tas mus li.
Kev tshuaj xyuas Probit ntawm 65 cov ntaub ntawv txog kev tuag ntsig txog koob tshuaj tau ua tiav siv Polo software (Polo Plus, LeOra Software, version 1.0) los xam 50% kev ua rau tuag taus (LC50), 90% kev ua rau tuag taus (LC90), qhov nqes hav, tus lej ntawm kev ua rau tuag taus, thiab 95% kev ua rau tuag taus. raws li qhov sib txawv ntawm kev ntseeg siab rau qhov sib piv ntawm cov koob tshuaj tuag taus rau cov kab sib txawv ntawm cov ntaub ntawv thiab cov kab sib txawv ntawm cov koob tshuaj tuag taus. Cov ntaub ntawv txog kev tuag taus yog raws li cov ntaub ntawv rov ua dua ntawm 180 tus kab menyuam uas raug rau txhua qhov kev kho mob. Kev tshuaj xyuas Probabilistic tau ua sib cais rau txhua pluas noj noob thiab txhua yam tshuaj lom neeg. Raws li qhov sib txawv ntawm 95% ntawm qhov sib piv ntawm cov koob tshuaj tuag taus, qhov lom ntawm cov pluas noj noob thiab cov tshuaj lom neeg rau cov kab menyuam yoov tshaj cum tau suav tias txawv heev, yog li qhov sib txawv ntawm kev ntseeg siab uas muaj tus nqi ntawm 1 tsis txawv heev, P = 0.0566.
Cov txiaj ntsig HPLC rau kev txiav txim siab ntawm cov glucosinolates tseem ceeb hauv cov hmoov noob defatted DFP, IG, PG thiab Ls tau teev nyob rau hauv Rooj 1. Cov glucosinolates tseem ceeb hauv cov hmoov noob uas tau sim sib txawv tsuas yog DFP thiab PG, uas ob qho tib si muaj myrosinase glucosinolates. Cov ntsiab lus myrosinin hauv PG siab dua li hauv DFP, 33.3 ± 1.5 thiab 26.5 ± 0.9 mg / g, raws li. Ls noob hmoov muaj 36.6 ± 1.2 mg / g glucoglycone, thaum IG noob hmoov muaj 38.0 ± 0.5 mg / g sinapine.
Cov kab menyuam ntawm Ae. Aedes aegypti yoov tshaj cum raug tua thaum kho nrog cov noob hmoov uas tsis muaj roj, txawm hais tias qhov ua tau zoo ntawm kev kho mob sib txawv nyob ntawm hom nroj tsuag. Tsuas yog DFP-NT tsis muaj tshuaj lom rau cov kab menyuam tom qab 24 thiab 72 teev ntawm kev raug (Rooj 2). Qhov tshuaj lom ntawm cov hmoov noob nquag nce ntxiv nrog kev nce ntxiv (Daim Duab 1A, B). Qhov tshuaj lom ntawm cov noob hmoov rau cov kab menyuam sib txawv ntau raws li 95% CI ntawm qhov piv txwv ntawm LC50 ntawm 24-teev thiab 72-teev kev ntsuam xyuas (Rooj 3). Tom qab 24 teev, qhov cuam tshuam ntawm cov noob hmoov Ls ntau dua li lwm cov tshuaj kho noob, nrog rau qhov ua haujlwm siab tshaj plaws thiab qhov tshuaj lom siab tshaj plaws rau cov kab menyuam (LC50 = 0.04 g / 120 ml dH2O). Cov kab me me tsis tshua muaj kev cuam tshuam rau DFP ntawm 24 teev piv rau IG, Ls thiab PG noob hmoov kho, nrog LC50 tus nqi ntawm 0.115, 0.04 thiab 0.08 g / 120 ml dH2O feem, uas yog siab dua li LC50 tus nqi. 0.211 g / 120 ml dH2O (Rooj 3). LC90 tus nqi ntawm DFP, IG, PG thiab Ls yog 0.376, 0.275, 0.137 thiab 0.074 g / 120 ml dH2O, feem (Rooj 2). Qhov siab tshaj plaws ntawm DPP yog 0.12 g / 120 ml dH2O. Tom qab 24 teev ntawm kev ntsuam xyuas, qhov nruab nrab ntawm cov kab me me tuag tsuas yog 12%, thaum qhov nruab nrab ntawm cov kab me me tuag ntawm IG thiab PG ncav cuag 51% thiab 82%, feem. Tom qab 24 teev ntawm kev soj ntsuam, qhov nruab nrab ntawm kev tuag ntawm cov kab rau qhov kev kho mob Ls noob ntau tshaj plaws (0.075 g/120 ml dH2O) yog 99% (Daim Duab 1A).
Cov kab nkhaus tuag tau kwv yees los ntawm cov tshuaj teb (Probit) ntawm Ae. Egyptian larvae (3rd instar larvae) rau cov noob hmoov nplej concentration 24 teev (A) thiab 72 teev (B) tom qab kev kho mob. Cov kab dotted sawv cev rau LC50 ntawm kev kho mob noob hmoov nplej. DFP Thlaspi arvense, DFP-HT Kub inactivated Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Thaum 72 teev kev ntsuam xyuas, cov nqi LC50 ntawm DFP, IG thiab PG noob hmoov yog 0.111, 0.085 thiab 0.051 g / 120 ml dH2O, raws li. Yuav luag txhua tus kab menyuam uas raug Ls noob hmoov tuag tom qab 72 teev ntawm kev raug, yog li cov ntaub ntawv tuag tsis sib xws nrog Probit kev tshuaj xyuas. Piv rau lwm cov noob hmoov, cov kab menyuam tsis tshua muaj kev cuam tshuam rau DFP noob hmoov kho thiab muaj cov nqi LC50 siab dua (Cov Lus Qhia 2 thiab 3). Tom qab 72 teev, cov nqi LC50 rau DFP, IG thiab PG noob hmoov kho tau kwv yees tias yog 0.111, 0.085 thiab 0.05 g / 120 ml dH2O, raws li. Tom qab 72 teev ntawm kev ntsuam xyuas, cov nqi LC90 ntawm DFP, IG thiab PG noob hmoov yog 0.215, 0.254 thiab 0.138 g / 120 ml dH2O, raws li. Tom qab 72 teev ntawm kev soj ntsuam, qhov nruab nrab ntawm cov kab me me tuag rau DFP, IG thiab PG noob hmoov kho mob ntawm qhov siab tshaj plaws ntawm 0.12 g / 120 ml dH2O yog 58%, 66% thiab 96%, raws li (Daim Duab 1B). Tom qab 72-teev kev soj ntsuam, PG noob hmoov tau pom tias muaj tshuaj lom ntau dua li IG thiab DFP noob hmoov.
Cov tshuaj isothiocyanates ua los ntawm cov khoom siv hluavtaws, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC) thiab 4-hydroxybenzylisothiocyanate (4-HBITC) tuaj yeem tua cov kab menyuam yoov tshaj cum tau zoo. Thaum 24 teev tom qab kho, BITC muaj tshuaj lom ntau dua rau cov kab menyuam nrog tus nqi LC50 ntawm 5.29 ppm piv rau 19.35 ppm rau AITC thiab 55.41 ppm rau 4-HBITC (Rooj 4). Piv rau AITC thiab BITC, 4-HBITC muaj tshuaj lom tsawg dua thiab tus nqi LC50 siab dua. Muaj qhov sib txawv tseem ceeb hauv kev lom ntawm cov kab menyuam yoov tshaj cum ntawm ob qho isothiocyanates loj (Ls thiab PG) hauv cov noob hmoov uas muaj zog tshaj plaws. Kev lom raws li qhov piv ntawm LC50 ntawm AITC, BITC, thiab 4-HBITC qhia txog qhov sib txawv ntawm cov lej suav uas 95% CI ntawm LC50 qhov piv ntawm 1 tsis suav nrog tus nqi ntawm 1 (P = 0.05, Rooj 4). Qhov siab tshaj plaws ntawm ob qho tib si BITC thiab AITC tau kwv yees tias tua 100% ntawm cov kab me me uas tau sim (Daim Duab 2).
Cov kab nkhaus tuag tau kwv yees los ntawm cov tshuaj teb (Probit) ntawm Ae. 24 teev tom qab kev kho mob, cov kab menyuam Iyiv (3rd instar larvae) tau mus txog cov tshuaj isothiocyanate concentrations. Cov kab dotted sawv cev rau LC50 rau kev kho mob isothiocyanate. Benzyl isothiocyanate BITC, allyl isothiocyanate AITC thiab 4-HBITC.
Kev siv cov tshuaj tua kab mob los ntawm cov nroj tsuag ua cov tshuaj tua kab tau ntev lawm. Ntau cov nroj tsuag tsim cov tshuaj lom neeg ntuj uas muaj cov haujlwm tua kab37. Lawv cov tshuaj lom neeg ua rau muaj lwm txoj hauv kev zoo dua li cov tshuaj tua kab uas muaj peev xwm zoo hauv kev tswj cov kab tsuag, suav nrog cov yoov tshaj cum.
Cov nroj tsuag mustard raug cog ua qoob loo rau lawv cov noob, siv ua txuj lom thiab yog qhov chaw muaj roj. Thaum cov roj mustard raug rho tawm ntawm cov noob lossis thaum mustard raug rho tawm los siv ua roj biofuel, 69 cov khoom seem yog cov noob hmoov defatted. Cov noob hmoov no khaws cia ntau yam ntawm nws cov khoom siv biochemical ntuj thiab cov enzymes hydrolytic. Qhov lom ntawm cov noob hmoov no yog vim muaj cov isothiocyanates55,60,61. Isothiocyanates yog tsim los ntawm hydrolysis ntawm glucosinolates los ntawm cov enzyme myrosinase thaum lub sijhawm hydrating ntawm noob hmoov38,55,70 thiab paub tias muaj cov teebmeem fungicidal, bactericidal, nematicidal thiab insecticidal, nrog rau lwm yam khoom suav nrog cov teebmeem sensory thiab chemotherapeutic khoom61,62,70. Ntau txoj kev tshawb fawb tau qhia tias cov nroj tsuag mustard thiab noob hmoov ua haujlwm zoo li fumigants tawm tsam cov av thiab cov zaub mov khaws cia kab tsuag57,59,71,72. Hauv kev tshawb fawb no, peb tau soj ntsuam qhov kev lom ntawm plaub-noob hmoov thiab nws peb yam khoom bioactive AITC, BITC, thiab 4-HBITC rau Aedes yoov tshaj cum larvae. Aedes aegypti. Ntxiv noob hmoov ncaj qha rau hauv dej uas muaj cov yoov tshaj cum larvae yuav ua rau cov txheej txheem enzymatic uas tsim cov isothiocyanates uas muaj tshuaj lom rau cov yoov tshaj cum larvae. Qhov biotransformation no tau pom ib feem los ntawm kev ua haujlwm larvicidal ntawm noob hmoov thiab poob ntawm kev ua haujlwm tua kab thaum cov noob mustard dwarf raug kho cua sov ua ntej siv. Kev kho cua sov yuav rhuav tshem cov hydrolytic enzymes uas ua rau glucosinolates ua haujlwm, yog li tiv thaiv kev tsim cov bioactive isothiocyanates. Qhov no yog thawj txoj kev tshawb fawb los lees paub cov khoom tua kab ntawm cov noob zaub qhwv tiv thaiv yoov tshaj cum hauv ib puag ncig dej.
Ntawm cov hmoov noob uas tau sim, cov hmoov noob watercress (Ls) yog cov tshuaj lom tshaj plaws, ua rau Aedes albopictus tuag ntau heev. Cov kab menyuam Aedes aegypti tau ua tiav tas li 24 teev. Peb cov hmoov noob seem (PG, IG thiab DFP) muaj kev ua haujlwm qeeb dua thiab tseem ua rau tuag ntau tom qab 72 teev ntawm kev kho tas li. Tsuas yog Ls noob hmoov muaj cov glucosinolates ntau heev, thaum PG thiab DFP muaj myrosinase thiab IG muaj glucosinolate ua glucosinolate tseem ceeb (Rooj 1). Glucotropaeolin raug hydrolyzed rau BITC thiab sinalbine raug hydrolyzed rau 4-HBITC61,62. Peb cov txiaj ntsig bioassay qhia tau tias ob qho tib si Ls noob hmoov thiab BITC hluavtaws muaj tshuaj lom heev rau cov kab menyuam yoov tshaj cum. Cov khoom tseem ceeb ntawm PG thiab DFP noob hmoov yog myrosinase glucosinolate, uas raug hydrolyzed rau AITC. AITC muaj txiaj ntsig zoo hauv kev tua cov kab menyuam yoov tshaj cum nrog tus nqi LC50 ntawm 19.35 ppm. Piv rau AITC thiab BITC, 4-HBITC isothiocyanate yog qhov tsis muaj tshuaj lom rau cov kab me. Txawm hais tias AITC tsis muaj tshuaj lom ntau dua li BITC, lawv cov nqi LC50 qis dua ntau cov roj tseem ceeb uas tau sim rau cov kab me32,73,74,75.
Peb cov hmoov noob cruciferous rau kev siv tawm tsam cov kab laug sab muaj ib qho tseem ceeb glucosinolate, suav txog ntau dua 98-99% ntawm tag nrho cov glucosinolates raws li txiav txim los ntawm HPLC. Cov nyiaj me me ntawm lwm cov glucosinolates tau pom, tab sis lawv cov qib tsawg dua 0.3% ntawm tag nrho cov glucosinolates. Watercress (L. sativum) noob hmoov muaj cov glucosinolates theem nrab (sinigrin), tab sis lawv qhov feem pua ​​yog 1% ntawm tag nrho cov glucosinolates, thiab lawv cov ntsiab lus tseem tsis tseem ceeb (kwv yees li 0.4 mg / g noob hmoov). Txawm hais tias PG thiab DFP muaj tib lub ntsiab glucosinolate (myrosin), qhov kev ua haujlwm larvicidal ntawm lawv cov noob noj sib txawv vim lawv cov nqi LC50. Txawv hauv kev lom rau powdery mildew. Kev tshwm sim ntawm Aedes aegypti larvae yuav yog vim qhov sib txawv ntawm myrosinase kev ua haujlwm lossis kev ruaj khov ntawm ob lub noob pub. Kev ua haujlwm ntawm Myrosinase ua lub luag haujlwm tseem ceeb hauv kev siv tau ntawm cov khoom hydrolysis xws li isothiocyanates hauv cov nroj tsuag Brassicaceae76. Cov ntawv ceeb toom yav dhau los los ntawm Pocock et al.77 thiab Wilkinson et al.78 tau qhia tias kev hloov pauv hauv kev ua haujlwm thiab kev ruaj khov ntawm myrosinase kuj tseem yuav cuam tshuam nrog cov yam ntxwv ntawm caj ces thiab ib puag ncig.
Cov ntsiab lus bioactive isothiocyanate xav tau raug suav raws li LC50 tus nqi ntawm txhua pluas noj noob ntawm 24 thiab 72 teev (Rooj 5) rau kev sib piv nrog cov ntawv thov tshuaj sib xws. Tom qab 24 teev, cov isothiocyanates hauv pluas noj noob muaj tshuaj lom ntau dua li cov tshuaj ntshiab. LC50 tus nqi xam raws li qhov ib feem ntawm ib lab (ppm) ntawm kev kho noob isothiocyanate qis dua LC50 tus nqi rau BITC, AITC, thiab 4-HBITC daim ntawv thov. Peb pom cov kab laug sab noj cov noob pellets (Daim Duab 3A). Yog li ntawd, cov kab laug sab yuav tau txais kev raug tshuaj lom ntau dua rau isothiocyanates los ntawm kev noj cov noob pellets. Qhov no tau pom tseeb tshaj plaws hauv kev kho noob IG thiab PG ntawm 24-teev raug, qhov twg LC50 concentration yog 75% thiab 72% qis dua li kev kho AITC thiab 4-HBITC ntshiab, raws li. Kev kho mob Ls thiab DFP muaj tshuaj lom ntau dua li isothiocyanate ntshiab, nrog LC50 tus nqi 24% thiab 41% qis dua, feem. Cov kab menyuam hauv kev kho mob tswj tau ua tiav pupated (Daim duab 3B), thaum feem ntau cov kab menyuam hauv kev kho noob tsis tau pupate thiab kev loj hlob ntawm cov kab menyuam tau qeeb heev (Daim duab 3B, D). Hauv Spodopteralitura, isothiocyanates cuam tshuam nrog kev loj hlob qeeb thiab kev loj hlob qeeb79.
Cov kab menyuam ntawm Ae. Aedes aegypti yoov tshaj cum tau raug rau cov hmoov noob Brassica tas li rau 24-72 teev. (A) Cov kab menyuam tuag nrog cov khoom me me ntawm cov noob hmoov hauv qhov ncauj (ua voj voog); (B) Kev kho mob tswj (dH20 yam tsis muaj ntxiv noob hmoov) qhia tau hais tias cov kab menyuam loj hlob zoo li qub thiab pib ua pupate tom qab 72 teev (C, D) Cov kab menyuam kho nrog noob hmoov; cov noob hmoov qhia txog qhov sib txawv hauv kev loj hlob thiab tsis ua pupate.
Peb tsis tau kawm txog cov txheej txheem ntawm cov teebmeem lom ntawm isothiocyanates rau cov kab menyuam yoov tshaj cum. Txawm li cas los xij, cov kev tshawb fawb yav dhau los hauv cov ntsaum liab hluav taws (Solenopsis invicta) tau qhia tias kev txwv ntawm glutathione S-transferase (GST) thiab esterase (EST) yog lub hauv paus tseem ceeb ntawm isothiocyanate bioactivity, thiab AITC, txawm tias ntawm kev ua haujlwm tsawg, kuj tuaj yeem txwv GST kev ua haujlwm. Cov ntsaum liab xa tuaj rau hauv cov concentration qis. Cov koob tshuaj yog 0.5 µg / ml80. Qhov sib piv, AITC inhibits acetylcholinesterase hauv cov kab laug sab laus (Sitophilus zeamais)81. Cov kev tshawb fawb zoo sib xws yuav tsum tau ua kom pom tseeb txog cov txheej txheem ntawm isothiocyanate kev ua haujlwm hauv cov kab menyuam yoov tshaj cum.
Peb siv kev kho mob DFP uas tsis ua haujlwm kub los txhawb qhov kev thov tias hydrolysis ntawm cov nroj tsuag glucosinolates los tsim cov isothiocyanates ua haujlwm ua lub tshuab rau kev tswj cov kab tsuag los ntawm cov hmoov nplej mustard. DFP-HT hmoov nplej tsis muaj tshuaj lom ntawm cov nqi thov sim. Lafarga et al. 82 tau tshaj tawm tias glucosinolates rhiab heev rau kev puas tsuaj ntawm qhov kub siab. Kev kho cua sov kuj xav tias yuav ua rau cov enzyme myrosinase hauv cov hmoov nplej noob tsis zoo thiab tiv thaiv kev hydrolysis ntawm glucosinolates los tsim cov isothiocyanates ua haujlwm. Qhov no kuj tau lees paub los ntawm Okunade et al. 75 tau qhia tias myrosinase rhiab heev rau qhov kub, qhia tias myrosinase kev ua haujlwm tau raug kaw tag nrho thaum mustard, mustard dub, thiab cov noob bloodroot raug rau qhov kub siab tshaj 80 ° C. Cov txheej txheem no yuav ua rau poob ntawm kev ua haujlwm tua kab ntawm cov hmoov nplej DFP uas tau kho cua sov.
Yog li ntawd, cov noob mustard hmoov thiab nws peb lub isothiocyanates tseem ceeb yog lom rau cov kab laug sab. Muab cov qhov sib txawv ntawm cov noob hmoov thiab kev kho tshuaj, kev siv cov noob hmoov yuav yog ib txoj hauv kev zoo ntawm kev tswj cov yoov tshaj cum. Muaj qhov xav tau los txheeb xyuas cov qauv tsim nyog thiab cov txheej txheem xa khoom zoo los txhim kho qhov ua tau zoo thiab kev ruaj khov ntawm kev siv cov hmoov noob. Peb cov txiaj ntsig qhia txog kev siv cov noob mustard hmoov ua lwm txoj hauv kev rau cov tshuaj tua kab. Cov thev naus laus zis no tuaj yeem dhau los ua lub cuab yeej tshiab rau kev tswj cov kab laug sab. Vim tias cov kab laug sab vam meej hauv cov dej thiab cov noob glucosinolates tau hloov pauv enzymatically mus rau isothiocyanates nquag thaum hydration, kev siv cov noob mustard hmoov hauv cov dej uas muaj yoov tshaj cum muaj peev xwm tswj tau zoo. Txawm hais tias cov kab laug sab ntawm isothiocyanates ua haujlwm sib txawv (BITC > AITC > 4-HBITC), xav tau kev tshawb fawb ntxiv los txiav txim siab seb kev sib xyaw cov noob hmoov nrog ntau glucosinolates synergistically ua rau muaj kev lom ntau ntxiv. Qhov no yog thawj txoj kev tshawb fawb los qhia txog cov teebmeem tua kab ntawm cov noob cruciferous defatted thiab peb bioactive isothiocyanates ntawm cov yoov tshaj cum. Cov txiaj ntsig ntawm txoj kev tshawb fawb no tau ua rau muaj kev hloov pauv tshiab los ntawm kev qhia tias cov noob zaub qhwv uas tau tshem roj, uas yog cov khoom seem ntawm kev rho roj los ntawm cov noob, yuav ua haujlwm ua tus neeg sawv cev tua kab mob zoo rau kev tswj cov yoov tshaj cum. Cov ntaub ntawv no tuaj yeem pab txhawb kev tshawb pom cov neeg sawv cev tswj cov nroj tsuag thiab lawv txoj kev loj hlob ua cov tshuaj tua kab mob pheej yig, siv tau zoo, thiab zoo rau ib puag ncig.
Cov ntaub ntawv tsim tawm rau txoj kev tshawb fawb no thiab cov kev tshuaj xyuas tau los ntawm tus sau ntawv sib xws yog tias muaj kev thov tsim nyog. Thaum kawg ntawm txoj kev tshawb fawb, txhua yam khoom siv hauv txoj kev tshawb fawb (kab thiab noob hmoov) raug rhuav tshem.