Kuti mogwira mtimakuletsa udzudzundi kuchepetsa kuchuluka kwa matenda omwe amanyamula, njira zoyendetsera bwino, zokhazikika komanso zosawononga chilengedwe m'malo mwa mankhwala ophera tizilombo.Tidawunika zakudya zambewu kuchokera ku Brassicaceae (banja la Brassica) ngati gwero la isothiocyanate yochokera ku mbewu yopangidwa ndi enzymatic hydrolysis ya biologically inactive glucosinolates kuti igwiritsidwe ntchito pakuwongolera Egypt Aedes (L., 1762).Chakudya chambewu chamafuta asanu (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 ndi Thlaspi arvense - mitundu itatu yayikulu yakuwonongeka kwamafuta mankhwala Kuti mudziwe kawopsedwe (LC50) wa allyl isothiocyanate, benzyl isothiocyanate ndi 4-hydroxybenzylisothiocyanate kupita ku Aedes aegypti mphutsi pa 24-hour exposure = 0.04 g/120 ml dH2O).Mtengo wa LC50 wa mpiru, mpiru woyera ndi mchira wa akavalo.chakudya chambewu chinali 0.05, 0.08 ndi 0.05 motsatira poyerekeza ndi allyl isothiocyanate (LC50 = 19.35 ppm) ndi 4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) inali yoopsa kwambiri ku mphutsi / 20 ml 1 pambuyo pa chithandizo cha 20 ml.Zotsatira izi zimagwirizana ndi kupanga ufa wambewu ya alfalfa.Kuchita bwino kwambiri kwa benzyl esters kumafanana ndi ma LC50 owerengeka.Kugwiritsa ntchito ufa wambewu kungapereke njira yabwino yopewera udzudzu.mphamvu ya ufa wa mbeu ya cruciferous ndi zigawo zake zazikulu za mankhwala motsutsana ndi mphutsi za udzudzu ndikuwonetsa momwe mankhwala achilengedwe mu ufa wa cruciferous ufa amatha kukhala ngati mankhwala otetezera zachilengedwe oteteza udzudzu.
Matenda oyambitsidwa ndi ma vector omwe amayambitsidwa ndi udzudzu wa Aedes akadali vuto lalikulu laumoyo wapadziko lonse lapansi.Kuchuluka kwa matenda oyambitsidwa ndi udzudzu kumafalikira kumadera1,2,3 ndikuyambiranso, zomwe zimadzetsa kuphulika kwa matenda oopsa4,5,6,7.Kufalikira kwa matenda pakati pa anthu ndi nyama (monga chikungunya, dengue, Rift Valley fever, yellow fever ndi Zika virus) sikunachitikepo.Dengue fever yokha imayika anthu pafupifupi 3.6 biliyoni pachiwopsezo chotenga matenda m'malo otentha, ndipo pafupifupi 390 miliyoni amadwala chaka chilichonse, zomwe zimapangitsa kuti 6,100-24,300 amafa pachaka8.Kuwonekeranso ndi kufalikira kwa kachilombo ka Zika ku South America kwakopa chidwi padziko lonse lapansi chifukwa cha kuwonongeka kwa ubongo komwe kumayambitsa ana obadwa ndi amayi omwe ali ndi kachilombo2.Kremer et al 3 akulosera kuti mitundu yosiyanasiyana ya udzudzu wa Aedes idzapitirirabe kukula ndipo podzafika 2050, theka la anthu padziko lonse lapansi adzakhala pa chiopsezo chotenga matenda oyambitsidwa ndi udzudzu.
Kupatulapo katemera wa dengue ndi yellow fever wopangidwa posachedwapa, katemera wolimbana ndi matenda ambiri ofalitsidwa ndi udzudzu sanapangidwebe9,10,11.Makatemera akadalipo ochepa ndipo amangogwiritsidwa ntchito m'mayesero azachipatala.Kuwongolera tizilombo toyambitsa udzudzu pogwiritsa ntchito mankhwala ophera udzudzu kwakhala njira yayikulu yothanirana ndi kufalikira kwa matenda oyambitsidwa ndi udzudzu12,13.Ngakhale kuti mankhwala ophera tizilombo ndi othandiza popha udzudzu, kugwiritsa ntchito mankhwala opha tizilombo kumakhudza kwambiri zamoyo zomwe sizili zolinga komanso zimawononga chilengedwe14,15,16.Chodetsa nkhaŵa kwambiri ndi chikhalidwe cha kuwonjezeka kwa udzudzu ku mankhwala ophera tizilombo17,18,19.Mavutowa okhudzana ndi mankhwala ophera tizilombo athandizira kufunafuna njira zogwirira ntchito komanso zowononga zachilengedwe zothana ndi tizilombo toyambitsa matenda.
Zomera zosiyanasiyana zapangidwa ngati magwero a phytopesticides pothana ndi tizirombo20,21.Zomera nthawi zambiri zimakhala zosagwirizana ndi chilengedwe chifukwa zimatha kuwonongeka ndi chilengedwe ndipo zimakhala ndi kawopsedwe kakang'ono kapena kocheperako kwa zamoyo zomwe sizili ndi zolinga monga nyama zoyamwitsa, nsomba ndi zamoyo zam'madzi20,22.Kukonzekera kwa zitsamba kumadziwika kuti kumapanga mitundu yosiyanasiyana ya bioactive mankhwala omwe ali ndi njira zosiyanasiyana zogwirira ntchito kuti athetse bwino magawo osiyanasiyana a moyo wa udzudzu23,24,25,26.Mafuta opangidwa ndi zomera monga mafuta ofunikira ndi zosakaniza zina zogwira ntchito za zomera zakhala zikudziwika bwino ndipo zatsegula njira ya zida zatsopano zothanirana ndi tizilombo toyambitsa udzudzu.Mafuta ofunikira, monoterpenes ndi sesquiterpenes amagwira ntchito ngati zothamangitsa, zoletsa kudyetsa ndi ovicides27,28,29,30,31,32,33.Mafuta ambiri a masamba amayambitsa imfa ya mphutsi za udzudzu, pupae ndi akuluakulu34,35,36, zomwe zimakhudza mitsempha, kupuma, endocrine ndi machitidwe ena ofunikira a tizilombo37.
Kafukufuku waposachedwa wapereka chidziwitso pakugwiritsa ntchito mbewu za mpiru ndi nthangala zake ngati magwero a bioactive mankhwala.Mbeu ya mpiru yayesedwa ngati biofumigant38,39,40,41 ndipo imagwiritsidwa ntchito ngati kusintha kwa dothi pofuna kupondereza udzu42,43,44 komanso kuwongolera tizilombo toyambitsa matenda obwera kunthaka45,46,47,48,49,50, zakudya za zomera.nematodes 41,51, 52, 53, 54 ndi tizilombo 55, 56, 57, 58, 59, 60. Ntchito ya fungicidal ya ufa wambewuyi imachokera ku zomera zoteteza mankhwala otchedwa isothiocyanates38,42,60.Muzomera, zinthu zotetezazi zimasungidwa m'maselo a zomera monga ma non-bioactive glucosinolates.Komabe, zomera zikawonongeka ndi kudyetsa tizilombo kapena tizilombo toyambitsa matenda, glucosinolates ndi hydrolyzed ndi myrosinase mu bioactive isothiocyanates55,61.Isothiocyanates ndi mankhwala osakanikirana omwe amadziwika kuti ali ndi antimicrobial ndi tizilombo toyambitsa matenda, ndipo mapangidwe awo, zochitika zamoyo ndi zomwe zili mkati zimasiyana kwambiri pakati pa Brassicaceae mitundu42,59,62,63.
Ngakhale ma isothiocyanate opangidwa kuchokera ku ufa wa mpiru amadziwika kuti ali ndi zochita zowononga tizilombo, zidziwitso zokhudzana ndi zamoyo zolimbana ndi ma vector ofunikira azachipatala zikusowa.Kafukufuku wathu adafufuza ntchito ya larvicidal ya ufa anayi wambewu wodetsedwa motsutsana ndi udzudzu wa Aedes.Mphutsi za Aedes aegypti.Cholinga cha kafukufukuyu chinali kuwunika momwe angagwiritsire ntchito ngati mankhwala othana ndi udzudzu osawononga chilengedwe.Zigawo zitatu zazikulu zamakemikolo a chakudya chambewu, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC), ndi 4-hydroxybenzylisothiocyanate (4-HBITC) adayesedwanso kuti ayese ntchito yachilengedwe ya zigawo zamankhwala izi pa mphutsi za udzudzu.Ili ndi lipoti loyamba lowunika momwe ufa wa mbewu za kabichi ndi zigawo zake zazikulu zolimbana ndi mphutsi za udzudzu.
Ma laboratory colonies a Aedes aegypti (Rockefeller strain) adasungidwa pa 26 ° C, 70% chinyezi wachibale (RH) ndi 10:14 h (L: D photoperiod).Azimayi okwatirana ankasungidwa m'makola apulasitiki (utali wa 11 cm ndi m'mimba mwake 9.5 cm) ndikudyetsedwa kudzera m'mabotolo pogwiritsa ntchito magazi a ng'ombe (HemoStat Laboratories Inc., Dixon, CA, USA).Kudyetsa magazi kunkachitika mwachizolowezi pogwiritsa ntchito kansalu kakang'ono ka magalasi (Chemglass, Life Sciences LLC, Vineland, NJ, USA) yolumikizidwa ndi chubu chosambira chamadzi (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) ndi kutentha. kuwongolera 37 °C.Tambasulani filimu ya Parafilm M pansi pa chipinda chilichonse chodyera chagalasi (malo a 154 mm2).Kenako chodyera chilichonse chinali kuikidwa pagululi pamwamba pomwe panali khola lomwe linali ndi yaikazi yokwerera.Pafupifupi 350–400 μl ya magazi a ng’ombe anawonjezedwa ku fungulo la magalasi pogwiritsa ntchito Pasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) ndipo nyongolotsi zazikuluzo zinaloledwa kukhetsa kwa ola limodzi.Azimayi oyembekezera adapatsidwa yankho la 10% la sucrose ndikuloledwa kuyikira mazira pa pepala lonyowa losefera lomwe lili m'makapu amtundu wa ultra-clear soufflé (1.25 fl oz size, Dart Container Corp., Mason, MI, USA).khola ndi madzi.Ikani pepala losefera lomwe lili ndi mazira muthumba lomata (SC Johnsons, Racine, WI) ndi kusunga pa 26°C.Mazirawa adaswedwa ndipo mphutsi pafupifupi 200-250 zidakwezedwa m'matayala apulasitiki okhala ndi zosakaniza za kalulu (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) ndi ufa wa chiwindi (MP Biomedicals, LLC, Solon, OH, USA).ndi minofu ya nsomba (TetraMin, Tetra GMPH, Meer, Germany) mu chiŵerengero cha 2:1:1.Mphutsi zakumapeto kwachitatu za instar zidagwiritsidwa ntchito poyesa bioassays yathu.
Mbeu za mbeu zomwe zagwiritsidwa ntchito mu kafukufukuyu zinapezedwa kuchokera ku magwero otsatirawa azamalonda ndi aboma: Brassica juncea (brown mustard-Pacific Gold) ndi Brassica juncea (white mustard-Ida Gold) kuchokera ku Pacific Northwest Farmers' Cooperative, Washington State, USA;(Garden Cress) kuchokera ku Kelly Seed and Hardware Co., Peoria, IL, USA ndi Thlaspi arvense (Field Pennycress-Elisabeth) kuchokera ku USDA-ARS, Peoria, IL, USA;Palibe mbewu zomwe zidagwiritsidwa ntchito mu kafukufukuyu zomwe zidathandizidwa ndi mankhwala ophera tizilombo.Mbeu zonse zidakonzedwa ndikugwiritsiridwa ntchito mu kafukufukuyu motsatira malamulo a m'deralo ndi dziko komanso motsatira malamulo onse a m'deralo ndi dziko.Kafukufukuyu sanayang'ane mitundu ya zomera za transgenic.
Brassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) mbewu zinapukutidwa kukhala ufa pogwiritsa ntchito mphero ya Retsch ZM200 ultracentrifugal (Retsch, Haan, Germany) yokhala ndi ma mesh 0.75 mm ndi Stainless rotor yachitsulo, mano 12, 10,000 rpm (Table 1).Mbeu yanthaka imasamutsidwa ku thimble yamapepala ndikutsuka ndi hexane mu chipangizo cha Soxhlet kwa 24 h.Chitsanzo cha mpiru wa mpiru wodetsedwa chinali kutentha kwa 100 ° C kwa 1 h kuti awononge myrosinase ndikuletsa hydrolysis ya glucosinolates kupanga biologically active isothiocyanates.Kutentha kwa horsetail mbewu ufa (DFP-HT) kunagwiritsidwa ntchito ngati kuwongolera koyipa mwa denaturing myrosinase.
Glucosinolate zomwe zili muzakudya zambewu zowonongeka zinatsimikiziridwa katatu pogwiritsa ntchito high-performance liquid chromatography (HPLC) molingana ndi protocol 64 yofalitsidwa kale.Mwachidule, 3 mL ya methanol inawonjezeredwa ku 250 mg ya ufa wambewu wodetsedwa.Aliyense chitsanzo anali sonicated mu madzi osamba kwa mphindi 30 ndipo anachoka mu mdima pa 23 ° C kwa 16 hours.1 mL aliquot ya organic wosanjikiza idasefedwa kudzera mu fyuluta ya 0.45 μm mu autosampler.Kuthamanga pa Shimadzu HPLC system (mapampu awiri a LC 20AD; SIL 20A autosampler; DGU 20As degasser; SPD-20A UV-VIS detector yowunikira pa 237 nm; ndi CBM-20A communication bus module), zomwe zili mu glucosinolate zambewu zambewu zinatsimikiziridwa. mu katatu.using Shimadzu LC Solution software version 1.25 (Shimadzu Corporation, Columbia, MD, USA).Mzerewu unali gawo la C18 Inertsil reverse phase (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA).Mikhalidwe yoyambirira yam'manja yam'manja idayikidwa pa 12% methanol / 88% 0.01 M tetrabutylammonium hydroxide m'madzi (TBAH; Sigma-Aldrich, St. Louis, MO, USA) ndi kuthamanga kwa 1 mL / min.Pambuyo jekeseni wa 15 μl wa zitsanzo, mikhalidwe yoyambirira idasungidwa kwa mphindi 20, ndiyeno chiŵerengero cha zosungunulira chinasinthidwa kukhala 100% methanol, ndi nthawi yowerengera zitsanzo za mphindi 65.Njira yokhotakhota (nM/mAb based) idapangidwa ndi kusungunuka kwanthawi yayitali kwa sinapine, glucosinolate ndi myrosin (Sigma-Aldrich, St. Louis, MO, USA) kuti ayerekezere kuchuluka kwa sulfure muzakudya zodetsedwa.glucosinolates.Magulu a Glucosinolate m'masampuli adayesedwa pa Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) pogwiritsa ntchito OpenLAB CDS ChemStation version (C.01.07 SR2 [255]) yokhala ndi gawo lomwelo ndikugwiritsa ntchito njira yomwe yafotokozedwa kale.Kuchuluka kwa glucosinolate kunatsimikiziridwa;kufanana pakati pa machitidwe a HPLC.
Allyl isothiocyanate (94%, stable) ndi benzyl isothiocyanate (98%) anagulidwa kuchokera ku Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA).4-Hydroxybenzylisothiocyanate inagulidwa kuchokera ku ChemCruz (Santa Cruz Biotechnology, CA, USA).Pamene enzymatically hydrolyzed ndi myrosinase, glucosinolates, glucosinolates, ndi glucosinolates kupanga allyl isothiocyanate, benzyl isothiocyanate, ndi 4-hydroxybenzylisothiocyanate, motero.
Ma bioassays a labotale adachitidwa molingana ndi njira ya Muturi et al.32 ndi zosintha.Zakudya zisanu zokhala ndi mafuta ochepa zinagwiritsidwa ntchito mu kafukufukuyu: DFP, DFP-HT, IG, PG ndi Ls.Mphutsi 20 zinayikidwa mu beaker ya njira zitatu yotaya 400 ml (VWR International, LLC, Radnor, PA, USA) yomwe ili ndi madzi opangidwa ndi 120 mL (dH2O).Mbeu zisanu ndi ziwiri za chakudya zinayesedwa kuti ziwopsezedwe ndi mphutsi ya udzudzu: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 ndi 0.12 g chakudya chambewu / 120 ml dH2O pa chakudya cha DFP, DFP-HT, IG ndi PG.Kuwunika koyambirira kukuwonetsa kuti ufa wambewu wa Ls wodetsedwa ndi wapoizoni kwambiri kuposa ufa wina wina wambewu womwe unayesedwa.Chifukwa chake, tidasintha magawo asanu ndi awiri a chithandizo cha Ls seed meal kuzinthu izi: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, ndi 0.075 g/120 mL dH2O.
Gulu loyang'anira losathandizidwa (dH20, palibe chowonjezera cha chakudya chambewu) linaphatikizidwa kuti liwunikire kufa kwa tizilombo mokhazikika poyesa.Kuyeza kwa toxicological bioassay pazakudya zilizonse kumaphatikizapo ma beaker atatu otsetsereka atatu (20 mphutsi zapakati pachitatu pa beaker), pazambiri zonse za 108.Zotengera zomwe zimasungidwa zimasungidwa kutentha (20-21 ° C) ndipo kufa kwa mphutsi kunalembedwa mkati mwa maola 24 ndi 72 akuwonetseredwa mosalekeza kwa chithandizo chamankhwala.Ngati thupi la udzudzu ndi zomangira zake sizikusuntha pobooledwa kapena kukhudza ndi spatula wopyapyala wachitsulo chosapanga dzimbiri, mphutsi za udzudzu zimatengedwa zakufa.Mphutsi zakufa nthawi zambiri zimakhala zosasunthika pamtunda kapena pamtunda pansi pa chidebe kapena pamwamba pa madzi.Kuyesera kunabwerezedwa katatu pamasiku osiyanasiyana pogwiritsa ntchito magulu osiyanasiyana a mphutsi, chifukwa cha mphutsi za 180 zomwe zimawonekera pamtundu uliwonse wa mankhwala.
Kuopsa kwa AITC, BITC, ndi 4-HBITC ku mphutsi za udzudzu kunayesedwa pogwiritsa ntchito njira yofananira ya bioassay koma ndi mankhwala osiyanasiyana.Konzani 100,000 ppm stock solutions pa mankhwala aliwonse powonjezera 100 µL ya mankhwalawo ku 900 µL ya absolute ethanol mu chubu cha 2-mL centrifuge ndikugwedeza kwa masekondi 30 kuti musakanize bwino.Kukhazikika kwamankhwala kunatsimikiziridwa kutengera ma bioassays athu oyambilira, omwe adapeza BITC kukhala yapoizoni kwambiri kuposa AITC ndi 4-HBITC.Kuti mudziwe kawopsedwe, 5 yokhazikika ya BITC (1, 3, 6, 9 ndi 12 ppm), 7 yokhazikika ya AITC (5, 10, 15, 20, 25, 30 ndi 35 ppm) ndi 6 ndende ya 4-HBITC (15). , 15, 20, 25, 30 ndi 35 ppm).30, 45, 60, 75 ndi 90 ppm).Chithandizo chowongolera chidayikidwa ndi 108 μL ya ethanol mtheradi, yomwe ndi yofanana ndi kuchuluka kwa mankhwalawo.Ma bioassays anabwerezedwa monga pamwambapa, kuwonetsa mphutsi zokwana 180 pa chithandizo chamankhwala.Kufa kwa mphutsi kunalembedwa pamndandanda uliwonse wa AITC, BITC, ndi 4-HBITC pambuyo pa maola 24 akuwonekera mosalekeza.
Kusanthula kwa probit kwa data yakufa yokhudzana ndi mlingo wa 65 kunachitidwa pogwiritsa ntchito pulogalamu ya Polo (Polo Plus, LeOra Software, version 1.0) kuti awerengere 50% yakupha (LC50), 90% lethal concentration (LC90), slope, lethal dose coefficient, ndi 95 % ndende yakupha.kutengera nthawi yachikhulupiriro pamiyezo yakupha ya mlingo wakupha wa ndende yosinthidwa chipika ndi ma curve a kufa kwa mlingo.Zomwe zimafa zimatengera kuphatikizika kwa mphutsi zokwana 180 zomwe zimawonekera pachiwonetsero chilichonse chamankhwala.Kusanthula kwachidziwitso kunkachitika padera pa chakudya chilichonse chambewu ndi chigawo chilichonse chamankhwala.Malingana ndi 95% nthawi yodalirika ya chiŵerengero chakupha, kuopsa kwa ufa wa mbewu ndi mankhwala opangidwa ndi mphutsi za udzudzu kumaganiziridwa kuti ndizosiyana kwambiri, kotero kuti nthawi yodalirika yokhala ndi mtengo wa 1 sinali yosiyana kwambiri, P = 0.0566.
Zotsatira za HPLC pakutsimikiza kwa glucosinolates zazikulu mu ufa wambewu wa DFP, IG, PG ndi Ls zalembedwa mu Table 1. Magulu akuluakulu a glucosinolate mu ufa wambewu omwe amayesedwa amasiyana mosiyana ndi DFP ndi PG, zomwe zonse zinali ndi myrosinase glucosinolates.Zomwe zili mu myrosinin mu PG zinali zapamwamba kuposa DFP, 33.3 ± 1.5 ndi 26.5 ± 0.9 mg/g, motsatira.Ls ufa wambewu unali ndi 36.6 ± 1.2 mg/g glucoglycone, pamene IG ufa wambewu unali ndi 38.0 ± 0.5 mg/g sinapine.
Mphutsi za Ae.Udzudzu wa Aedes aegypti unkaphedwa ukathiridwa ndi ufa wothira mafuta, ngakhale mphamvu ya mankhwalawa imasiyanasiyana malinga ndi mitundu ya mbewu.DFP-NT yokhayo inalibe poizoni ku mphutsi za udzudzu pambuyo pa maola 24 ndi 72 (Table 2).The kawopsedwe wa yogwira mbewu ufa chinawonjezeka ndi kuwonjezeka ndende (mkuyu. 1A, B).Kuopsa kwa ufa wambewu ku mphutsi za udzudzu kumasiyana kwambiri kutengera 95% CI ya chiwopsezo chakupha cha LC50 pamayendedwe a maola 24 ndi maola 72 (Table 3).Pambuyo pa maola 24, mphamvu ya poizoni ya Ls seed meal inali yayikulu kuposa njira zina zochiritsira zambewu, zokhala ndi zochitika zapamwamba kwambiri komanso kawopsedwe wamkulu wa mphutsi (LC50 = 0.04 g/120 ml dH2O).Mphutsi sizinali zokhudzidwa kwambiri ndi DFP pa maola 24 poyerekeza ndi mankhwala a IG, Ls ndi PG a ufa wambewu, okhala ndi LC50 values ya 0.115, 0.04 ndi 0.08 g/120 ml dH2O motsatana, omwe anali okwera kwambiri kuposa mtengo wa LC50.0,211 g/120 ml dH2O (Table 3).Makhalidwe a LC90 a DFP, IG, PG ndi Ls anali 0.376, 0.275, 0.137 ndi 0.074 g/120 ml dH2O, motsatana (Table 2).Kuchuluka kwa DPP kunali 0.12 g/120 ml dH2O.Pambuyo pakuwunika kwa maola 24, kuchuluka kwa kufa kwa mphutsi kunali 12% yokha, pomwe kufa kwa IG ndi PG mphutsi kudafika 51% ndi 82%, motsatana.Pambuyo maola 24 kuwunika, pafupifupi imfa mphutsi kwa mkulu ndende ya Ls mbewu chakudya mankhwala (0.075 g/120 ml dH2O) anali 99% (mkuyu. 1A).
Mapiritsi a imfa adayesedwa kuchokera ku kuyankha kwa mlingo (Probit) ya Ae.Mphutsi za ku Aigupto (mphutsi za 3rd instar) ku ndende yambewu yambewu maola 24 (A) ndi maola 72 (B) pambuyo pa chithandizo.Mzere wamadontho umayimira LC50 ya chithandizo cha chakudya chambewu.DFP Thlaspi arvense, DFP-HT Heat inactivated Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Pakuwunika kwa maola 72, mikhalidwe ya LC50 ya DFP, IG ndi PG mbewu yambewu inali 0.111, 0.085 ndi 0.051 g/120 ml dH2O, motsatana.Pafupifupi mphutsi zonse zomwe zimakhudzidwa ndi chakudya cha Ls zinafa pambuyo pa maola 72, kotero kuti chiwerengero cha imfa sichinali chogwirizana ndi kusanthula kwa Probit.Poyerekeza ndi chakudya china chambewu, mphutsi sizimamva bwino kwambiri ndi chithandizo cha mbewu za DFP ndipo zinali ndi ma LC50 apamwamba kwambiri (Matebulo 2 ndi 3).Pambuyo pa maola 72, milingo ya LC50 pazakudya za DFP, IG ndi PG idayerekezedwa kukhala 0.111, 0.085 ndi 0.05 g/120 ml dH2O, motsatana.Pambuyo pakuwunika kwa maola 72, mikhalidwe ya LC90 ya ufa wambewu wa DFP, IG ndi PG inali 0.215, 0.254 ndi 0.138 g/120 ml dH2O, motsatana.Pambuyo pa kuwunika kwa maola 72, kuchuluka kwa kufa kwa mphutsi kwa DFP, IG ndi PG mankhwala ambewu yambewu pamlingo wopitilira 0,12 g/120 ml dH2O kunali 58%, 66% ndi 96%, motsatana (mkuyu 1 B).Pambuyo pakuwunika kwa maola 72, chakudya chambewu cha PG chidapezeka kuti ndi chapoizoni kuposa IG ndi DFP mbewu yambewu.
Synthetic isothiocyanates, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC) ndi 4-hydroxybenzylisothiocyanate (4-HBITC) amatha kupha mphutsi za udzudzu.Pamaola 24 mutalandira chithandizo, BITC inali yapoizoni kwambiri ku mphutsi zamtengo wapatali za LC50 za 5.29 ppm poyerekeza ndi 19.35 ppm ya AITC ndi 55.41 ppm ya 4-HBITC (Table 4).Poyerekeza ndi AITC ndi BITC, 4-HBITC ili ndi kawopsedwe kochepa komanso mtengo wapamwamba wa LC50.Pali kusiyana kwakukulu kwa kawopsedwe ka mphutsi ya udzudzu wa ma isothiocyanates awiri akuluakulu (Ls ndi PG) muzakudya zamphamvu kwambiri zambewu.Kuopsa kotengera kuchuluka kwa mlingo wakupha wa LC50 pakati pa AITC, BITC, ndi 4-HBITC kunawonetsa kusiyana kwa ziwerengero kotero kuti 95% CI ya chiŵerengero cha LC50 chakupha sichinaphatikizepo mtengo wa 1 (P = 0.05, Table 4).Kuchuluka kwambiri kwa BITC ndi AITC kunkapha 100% ya mphutsi zomwe zayesedwa (Chithunzi 2).
Mapiritsi a imfa adayesedwa kuchokera ku kuyankha kwa mlingo (Probit) ya Ae.Maola a 24 atalandira chithandizo, mphutsi za ku Egypt (3rd instar larvae) zinafika pakupanga isothiocyanate.Mzere wamadontho umayimira LC50 ya chithandizo cha isothiocyanate.Benzyl isothiocyanate BITC, allyl isothiocyanate AITC ndi 4-HBITC.
Kugwiritsa ntchito mankhwala ophera tizilombo ngati tizilombo toyambitsa udzudzu kwaphunziridwa kalekale.Zomera zambiri zimapanga mankhwala achilengedwe omwe ali ndi mankhwala ophera tizilombo37.Mankhwalawa amapereka njira yowoneka bwino yopangira mankhwala ophera tizilombo omwe ali ndi kuthekera kwakukulu kothana ndi tizirombo, kuphatikiza udzudzu.
Zomera za mpiru zimabzalidwa ngati mbewu zambewu zawo, zimagwiritsidwa ntchito ngati zonunkhira komanso gwero lamafuta.Mafuta a mpiru akachotsedwa mu njere kapena mpiru akachotsedwa kuti agwiritsidwe ntchito ngati mafuta achilengedwe, 69 chotulukapo chake ndi ufa wambewu wodetsedwa.Chakudya chambewuchi chimakhalabe ndi zinthu zambiri zachilengedwe zachilengedwe komanso ma hydrolytic enzymes.Kuopsa kwa chakudya chambewuchi kumabwera chifukwa chopanga isothiocyanates55,60,61.Isothiocyanates amapangidwa ndi hydrolysis wa glucosinolates ndi enzyme myrosinase pa hydration wa mbewu chakudya38,55,70 ndipo amadziwika kuti ndi fungicidal, bactericidal, nematicidal ndi insecticidal zotsatira, komanso katundu wina kuphatikizapo zotsatira za mankhwala zomverera ndi chemotherapeutic katundu61,62, 70.Kafukufuku wambiri wasonyeza kuti mbewu za mpiru ndi chakudya chambewu zimagwira ntchito bwino ngati zofukiza zowononga nthaka ndi tizirombo tosungidwa m'zakudya57,59,71,72.M'kafukufukuyu, tawunika kawopsedwe ka ufa wambewu zinayi ndi zinthu zake zitatu zokhala ndi bioactive AITC, BITC, ndi 4-HBITC kupita ku mphutsi za udzudzu wa Aedes.Aedes aegypti.Kuonjezera chakudya chambewu kumadzi omwe ali ndi mphutsi za udzudzu kukuyembekezeka kuyambitsa njira za enzymatic zomwe zimapanga isothiocyanates zomwe zimakhala zoopsa ku mphutsi za udzudzu.Biotransformation iyi idawonetsedwa mwa zina ndi zomwe zidachitika pazakudya zambewu komanso kuwonongeka kwa tizirombo pomwe ufa wocheperako wambewu yampiru udatenthedwa usanagwiritsidwe ntchito.Chithandizo cha kutentha chikuyembekezeka kuwononga michere ya hydrolytic yomwe imayambitsa glucosinolates, potero kulepheretsa mapangidwe a bioactive isothiocyanates.Aka ndi kafukufuku woyamba kutsimikizira kuti ufa wa mbewu ya kabichi ndi udzudzu m'madzi.
Pakati pa ufa wambewu womwe unayesedwa, madzi amadzimadzi a ufa (Ls) anali oopsa kwambiri, omwe amachititsa kuti Aedes albopictus afe.Mphutsi za Aedes aegypti zinkakonzedwa mosalekeza kwa maola 24.Mafuta atatu otsala a mbewu (PG, IG ndi DFP) anali ndi ntchito pang'onopang'ono ndipo adayambitsabe kufa kwakukulu pambuyo pa maola 72 akuchipatala mosalekeza.Chakudya cha Ls chokha chinali ndi ma glucosinolates ambiri, pomwe PG ndi DFP zinali ndi myrosinase ndipo IG inali ndi glucosinolate monga glucosinolate yayikulu (Table 1).Glucotropaeolin imapangidwa ndi hydrolyzed kupita ku BITC ndipo sinalbine imasinthidwa kukhala 4-HBITC61,62.Zotsatira zathu za bioassay zikuwonetsa kuti zonse ziwiri za Ls seed meal ndi synthetic BITC ndizowopsa ku mphutsi za udzudzu.Chigawo chachikulu cha chakudya cha PG ndi DFP ndi myrosinase glucosinolate, yomwe imapangidwa ndi hydrolyzed kukhala AITC.AITC imathandiza kupha mphutsi za udzudzu ndi mtengo wa LC50 wa 19.35 ppm.Poyerekeza ndi AITC ndi BITC, 4-HBITC isothiocyanate ndiyowopsa kwambiri ku mphutsi.Ngakhale AITC ilibe poizoni pang'ono kuposa BITC, ma LC50 awo ndi otsika kuposa mafuta ambiri ofunikira omwe amayesedwa pa mphutsi za udzudzu32,73,74,75.
Ufa wathu wambewu wa cruciferous womwe umagwiritsidwa ntchito polimbana ndi mphutsi za udzudzu uli ndi glucosinolate imodzi yaikulu, yomwe imawerengera 98-99% ya glucosinolates yonse monga momwe HPLC yatsimikizira.Kufufuza kwa ma glucosinolates ena kunapezeka, koma milingo yawo inali yochepera 0.3% ya ma glucosinolates onse.Watercress (L. sativum) ufa wambewu uli ndi glucosinolates yachiwiri (sinigrin), koma gawo lawo ndi 1% ya glucosinolates onse, ndipo zomwe zili nazo zimakhalabe zochepa (pafupifupi 0.4 mg / g ufa wambewu).Ngakhale PG ndi DFP zili ndi glucosinolate (myrosin), zomwe zimachitika pakudya kwa mbewu zawo zimasiyana kwambiri chifukwa cha LC50.Zimasiyanasiyana potengera powdery mildew.Kutuluka kwa mphutsi za Aedes aegypti kungakhale chifukwa cha kusiyana kwa ntchito ya myrosinase kapena kukhazikika pakati pa zakudya ziwiri zambewu.Ntchito ya Myrosinase imakhala ndi gawo lofunikira pakupezeka kwazinthu za hydrolysis monga isothiocyanates mu zomera za Brassicaceae76.Malipoti am'mbuyomu a Pocock et al.77 ndi Wilkinson et al.78 awonetsa kuti kusintha kwa ntchito ya myrosinase ndi kukhazikika kungagwirizanenso ndi majini ndi chilengedwe.
Zomwe zimayembekezeredwa za isothiocyanate za bioactive zidawerengedwa kutengera milingo ya LC50 ya chakudya chilichonse chambewu pa maola 24 ndi 72 (Table 5) poyerekeza ndi kugwiritsa ntchito mankhwala ofanana.Pambuyo pa maola 24, ma isothiocyanates muzakudya zambewu anali owopsa kwambiri kuposa mankhwala oyera.Miyezo ya LC50 yowerengeredwa kutengera magawo miliyoni (ppm) amankhwala ambewu ya isothiocyanate anali otsika kuposa ma LC50 pamitengo ya BITC, AITC, ndi 4-HBITC.Tidawona mphutsi zikudya mbewu zambewu (Chithunzi 3A).Chifukwa chake, mphutsi zimatha kukhudzidwa kwambiri ndi ma isothiocyanates oopsa pomeza ma pellets ambewu.Izi zidawonekera kwambiri pazakudya za IG ndi PG pakuwonetsa kwa 24-h, pomwe LC50 yokhazikika inali 75% ndi 72% yotsika kuposa mankhwala a AITC ndi 4-HBITC, motsatana.Mankhwala a Ls ndi DFP anali oopsa kwambiri kuposa isothiocyanate yoyera, yokhala ndi LC50 24% ndi 41% kutsika, motsatana.Mphutsi mu ulamuliro mankhwala bwinobwino pupated (mkuyu. 3B), pamene ambiri mphutsi mu mbewu chakudya mankhwala sanali pupate ndi mphutsi chitukuko anali kwambiri anachedwa (mkuyu. 3B, D).Mu Spodopteralitura, isothiocyanates imalumikizidwa ndi kuchepa kwa kukula komanso kuchedwa kwachitukuko79.
Mphutsi za Ae.Udzudzu wa Aedes aegypti unkawonekera mosalekeza ku ufa wa mbewu ya Brassica kwa maola 24-72.(A) Mphutsi zakufa zokhala ndi tinthu tating'ono ta ufa wambewu m'kamwa (zozungulira);(B) Kuchiza (dH20 popanda chakudya chowonjezera) kumasonyeza kuti mphutsi zimakula bwino ndipo zimayamba kuphulika pambuyo pa maola 72 (C, D) Mphutsi zomwe zimagwiritsidwa ntchito ndi mbewu;mbewu ufa anasonyeza kusiyana chitukuko ndipo sanali pupate.
Sitinaphunzire njira ya poizoni wa isothiocyanates pa mphutsi za udzudzu.Komabe, kafukufuku wam'mbuyomu mu nyerere zofiira (Solenopsis invicta) awonetsa kuti kuletsa kwa glutathione S-transferase (GST) ndi esterase (EST) ndiyo njira yayikulu ya isothiocyanate bioactivity, ndipo AITC, ngakhale pa ntchito yochepa, imathanso kuletsa ntchito ya GST. .nyerere zofiira zochokera kunja m'madera otsika.Mlingo ndi 0.5 µg/ml80.Mosiyana ndi zimenezi, AITC imaletsa acetylcholinesterase m'magulu akuluakulu a chimanga (Sitophilus zeamais)81.Maphunziro ofananawo ayenera kuchitidwa kuti afotokoze momwe ntchito ya isothiocyanate imagwirira ntchito mu mphutsi za udzudzu.
Timagwiritsa ntchito chithandizo cha DFP chopanda kutentha kuti tithandizire lingaliro lakuti hydrolysis ya zomera glucosinolates kupanga reactive isothiocyanates imakhala njira yothetsera mphutsi ya udzudzu ndi ufa wa mpiru.Zakudya zambewu za DFP-HT sizinali zapoizoni pamitengo yoyesedwa yoyesedwa.Lafarga et al.82 inanena kuti glucosinolates amakhudzidwa ndi kuwonongeka pa kutentha kwakukulu.Kuchiza kwa kutentha kumayembekezeredwanso kusokoneza enzyme ya myrosinase muzakudya zambewu ndikuletsa hydrolysis ya glucosinolates kupanga isothiocyanates yogwira ntchito.Izi zidatsimikiziridwanso ndi Okunade et al.75 inasonyeza kuti myrosinase ndi yokhudzidwa ndi kutentha, kusonyeza kuti ntchito ya myrosinase inazimitsidwa pamene mpiru, mpiru wakuda, ndi njere za bloodroot zinawonetsedwa ndi kutentha pamwamba pa 80 °.C. Njirazi zingayambitse kutaya kwa tizilombo toyambitsa matenda a ufa wambewu wa DFP wotenthedwa.
Chifukwa chake, ufa wambewu ya mpiru ndi ma isothiocyanate ake akuluakulu atatu ndi owopsa ku mphutsi za udzudzu.Poganizira kusiyana kumeneku pakati pa chakudya chambewu ndi mankhwala ochiritsira, kugwiritsa ntchito ufa wambewu kungakhale njira yabwino yoletsera udzudzu.M'pofunika kuzindikira mapangidwe oyenera ndi njira zoperekera zoperekera bwino kuti zikhale zogwira mtima komanso zokhazikika za kugwiritsa ntchito ufa wambewu.Zotsatira zathu zikuwonetsa kugwiritsa ntchito ufa wa mpiru m'malo mwa mankhwala ophera tizilombo.Ukadaulowu utha kukhala chida chanzeru chowongolera ma vector a udzudzu.Chifukwa mphutsi za udzudzu zimakula bwino m'madzi ndipo ma glucosinolates ambewu amasandulika enzymatically kukhala isothiocyanates pa hydration, kugwiritsa ntchito ufa wa mpiru m'madzi odzaza ndi udzudzu kumapereka mphamvu zowonongeka.Ngakhale ntchito ya larvicidal ya isothiocyanates imasiyanasiyana (BITC> AITC> 4-HBITC), kafukufuku wochulukirapo amafunika kudziwa ngati kuphatikiza ufa wambewu ndi ma glucosinolate angapo kumawonjezera kawopsedwe.Uwu ndi kafukufuku woyamba kuwonetsa zotsatira zowononga tizilombo ta cruciferous seed meal ndi ma bioactive isothiocyanates atatu pa udzudzu.Zotsatira za kafukufukuyu zikuwonetsa kuti ufa wambewu wa kabichi wodetsedwa, womwe umachokera ku njere zamafuta, utha kukhala ngati chida chothandiza polimbana ndi udzudzu.Chidziwitsochi chingathandize kupititsa patsogolo kupezeka kwa maantibiotiki a zomera ndi chitukuko chake monga mankhwala ophera tizilombo otsika mtengo, ogwira ntchito, komanso osamalira chilengedwe.
Ma data omwe apangidwa pa kafukufukuyu ndi zotsatira zake akupezeka kuchokera kwa mlembi wogwirizana ndi zomwe akufuna.Pamapeto pa phunzirolo, zida zonse zomwe zidagwiritsidwa ntchito pophunzira (tizilombo ndi ufa wambewu) zidawonongeka.
Nthawi yotumiza: Jul-29-2024