I ka ponohoʻomalu namua e ho'ēmi i ka nui o nā maʻi a lākou e lawe nei, pono nā mea ʻokoʻa ʻē aʻe e pili ana i nā lāʻau pesticides kemika.Ua loiloi mākou i nā huaʻai mai kekahi Brassicaceae (ʻohana Brassica) ma ke ʻano he kumu o nā isothiocyanates i loaʻa i nā mea kanu i hana ʻia e ka hydrolysis enzymatic o nā glucosinolates biologically inactive no ka hoʻohana ʻana i ka mālama ʻana i ka Aedes ʻAigupita (L., 1762).ʻElima-defatted hua ʻai (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 a me Thlaspi arvense - ʻekolu mau ʻano nui o ka thermal inactivation a me ka enzymatic degradation. nā huahana No ka hoʻoholo ʻana i nā mea ʻawaʻawa (LC50) o allyl isothiocyanate, benzyl isothiocyanate a me 4-hydroxybenzylisothiocyanate iā Aedes aegypti larvae ma 24-hola hoʻolaha = 0.04 g/120 ml dH2O).Nā waiwai LC50 no ka sinapi, ka sinapi keʻokeʻo a me ka huelo lio.ʻO kaʻai hua he 0.05, 0.08 a me 0.05 i hoʻohālikelikeʻia me allyl isothiocyanate (LC50 = 19.35 ppm) a me 4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) uaʻoi aku kaʻawaʻawa i nā larvae ma o 24 mau hola g / 200 ma hope o ka lāʻau.Kūlike kēia mau hopena me ka hana ʻana i ka ʻai hua alfalfa.ʻO ka maikaʻi kiʻekiʻe o nā benzyl esters e pili ana i nā helu LC50 i helu ʻia.ʻO ka hoʻohana ʻana i ka ʻai hua hiki ke hāʻawi i kahi ala maikaʻi no ka pale ʻana i ka namu.ka maikaʻi o ka pauka hua cruciferous a me kāna mau mea kemika nui e kūʻē i nā ʻōpala namu a hōʻike i ke ʻano o nā pūhui kūlohelohe i loko o ka pauka hua cruciferous hiki ke lilo i larvicide maikaʻi maikaʻi no ka namu.
ʻO nā maʻi i lawe ʻia e ka namu Aedes he pilikia nui o ke olakino lehulehu.Hoʻolaha ka maʻi i lawe ʻia e ka makika ma ka ʻāina1,2,3 a puka hou mai, e alakaʻi ana i nā maʻi maʻi nui4,5,6,7.ʻO ka laha ʻana o nā maʻi ma waena o nā kānaka a me nā holoholona (e laʻa, chikungunya, dengue, Rift Valley fever, yellow fever a me Zika virus) ʻaʻole i loaʻa ma mua.ʻO ka maʻi maʻi Dengue wale nō ka mea e loaʻa ai ma kahi o 3.6 biliona mau kānaka i ka pilikia o ka maʻi i loko o ka ʻāina, me ka manaʻo ʻia he 390 miliona mau maʻi e loaʻa ana i kēlā me kēia makahiki, a he 6,100-24,300 mau make i kēlā me kēia makahiki8.ʻO ka puka hou ʻana a me ka laha ʻana o ka maʻi Zika ma ʻAmelika Hema ua hoʻohuli ʻia ka manaʻo o ka honua ma muli o ka pōʻino o ka lolo i hana ʻia i nā keiki i hānau ʻia e nā wahine maʻi.Ua wānana ʻo Kremer et al 3 e hoʻonui ʻia ana ka nui o nā makika Aedes a hiki i ka makahiki 2050, hiki i ka hapalua o ka heluna o ka honua ke loaʻa i ka maʻi e nā arbovirus e lawe ʻia e nā makika.
Me ka ʻokoʻa o nā lāʻau lapaʻau i kūkulu hou ʻia e kūʻē i ka dengue a me ka maʻi melemele, ʻaʻole i hoʻomohala ʻia nā lāʻau lapaʻau e kūʻē i ka hapa nui o nā maʻi i lawe ʻia e ka makika9,10,11.Loaʻa nā lāʻau lapaʻau ma nā helu liʻiliʻi a hoʻohana wale ʻia i nā hoʻokolohua lapaʻau.ʻO ka hoʻomalu ʻana i nā mea namu me ka hoʻohana ʻana i nā insecticides synthetic he kumu koʻikoʻi ia e pale ai i ka laha ʻana o nā maʻi i lawe ʻia e ka makika12,13.ʻOiai he maikaʻi nā pesticides synthetic i ka pepehi ʻana i nā makika, ʻo ka hoʻohana mau ʻana i nā pesticides synthetic e hoʻopilikia maikaʻi ʻole i nā mea ola ʻole a hoʻohaumia i ke kaiapuni14,15,16.ʻOi aku ka weliweli ʻo ke ʻano o ka hoʻonui ʻana i ke kūʻē ʻana o ka makika i nā lāʻau insecticides17,18,19.ʻO kēia mau pilikia e pili ana i nā pesticides ua hoʻonui i ka ʻimi ʻana i nā mea ʻē aʻe kūpono a pili i ke kaiapuni no ka hoʻopaʻa ʻana i nā vector maʻi.
Ua hoʻomohala ʻia nā mea kanu like ʻole i kumu o nā phytopesticides no ka hoʻomalu ʻana i nā pest20,21.ʻO nā mea kanu ka mea maʻamau i ke kaiapuni no ka mea hiki ke hoʻoheheʻe ʻia a he haʻahaʻa a haʻahaʻa ʻole paha ka ʻona i nā mea ola ʻole e like me nā mammals, nā iʻa a me nā amphibians20,22.ʻIke ʻia ka hoʻomākaukau ʻana i nā lāʻau lapaʻau e hana i nā ʻano mea like ʻole me nā ʻano hana like ʻole e hoʻomalu pono i nā ʻano ola like ʻole o nā makika23,24,25,26.ʻO nā pūhui i loaʻa i nā mea kanu e like me nā ʻaila koʻikoʻi a me nā mea kanu ikaika ʻē aʻe ua loaʻa ka manaʻo a hoʻokaʻawale i ke ala no nā mea hana hou e kāohi ai i nā namu.ʻO nā aila koʻikoʻi, monoterpenes a me sesquiterpenes e hana ma ke ʻano he repellents, hānai i nā deterrents a me nā ovicides27,28,29,30,31,32,33.ʻO ka nui o nā aila meaʻai ke kumu i ka make ʻana o ka makika, pupae a me nā mākua34,35,36, e pili ana i ka puʻuwai, hanu, endocrine a me nā ʻōnaehana koʻikoʻi ʻē aʻe o nā pepeke37.
Ua hāʻawi nā haʻawina hou i ka ʻike e pili ana i ka hoʻohana ʻana i nā mea kanu sinapi a me kā lākou mau hua ma ke ʻano he kumu o nā pūhui bioactive.Ua ho'āʻo ʻia ka ʻai hua sinapi ma ke ʻano he biofumigant38,39,40,41 a hoʻohana ʻia ma ke ʻano he hoʻololi lepo no ka hoʻopau ʻana i ka weed42,43,44 a me ka hoʻomalu ʻana i nā pathogens mea kanu i lawe ʻia i ka lepo45,46,47,48,49,50, meaʻai meaʻai.nematodes 41,51, 52, 53, 54 a me nā pests 55, 56, 57, 58, 59, 60. ʻO ka hana fungicidal o kēia mau mea kanu pauda ua pili i nā mea kanu pale i kapa ʻia isothiocyanates38,42,60.I loko o nā mea kanu, mālama ʻia kēia mau mea pale i loko o nā mea kanu i ke ʻano o nā glucosinolates non-bioactive.Eia nō naʻe, ke hoʻopōʻino ʻia nā mea kanu e ka hānai ʻana a i ʻole ka maʻi pathogen, hoʻoheheʻe ʻia nā glucosinolates e myrosinase i bioactive isothiocyanates55,61.ʻO Isothiocyanates he mau mea hoʻoheheʻe ʻia i ʻike ʻia he ākea ākea ka antimicrobial a me ka hana insecticidal, a ʻokoʻa ko lākou ʻano, hana ola a me nā ʻike ma waena o nā ʻano Brassicaceae42,59,62,63.
ʻOiai ua ʻike ʻia nā isothiocyanates i loaʻa mai ka ʻai hua sinapi he hana insecticidal, nele ka ʻikepili e pili ana i ka hana olaola e kūʻē i nā vectors arthropod koʻikoʻi.Ua nānā kā mākou noiʻi ʻana i ka hana larvicidal o ʻehā pauka ʻanoʻano defatted e kūʻē i nā makika Aedes.ʻO nā ʻanuʻu o Aedes aegypti.ʻO ka pahuhopu o ke aʻo ʻana ʻo ia ka loiloi ʻana i kā lākou hiki ke hoʻohana ʻia ma ke ʻano he biopesticides pili i ke kaiapuni no ka pale namu.ʻEkolu mau mea kemika nui o ka ʻai hua, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC), a me 4-hydroxybenzylisothiocyanate (4-HBITC) i hoʻāʻo pū ʻia e hoʻāʻo i ka hana olaola o kēia mau mea kemika ma nā ʻano namu.ʻO kēia ka hōʻike mua e loiloi i ka maikaʻi o nā pauka hua kāpeti ʻehā a me kā lākou mau mea kemika nui e kūʻē i nā ʻōpala namu.
Ua mālama ʻia nā ʻāpana Laboratory o Aedes aegypti (Rockefeller strain) ma 26°C, 70% humidity pili (RH) a me 10:14 h (L:D photoperiod).Hoʻokomo ʻia nā wahine male i loko o nā hīnaʻi plastik (ke kiʻekiʻe 11 knm a me ke anawaena 9.5 knm) a hānai ʻia ma o kahi ʻōnaehana hānai omole me ka hoʻohana ʻana i ke koko bovine citrated (HemoStat Laboratories Inc., Dixon, CA, USA).Hoʻokō ʻia ka hānai koko e like me ka mea maʻamau me ka hoʻohana ʻana i kahi mea hānai aniani multi-membrane (Chemglass, Life Science LLC, Vineland, NJ, USA) i hoʻopili ʻia i kahi paipu ʻauʻau wai kahe (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) me ka mahana. mana 37 °C.E kau i kahi kiʻiʻoniʻoni o Parafilm M ma lalo o kēlā me kēia keʻena hānai aniani (ʻāpana 154 mm2).A laila kau ʻia kēlā me kēia mea hānai ma luna o ka mākia luna e uhi ana i ka hīnaʻi i loko o ka wahine male.Ma kahi o 350–400 μl o ke koko bipi i hoʻokomo ʻia i loko o kahi ʻōhua hānai aniani me ka hoʻohana ʻana i kahi paipu Pasteur (Fisherbrand, Fisher Scientific, Waltham, MA, USA) a ua ʻae ʻia nā ilo makua e hoʻokahe no hoʻokahi hola.Hāʻawi ʻia nā wahine hāpai i ka wai 10% sucrose a ʻae ʻia e waiho i nā hua ma luna o ka pepa kānana maʻemaʻe i laina ʻia i loko o nā kīʻaha soufflé ultra-clear (1.25 fl oz ka nui, Dart Container Corp., Mason, MI, USA).hale me ka wai.E kau i ka pepa kānana i loko o kahi ʻeke i sila ʻia (SC Johnsons, Racine, WI) a mālama i ka 26°C.Ua paʻi ʻia nā hua a ma kahi o 200-250 larvae i hānai ʻia i loko o nā pā plastik i loaʻa kahi hui ʻana o ka rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) a me ka pauka ate (MP Biomedicals, LLC, Solon, OH, USA).a me ka fillet iʻa (TetraMin, Tetra GMPH, Meer, Kelemānia) i ka ratio o 2:1:1.Ua hoʻohana ʻia nā larvae instar hope ʻekolu i kā mākou bioassays.
Ua loaʻa nā mea kanu i hoʻohana ʻia ma kēia noiʻi ʻana mai nā kumu kūʻai kālepa a me ke aupuni: Brassica juncea (brown mustard-Pacific Gold) a me Brassica juncea (white mustard-Ida Gold) mai ka Pacific Northwest Farmers' Cooperative, Washington State, USA;(Garden Cress) mai Kelly Seed and Hardware Co., Peoria, IL, USA a me Thlaspi arvense (Field Pennycress-Elisabeth) mai USDA-ARS, Peoria, IL, USA;ʻAʻohe o nā hua i hoʻohana ʻia i ke aʻo ʻana i mālama ʻia me nā pesticides.Ua hoʻohana ʻia a hoʻohana ʻia nā mea kanu a pau i loko o kēia haʻawina e like me nā lula kūloko a me ka ʻāina a me ka hoʻokō ʻana i nā lula pili kūloko a me nā aupuni āpau.ʻAʻole i nānā kēia haʻawina i nā ʻano mea kanu transgenic.
ʻO nā hua Brassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) hua ʻia i ka pauka maikaʻi me ka Retsch ZM200 ultracentrifugal mill (Retsch, Haan, Kelemānia) i hoʻolako ʻia me kahi 0.75 mm mesh a me Stainless. rotor kila, 12 niho, 10,000 rpm (Papa 1).Hoʻololi ʻia ka pauka hua ʻāina i kahi thimble pepa a hoʻopau ʻia me ka hexane i loko o kahi mea hana Soxhlet no 24 mau hola.Ua mālama ʻia kahi subsample o ka sinapi māla i hoʻomaʻemaʻe ʻia ma 100 °C no 1 h e hoʻopau i ka myrosinase a pale i ka hydrolysis o nā glucosinolates e hana i nā isothiocyanates biologically active.Ua hoʻohana ʻia ka pauka hua liʻiliʻi i mālama ʻia me ka wela (DFP-HT) ma ke ʻano he mana maikaʻi ʻole ma ka denaturing myrosinase.
Hoʻoholo ʻia ka maʻiʻo Glucosinolate o ka ʻai hua defatted ma ke ʻano ʻekolu me ka hoʻohana ʻana i ka chromatography wai hana kiʻekiʻe (HPLC) e like me kahi protocol 64 i paʻi mua ʻia.ʻO ka pōkole, ua hoʻohui ʻia he 3 mL o ka methanol i kahi laʻana 250 mg o ka pauka hua defatted.Kēlā me kēia hāpana i sonicated i loko o ka wai auau no 30 minuke a waiho i loko o ka pouli ma 23 ° C no 16 hola.Ua kānana ʻia kahi 1 mL aliquot o ka papa organik ma o kahi kānana 0.45 μm i loko o kahi autosampler.Ke holo nei ma kahi ʻōnaehana Shimadzu HPLC (ʻelua LC 20AD pumps; SIL 20A autosampler; DGU 20As degasser; SPD-20A UV-VIS detector no ka nānā ʻana ma 237 nm; a me CBM-20A communication bus module), ua hoʻoholo ʻia ka ʻike glucosinolate o ka ʻai hua. ma ka papakolu .me ka hoʻohana ʻana iā Shimadzu LC Solution software version 1.25 (Shimadzu Corporation, Columbia, MD, USA).ʻO ke kolamu he kolamu C18 Inertsil reverse phase (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA).Ua hoʻonohonohoʻia nā kūlana hoʻolālā pūnaewele mua ma 12% methanol / 88% 0.01 M tetrabutylammonium hydroxide i loko o ka wai (TBAH; Sigma-Aldrich, St. Louis, MO, USA) me ke kahe o 1 mL / min.Ma hope o ka injection o 15 μl o ka laʻana, ua mālama ʻia nā kūlana mua no 20 mau minuke, a laila ua hoʻoponopono ʻia ka lākiō solvent i 100% methanol, me ka manawa hōʻike hōʻike holoʻokoʻa o 65 mau minuke.Ua hoʻokumu ʻia kahi ʻāpana maʻamau (nM/mAb) e nā dilutions serial o nā maʻamau sinapine, glucosinolate a me myrosin hou i hoʻomākaukau ʻia (Sigma-Aldrich, St. Louis, MO, USA) no ka hoʻohālikelike ʻana i ka maʻi sulfur o ka ʻai hua defatted.glucosinolates.Ua ho'āʻoʻia nā manaʻo Glucosinolate i nā laʻana ma kahi Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) me ka hoʻohanaʻana i ka OpenLAB CDS ChemStation version (C.01.07 SR2 [255]) i hoʻolakoʻia me ke kolamu like a me ka hoʻohanaʻana i keʻano i wehewehe muaʻia.Ua hoʻoholo ʻia ka ʻike ʻana o Glucosinolate;e hoʻohālikelike ma waena o nā ʻōnaehana HPLC.
Ua kūʻai ʻia ʻo Allyl isothiocyanate (94%, stable) a me benzyl isothiocyanate (98%) mai Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA).Ua kūʻai ʻia ʻo 4-Hydroxybenzylisothiocyanate mai ChemCruz (Santa Cruz Biotechnology, CA, USA).Ke hoʻoheheʻe ʻia ka enzymatically e myrosinase, nā glucosinolates, glucosinolates, a me nā glucosinolates e hana i allyl isothiocyanate, benzyl isothiocyanate, a me 4-hydroxybenzylisothiocyanate, kēlā me kēia.
Ua hana ʻia nā bioassays Laboratory e like me ke ʻano o Muturi et al.32 me nā hoʻololi.ʻElima mau meaʻai momona haʻahaʻa i hoʻohana ʻia i ka haʻawina: DFP, DFP-HT, IG, PG a me Ls.Ua hoʻokomo ʻia he iwakālua larvae i loko o ka beaker ʻekolu-way beaker 400 mL (VWR International, LLC, Radnor, PA, USA) me 120 mL wai deionized (dH2O).Ua hoʻāʻo ʻia ʻehiku mau ʻano ʻai ʻai no ka ʻawaʻawa o ka namu: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 a me 0.12 g ʻai hua/120 ml dH2O no ka ʻai hua DFP, DFP-HT, IG a me PG.Hōʻike nā bioassays mua he ʻoi aku ka ʻawaʻawa o ka palaoa hua Ls i hoʻopau ʻia ma mua o ʻehā mau palaoa hua ʻē aʻe i hoʻāʻo ʻia.No laila, ua hoʻololi mākou i nā ʻano hoʻomaʻamaʻa ʻehiku o ka ʻai hua Ls i kēia mau ʻano: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, a me 0.075 g/120 mL dH2O.
Ua hoʻokomo ʻia kahi pūʻulu hoʻomalu i mālama ʻole ʻia (dH20, ʻaʻohe kumu ʻai meaʻai) e nānā i ka make maʻamau o nā pepeke ma lalo o nā kūlana hoʻāʻo.ʻO nā bioassays lāʻau make no kēlā me kēia ʻai hua, ua loaʻa i ʻekolu mau ʻano ipu ʻekolu-slope (20 hope ʻekolu instar larvae no ka beaker), no ka huina o 108 vial.Ua mālama ʻia nā pahu i mālama ʻia ma ka lumi wela (20-21°C) a ua hoʻopaʻa ʻia ka make ʻana o nā larval i loko o 24 a me 72 mau hola o ka hoʻomau mau ʻana i nā ʻano lapaʻau.Inā ʻaʻole e neʻe ke kino o ka makika a me nā mea hoʻohui i ka wā e hou ʻia ai a pā ʻia paha me kahi moʻi kila kuhili ʻole, ua manaʻo ʻia ua make ka namu.Noho mau nā ʻōpala make me ka neʻe ʻole ma ke kua a i ʻole ka ventral ma lalo o ka ipu a i ʻole ma luna o ka wai.Ua hana hou ʻia ka hoʻokolohua ʻekolu manawa i nā lā like ʻole me ka hoʻohana ʻana i nā pūʻulu ʻokoʻa o nā larvae, no ka huina o 180 larvae i hōʻike ʻia i kēlā me kēia ʻano lapaʻau.
Ua helu ʻia ka ʻona o AITC, BITC, a me 4-HBITC i ka namu i ka hoʻohana ʻana i ke kaʻina bioassay like akā me nā lāʻau lapaʻau like ʻole.E hoʻomākaukau i 100,000 ppm mea hoʻonā waiwai no kēlā me kēia kemika ma ka hoʻohui ʻana i 100 µL o ke kemika i 900 µL o ka ethanol piha i loko o kahi paipu centrifuge 2-mL a lulu no 30 kekona e hui pono.Ua hoʻoholo ʻia nā manaʻo lapaʻau e pili ana i kā mākou bioassays mua, i ʻike ʻia ʻoi aku ka ʻona o ka BITC ma mua o AITC a me 4-HBITC.No ka hoʻoholo ʻana i nā mea ʻawaʻawa, 5 mau ʻāpana o BITC (1, 3, 6, 9 a me 12 ppm), 7 mau ʻāpana o AITC (5, 10, 15, 20, 25, 30 a me 35 ppm) a me 6 mau ʻāpana o 4-HBITC (15). , 15, 20, 25, 30 a me 35 ppm).30, 45, 60, 75 a me 90 ppm).Hoʻokomo ʻia ka lāʻau hoʻomalu me 108 μL o ka ethanol piha, ʻo ia ka like me ka nui o ka nui o ka lāʻau lapaʻau.Ua hana hou ʻia nā bioassays e like me ka mea i luna, e hōʻike ana i ka huina o 180 larvae i kēlā me kēia neʻe lapaʻau.Ua hoʻopaʻa ʻia ka make larval no kēlā me kēia neʻe o AITC, BITC, a me 4-HBITC ma hope o 24 mau hola o ka hoʻomau mau ʻana.
Ua hoʻohana ʻia ka ʻikepili probit o 65 mau ʻikepili make e pili ana i ka make me ka hoʻohana ʻana i ka polokalamu Polo (Polo Plus, LeOra Software, version 1.0) e helu i ka 50% lethal concentration (LC50), 90% lethal concentration (LC90), slope, lethal dose coefficient, a me 95 % hoʻopaʻapaʻa make.e pili ana i nā wā hilinaʻi no nā lākiō lethal dose no ka log-transformed concentration and dose-mortality curves.Hoʻokumu ʻia ka ʻikepili make ma luna o ka ʻikepili hoʻopiʻi hui ʻia o 180 larvae i hōʻike ʻia i kēlā me kēia ʻano o ka mālama ʻana.Hoʻokaʻawale ʻia nā loiloi probabilistic no kēlā me kēia ʻai hua a me kēlā me kēia mea kemika.Ma muli o ka 95% o ka hilinaʻi manawa o ka lethal dose ratio, ua manaʻo ʻia he ʻokoʻa loa ka ʻawaʻawa o ka ʻai hua a me nā mea kemika i ka namu larvae, no laila ʻaʻole ʻokoʻa loa ka wā hilinaʻi i loaʻa ka waiwai o 1, P = 0.0566.
ʻO nā hualoaʻa HPLC no ka hoʻoholoʻana i nā glucosinolates nui i loko o ka palaoa hua defatted DFP, IG, PG a me Ls ua heluʻia ma ka Papa 1. ʻO nā glucosinolates nui i loko o ka palaoa hua i ho'āʻoʻia heʻano likeʻole me ka DFP a me ka PG, i loko o lāuaʻelua myrosinase glucosinolates.ʻOi aku ka nui o ka myrosinin ma PG ma mua o ka DFP, 33.3 ± 1.5 a me 26.5 ± 0.9 mg / g.ʻO ka pauka hua Ls he 36.6 ± 1.2 mg / g glucoglycone, aʻo ka pauka hua IG he 38.0 ± 0.5 mg / g sinapine.
ʻO nā ʻanuʻu o Ae.Ua pepehi ʻia nā makika Aedes aegypti i ka mālama ʻia ʻana me ka ʻai ʻanoʻano defatted, ʻoiai ʻokoʻa ka maikaʻi o ka mālama ʻana ma muli o ke ʻano mea kanu.ʻO DFP-NT wale nō ʻaʻole i ʻawaʻawa i nā larvae namu ma hope o 24 a me 72 mau hola o ka hoʻolaha ʻana (Table 2).Ua hoʻonui ʻia ka ʻawaʻawa o ka pauka hua hana me ka hoʻonui ʻana i ka ʻike (Fig. 1A, B).Ua ʻano like ʻole ka ʻona o ka ʻai hua i ka namu larvae ma muli o ka 95% CI o ka lakene lethal dose o nā waiwai LC50 ma nā loiloi 24-hola a me 72-hola (Table 3).Ma hope o 24 mau hola, ʻoi aku ka nui o ka hopena ʻona o ka ʻai hua Ls ma mua o nā lāʻau lapaʻau ʻai hua ʻē aʻe, me ka hana kiʻekiʻe loa a me nā mea ʻawaʻawa kiʻekiʻe loa i nā larvae (LC50 = 0.04 g/120 ml dH2O).ʻAʻole i maʻalahi ka larvae i ka DFP i nā hola 24 i hoʻohālikelike ʻia me IG, Ls a me PG seed powder treatments, me nā waiwai LC50 o 0.115, 0.04 a me 0.08 g/120 ml dH2O pākahi, ʻoi aku ka kiʻekiʻe ma mua o ka waiwai LC50.0.211 g/120 ml dH2O (Papa 3).ʻO nā waiwai LC90 o DFP, IG, PG a me Ls he 0.376, 0.275, 0.137 a me 0.074 g/120 ml dH2O, i kēlā me kēia (Papa 2).ʻO ka manaʻo kiʻekiʻe loa o DPP he 0.12 g/120 ml dH2O.Ma hope o 24 mau hola o ka loiloi, ʻo 12% wale nō ka nui o ka make larval, aʻo ka awelika o ka make o IG a me PG larvae i hiki i 51% a me 82%, kēlā me kēia.Ma hope o 24 mau hola o ka loiloi, ʻo ka awelika o ka make larval no ka kiʻekiʻe kiʻekiʻe o ka Ls seed meal treatment (0.075 g / 120 ml dH2O) he 99% (Fig. 1A).
Ua manaʻo ʻia nā ʻōkuhi make mai ka pane ʻana o ka dose (Probit) o Ae.ʻO ʻAigupita (3rd instar larvae) i ka ʻai ʻana i nā hua 24 hola (A) a me 72 hola (B) ma hope o ka mālama ʻana.Hōʻike ka laina kiko i ka LC50 o ka mālama ʻana i nā hua.DFP Thlaspi arvense, DFP-HT Heat inactivated Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Ma ka loiloi 72-hola, ʻo nā waiwai LC50 o DFP, IG a me PG ʻai hua ʻai he 0.111, 0.085 a me 0.051 g/120 ml dH2O, kēlā me kēia.Aneane pau loa na larvae i hoikeia i ka ai hua Ls i make ma hope o 72 h o ka ike ana, no laila, ua like ole ka ikepili make me ka anana Probit.Ke hoʻohālikelike ʻia me nā meaʻai hua ʻē aʻe, ʻaʻole i maʻalahi ka larvae i ka mālama ʻana i ka ʻai hua DFP a loaʻa i nā helu LC50 kiʻekiʻe (Papa 2 a me 3).Ma hope o 72 mau hola, ua manaʻo ʻia nā koina LC50 no ka DFP, IG a me PG i ka mālama ʻana i ka meaʻai hua 0.111, 0.085 a me 0.05 g/120 ml dH2O, kēlā me kēia.Ma hope o 72 mau hola o ka loiloi, ʻo nā waiwai LC90 o DFP, IG a me PG nā pauka hua he 0.215, 0.254 a me 0.138 g/120 ml dH2O, kēlā me kēia.Ma hope o 72 mau hola o ka loiloi, ʻo ka awelika o ka make larval no ka DFP, IG a me ka PG seed meal treatments ma kahi kiʻekiʻe o 0.12 g / 120 ml dH2O he 58%, 66% a me 96% (Fig. 1B).Ma hope o 72-hola loiloi, ʻike ʻia ka ʻai hua PG i ʻoi aku ka ʻona ma mua o ka ʻai hua IG a me DFP.
Synthetic isothiocyanates, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC) a me 4-hydroxybenzylisothiocyanate (4-HBITC) hiki ke pepehi pono i na makika.Ma 24 mau hola ma hope o ka mālama ʻana, ʻoi aku ka ʻona o ka BITC i nā larvae me kahi waiwai LC50 o 5.29 ppm i hoʻohālikelike ʻia me 19.35 ppm no AITC a me 55.41 ppm no 4-HBITC (Table 4).Hoʻohālikelike ʻia me AITC a me BITC, ʻo 4-HBITC ka haʻahaʻa haʻahaʻa a me ka waiwai LC50 kiʻekiʻe.Aia nā ʻokoʻa nui o ka ʻawaʻawa o ka namu o nā isothiocyanates nui ʻelua (Ls a me PG) i ka ʻai hua ʻoi loa.ʻO ka lāʻau make e pili ana i ka lakene lethal dose o nā waiwai LC50 ma waena o AITC, BITC, a me 4-HBITC i hōʻike i kahi ʻokoʻa helu e like me ka 95% CI o ka LC50 lethal dose ratio ʻaʻole i hoʻokomo i kahi waiwai o 1 (P = 0.05, Table. 4).Ua manaʻo ʻia nā koʻikoʻi kiʻekiʻe loa o BITC a me AITC e pepehi i 100% o nā larvae i hoʻāʻo ʻia (Figure 2).
Ua manaʻo ʻia nā ʻōkuhi make mai ka pane ʻana o ka dose (Probit) o Ae.He 24 mau hola ma hope o ka mālama ʻana, ua hiki i nā ʻōpala ʻAigupita (3rd instar larvae) i ka synthetic isothiocyanate concentrations.Hōʻike ka laina kiko i ka LC50 no ka mālama isothiocyanate.Benzyl isothiocyanate BITC, allyl isothiocyanate AITC a me 4-HBITC.
Ua aʻo lōʻihi ʻia ka hoʻohana ʻana i nā lāʻau biopesticides ma ke ʻano he mea hoʻokele namu.Nui nā mea kanu e hoʻopuka i nā kemika kūlohelohe i loaʻa ka hana insecticidal37.Hāʻawi ko lākou mau pūhui bioactive i kahi koho ʻē aʻe i nā insecticides synthetic me ka mana nui i ke kāohi ʻana i nā pests, me nā makika.
Hoʻoulu ʻia nā mea kanu ma ke ʻano he mea kanu no kā lākou mau hua, hoʻohana ʻia i mea ʻala a i kumu aila.Ke lawe ʻia ka ʻaila sinapi mai nā ʻanoʻano a i ʻole ka lawe ʻia ʻana o ka sinapi no ka hoʻohana ʻana ma ke ʻano he biofuel, 69 ʻo ia ka mea ʻai i hoʻopau ʻia i ka momona.Mālama kēia ʻai hua i ka nui o kāna mau ʻāpana biochemical maoli a me nā enzyme hydrolytic.Hoʻopili ʻia ka ʻona o kēia ʻai hua i ka hana ʻana o isothiocyanates55,60,61.Hoʻokumu ʻia nā Isothiocyanates e ka hydrolysis o glucosinolates e ka myrosinase enzyme i ka wā o ka hydration o ka ʻai hua38,55,70 a ʻike ʻia he fungicidal, bactericidal, nematicidal a me nā hopena insecticidal, a me nā waiwai ʻē aʻe e pili ana i nā hopena sensory kemika a me nā waiwai chemotherapeutic61,62, 70.Ua hōʻike ʻia kekahi mau haʻawina e hana maikaʻi nā mea kanu sinapi a me ka ʻai hua ma ke ʻano he fumigants e kūʻē i ka lepo a me nā meaʻai meaʻai mālama ʻia57,59,71,72.I loko o kēia haʻawina, ua loiloi mākou i ka ʻona o ka ʻai ʻehā hua a me kāna mau huahana bioactive ʻekolu AITC, BITC, a me 4-HBITC i nā ʻōpala namu Aedes.Aedes aegypti.ʻO ka hoʻohui pono ʻana i ka ʻai hua i ka wai i loaʻa i nā ʻōpala makika e manaʻo ʻia e hoʻāla i nā kaʻina enzymatic e hoʻohua ai i ka isothiocyanates i mea ʻawaʻawa i nā ʻōpala namu.Ua hōʻike ʻia kēia biotransformation e ka hana larvicidal i ʻike ʻia o ka ʻai hua a me ka nalo ʻana o ka hana insecticidal i ka wā i mālama ʻia ai ka ʻai hua sinapi dwarf ma mua o ka hoʻohana ʻana.Manaʻo ʻia ka hoʻomaʻamaʻa wela e luku i nā enzyme hydrolytic e hoʻāla ai i nā glucosinolates, a laila e pale ai i ka hoʻokumu ʻana o isothiocyanates bioactive.ʻO kēia ka noiʻi mua e hōʻoia i nā waiwai insecticidal o ka pauka hua kāpeti e kūʻē i nā makika i loko o kahi kaiaola.
Ma waena o nā pauka hua i hoʻāʻo ʻia, ʻo ka pauda hua manu (Ls) ka mea ʻawaʻawa loa, e make nui ai ka Aedes albopictus.Hoʻomaʻamaʻa mau ʻia nā ʻōpala Aedes aegypti no 24 mau hola.ʻO nā pauka hua ʻekolu i koe (PG, IG a me DFP) ua ʻoi aku ka lohi a ua make nui ma hope o 72 mau hola o ka mālama mau ʻana.ʻO ka ʻai hua Ls wale nō ka nui o nā glucosinolates, ʻoiai ʻo PG a me DFP i loaʻa i ka myrosinase a me IG i loko o ka glucosinolate ma ke ʻano he glucosinolate nui (Table 1).Hoʻopili ʻia ʻo Glucotropaeolin i ka BITC a ua hydrolyzed ka sinalbine i 4-HBITC61,62.Hōʻike kā mākou mau hopena bioassay he mea ʻawaʻawa loa ka ʻai hua Ls a me ka BITC synthetic i nā ʻōpala namu.ʻO ka mea nui o ka ʻai hua PG a me DFP ʻo ka myrosinase glucosinolate, i hydrolyzed i AITC.He mea maikaʻi ʻo AITC i ka pepehi ʻana i nā ʻōpala makika me ka waiwai LC50 o 19.35 ppm.Ke hoʻohālikelike ʻia me AITC a me BITC, ʻo 4-HBITC isothiocyanate ka mea ʻawaʻawa loa i nā larvae.ʻOiai ʻoi aku ka liʻiliʻi o ka AITC ma mua o BITC, ʻoi aku ka haʻahaʻa o kā lākou LC50 ma mua o ka nui o nā aila pono i hoʻāʻo ʻia ma nā larvae namu32,73,74,75.
ʻO kā mākou pauka hua cruciferous no ka hoʻohana ʻana e kūʻē i nā larvae namu he hoʻokahi glucosinolate nui, ʻoi aku ka nui o 98-99% o ka nui o nā glucosinolates e like me ka HPLC i hoʻoholo ai.Ua ʻike ʻia ka nui o nā glucosinolates ʻē aʻe, akā ua emi ko lākou mau pae ma mua o 0.3% o ka nui o nā glucosinolates.ʻO ka pauka hua wai (L. sativum) he mau glucosinolates lua (sinigrin), akā ʻo ko lākou ʻāpana he 1% o ka nui o nā glucosinolates, a ʻaʻole nui ko lākou ʻike (e pili ana i 0.4 mg/g pauka hua).ʻOiai aia ka PG a me DFP i ka glucosinolate nui (myrosin), ʻokoʻa ka hana larvicidal o kā lākou ʻai hua ma muli o kā lākou mau waiwai LC50.He ʻokoʻa ka ʻona a hiki i ka pauda.Ma muli paha o ka ʻokoʻa ʻana o ka hana myrosinase a i ʻole ka paʻa ʻana ma waena o nā meaʻai ʻelua.He kuleana koʻikoʻi ka hana Myrosinase i ka bioavailability o nā huahana hydrolysis e like me isothiocyanates i nā mea kanu Brassicaceae76.ʻO nā hōʻike mua a Pocock et al.77 a me Wilkinson et al.78 i hōʻike i ka hoʻololi ʻana i ka hana myrosinase a me ke kūpaʻa e pili pū kekahi me nā mea genetic a me nā mea kūlohelohe.
Ua helu ʻia ka ʻike bioactive isothiocyanate e pili ana i nā waiwai LC50 o kēlā me kēia pāʻina hua ma 24 a me 72 mau hola (Papa 5) no ka hoʻohālikelike ʻana me nā noi kemika pili.Ma hope o 24 mau hola, ʻoi aku ka ʻona o ka isothiocyanates i loko o ka ʻai hua ma mua o nā pūhui maʻemaʻe.Ua helu ʻia nā waiwai LC50 e pili ana i nā ʻāpana o ka miliona (ppm) o ka mālama ʻana i nā hua isothiocyanate ma mua o nā waiwai LC50 no nā noi BITC, AITC, a me 4-HBITC.Ua ʻike mākou i ka ʻai ʻana i nā ʻanoʻano pellet (Figure 3A).No laila, hiki ke loa'a i nā larvae ka 'ike nui aku i ka isothiocyanates 'awa'awa ma o ka 'ai 'ana i nā pellet mea'ai.Ua ʻike nui ʻia kēia ma ka mālama ʻana i ka ʻai hua IG a me PG ma ka hōʻike ʻana o 24-h, kahi o 75% a me 72% ka haʻahaʻa o LC50 ma mua o ka mālama AITC maʻemaʻe a me 4-HBITC, kēlā me kēia.ʻOi aku ka ʻona o nā lāʻau Ls a me DFP ma mua o ka isothiocyanate maʻemaʻe, me nā waiwai LC50 24% a me 41% haʻahaʻa.ʻO nā larvae i loko o ka lāʻau hoʻomalu i hoʻopōmaikaʻi maikaʻi ʻia (Fig. 3B), ʻoiai ʻo ka hapa nui o ka mālama ʻana i ka meaʻai ʻaʻole i pupate a ua lohi nui ka ulu ʻana o nā larval (Fig. 3B,D).Ma Spodopteralitura, pili ka isothiocyanates me ka hoʻopaneʻe ulu a me ka lohi o ka ulu ʻana79.
ʻO nā ʻanuʻu o Ae.Hōʻike mau ʻia nā makika Aedes aegypti i ka pauka hua Brassica no 24-72 hola.(A) ʻO nā larva make me nā ʻāpana o ka ʻai hua ma nā waha (poʻe ʻia);(B) Hōʻike ka lāʻau hoʻomalu (dH20 me ka ʻole o ka ʻai hua) e ulu maʻamau a hoʻomaka ka pupa ma hope o 72 hola (C, D) mālama ʻia nā ʻōpala me ka ʻai hua;ua hōʻike ka ʻai hua i nā ʻokoʻa o ka ulu ʻana a ʻaʻole i pupa.
ʻAʻole mākou i aʻo i ke ʻano o nā hopena ʻawaʻawa o ka isothiocyanates ma luna o nā larvae namu.Eia naʻe, ua hōʻike ʻia nā haʻawina mua i nā ʻanuʻu ahi ʻulaʻula (Solenopsis invicta) ʻo ka inhibition o ka glutathione S-transferase (GST) a me ka esterase (EST) ke kumu nui o ka bioactivity isothiocyanate, a me AITC, ʻoiai ma ka haʻahaʻa haʻahaʻa, hiki ke kāohi i ka hana GST. .ʻulaʻula i lawe ʻia mai i nā mea ahi ma nā haʻahaʻa haʻahaʻa.ʻO 0.5 µg/ml80 ka nui.I ka hoʻokaʻawale ʻana, ke kaʻe nei ʻo AITC i ka acetylcholinesterase i nā weevil kulina makua (Sitophilus zeamais)81.Pono e hoʻokō ʻia nā haʻawina like e wehewehe i ke ʻano o ka hana isothiocyanate i loko o nā larvae namu.
Hoʻohana mākou i ka lāʻau DFP i hoʻohana ʻole ʻia i ka wela no ke kākoʻo ʻana i ka hydrolysis o nā mea kanu glucosinolates e hana i ka isothiocyanates reactive ma ke ʻano he hana no ka hoʻomalu ʻana i ka namu e ka ʻai hua sinapi.ʻAʻole ʻawaʻawa ka ʻai hua DFP-HT i nā helu noi i hoʻāʻo ʻia.Lafarga et al.Ua hōʻike ʻo 82 e pili ana nā glucosinolates i ka hoʻohaʻahaʻa ʻana i nā wela kiʻekiʻe.Manaʻo ʻia ka mālama ʻana i ka wela e hoʻopau i ka enzyme myrosinase i ka ʻai hua a pale i ka hydrolysis o nā glucosinolates e hana i ka isothiocyanates reactive.Ua hōʻoia pū ʻia kēia e Okunade et al.Ua hōʻike ʻo 75 he ʻano wela ka myrosinase, e hōʻike ana ua hoʻopau ʻia ka hana myrosinase i ka wā i ʻike ʻia ai ka sinapi, ka sinapi ʻeleʻele, a me nā ʻanoano aʻa koko i nā mahana ma luna o 80 °.C. Hiki i kēia mau hana ke lilo i ka hana insecticidal o ka ʻai hua DFP i mālama ʻia me ka wela.
No laila, ʻo ka ʻai hua sinapi a me kāna mau isothiocyanates nui ʻekolu he mea ʻawaʻawa ia i nā elo namu.Ma muli o kēia mau ʻokoʻa ma waena o ka ʻai hua a me nā lāʻau lapaʻau kemika, ʻo ka hoʻohana ʻana i ka ʻai ʻanoʻano he ala maikaʻi ia o ka namu.Pono e ʻike i nā ʻano hana kūpono a me nā ʻōnaehana hoʻopuka maikaʻi e hoʻomaikaʻi i ka maikaʻi a me ke kūpaʻa o ka hoʻohana ʻana i nā pauka hua.Hōʻike kā mākou hopena i ka hoʻohana ʻana i ka ʻai hua sinapi ma ke ʻano he ʻokoʻa i nā pesticides synthetic.Hiki i kēia ʻenehana ke lilo i mea hana hou no ka hoʻomalu ʻana i nā namu.Ma muli o ka ulu ʻana o ka namu i loko o ka wai a ua hoʻololi ʻia nā glucosinolates meaʻai meaʻai i isothiocyanates ikaika ma ka hydration, hiki i ka hoʻohana ʻana i ka ʻai hua sinapi i loko o ka wai i hoʻopili ʻia e nā makika.ʻOiai ʻokoʻa ka hana larvicidal o isothiocyanates (BITC > AITC > 4-HBITC), pono hou ka noiʻi ʻana no ka hoʻoholo ʻana inā hoʻohui ʻia ka ʻai hua me nā glucosinolates he nui i ka hoʻonui ʻana i ka ʻona.ʻO kēia ka noiʻi mua e hōʻike i nā hopena insecticidal o ka ʻai hua cruciferous defatted a me ʻekolu isothiocyanates bioactive ma nā makika.ʻO nā hualoaʻa o kēia noiʻi ʻana i wāwahi i ka ʻāina hou ma ka hōʻike ʻana i ka ʻai hua kāpeti i hoʻopau ʻia, kahi hopena o ka ʻaila ʻaila mai nā ʻanoʻano, hiki ke lilo ma ke ʻano he lāʻau larvicidal maikaʻi no ka hoʻomake ʻana i ka makika.Hiki i kēia ʻike ke kōkua hou aku i ka loaʻa ʻana o nā mea kanu lāʻau biocontrol a me kā lākou hoʻomohala ʻana ma ke ʻano he biopesticides maʻalahi, kūpono, a me ke kaiapuni.
Loaʻa nā ʻikepili i hana ʻia no kēia noiʻi a me nā loiloi hopena mai ka mea kākau pili ma ke noi kūpono.I ka pau ʻana o ke aʻo ʻana, ua luku ʻia nā mea a pau i hoʻohana ʻia i ke aʻo ʻana (nā inikini a me ka ʻai hua).
Ka manawa hoʻouna: Iulai-29-2024