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 ka ʻona (LC50) o allyl isothiocyanate, benzyl isothiocyanate a me 4-hydroxybenzylisothiocyanate i 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 i nā wahine hāpai i kahi solution 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 vector 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