ʻO ka pine nematode kahi endoparasite migratory quarantine i ʻike ʻia e hoʻopau i ka hoʻokele waiwai nui i nā kaiaola kaiaola paina. Hōʻike ka haʻawina i kēia manawa i ka hana nematicidal o nā indoles halogenated e kūʻē i nā nematodes pine a me kā lākou hana. ʻO nā hana nematicidal o 5-iodoindole a me avermectin (ka mana maikaʻi) e kūʻē i nā nematodes pine ua like a kiʻekiʻe ma nā haʻahaʻa haʻahaʻa (10 μg / mL). ʻO 5-iodoindole i hoʻemi i ka fecundity, ka hana hānau, embryonic a me ka larval make, a me ka locomotor behavior. Kākoʻo ʻia ka manaʻo o ka 5-iodoindole, e like me avermectin, e hoʻopaʻa paʻa i ke kahua hoʻokipa. 5-Iodoindole kekahi i hoʻokomo i nā ʻano hoʻololi phenotypic i loko o nā nematodes, me ka hāʻule ʻana a me ka hoʻonui ʻana i ka vacuolization. Ke hōʻike nei kēia mau hopena e hiki i nā vacuoles ke hana i ka nematode methylation-mediated make. ʻO ka mea nui, ʻaʻohe mea ʻawaʻawa ʻo 5-iodoindole i nā ʻano mea kanu ʻelua (kāpī a me radish). No laila, ke hōʻike nei kēia haʻawina e hiki i ka hoʻohana ʻana i ka iodoindole ma lalo o nā kūlana kaiapuni ke hoʻomalu i ka hōʻeha pine.
No nā nematodes lāʻau paina (PWN), nematodes endoparasitic nematode neʻe i ʻike ʻia e hoʻopōʻino nui i ke kaiaola i nā kaiaola ulu lāʻau paina1. ʻO ka maʻi wilt Pine (PWD) ma muli o ka nematode lāʻau paina ke lilo nei i pilikia koʻikoʻi ma kekahi mau ʻāina, ʻo ia hoʻi ʻo ʻAsia a me ʻEulopa, a ma ʻAmelika ʻĀkau, hoʻopau ka nematode i nā ʻano pine i hoʻokomo ʻia1,2. He pilikia hoʻokele waiwai nui ka emi ʻana o ka lāʻau pine, a hopohopo ka manaʻolana o ka laha ʻana o ka honua. Hoʻouka pinepine ʻia nā ʻano paina e ka nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, a me Pinus radiata4. ʻO ka nematode pine he maʻi koʻikoʻi hiki ke pepehi i nā lāʻau pine i loko o nā pule a i ʻole mau mahina o ka maʻi. Eia hou, he mea ma'amau ka ma'i nematode pine i loko o nā kaiaola like 'ole, no laila, ua ho'okumu 'ia nā kaulahao ma'i mau.
ʻO Bursaphelenchus xylophilus kahi nematode kanu-parasitic quarantine no ka superfamily Aphelenchoidea a me ka clade 102.5. ʻAi ka nematode i nā haʻani a hana hou i loko o nā ʻiʻo lāʻau o nā lāʻau paina, e ulu ana i ʻehā mau ʻāpana larval: L1, L2, L3, L4 a me kahi kanaka makua1,6. Ma lalo o nā kūlana o ka nele i ka meaʻai, hele ka pine nematode i loko o kahi pae larval kūikawā - dauer, ka mea e hoʻopiʻi i kāna vector - ka pine bark beetle (Monochamus alternatus) a hoʻololi ʻia i nā lāʻau paina olakino. I loko o nā pūʻali olakino, neʻe koke nā nematodes ma o nā ʻiʻo mea kanu a hānai i nā cell parenchymatous, kahi e alakaʻi ai i ka nui o ka hypersensitivity reactions, pine wilting a me ka make i loko o hoʻokahi makahiki ma hope o ka maʻi1,7,8.
He mea paʻakikī ka mālama olaola o nā nematodes pine, me nā hana quarantine e pili ana i ke kenekulia 20. ʻO nā hoʻolālā o kēia manawa no ka hoʻomalu ʻana i nā nematodes pine e pili ana i ka mālama kemika, me ka fumigation wahie a me ka hoʻokomo ʻana i nā nematicides i loko o nā kumu lāʻau. ʻO nā nematicides i hoʻohana pinepine ʻia he avermectin a me avermectin benzoate, no ka ʻohana avermectin. ʻO kēia mau kemika makamae he mea maikaʻi loa ia e kūʻē i nā ʻano nematode he nui a ua manaʻo ʻia he palekana no ke kaiapuni9. Eia nō naʻe, ʻo ka hoʻohana pinepine ʻana i kēia mau nematicides e manaʻo ʻia e hana i ke kaomi koho e hiki ke alakaʻi i ka puka ʻana mai o nā nematodes pine kūpaʻa, e like me ka mea i hōʻike ʻia no kekahi mau pepeke pests, e like me Leptinotarsa decemlineata, Plutella xylostella a me nā nematodes Trichostrongylus colubriformis a me Ostertagia circumcinducta i hoʻomohala ʻia. avermectins10,11,12. No laila, pono e aʻo mau ʻia nā kumu kūʻē a nānā mau ʻia nā nemicides e ʻimi i nā ʻano hana ʻokoʻa, kumu kūʻai a me nā ʻano pili i ke kaiapuni e hoʻomalu i ka PVD. I nā makahiki i hala iho nei, ua manaʻo kekahi mau mea kākau i ka hoʻohana ʻana i nā mea kanu mea kanu, nā aila pono a me nā volatiles ma ke ʻano he nematode control agents13,14,15,16.
Ua hōʻike koke mākou i ka hana nematicidal o indole, kahi mole hōʻailona intercellular a interkingdom, ma Caenorhabditis elegans 17. ʻO Indole kahi hōʻailona intracellular ākea i ka microbial ecology, e kāohi ana i nā hana he nui e pili ana i ka microbial physiology, spore formation, plasmid stability, drug resistance, biofilm formation, and virulence 18, 19. ʻAʻole i aʻo ʻia ka hana o ka indole a me kāna mau derivatives e kūʻē i nā nematodes pathogenic. I loko o kēia haʻawina, ua noiʻi mākou i ka hana nematicidal o 34 indoles e kūʻē i nā nematodes pine a wehewehe i ke ʻano o ka hana o ka 5-iodoindole ikaika loa e hoʻohana ana i ka microscopy, time-lapse kiʻi, a me nā hoʻokolohua docking molecular, a loiloi i kona hopena ʻona i nā mea kanu me ka hoʻohana ʻana i ka hoʻāʻo germination hua.
Ua hōʻike mua ʻia nā kiʻekiʻe kiʻekiʻe (> 1.0 mM) o indole i ka hopena nematicidal ma nā nematodes17. Ma hope o ka mālama ʻana i ka B. xylophilus (ke ola hui pū ʻia) me ka indole a i ʻole 33 mau ʻano indole derivatives ma 1 mM, ua ana ʻia ka make ʻana o B. xylophilus ma ka helu ʻana i nā nematodes ola a make i loko o nā pūʻulu mana a mālama ʻia. ʻElima indoles i hōʻike i ka hana nematicidal koʻikoʻi; ʻO ke ola o ka hui hoʻomalu ʻole i mālama ʻia he 95 ± 7% ma hope o 24 mau hola. ʻO nā indoles 34 i ho'āʻoʻia,ʻo 5-iodoindole a me 4-fluoroindole ma 1 mM ka mea i make ai ka 100%, akāʻo 5,6-difluoroindigo, methylindole-7-carboxylate, a me 7-iodoindole ka mea i make ma kahi o 50% o ka make (Table 1).
Ka hopena o ka 5-iodoindole i ka hoʻokumu ʻana o ka vacuole a me ka metabolism o ka nematode lāʻau pine. (A) Ka hopena o ka avermectin a me ka 5-iodoindole i nā nematodes kāne makua, (B) L1 stage nematode hua a me (C) metabolism o B. xylophilus, (i) ʻaʻole i ʻike ʻia nā vacuoles i ka hola 0, ua loaʻa i ka mālama ʻana i (ii) vacuoles, (iii) hōʻiliʻili o nā vacuoles he nui, (iv) fusions o nā vacuoleswell. (vi) hoʻokumu ʻana i nā vacuole nunui. Hōʻike nā pua ʻulaʻula i ka pehu ʻana o nā vacuoles, hōʻike nā pua uliuli i ka hui ʻana o nā vacuoles a ʻo nā pua ʻeleʻele e hōʻike ana i nā vacuoles nunui. Paʻa unahi = 50 μm.
Eia hou, ua wehewehe kēia haʻawina i ke kaʻina hana o ka make methane-induced i loko o nā nematodes pine (Figure 4C). ʻO ka make methanogenic he ʻano apoptotic ʻole o ka make cell e pili ana me ka hōʻiliʻili ʻana o nā vacuoles cytoplasmic kaulana27. ʻO nā hemahema morphological i ʻike ʻia i loko o nā nematodes pine e pili kokoke ana i ke ʻano o ka make methane-induced. Ua hōʻike ʻia ka nānā ʻana i ka microscopic i nā manawa like ʻole ua hana ʻia nā vacuoles nunui ma hope o 20 mau hola o ka ʻike ʻana i ka 5-iodoindole (0.1 mM). Ua ʻike ʻia nā vacuoles microscopic ma hope o 8 mau hola o ka mālama ʻana, a ua hoʻonui ʻia ko lākou helu ma hope o 12 mau hola. Ua ʻike ʻia kekahi mau vacuoles nui ma hope o 14 mau hola. Ua ʻike maopopo ʻia kekahi mau vacuoles fused ma hope o 12-16 h o ka mālama ʻana, e hōʻike ana ʻo ka hui ʻana o ka vacuole ke kumu o ka mīkini make methanogenic. Ma hope o 20 mau hola, ua ʻike ʻia kekahi mau vacuoles nunui a puni ka ilo. Hōʻike kēia mau ʻike i ka hōʻike mua o metuosis ma C. elegans.
I loko o nā ilo 5-iodoindole-treated, ua ʻike pū ʻia ka hoʻohui ʻana o ka vacuole a me ka rupture (Fig. 5), e like me ka mea i ʻike ʻia e ka worm bending a me ka hoʻokuʻu ʻana i ka vacuole i loko o ke kaiapuni. Ua ʻike pū ʻia ka hoʻohaunaele ʻana o ka vacuole i loko o ka membrane pūpū hua, ka mea i mālama mau ʻia e L2 i ka wā o ka hatching (Supplementary Fig. S2). Ke kākoʻo nei kēia mauʻike i ke komoʻana o ka hoʻouluʻana o ka wai a me ka hemahema o ka osmoregulatory, a me ka hōʻeha cell reversible (RCI), i ke kaʻina hana o ka vacuole a me ka suppuration (Fig. 5).
I ka hypothesizing i ka hana o iodine i ka hana vacuole i ikeia, ua noii makou i ka hana nematicidal o ka sodium iodide (NaI) a me ka potassium iodide (KI). Eia nō naʻe, ma ka hoʻopaʻa ʻana (0.1, 0.5 a i ʻole 1 mM), ʻaʻole lākou i pili i ke ola nematode a i ʻole ka hoʻokumu ʻana o ka vacuole (Supplementary Fig. S5), ʻoiai ʻo 1 mM KI he hopena nematicidal liʻiliʻi. Ma kekahiʻaoʻao, 7-iodoindole (1 a 2 mM), e like me 5-iodoindole, hoʻoulu i nā vacuoles he nui a me nā hoʻololi o ka hale (Supplementary Fig. S6). Hōʻike nā iodoindoles ʻelua i nā hiʻohiʻona phenotypic like i nā nematodes pine, akā ʻaʻole ʻo NaI a me KI. ʻO ka mea e mahalo ai, ʻaʻole i hoʻokomo ʻo indole i ka hoʻokumu ʻana o ka vacuole i B. xylophilus i nā ʻike i hoʻāʻo ʻia (ʻaʻole i hōʻike ʻia ka ʻikepili). No laila, ua hōʻoia nā hopena i ke kuleana o ka indole-iodine complex no ka vacuolization a me ka metabolism o B. xylophilus.
Ma waena o nā indoles i ho'āʻoʻia no ka hana nematicidal, ua loaʻa i ka 5-iodoindole ka helu helu slip kiʻekiʻe loa o -5.89 kcal / mol, a ma hope o 7-iodoindole (-4.48 kcal / mol), 4-fluoroindole (-4.33), a me indole (-4.03) (Figure 6). ʻO ka hoʻopaʻa ʻana o ka hydrogen iwi kuamoʻo ikaika o 5-iodoindole i ka leucine 218 e hoʻopaʻa i kona hoʻopaʻa ʻana, aʻo nā mea indole derivatives ʻē aʻe e hoʻopaʻa i ka serine 260 ma o nā kaulahao hydrogen ʻaoʻao. Ma waena o nā iodoindoles hoʻohālike ʻē aʻe, loaʻa i ka 2-iodoindole ka waiwai hoʻopaʻa o -5.248 kcal/mol, ʻo ia ka mea ma muli o kāna paʻa hydrogen nui me ka leucine 218. ʻO nā mea paʻa ʻē aʻe i ʻike ʻia ʻo 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), a me (6-2.6. S8). ʻO ka hapa nui o nā indoles halogenated a me nā indole ponoʻī, koe wale nō ka 5-iodoindole a me 2-iodoindole, e hana i kahi paʻa me ka serine 260. ʻO ka ʻoiaʻiʻo o ka hoʻopaʻa ʻana o ka hydrogen me ka leucine 218 he hōʻike ia i ka hoʻopaʻa ʻana i ka receptor-ligand maikaʻi, e like me ka nānā ʻana no ka ivermectin (Supplementary Fig. S7), hōʻoia i ka 5-iodole a me 5-iodole like me 5-iodole. e hoʻopaʻa paʻa i ka pūnaewele ikaika o ka mea loaʻa GluCL ma o ka leucine 218 (Fig. 6 a me Supplementary Fig. S8). Manaʻo mākou e koi ʻia kēia hoʻopaʻa ʻana no ka mālama ʻana i ke ʻano o ka pore hāmama o ka GluCL complex a me ka hoʻopaʻa paʻa ʻana i ka wahi hana o ka mea hoʻokipa GluCL, 5-iodoindole, 2-iodoindole, avermectin a me ivermectin no laila e mālama i ke kahawai ion e wehe a ʻae i ka lawe ʻana i ka wai.
ʻO ka hoʻopaʻa molekala o ka indole a me ka indole halogenated i GluCL. Hoʻopaʻa paʻa o (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, a me (D) 5-iodoindole ligands i ka pae hana o GluCL. Hōʻike ʻia ka protein e kahi lipine, a hōʻike ʻia nā paʻa hydrogen iwi kuamoʻo e like me nā laina kiko melemele. (A′), (B′), (C′), a me (D′) hōʻike i ka pilina o nā ligands pili me nā koena amino acid e hoʻopuni ana, a ua hōʻike ʻia nā kaulahao hydrogen ʻaoʻao e nā pua kiko ʻulaʻula.
Ua hana ʻia nā hoʻokolohua e loiloi i ka hopena ʻawaʻawa o 5-iodoindole i ka germination o ke kāpeti a me nā kumulāʻau radish. ʻO 5-iodoindole (0.05 a i ʻole 0.1 mM) a i ʻole avermectin (10 μg/mL) he mea liʻiliʻi a ʻaʻohe hopena i ka germination mua a me ka puka ʻana o ka mea kanu (Figure 7). Eia hou, ʻaʻohe ʻokoʻa koʻikoʻi i ʻike ʻia ma waena o ka helu germination o nā mana i mālama ʻole ʻia a me nā hua i mālama ʻia me 5-iodoindole a i ʻole avermectin. ʻO ka hopena o ka elongation taproot a me ka helu o nā aʻa lateral i hoʻokumu ʻia he mea liʻiliʻi, ʻoiai ʻo 1 mM (10 mau manawa o kona ʻano ikaika) o 5-iodoindole i hoʻopaneʻe iki i ka ulu ʻana o nā aʻa lateral. Hōʻike kēia mau hopena i ka 5-iodoindole ʻaʻole ʻona i nā mea kanu a ʻaʻole ia e keʻakeʻa i nā kaʻina hana hoʻomohala i nā mea i aʻo ʻia.
Ka hopena o ka 5-iodoindole i ka ulu ʻana o nā hua. ʻO ka germination, ka ulu ʻana a me ke aʻa ʻaoʻao o nā hua B. oleracea a me R. raphanistrum ma luna o Murashige a me Skoog agar media me ka avermectin a i ʻole 5-iodoindole. Hoʻopaʻa ʻia ka germination ma hope o 3 mau lā o ka incubation ma 22 ° C.
Hōʻike kēia haʻawina i kekahi mau hihia o ka pepehi ʻana i nā nematode e nā indoles. ʻO ka mea nui, ʻo kēia ka hōʻike mua o ka iodoindole inducing methylation (he kaʻina hana ma muli o ka hōʻiliʻili ʻana o nā vacuoles liʻiliʻi e hui mālie i loko o nā vacuoles nunui, a hiki i ka hemo ʻana o ka membrane a make) i loko o nā nila paina, me ka iodoindole e hōʻike ana i nā waiwai nematicidal koʻikoʻi e like me nā nematicide avermectin kalepa.
Ua hōʻike mua ʻia nā Indoles e hoʻokō i nā hana hōʻailona he nui i nā prokaryotes a me nā eukaryotes, me ka biofilm inhibition / formation, bacterial survival, and pathogenicity19,32,33,34. I kēia mau lā, ʻo nā hopena lapaʻau kūpono o nā indoles halogenated, indole alkaloids, a me nā derivatives semisynthetic indole i hoʻopaʻa i ka hoihoi noiʻi nui35,36,37. No ka laʻana, ua hōʻike ʻia nā indoles halogenated e pepehi i nā cell Escherichia coli a me Staphylococcus aureus37 mau loa. Eia hou, he mea ʻepekema ʻepekema ke aʻo ʻana i ka maikaʻi o nā indoles halogenated e kūʻē i nā ʻano ʻano, genera, a me nā aupuni ʻē aʻe, a ʻo kēia haʻawina he ʻanuʻu i ka hoʻokō ʻana i kēia pahuhopu.
Maʻaneʻi, hāʻawi mākou i kahi hana no ka 5-iodoindole-induced lethality i C. elegans ma muli o ka hōʻeha cell reversible (RCI) a me ka methylation (Nā Kiʻi 4C a me 5). ʻO nā hoʻololi Edematous e like me ka puffiness a me ka degeneration vacuolar nā hōʻailona o RCI a me ka methylation, i hōʻike ʻia e like me nā vacuoles nunui i ka cytoplasm48,49. Hoʻopilikia ʻo RCI i ka hana ʻana o ka ikehu ma o ka hoʻemi ʻana i ka hana ATP, e hoʻopōʻino ai i ka pauma ATPase, a i ʻole ke hoʻopau ʻana i nā membrane cell a hoʻoulu i ka wikiwiki o Na +, Ca2 +, a me ka wai50,51,52. Puka mai nā vacuoles Intracytoplasmic i loko o nā pūnae holoholona ma muli o ka hōʻiliʻili ʻana o ka wai i loko o ka cytoplasm ma muli o ke komo ʻana o Ca2+ a me ka wai53. ʻO ka mea e mahalo ai, hiki ke hoʻohuli ʻia kēia ʻano hana o ka pōʻino cell inā he manawa pōkole a hoʻomaka nā cell e hana i ka ATP no kekahi manawa, akā inā hoʻomau a ʻoi paha ka pōʻino, make nā cell.54 Hōʻike kā mākou ʻike i nā nematodes i mālama ʻia me 5-iodoindole hiki ʻole ke hoʻihoʻi i ka biosynthesis maʻamau ma hope o ka ʻike ʻana i nā kūlana koʻikoʻi.
ʻO ka methylation phenotype i hoʻokomoʻia e 5-iodoindole i loko o B. xylophilus ma muli paha o ka loaʻaʻana o ka iodine a me kona mahele mole, no ka mea, ua emi iho ka hopena o ka 7-iodoindole i ka B. xylophilus ma mua o ka 5-iodoindole (Papa 1 a me ke Kii S6). Ua kūlike kēia mau hopena me nā haʻawina o Maltese et al. (2014), nāna i hōʻike i ka translocation o ka pyridyl nitrogen moiety i indole mai ka para- a hiki i ka meta-kūlana hoʻopau vacuolization, ulu inhibition, a me cytotoxicity i U251 mau keena, e manao ana i ka pilina o ka mole me kahi kiko'ī wahi hana i loko o ka protein mea critical27,44,45. Ke kākoʻo pū nei nā pilina ma waena o indole a halogenated indoles a me GluCL i ʻike ʻia ma kēia haʻawina, ʻoiai ua ʻike ʻia ʻo 5- a me 2-iodoindole e hoʻopaʻa ikaika i nā mea loaʻa GluCL ma mua o nā indoles ʻē aʻe i nānā ʻia (Figure 6 a me Supplementary Figure S8). Ua ʻike ʻia ka iodine ma ka lua a i ʻole ka ʻelima o ka indole e hoʻopaʻa i ka leucine 218 o ka mea hoʻokipa GluCL ma o nā paʻa hydrogen iwi kuamoʻo, aʻo nā indoles halogenated ʻē aʻe a me ka indole ponoʻī e hana i nā kaulahao hydrogen ʻaoʻao nāwaliwali me ka serine 260 (Figure 6). No laila ke manaʻo nei mākou he kuleana koʻikoʻi ka localization o ka halogen i ka hoʻokomo ʻana i ka degeneration vacuolar, akā ʻo ka paʻa paʻa ʻana o 5-iodoindole e hoʻomau i ka wehe ʻana o ke kahawai ion, no laila e ʻae ai i ka komo ʻana o ka wai a me ka haki ʻana o ka vacuole. Eia naʻe, e hoʻoholo ʻia ke ʻano kikoʻī o ka hana o 5-iodoindole.
Ma mua o ka hoʻohana pono ʻana o 5-iodoindole, pono e nānā ʻia kona hopena ʻona i nā mea kanu. Ua hōʻike ʻia kā mākou mau hoʻokolohua hoʻoulu ʻana i nā hua ʻaʻohe hopena maikaʻi ʻole o ka 5-iodoindole i ka hoʻoulu ʻana i nā hua a i ʻole nā kaʻina hana hoʻomohala ma hope o nā mea i aʻo ʻia (Figure 7). No laila, hāʻawi kēia haʻawina i kumu no ka hoʻohana ʻana i ka 5-iodoindole i loko o ke kaiaola e hoʻomalu i ka pōʻino o nā nematodes pine i nā lāʻau pine.
Ua hōʻike ʻia nā hōʻike mua e hōʻike ana ka lāʻau indole i kahi ala kūpono e hoʻoponopono ai i ka pilikia o ka pale ʻana i ka antibiotic a me ka holomua o ka maʻi maʻi55. Eia kekahi, loaʻa i nā indoles nā hana antibacterial, anticancer, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative a me antituberculosis hana a hiki ke lilo i kumu hoʻohiki no ka hoʻomohala lāʻau 56,57. Hōʻike kēia haʻawina no ka manawa mua i ka hoʻohana ʻana i ka iodine ma ke ʻano he antiparasitic a me anthelmintic agent.
Ua ʻike ʻia ʻo Avermectin i ʻekolu mau makahiki i hala aku nei a lanakila i ka Nobel Prize ma 2015, a ke hoʻomau nei ka hoʻohana ʻana ma ke ʻano he anthelmintic. Eia nō naʻe, ma muli o ka ulu wikiwiki ʻana o ke kūʻē ʻana i nā avermectins i nā nematodes a me nā pests insect, pono kahi hoʻolālā ʻokoʻa, haʻahaʻa haʻahaʻa, a me ke kaiapuni e hoʻomalu i ka maʻi PWN i nā lāʻau paina. Hōʻike pū kēia haʻawina i ke ʻano hana e pepehi ai ʻo 5-iodoindole i nā nematodes pine a he haʻahaʻa ko 5-iodoindole i nā mea ʻawaʻawa i nā mea kanu, e wehe ana i nā manaʻo maikaʻi no kāna noi ʻoihana e hiki mai ana.
Ua'āponoʻia nā hoʻokolohua a pau e ke Kōmike Kūikawā o ke Kulanuiʻo Yeungnam, Gyeongsan, Korea, a ua hanaʻia nāʻano hana e like me nā alakaʻi a ke Kōmike Ethics o ke Kulanuiʻo Yeungnam.
Hana ʻia nā hoʻokolohua hoʻoulu hua manu me ka hoʻohana ʻana i nā kaʻina hana paʻa43. No ka helu ʻana i nā helu hatching (HR), ua hoʻoneʻe ʻia nā nematodes makua 1 mau lā (ma kahi o 100 wahine a me 100 kāne) i nā kīʻaha Petri i loaʻa ka haʻuki a ʻae ʻia e ulu no 24 mau hola. A laila hoʻokaʻawale ʻia nā hua manu a mālama ʻia me 5-iodoindole (0.05 mM a me 0.1 mM) a i ʻole avermectin (10 μg/ml) ma ke ʻano he hoʻokuʻu ʻia i loko o ka wai distilled sterile. Ua hoʻoneʻe ʻia kēia mau mea hoʻokuʻu (500 μl; ma kahi o 100 hua) i nā pūnāwai o kahi pā moʻomeheu 24-well tissue a hoʻomoʻa ʻia ma 22 °C. Hana ʻia nā helu L2 ma hope o 24 mau hola o ka incubation akā ua manaʻo ʻia ua make inā ʻaʻole neʻe nā cell i ka wā i hoʻoulu ʻia me kahi uwea platinum maikaʻi. Ua hana ʻia kēia hoʻokolohua ma nā ʻanuʻu ʻelua, kēlā me kēia me ʻeono hana hou. Ua hui pū ʻia nā ʻikepili mai nā hoʻokolohua ʻelua a hōʻike ʻia. Ua helu ʻia ka pākēneka o HR penei:
Ua helu ʻia ka make ʻana o ka larval me ka hoʻohana ʻana i nā kaʻina hana i kūkulu mua ʻia. Ua hōʻiliʻili ʻia nā hua Nematode a ua hui pū ʻia nā embryos ma o ka paʻi ʻana i loko o ka wai i hoʻomaʻemaʻe ʻia no ka hoʻohua ʻana i nā larvae pae L2. Ua mālama ʻia nā larvae synchronized (ma kahi o 500 nematodes) me 5-iodoindole (0.05 mM a me 0.1 mM) a i ʻole avermectin (10 μg / ml) a hānai ʻia ma nā papa B. cinerea Petri. Ma hope o 48 mau hola o ka incubation ma 22 ° C, ua hōʻiliʻili ʻia nā nematodes i loko o ka wai distilled sterile a nānā ʻia no ke alo o nā pae L2, L3, a me L4. ʻO ka hiki ʻana mai o nā pae L3 a me L4 i hōʻike ʻia i ka hoʻololi ʻana i ka larval, akā ʻo ka hele ʻana o ka pae L2 i hōʻike ʻole i ka loli. Ua loaʻa nā kiʻi me ka hoʻohana ʻana i ka iRiS™ Digital Cell Imaging System. Ua hana ʻia kēia hoʻokolohua ma nā ʻanuʻu ʻelua, kēlā me kēia me ʻeono hana hou. Ua hui pū ʻia nā ʻikepili mai nā hoʻokolohua ʻelua a hōʻike ʻia.
Ua loiloi ʻia ka ʻona o 5-iodoindole a me avermectin i nā hua me ka hoʻohana ʻana i nā hoʻāʻo germination ma nā pāpaʻa Murashige a me Skoog. min, a holoi ʻia ʻelima manawa me 1 ml wai sterile. A laila kaomi ʻia nā ʻanoʻano i hoʻomaʻemaʻe ʻia ma luna o nā pā germination agar me 0.86 g/l (0.2X) Murashige a me Skoog medium a me 0.7% bacteriological agar me 5-iodoindole a i ʻole avermectin. Hoʻopili ʻia nā papa ma 22 ° C, a lawe ʻia nā kiʻi ma hope o 3 mau lā o ka incubation. Ua hana ʻia kēia hoʻokolohua ma nā ʻanuʻu ʻelua, ʻeono mau hana hou ʻana o kēlā me kēia.
Ka manawa hoʻouna: Feb-26-2025