The pine nematode ke sebaka se arolelanoang ho falla se tsejoang ka ho baka tahlehelo e kholo ea moruo tikolohong ea meru ea phaene. Boithuto ba hona joale bo lekola ts'ebetso ea nemacidal ea li-indoles tsa halogenated khahlano le li-pine nematodes le mokhoa oa tsona oa ts'ebetso. Mesebetsi ea nemacidal ea 5-iodoindole le avermectin (taolo e ntle) khahlanong le li-nematode tsa phaene li ne li tšoana 'me li phahame maemong a tlase (10 μg/mL). 5-iodoindole e fokolitse fecundity, ts'ebetso ea ho ikatisa, lefu la embryonic le larval, le boitšoaro ba li-locomotor. Litšebelisano tsa limolek'hule tsa li-ligands tse nang le invertebrate-specific glutamate-gated chloride channel receptors li tšehetsa khopolo ea hore 5-iodoindole, joaloka avermectin, e tlama ka thata sebakeng se sebetsang sa receptor. 5-Iodoindole e boetse e bakile ho holofala ha phenotypic ho li-nematode, ho kenyelletsa le ho putlama ha setho se sa tloaelehang / ho honyela le ho eketseha ha vacuolization. Liphetho tsena li fana ka maikutlo a hore li-vacuoles li ka bapala karolo ea lefu la nematode methylation-mediated. Habohlokoa, 5-iodoindole e ne e se chefo ho mefuta e 'meli ea limela (k'habeche le radish). Kahoo, thuto ena e bontša hore tšebeliso ea iodoindole tlas'a maemo a tikoloho e ka laola kotsi ea pine wilt.
Pine wood nematode (Bursaphelenchus xylophilus) ke ya pine wood nematodes (PWN), migratory endoparasitic nematode tse tsejwang ka ho baka tshenyo e kgolo ya tikoloho tikolohong ya meru ya phaene1. Pine wilt disease (PWD) e bakoang ke pine wood nematode e fetoha bothata bo tebileng lik'honthinenteng tse 'maloa, ho kenyeletsa Asia le Europe,' me Amerika Leboea, nematode e senya mefuta ea phaene e hlahisitsoeng1,2. Ho theoha ha sefate sa phaene ke bothata bo boholo ba moruo, 'me tebello ea ho ata ha eona lefatšeng ka bophara ea tšoenya3. Mefuta e latelang ea phaene e atisa ho hlaseloa ke nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, le Pinus radiata4. Pine nematode ke lefu le kotsi le ka bolaeang lifate tsa phaene nakong ea libeke kapa likhoeli tsa tšoaetso. Ho phaella moo, ho qhoma ha li-pine nematode ho tloaelehile libakeng tse sa tšoaneng tsa tikoloho, kahoo ho thehiloe liketane tse tsitsitseng tsa ts'oaetso1.
Bursaphelenchus xylophilus ke nematode ya semela se arolang batho ka thoko ho ba lelapa la superfamily Aphelenchoidea le clade 102.5. Nematode e ja li-fungus 'me e ikatisa ka har'a lehong la lifate tsa phaene, e fetoha mekhahlelo e mene e fapaneng ea larval: L1, L2, L3, L4 le motho e moholo1,6. Tlas'a maemo a khaello ea lijo, nematode ea phaene e fetela sethaleng se khethehileng sa larval - dauer, e senyang vector ea eona - beetle ea pine bark (Monochamus alternatus) 'me e fetisetsoa lifateng tsa phaene tse phetseng hantle. Libakeng tse phelang hantle, li-nematode li falla ka potlako ka har'a lisele tsa limela 'me li ja lisele tsa parenchymatous, e leng se lebisang ho liphetoho tse ngata tsa hypersensitivity, pine wilting le lefu nakong ea selemo ka mor'a tšoaetso1,7,8.
Taolo ea baeloji ea li-pine nematodes haesale e le phephetso, ka mehato ea ho behella batho ka thoko ho tloha lekholong la bo20 la lilemo. Maano a hajoale a ho laola li-nematode tsa phaene haholo-holo a kenyelletsa kalafo ea lik'hemik'hale, ho kenyeletsoa ho chesoa ha lehong le ho kenngoa ha li-nematicides likutung tsa lifate. Li-nematicide tse sebelisoang haholo ke avermectin le avermectin benzoate, tseo e leng tsa lelapa la avermectin. Lik'hemik'hale tsena tse turang li sebetsa haholo khahlano le mefuta e mengata ea li-nematode 'me li nkoa li sireletsehile tikolohong9. Leha ho le joalo, tšebeliso e pheta-phetoang ea li-nematicides tsena e lebeletsoe ho theha khatello ea khetho e tla lebisa ho hlaha ha li-nematode tsa phaene tse hanyetsanang, joalokaha ho bontšitsoe bakeng sa likokoanyana tse ngata tse senyang lijalo, tse kang Leptinotarsa decemlineata, Plutella xylostella le nematodes Trichostrongylus colubriformis le Ostertagiaverct 12 butle-butle e tsoetse pele ho hanyetsa. Ka hona, mekhoa ea ho hanyetsa e hloka ho ithutoa khafetsa le ho hlahlobisisoa li-nematicides ka nako e telele ho fumana mekhoa e meng, e baballang chelete le e baballang tikoloho ho laola PVD. Lilemong tse mashome tsa morao tjena, bangoli ba 'maloa ba hlahisitse tšebeliso ea lihlahisoa tsa limela, oli ea bohlokoa le li-volatiles e le li-agent tsa ho laola nematode13,14,15,16.
Haufinyane tjena re bonts'itse ts'ebetso ea nematicidal ea indole, molek'hule ea lipontšo tsa intercellular le interkingdom, ho Caenorhabditis elegans 17. Indole ke pontšo e atileng ea intracellular ho ecology ea likokoana-hloko, e laolang mesebetsi e mengata e amang physiology ea likokoana-hloko, sebopeho sa spore, botsitso ba plasmid, ho hanyetsa lithethefatsi, ho thehoa ha biofilm, le likokoana-hloko 18, 19. Ts'ebetso ea indole le lihlahisoa tsa eona khahlano le li-nematode tse ling tsa pathogenic ha e so ithutoe. Thutong ena, re batlisitse ts'ebetso ea nemacidal ea li-indoles tse 34 khahlano le li-pine nematodes mme ra hlakisa mokhoa oa ts'ebetso ea 5-iodoindole e matla ka ho fetesisa e sebelisang microscopy, ho nka lifoto ka nako e telele, le liteko tsa ho emisa limolek'hule, mme ra lekola litlamorao tsa chefo ho limela tse sebelisang tlhahlobo ea ho mela ha peo.
Likhakanyo tse phahameng (> 1.0 mM) tsa indole li tlalehiloe pele li na le phello ea nemacidal ho nematodes17. Ka mor'a phekolo ea B. xylophilus (mekhahlelo e tsoakiloeng ea bophelo) e nang le indole kapa lihlahisoa tse fapaneng tsa 33 tsa indole ho 1 mm, lefu la B. xylophilus le ne le lekanngoa ka ho bala li-nematode tse phelang le tse shoeleng lihlopheng tsa taolo le tse phekoloang. Li-indoles tse hlano li bontšitse ts'ebetso ea bohlokoa ea nemacidal; ho pholoha ha sehlopha sa taolo e sa phekoloeng e ne e le 95 ± 7% ka mor'a lihora tse 24. Har'a li-indoles tse 34 tse lekiloeng, 5-iodoindole le 4-fluoroindole ho 1 mmM li bakile lefu la 100%, athe 5,6-difluoroindigo, methylindole-7-carboxylate, le 7-iodoindole e bakile hoo e ka bang 50% ea lefu (Letlapa la 1).
Phello ea 5-iodoindole ho thehoeng ha vacuole le metabolism ea pine wood nematode. (A) Phello ea avermectin le 5-iodoindole ho li-nematode tsa banna ba baholo, (B) L1 sethaleng sa mahe a nematode le (C) metabolism ea B. xylophilus, (i) li-vacuoles ha lia ka tsa hlokomeloa ka 0 h, phekolo e ile ea fella ka (ii) li-vacuoles, (iii) ho bokella li-vacuoles tse ngata, li-vacuoles tse ngata, liv) le (vi) ho thehoa ha li-vacuole tse kholo. Metsu e khubelu e bontša ho ruruha ha li-vacuoles, metsu e putsoa e bontša ho kopanngoa ha li-vacuoles le metsu e ntšo e bontša li-vacuoles tse khōlō. Scale bar = 50 μm.
Ho phaella moo, thuto ena e boetse e hlalosa mokhoa o latellanang oa lefu le bakoang ke methane ho li-nematodes tsa phaene (Setšoantšo sa 4C). Lefu la Methanogenic ke mofuta o seng oa apoptotic oa lefu la sele le amanang le ho bokellana ha li-cytoplasmic vacuoles tse hlahelletseng27. Mefokolo ea morphological e bonoang ho nematode ea phaene e bonahala e amana haufi-ufi le mochini oa lefu le bakoang ke methane. Tlhahlobo ea microscopic ka linako tse fapaneng e bontšitse hore li-vacuoles tse kholo li entsoe ka mor'a lihora tse 20 tsa ho pepesehela 5-iodoindole (0.1 mM). Li-vacuole tsa microscopic li ile tsa bonoa kamora lihora tse 8 tsa kalafo, mme palo ea tsona e ile ea eketseha kamora lihora tse 12. Li-vacuoles tse 'maloa tse kholo li ile tsa bonoa ka mor'a lihora tse 14. Li-vacuole tse 'maloa tse kopantsoeng li ne li bonahala ka ho hlaka ka mor'a 12-16 h ea phekolo, e leng se bontšang hore fusion ea vacuole ke motheo oa mochine oa lefu la methanogenic. Ka mor'a lihora tse 20, li-vacuoles tse ngata tse khōlō li ile tsa fumanoa ho pholletsa le sebōkō. Maikutlo ana a emela tlaleho ea pele ea metuosis ho C. elegans.
Ho liboko tse tšoaroang ke 5-iodoindole, ho kopana ha vacuole le ho phatloha ho ile ha boela ha hlokomeloa (setšoantšo sa 5), joalokaha ho pakoa ke ho kobeha ha sebōkō le ho lokolloa ha vacuole tikolohong. Ho senyeha ha vacuole ho ile ha boela ha hlokomeloa ka lera la lekhapetla la mahe, leo ka tloaelo le bolokiloeng le tiile ke L2 nakong ea ho qhotsoa (Supplementary Fig. S2). Litlhokomeliso tsena li tšehetsa ho kenya letsoho ha ho bokellana ha mokelikeli le ho hlōleha ha osmoregulatory, hammoho le kotsi e ka fetoloang ea lisele (RCI), nakong ea ho thehoa ha vacuole le suppuration (setšoantšo sa 5).
Ha re nahana ka karolo ea iodine ho thehoeng ha vacuole, re ile ra batlisisa tšebetso ea nemacidal ea sodium iodide (NaI) le potassium iodide (KI). Leha ho le joalo, lihlopheng (0.1, 0.5 kapa 1 mM), ha lia ka tsa ama bophelo ba nematode kapa sebopeho sa vacuole (Supplementary Fig. S5), le hoja 1 mM KI e ne e e-na le phello e fokolang ea nematicidal. Ka lehlakoreng le leng, 7-iodoindole (1 kapa 2 mM), joalo ka 5-iodoindole, e ile ea etsa hore li-vacuoles tse ngata li senyehe le ho senyeha ha meralo (Supplementary Fig. S6). Li-iodoindoles tse peli li bontšitse litšobotsi tse tšoanang tsa phenotypic ho li-pine nematodes, athe NaI le KI ha lia ka tsa etsa joalo. Hoa thahasellisa hore indole ha ea ka ea etsa hore ho be le vacuole ho thehoa B. xylophilus ka litekanyetso tse lekiloeng (data e sa bontšoang). Ka hona, liphetho li netefalitse hore motsoako oa indole-iodine o ikarabella bakeng sa vacuolization le metabolism ea B. xylophilus.
Har'a li-indoles tse lekiloeng bakeng sa ts'ebetso ea nematicidal, 5-iodoindole e ne e e-na le index e phahameng ka ho fetisisa ea -5.89 kcal / mol, e lateloa ke 7-iodoindole (-4.48 kcal / mol), 4-fluoroindole (-4.33), le indole (-4.03) (Setšoantšo sa 6). Khokahano e matla ea mokokotlo oa hydrogen ea 5-iodoindole ho leucine 218 e tsitsisa tlamo ea eona, athe tse ling kaofela tse tsoang ho indole li tlamella ho serine 260 ka li-side chain hydrogen bonds. Har'a tse ling tsa li-iodoindoles tsa mohlala, 2-iodoindole e na le boleng bo tlamang -5.248 kcal / mol, e leng ka lebaka la tlamo ea eona e kholo ea hydrogen le leucine 218. Litlamo tse ling tse tsejoang li kenyelletsa 3-iodoindole (-4.3 kcal / mol), 4-iodoindole (-4.0 kcal 6. kcal / mol) (Setšoantšo sa Tlatsetso S8). Boholo ba li-indoles tse nang le li-halogenated le indole ka boeona, ntle le 5-iodoindole le 2-iodoindole, li theha maqhama le serine 260. Taba ea hore hydrogen bonding le leucine 218 ke pontšo ea ho tlama ha li-receptor-ligand hantle, joalokaha ho hlokometsoe ivermectin (Supplementary Fig. 2-iodoindole, joaloka ivermectin, e tlama ka thata sebakeng se sebetsang sa GluCL receptor ka leucine 218 (setšoantšo sa 6 le Supplementary Fig. S8). Re etsa tlhahiso ea hore tlamahano ena e hlokeha ho boloka sebopeho sa pore se bulehileng sa GluCL complex le hore ka ho tlama ka thata sebakeng se sebetsang sa GluCL receptor, 5-iodoindole, 2-iodoindole, avermectin le ivermectin kahoo ho boloka mocha oa ion o bulehile le ho lumella ho kenngoa ha metsi.
Ho emisa limolek'hule tsa indole le halogenated indole ho GluCL. Litloaelo tse tlamang tsa (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, le (D) 5-iodoindole ligands sebakeng se sebetsang sa GluCL. Protheine e emeloa ke lente, 'me li-bond tsa hydrogen tsa mokokotlo li bontšoa e le mela e mosehla ea matheba. (A′), (B′), (C′), le (D′) li bonts'a litšebelisano tsa li-ligand tse tsamaellanang le masalla a amino acid a pota-potileng, 'me li-bond tsa hydrogen tsa mahlakoreng li bonts'oa ka metsu e matheba a pinki.
Liteko li ile tsa etsoa ho lekola phello e chefo ea 5-iodoindole ho mela ha k'habeche le peō ea radish. 5-iodoindole (0.05 kapa 0.1 mM) kapa avermectin (10 μg/mL) e bile le phello e fokolang kapa e se nang eona ho mela ea pele le ho hlaha ha semela (Setšoantšo sa 7). Ho phaella moo, ha ho phapang e khōlō e ileng ea fumanoa pakeng tsa sekhahla sa ho mela ha taolo e sa phekoloeng le lipeo tse tšoaroang ka 5-iodoindole kapa avermectin. Phello ea taproot elongation le palo ea metso ea lateral e thehiloeng e ne e se letho, le hoja 1 mm (makhetlo a 10 a mahloriso a eona a sebetsang) ea 5-iodoindole e liehile hanyenyane nts'etsopele ea metso ea lateral. Liphetho tsena li bonts'a hore 5-iodoindole ha e na chefo ho lisele tsa semela mme ha e kena-kenane le mekhoa ea nts'etsopele ea limela maemong a ithutoang.
Phello ea 5-iodoindole ho mela ha peo. Ho mela, ho mela le ho mela ka mahlakoreng a peo ea B. oleracea le R. raphanistrum ho Murashige le Skoog agar medium e nang le avermectin kapa 5-iodoindole kapa ntle le eona. Ho mela ho tlalehiloe ka mor'a matsatsi a 3 a ho futhumala ho 22°C.
Phuputso ena e tlaleha liketsahalo tse 'maloa tsa ho bolaoa ha li-nematode ke li-indoles. Habohlokoa, ena ke tlaleho ea pele ea iodoindole inducing methylation (ts'ebetso e bakoang ke ho bokella li-vacuoles tse nyenyane tse kopanang butle-butle ho li-vacuoles tse khōlō, qetellong li lebisa ho phatloheng le lefu) ka linale tsa phaene, ka iodoindole e bonts'a thepa ea bohlokoa ea nematicide e tšoanang le ea nematicide avermectin ea khoebo.
Li-Indoles li kile tsa tlalehoa hore li na le mesebetsi e mengata ea lipontšo ho prokaryotes le eukaryote, ho kenyelletsa le ho thibela likokoana-hloko / ho thehoa, ho phela ha baktheria, le pathogenicity19,32,33,34. Haufinyane tjena, litlamorao tse ka bang teng tsa kalafo tsa halogenated indoles, indole alkaloids, le semisynthetic indole derivatives li hohetse thahasello e kholo ea lipatlisiso35,36,37. Ka mohlala, li-indoles tsa halogenated li bontšitsoe ho bolaea Escherichia coli e tsitsitseng le lisele tsa Staphylococcus aureus37. Ho feta moo, ho na le thahasello ea saense ho ithuta ka katleho ea li-indoles tse nang le li-halogenated khahlanong le mefuta e meng, meloko le mebuso, 'me thuto ena ke mohato oa ho finyella sepheo sena.
Mona, re etsa tlhahiso ea mokhoa oa ho bolaea 5-iodoindole-induced lethality C. elegans e thehiloeng ho reversible cell injury (RCI) le methylation (Figures 4C le 5). Liphetoho tse senyang tse kang puffiness le vacuolar degeneration ke matšoao a RCI le methylation, e bonahatsoang e le li-vacuoles tse khōlō ho cytoplasm48,49. RCI e sitisa tlhahiso ea matla ka ho fokotsa tlhahiso ea ATP, e bakang ho hlōleha ha pompo ea ATPase, kapa ho senya lisele tsa lisele le ho baka phallo e potlakileng ea Na +, Ca2 +, le metsi50,51,52. Li-vacuoles tsa Intracytoplasmic li hlaha liseleng tsa liphoofolo ka lebaka la ho bokellana ha metsi ka har'a cytoplasm ka lebaka la ho phalla ha Ca2 + le metsi53. Hoa thahasellisa hore mochine ona oa tšenyo ea lisele o ka tsosolosoa haeba tšenyo e le ea nakoana 'me lisele li qala ho hlahisa ATP ka nako e itseng, empa haeba tšenyo e ntse e tsoela pele kapa e mpefala, lisele li shoa.54 Maikutlo a rona a bontša hore li-nematode tse tšoaroang ka 5-iodoindole ha li khone ho tsosolosa biosynthesis e tloaelehileng ka mor'a ho pepesehela maemo a khatello ea kelello.
The methylation phenotype e hlahisitsoeng ke 5-iodoindole ho B. xylophilus e ka 'na ea e-ba ka lebaka la boteng ba iodine le ho aroloa ha molek'hule, kaha 7-iodoindole e ne e e-na le phello e fokolang ea ho thibela B. xylophilus ho feta 5-iodoindole (Letlapa la 1 le Supplementary Figure S6). Liphetho tsena li lumellana hanyane le lithuto tsa Malta et al. (2014), ea tlalehileng hore phetisetso ea pyridyl nitrogen moiety ka indole ho tloha para- ho ea ho meta-position e felisitse vacuolization, inhibition ea kholo, le cytotoxicity liseleng tsa U251, a fana ka maikutlo a hore tšebelisano ea molek'hule e nang le sebaka se itseng se sebetsang protheine ke ea bohlokoa27,44,45. Litšebelisano pakeng tsa li-indoles tsa indole kapa tsa halogenated le li-receptor tsa GluCL tse hlokometsoeng thutong ena le tsona li tšehetsa khopolo ena, kaha 5- le 2-iodoindole li fumanoe li tlama li-receptor tsa GluCL ka matla ho feta li-indoles tse ling tse hlahlobiloeng (Setšoantšo sa 6 le Supplementary Figure S8). Iodine sebakeng sa bobeli kapa sa bohlano sa indole e ile ea fumanoa e tlama ho leucine 218 ea receptor ea GluCL ka mokokotlo oa hydrogen bond, athe li-indoles tse ling tse halogenated le indole ka boeona li theha maqhama a fokolang a mahlakoreng a hydrogen le serine 260 (Setšoantšo sa 6). Ka hona re nahana hore sebaka sa sebaka sa halogen se phetha karolo ea bohlokoa ho kenngoa ha vacuolar degeneration, athe ho tlama ka thata ha 5-iodoindole ho boloka mocha oa ion o bulehile, kahoo o lumella ho phalla ha metsi ka potlako le ho phatloha ha vacuole. Leha ho le joalo, mokhoa o qaqileng oa ts'ebetso ea 5-iodoindole o ntse o lokela ho khethoa.
Pele o sebelisa 5-iodoindole, phello ea chefo ea eona ho limela e lokela ho hlahlojoa. Liteko tsa rona tsa ho mela ha peo li bontšitse hore 5-iodoindole e ne e se na phello e mpe ho mela ha peo kapa mekhoa ea tsoelo-pele e latelang ho tse ithutoang (Setšoantšo sa 7). Kahoo, thuto ena e fana ka motheo oa tšebeliso ea 5-iodoindole tikolohong ea tikoloho ho laola kotsi ea pine nematodes ho lifate tsa phaene.
Litlaleho tse fetileng li bontšitse hore phekolo e thehiloeng ho indole e emela mokhoa o ka bang teng oa ho rarolla bothata ba ho hanyetsa lithibela-mafu le tsoelo-pele ea mofetše55. Ho phaella moo, li-indoles li na le li-antibacterial, anticancer, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative le antituberculosis mesebetsi 'me e ka sebetsa e le motheo o tšepisang bakeng sa nts'etsopele ea lithethefatsi56,57. Phuputso ena e fana ka maikutlo a lekhetlo la pele tšebeliso e ka bang teng ea iodine e le moemeli oa antiparasitic le anthelmintic.
Avermectin e ile ea sibolloa lilemong tse mashome a mararo tse fetileng 'me ea hapa Khau ea Nobel ka 2015,' me tšebeliso ea eona e le anthelmintic e ntse e tsoela pele ka mafolofolo. Leha ho le joalo, ka lebaka la tsoelo-pele e potlakileng ea ho hanyetsa li-avermectins ho li-nematodes le likokoanyana tse senyang lijalo, ho hlokahala leano le leng, le theko e tlaase le le sireletsang tikoloho ho laola tšoaetso ea PWN lifate tsa phaene. Phuputso ena e boetse e tlaleha mokhoa oo ka oona 5-iodoindole e bolaeang li-nematode tsa phaene le hore 5-iodoindole e na le chefo e tlaase ho lisele tsa semela, e leng se bulang menyetla e metle ea ts'ebeliso ea eona ea nakong e tlang ea khoebo.
Liteko tsohle li ile tsa amoheloa ke Komiti ea Boitšoaro ea Univesithi ea Yeungnam, Gyeongsan, Korea, 'me mekhoa e entsoe ho latela tataiso ea Komiti ea Boitšoaro ea Univesithi ea Yeungnam.
Liteko tsa ho qhomisa lehe li ile tsa etsoa ho sebelisoa mekhoa e hlophisitsoeng43. Ho hlahloba litekanyetso tsa ho qhotsoa (HR), li-nematode tsa batho ba baholo ba letsatsi le le leng (hoo e ka bang tse 100 tse tšehali le tse tona tse 100) li ile tsa fetisetsoa ho lijana tsa Petri tse nang le fungus 'me tsa lumelloa ho hōla bakeng sa 24 h. Mahe ka nako eo a ne a arotsoe 'me a phekoloa ka 5-iodoindole (0.05 mM le 0.1 mM) kapa avermectin (10 μg/ml) e le ho emisoa metsing a sa hloekang a sa hloekang. Li-suspension tsena (500 μl; mahe a ka bang 100) li ile tsa fetisetsoa lilibeng tsa sejana se nang le liliba tse 24 'me tsa kenngoa ka 22 °C. Lipalo tsa L2 li entsoe ka mor'a lihora tse 24 tsa incubation empa li ne li nkoa li shoele haeba lisele li sa sisinyehe ha li hlohlelletsoa ka terata e ntle ea platinum. Teko ena e entsoe ka mekhahlelo e 'meli, e' ngoe le e 'ngoe e na le makhetlo a tšeletseng. Lintlha tse tsoang litekong ka bobeli li ile tsa kopanngoa 'me tsa hlahisoa. Peresente ea HR e baloa ka tsela e latelang:
Ho shoa ha Larval ho ile ha hlahlojoa ho sebelisoa mekhoa e entsoeng pele. Mahe a Nematode a ile a bokelloa 'me mahe a emolisitsoeng a ile a amahanngoa ka ho qhotsoa ka metsing a hloekisitsoeng a hloekileng ho hlahisa li-larvae tsa L2. Li-larvae tse lumellanang (hoo e ka bang 500 nematodes) li ne li phekoloa ka 5-iodoindole (0.05 mM le 0.1 mM) kapa avermectin (10 μg/ml) 'me li hōliselitsoe lipoleiting tsa B. cinerea Petri. Kamora 48 h of incubation at 22 °C, nematode di ile tsa bokellwa metsing a hlwekileng a sa hlwekang mme tsa hlahlojwa bakeng sa boteng ba methati ya L2, L3, le L4. Ho ba teng ha mekhahlelo ea L3 le L4 ho ne ho bontša phetoho ea larval, athe boteng ba sethala sa L2 bo ne bo bontša hore ha ho na phetoho. Litšoantšo li fumanoe ho sebelisoa iRiS™ Digital Cell Imaging System. Teko ena e entsoe ka mekhahlelo e 'meli, e' ngoe le e 'ngoe e na le makhetlo a tšeletseng. Lintlha tse tsoang litekong ka bobeli li ile tsa kopanngoa 'me tsa hlahisoa.
Chefo ea 5-iodoindole le avermectin ho peo e ile ea hlahlojoa ho sebelisoa liteko tsa ho mela ka lipoleiti tsa Murashige le Skoog agar. hypochlorite) bakeng sa metsotso e 15, 'me e hlatsoe ka makhetlo a mahlano ka 1 ml ea metsi a sa hloekang. Lipeo tse hloekisitsoeng li ile tsa hatelloa holim'a lipoleiti tsa agar tse nang le 0.86 g/l (0.2X) Murashige le Skoog medium le 0.7% ea bacteriological agar e nang le 5-iodoindole kapa avermectin kapa ntle le eona. Joale lipoleiti li ne li e-na le mocheso oa 22 ° C, 'me litšoantšo li nkuoe ka mor'a matsatsi a 3 a ho fuama. Teko ena e ile ea etsoa ka mekhahlelo e 'meli, e 'ngoe le e 'ngoe ea tsona e ne e pheta-pheta tse tšeletseng.
Nako ea poso: Feb-26-2025