Nematode ea phaene ke endoparasite e fallang e tsejoang ka ho baka tahlehelo e kholo ea moruo litsing tsa tikoloho tsa meru ea phaene. Phuputso ea hona joale e hlahloba ts'ebetso ea ho bolaea likokoana-hloko tsa li-indole tse nang le halogenated khahlanong le li-nematode tsa phaene le mokhoa oa tsona oa ts'ebetso. Mesebetsi ea ho bolaea likokoana-hloko ea 5-iodoindole le avermectin (taolo e ntle) khahlanong le li-nematode tsa phaene e ne e tšoana 'me e le holimo maemong a tlase (10 μg/mL). 5-iodoindole e fokolitse ho bola, ts'ebetso ea ho ikatisa, lefu la lesea le sa tsoa emoloa le larval, le boitšoaro ba locomotor. Litšebelisano tsa limolek'hule tsa ligand tse nang le li-receptor tsa chloride tse nang le glutamate e khethehileng e tšehetsang khopolo ea hore 5-iodoindole, joalo ka avermectin, e tlama ka thata sebakeng se sebetsang sa receptor. 5-Iodoindole e boetse e bakile ho fetoha ho fapaneng ha phenotypic ho li-nematode, ho kenyeletsoa ho putlama/ho fokotseha ho sa tloaelehang ha litho le ho eketseha ha vacuolization. Liphetho tsena li bontša hore li-vacuole li ka bapala karolo lefung le bakoang ke methylation ea nematode. Habohlokoa, 5-iodoindole e ne e se chefo ho mefuta ka bobeli ea limela (k'habeche le radish). Kahoo, phuputso ena e bontša hore tšebeliso ea iodoindole tlas'a maemo a tikoloho e ka laola kotsi ea ho bola ha phaene.
Nematode ea patsi ea phaene (Bursaphelenchus xylophilus) ke ea nematode ea patsi ea phaene (PWN), nematode ea endoparasitic e fallang e tsejoang ka ho baka tšenyo e kholo ea tikoloho ho tikoloho ea meru ea phaene1. Lefu la ho bola ha phaene (PWD) le bakoang ke nematode ea patsi ea phaene le ntse le fetoha bothata bo boholo lik'honthinenteng tse 'maloa, ho kenyeletsoa Asia le Europe,' me Amerika Leboea, nematode e senya mefuta ea phaene e kentsoeng1,2. Ho fokotseha ha lifate tsa 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 hlaseloa haholo ke nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, le Pinus radiata4. Nematode ea phaene ke lefu le tebileng le ka bolaeang lifate tsa phaene ka mor'a libeke kapa likhoeli tsa tšoaetso. Ho phaella moo, ho qhoma ha nematode ea phaene ho tloaelehile libakeng tse fapaneng tsa tikoloho, kahoo liketane tse sa khaotseng tsa tšoaetso li thehiloe1.
Bursaphelenchus xylophilus ke nematode ea limela tse phelang ka thoko e leng ea lelapa le leholo la Aphelenchoidea le clade 102.5. Nematode e ja li-fungus 'me e ikatisa ka har'a lisele tsa patsi tsa lifate tsa phaene, e fetoha mekhahlelo e mene e fapaneng ea liboko: L1, L2, L3, L4 le motho e moholo1,6. Tlas'a maemo a khaello ea lijo, nematode ea phaene e fetela mohatong o ikhethileng oa liboko - dauer, e bolaeang vector ea eona - kokoanyana ea makhapetla a phaene (Monochamus alternatus) 'me e fetisetsoa lifateng tse phetseng hantle tsa phaene. Ho beng ba phetseng hantle, liboko li falla kapele ka har'a lisele tsa limela 'me li ja lisele tse nang le li-parenchymatous, e leng se lebisang ho likarabo tse ngata tsa hypersensitivity, ho pona ha phaene le lefu nakong ea selemo kamora tšoaetso1,7,8.
Taolo ea likokoana-hloko tsa phaene nematode e bile phephetso ka nako e telele, ka mehato ea ho itšehla thajana e qalileng lekholong la bo20 la lilemo. Maano a hajoale a ho laola nematode tsa phaene a kenyelletsa haholo-holo mekhoa ea phekolo ea lik'hemik'hale, ho kenyeletsoa le ho tsubella lehong le ho kenya nematicides likutung tsa lifate. Nematicides tse sebelisoang haholo ke avermectin le avermectin benzoate, tse leng tsa lelapa la avermectin. Lik'hemik'hale tsena tse turang li sebetsa hantle haholo khahlanong le mefuta e mengata ea nematode 'me li nkoa li sireletsehile tikolohong9. Leha ho le joalo, tšebeliso e pheta-phetoang ea nematicides tsena e lebelletsoe ho baka khatello ea khetho e tla lebisa ho hlaheng ha nematode tsa phaene tse hanyetsanang, joalo ka ha ho bontšitsoe bakeng sa likokoanyana tse 'maloa tse senyang likokoanyana, tse kang Leptinotarsa decemlineata, Plutella xylostella le nematode Trichostrongylus colubriformis le Ostertagia circumcincta, tse ntseng li hola butle-butle ho hanyetsa avermectins10,11,12. Ka hona, mekhoa ea ho hanyetsa e hloka ho ithutoa khafetsa le nematicides e hlahlojoa khafetsa ho fumana mekhoa e meng, e theko e tlaase le e mosa tikolohong ho laola PVD. Lilemong tse mashome tsa morao tjena, bangoli ba 'maloa ba sisintse tšebeliso ea li-extracts tsa limela, oli ea bohlokoa le li-volatine e le li-agent tsa taolo ea nematode13,14,15,16.
Haufinyane re bontšitse ts'ebetso ea ho bolaea likokoana-hloko ea indole, molek'hule ea lipontšo tsa li-intercellular le interkingdom, ho Caenorhabditis elegans 17. Indole ke lets'oao le pharaletseng la kahare ho li-microbial ecology, le laolang mesebetsi e mengata e amang fisioloji ea likokoana-hloko, ho thehoa ha spore, botsitso ba plasmid, ho hanyetsa lithethefatsi, ho thehoa ha biofilm, le ho ba kotsi ha 18, 19. Ts'ebetso ea indole le lintho tse tsoang ho eona khahlanong le li-nematode tse ling tse bakang pathogenic ha e so ithutoe. Thutong ena, re batlisisitse ts'ebetso ea ho bolaea likokoana-hloko ea li-indole tse 34 khahlanong le li-nematode tsa phaene 'me ra hlakisa mokhoa oa ts'ebetso oa 5-iodoindole e matla ka ho fetisisa re sebelisa microscopy, ho nka lifoto tsa nako e telele, le liteko tsa ho emisa limolek'hule, 'me ra lekola litlamorao tsa eona tse chefo limela re sebelisa teko ea ho mela ha peo.
Ho tlalehiloe hore maemo a phahameng a indole (>1.0 mM) a na le phello ea ho bolaea likokoana-hloko tsa nematode17. Kamora kalafo ea B. xylophilus (mehato e tsoakiloeng ea bophelo) ka indole kapa li-derivatives tse 33 tse fapaneng tsa indole ho 1 mM, lefu la B. xylophilus le ile la lekanngoa ka ho bala likokoana-hloko tse phelang le tse shoeleng lihlopheng tsa taolo le tse phekoloang. Li-indole tse hlano li bontšitse ts'ebetso ea bohlokoa ea ho bolaea likokoanyana tsa nematode; ho pholoha ha sehlopha sa taolo se sa phekoloang e ne e le 95 ± 7% kamora lihora tse 24. Ho li-indole tse 34 tse lekiloeng, 5-iodoindole le 4-fluoroindole ho 1 mM li bakile lefu la 100%, athe 5,6-difluoroindigo, methylindole-7-carboxylate, le 7-iodoindole li bakile lefu la hoo e ka bang 50% (Tafole ea 1).
Tšusumetso ea 5-iodoindole ho thehoeng ha vacuole le metabolism ea nematode ea patsi ea phaene. (A) Tšusumetso ea avermectin le 5-iodoindole ho nematode ea banna ba baholo, (B) mahe a nematode a mohato oa L1 le (C) metabolism ea B. xylophilus, (i) vacuole ha ea ka ea bonoa ka hora ea 0, kalafo e bakile (ii) vacuole, (iii) ho bokellana ha vacuole tse ngata, (iv) ho ruruha ha vacuole, (v) ho kopana ha vacuole le (vi) ho thehoa ha vacuole tse kholo. Metsu e khubelu e bontša ho ruruha ha vacuole, metsu e putsoa e bontša ho kopana ha vacuole le metsu e ntšo e bontša vacuole tse kholo. Sekala sa bar = 50 μm.
Ho phaella moo, phuputso ena e boetse e hlalositse ts'ebetso e latellanang ea lefu le bakoang ke methane ho li-nematode tsa phaene (Setšoantšo sa 4C). Lefu la methanogenic ke mofuta o sa tsoeheng oa lefu la sele o amanang le ho bokellana ha li-vacuole tse hlahelletseng tsa cytoplasmic27. Liphoso tsa sebopeho tse bonoang ho li-nematode tsa phaene li bonahala li amana haufi-ufi le mokhoa oa lefu le bakoang ke methane. Tlhahlobo ea microscopic ka linako tse fapaneng e bontšitse hore li-vacuole tse kholo li thehiloe kamora lihora tse 20 tsa ho pepesehela 5-iodoindole (0.1 mM). Li-vacuole tse nyenyane li ile tsa bonoa kamora lihora tse 8 tsa kalafo, 'me palo ea tsona ea eketseha kamora lihora tse 12. Li-vacuole tse 'maloa tse kholo li ile tsa bonoa kamora lihora tse 14. Li-vacuole tse 'maloa tse kopantsoeng li ile tsa bonahala ka ho hlaka kamora lihora tse 12-16 tsa kalafo, e leng se bontšang hore ho kopana ha li-vacuole ke motheo oa mokhoa oa lefu la methanogenic. Kamora lihora tse 20, li-vacuole tse 'maloa tse kholo li ile tsa fumanoa ho pholletsa le seboko. Litemoso tsena li emela tlaleho ea pele ea metuosis ho C. elegans.
Libokong tse phekotsoeng ka iodoindole tse 5, ho bokellana ha vacuole le ho phatloha ho ile ha bonoa (Setšoantšo sa 5), joalo ka ha ho pakoa ke ho kobeha ha liboko le ho lokolloa ha vacuole tikolohong. Ho sitisoa ha vacuole ho ile ha bonoa le ka har'a lera la khetla ea lehe, leo hangata le bolokoang le sa senyeha ke L2 nakong ea ho qhotsoa (Setšoantšo se Tlatsetsang S2). Litlhokomeliso tsena li tšehetsa ho kenella ha ho bokellana ha mokelikeli le ho hloleha ha osmoregulatory, hammoho le kotsi ea sele e ka khutlisetsoang morao (RCI), ts'ebetsong ea ho thehoa ha vacuole le ho phunya (Setšoantšo sa 5).
Ha re nahana ka karolo ea iodine sebopehong sa vacuole se bonoang, re batlisisitse ts'ebetso ea nematicidal ea sodium iodide (NaI) le potassium iodide (KI). Leha ho le joalo, maemong a (0.1, 0.5 kapa 1 mM), ha lia ka tsa ama ho phela ha nematode kapa sebopeho sa vacuole (Setšoantšo se Tlatsetsang S5), leha 1 mM KI e bile le phello e nyane ea nematicidal. Ka lehlakoreng le leng, 7-iodoindole (1 kapa 2 mM), joalo ka 5-iodoindole, e bakile li-vacuole tse ngata le ho fetoha ha sebopeho (Setšoantšo se Tlatsetsang S6). Li-iodoindole tse peli li bontšitse litšobotsi tse tšoanang tsa phenotypic ho li-nematode tsa phaene, athe NaI le KI ha lia ka tsa etsa joalo. Hoa thahasellisa hore indole ha ea ka ea baka sebopeho sa vacuole ho B. xylophilus maemong a lekiloeng (data ha e bontšoe). Kahoo, liphetho li tiisitse hore motsoako oa indole-iodine o ikarabella bakeng sa vacuolization le metabolism ea B. xylophilus.
Har'a li-indole tse lekiloeng bakeng sa ts'ebetso ea ho bolaea likokoana-hloko, 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). Kamano e matla ea hydrogen ea mokokotlo oa 5-iodoindole ho leucine 218 e tsitsisa ho tlamahana ha eona, athe li-derivatives tse ling tsohle tsa indole li tlamahana le serine 260 ka litlamo tsa hydrogen tsa ketane ea lehlakore. Har'a li-iodoindole tse ling tse entsoeng mohlala, 2-iodoindole e na le boleng ba ho tlama ba -5.248 kcal/mol, e leng se bakoang ke tlamo ea eona ea mantlha ea haedrojene le leucine 218. Litlamo tse ling tse tsejoang li kenyelletsa 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), le 6-fluoroindole (-2.6 kcal/mol) (Setšoantšo se Tlatsetsang S8). Bongata ba li-indole tse nang le halogenated le indole ka boeona, ntle le 5-iodoindole le 2-iodoindole, li theha tlamo le serine 260. Taba ea hore tlamo ea haedrojene le leucine 218 e bontša ho tlamahana hantle ha receptor-ligand, joalo ka ha ho bonoe bakeng sa ivermectin (Setšoantšo se Tlatsetsang S7), e tiisa hore 5-iodoindole le 2-iodoindole, joalo ka ivermectin, li tlamahana ka thata sebakeng se sebetsang sa receptor ea GluCL ka leucine 218 (Setšoantšo sa 6 le Setšoantšo se Tlatsetsang S8). Re sisinya hore tlamo ena ea hlokahala ho boloka sebopeho sa masoba a bulehileng sa motsoako oa GluCL le hore ka ho tlamahana ka thata sebakeng se sebetsang sa receptor ea GluCL, 5-iodoindole, 2-iodoindole, avermectin le ivermectin ka hona li boloka kanale ea ion e bulehile 'me li lumella ho monngoa ha metsi.
Ho koalloa ha limolek'hule tsa indole le halogenated indole ho GluCL. Litsela tsa ho tlamahanya tsa (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, le (D) 5-iodoindole ligands sebakeng se sebetsang sa GluCL. Protheine e emetsoe ke ribone, 'me litlamo tsa haedrojene tsa mokokotlo li bontšoa e le mela e mosehla e nang le matheba. (A′), (B′), (C′), le (D′) li bontša tšebelisano ea ligands tse tsamaellanang le masala a amino acid a potolohileng, 'me litlamo tsa haedrojene tsa ketane ea lehlakore li bontšoa ka metsu e pinki e nang le matheba.
Liteko li entsoe ho lekola phello e chefo ea 5-iodoindole ho meleng ha peo ea k'habeche le radish. 5-iodoindole (0.05 kapa 0.1 mM) kapa avermectin (10 μg/mL) e bile le phello e nyane kapa ha e na phello ho meleng ha pele le ho hlaheng ha semela (Setšoantšo sa 7). Ho phaella moo, ha ho na phapang e kholo e fumanoeng pakeng tsa sekhahla sa ho mela ha taolo e sa phekoloang le peo e phekoloang ka 5-iodoindole kapa avermectin. Tšusumetso ho atoloheng ha motso o teteaneng le palo ea metso e ka thoko e entsoeng e ne e se ea bohlokoa, leha 1 mM (makhetlo a 10 a eona a sebetsang) ea 5-iodoindole e liehisitse hanyane nts'etsopele ea metso e ka thoko. Liphetho tsena li bontša hore 5-iodoindole ha e na chefo ho lisele tsa limela 'me ha e sitise lits'ebetso tsa nts'etsopele ea limela maemong a ithutiloeng.
Tšusumetso ea 5-iodoindole ho meleng ha peo. Ho mela, ho mela le ho mela ha metso ka lehlakoreng la peo ea B. oleracea le R. raphanistrum holim'a Murashige le Skoog agar medium ka kapa ntle le avermectin kapa 5-iodoindole. Ho mela ho tlalehiloe kamora matsatsi a 3 a ho holisa peo mochesong oa 22°C.
Phuputso ena e tlaleha linyeoe tse 'maloa tsa ho bolaoa ha nematode ke li-indoles. Habohlokoa, ena ke tlaleho ea pele ea methylation e bakang iodoindole (ts'ebetso e bakoang ke ho bokellana ha li-vacuole tse nyane tse kopanang butle-butle ho ba li-vacuole tse kholo, qetellong li lebisa ho phatloha ha lera le lefu) ka linaleng tsa phaene, 'me iodoindole e bontša thepa ea bohlokoa ea ho bolaea nematode e tšoanang le ea avermectin ea khoebo ea nematicide.
Ho kile ha tlalehoa hore li-Indole li na le mesebetsi e mengata ea lipontšo ho li-prokaryotes le li-eukaryotes, ho kenyeletsoa thibelo/tlhaho ea biofilm, ho pholoha ha baktheria, le ho baka mafu19,32,33,34. Haufinyane tjena, litlamorao tse ka bang teng tsa phekolo ea li-indole tse nang le halogenated, li-alkaloid tsa indole, le li-derivatives tsa indole tse entsoeng ka semisynthetic li hohetse thahasello e kholo ea lipatlisiso35,36,37. Mohlala, li-indole tse nang le halogenated li bontšitsoe li bolaea lisele tse sa feleng tsa Escherichia coli le Staphylococcus aureus37. Ho phaella moo, hoa thahasellisa saense ho ithuta ka katleho ea li-indole tse nang le halogenated khahlanong le mefuta e meng, meloko le mebuso, 'me thuto ena ke mohato o lebisang ho fihlelleng sepheo sena.
Mona, re sisinya mokhoa oa ho bolaea ka lebaka la 5-iodoindole ho C. elegans ho latela kotsi ea lisele e ka khutlisetsoang morao (RCI) le methylation (Litšoantšo tsa 4C le 5). Liphetoho tse bakoang ke ho ruruha tse kang ho ruruha le ho senyeha ha vacuolar ke matšoao a RCI le methylation, tse bonahalang e le li-vacuoles tse kholo ka har'a cytoplasm48,49. RCI e sitisa tlhahiso ea matla ka ho fokotsa tlhahiso ea ATP, e bakang ho hloleha ha pompo ea ATPase, kapa e sitisa lera la sele le ho baka phallo e potlakileng ea Na+, Ca2+, le water50,51,52. Li-vacuoles tsa intracytoplasmic li hlaha liseleng tsa liphoofolo ka lebaka la ho bokellana ha mokelikeli ka har'a cytoplasm ka lebaka la phallo ea Ca2+ le water53. Hoa thahasellisa hore mokhoa ona oa tšenyo ea sele o ka fetoloa haeba tšenyo e le ea nakoana 'me lisele li qala ho hlahisa ATP ka nako e itseng, empa haeba tšenyo e tsoela pele kapa e mpefala, lisele lia shoa.54 Lintho tseo re li hlokometseng li bontša hore li-nematode tse phekoloang ka 5-iodoindole li sitoa ho tsosolosa biosynthesis e tloaelehileng kamora ho pepesehela maemo a khatello.
Mofuta oa methylation o bakoang ke 5-iodoindole ho B. xylophilus e ka 'na ea bakoa ke boteng ba iodine le kabo ea eona ea limolek'hule, kaha 7-iodoindole e bile le phello e fokolang ea thibelo ho B. xylophilus ho feta 5-iodoindole (Tafole ea 1 le Setšoantšo se Tlatsetsang S6). Liphetho tsena li lumellana hanyane le lithuto tsa Maltese et al. (2014), ba tlalehileng hore phetisetso ea karolo ea naetrojene ea pyridyl ka har'a indole ho tloha para- ho ea ho meta-position e felisitse vacuolization, thibelo ea kholo, le cytotoxicity liseleng tsa U251, e leng se bontšang hore tšebelisano ea molek'hule le sebaka se itseng se sebetsang ka har'a protheine ke ea bohlokoa27,44,45. Litšebelisano lipakeng tsa indole kapa li-indole tse nang le halogenated le li-receptor tsa GluCL tse bonoeng thutong ena le tsona li tšehetsa khopolo ena, kaha 5- le 2-iodoindole li fumanoe li tlama ho li-receptor tsa GluCL ka matla ho feta li-indole tse ling tse hlahlobiloeng (Setšoantšo sa 6 le Setšoantšo se Tlatsetsang S8). Iodine e boemong ba bobeli kapa ba bohlano ba indole e fumanoe e tlama ho leucine 218 ea receptor ea GluCL ka litlamo tsa hydrogen tsa mokokotlo, athe li-indole tse ling tse nang le halogen le indole ka boeona li theha litlamo tse fokolang tsa hydrogen tsa ketane ea lehlakore le serine 260 (Setšoantšo sa 6). Ka hona re nahana hore sebaka sa halogen se bapala karolo ea bohlokoa ho qaleng ha ho senyeha ha vacuolar, athe tlamo e tiileng ea 5-iodoindole e boloka kanale ea ion e bulehile, ka hona e lumella phallo e potlakileng ea mokelikeli le ho phatloha ha vacuole. Leha ho le joalo, mokhoa o qaqileng oa ts'ebetso ea 5-iodoindole o ntse o lokela ho fumanoa.
Pele ho sebelisoa 5-iodoindole ka mokhoa o sebetsang, phello ea eona e chefo limela e lokela ho hlahlojoa. Liteko tsa rona tsa ho mela ha peo li bontšitse hore 5-iodoindole ha e na phello e mpe ho mela ha peo kapa lits'ebetsong tse latelang tsa nts'etsopele maemong a ithutiloeng (Setšoantšo sa 7). Kahoo, phuputso ena e fana ka motheo oa tšebeliso ea 5-iodoindole tikolohong ea tikoloho ho laola kotsi ea li-nematode tsa phaene lifateng 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 kankere55. Ho phaella moo, indoles e na le mesebetsi ea antibacterial, anticancer, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative le antituberculosis 'me e ka sebetsa e le motheo o tšepisang oa nts'etsopele ea meriana56,57. Phuputso ena e fana ka maikutlo a hore ka lekhetlo la pele tšebeliso e ka bang teng ea iodine e le moemeli oa antiparasitic le anthelmintic.
Avermectin e fumanoe lilemong tse mashome a mararo tse fetileng mme e hapile Khau ea Nobel ka 2015, 'me tšebeliso ea eona e le moriana o thibelang likokoana-hloko e ntse e tsoela pele ka mafolofolo. Leha ho le joalo, ka lebaka la nts'etsopele e potlakileng ea ho hanyetsa li-avermectin ho li-nematode le likokoanyana tse senyang lijalo, ho hlokahala leano le leng, le theko e tlase, le le sireletsang tikoloho ho laola tšoaetso ea PWN lifateng tsa phaene. Phuputso ena e boetse e tlaleha mokhoa oo 5-iodoindole e bolaeang li-nematode tsa phaene ka oona le hore 5-iodoindole e na le chefo e tlase liseleng tsa limela, e leng se bulang menyetla e metle bakeng sa ts'ebeliso ea eona ea khoebo nakong e tlang.
Liteko tsohle li amohetsoe ke Komiti ea Boitšoaro ea Univesithi ea Yeungnam, Gyeongsan, Korea, 'me mekhoa e entsoe ho latela litataiso tsa Komiti ea Boitšoaro ea Univesithi ea Yeungnam.
Liteko tsa ho behela mahe li entsoe ho sebelisoa mekhoa e thehiloeng43. Ho lekola sekhahla sa ho behela mahe (HR), li-nematode tse kholo tsa letsatsi le le leng (tse tšehali tse ka bang 100 le tse tona tse 100) li ile tsa fetisetsoa lijong tsa Petri tse nang le fungus 'me tsa lumelloa ho hola lihora tse 24. Mahe a ile a aroloa 'me a phekoloa ka 5-iodoindole (0.05 mM le 0.1 mM) kapa avermectin (10 μg/ml) e le ho behela metsing a hloekisitsoeng a hloekileng. Ho behela mahe ana (500 μl; mahe a ka bang 100) ho ile ha fetisetsoa lilibeng tsa poleiti ea ho hōlisa lisele tse 24 'me ha beheloa mochesong oa 22 °C. Lipalo tsa L2 li entsoe kamora lihora tse 24 tsa ho behela mahe empa li ne li nkoa li shoele haeba lisele li sa sisinyehe ha li tsosoa ka terata e ntle ea platinum. Teko ena e entsoe ka mekhahlelo e 'meli, e 'ngoe le e 'ngoe e e-na le makhetlo a tšeletseng a pheta-phetoang. 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 liboko ho ile ha hlahlojoa ho sebelisoa mekhoa e neng e se e qapiloe pele. Mahe a Nematode a ile a bokelloa 'me mahe a embryo a ile a kopanngoa ka ho qhotsoa ka metsing a hloekisitsoeng a hloekileng ho hlahisa liboko tsa mohato oa L2. Liboko tse kopantsoeng (liboko tse ka bang 500 tsa nematode) li ile tsa phekoloa ka 5-iodoindole (0.05 mM le 0.1 mM) kapa avermectin (10 μg/ml) 'me tsa holisoa holim'a lipoleiti tsa B. cinerea Petri. Kamora lihora tse 48 tsa ho beoa mochesong oa 22 °C, liboko li ile tsa bokelloa ka metsing a hloekisitsoeng a hloekileng 'me tsa hlahlojoa bakeng sa boteng ba mekhahlelo ea L2, L3, le L4. Boteng ba mekhahlelo ea L3 le L4 bo bontšitse phetoho ea liboko, athe boteng ba mohato oa L2 bo bontšitse phetoho. Litšoantšo li ile tsa fumanoa ho sebelisoa Sistimi ea ho Itšoaea Lisele tsa Dijithale ea iRiS™. Teko ena e entsoe ka mekhahlelo e 'meli, e 'ngoe le e 'ngoe ka makhetlo a tšeletseng a pheta-phetoang. 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 ka ho sebelisa liteko tsa ho mela lipoleiting tsa Murashige le Skoog agar.62 Peo ea B. oleracea le R. raphanistrum li ile tsa qala ho kolobisoa ka metsing a hloekisitsoeng a sterile ka letsatsi le le leng, tsa hlatsuoa ka 1 ml ea ethanol ea 100%, tsa hloekisoa ka 1 ml ea 50% ea bleach ea khoebo (3% sodium hypochlorite) metsotso e 15, 'me tsa hlatsuoa ka makhetlo a mahlano ka 1 ml ea metsi a hloekileng. Peo e hloekisitsoeng e ile ea hatelloa lipoleiting tsa agar ea ho mela tse nang le 0.86 g/l (0.2X) Murashige le Skoog medium le 0.7% bacteriological agar e nang le kapa e se nang 5-iodoindole kapa avermectin. Lipoleiti li ile tsa beoa ka har'a 22 °C, 'me litšoantšo tsa nkuoa kamora matsatsi a 3 a ho beoa. Teko ena e entsoe ka mekhahlelo e 'meli, eo e 'ngoe le e 'ngoe ea eona e bileng le makhetlo a tšeletseng a pheta-phetoang.
Nako ea poso: Hlakola-26-2025