Tus kab mob ntoo thuv yog ib qho kab mob endoparasite uas paub tias ua rau muaj kev puas tsuaj loj heev rau cov ecosystem ntawm cov hav zoov ntoo thuv. Daim ntawv tshawb fawb tam sim no tshuaj xyuas cov haujlwm nematicidal ntawm halogenated indoles tawm tsam cov kab mob ntoo thuv thiab lawv cov txheej txheem ua haujlwm. Cov haujlwm nematicidal ntawm 5-iodoindole thiab avermectin (kev tswj hwm zoo) tawm tsam cov kab mob ntoo thuv zoo sib xws thiab siab ntawm qhov tsawg tsawg (10 μg / mL). 5-iodoindole txo qhov muaj menyuam, kev ua haujlwm yug me nyuam, kev tuag ntawm cov menyuam thiab cov menyuam yaus, thiab tus cwj pwm locomotor. Kev sib cuam tshuam molecular ntawm ligands nrog invertebrate-specific glutamate-gated chloride channel receptors txhawb nqa lub tswv yim tias 5-iodoindole, zoo li avermectin, khi nruj nreem rau qhov chaw ua haujlwm ntawm tus receptor. 5-Iodoindole kuj ua rau muaj ntau yam kev hloov pauv phenotypic hauv nematodes, suav nrog kev puas tsuaj ntawm lub cev tsis zoo / shrinkage thiab nce vacuolization. Cov txiaj ntsig no qhia tias vacuoles yuav ua lub luag haujlwm hauv kev tuag ntawm nematode methylation-mediated. Qhov tseem ceeb, 5-iodoindole tsis muaj tshuaj lom rau ob hom nroj tsuag (zaub qhwv thiab radish). Yog li, txoj kev tshawb fawb no qhia tau tias kev siv iodoindole nyob rau hauv ib puag ncig tuaj yeem tswj kev raug mob ntawm cov ntoo thuv.
Kab mob nematode ntoo thuv (Bursaphelenchus xylophilus) yog ib hom kab mob nematode ntoo thuv (PWN), uas yog cov kab mob nematode uas kis tau rau lwm qhov chaw hauv hav zoov thuv uas paub tias ua rau muaj kev puas tsuaj loj heev rau cov ecosystem ntawm hav zoov thuv1. Kab mob ploog ntoo thuv (PWD) uas tshwm sim los ntawm kab mob nematode ntoo thuv tab tom ua teeb meem loj heev rau ntau lub teb chaws, suav nrog Asia thiab Europe, thiab hauv North America, kab mob nematode rhuav tshem cov hom ntoo thuv uas tau qhia1,2. Kev poob qis ntawm tsob ntoo thuv yog teeb meem loj rau kev lag luam, thiab qhov kev cia siab ntawm nws txoj kev kis thoob ntiaj teb yog qhov txhawj xeeb3. Cov hom ntoo thuv hauv qab no feem ntau raug kab mob nematode tawm tsam: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, thiab Pinus radiata4. Kab mob nematode ntoo thuv yog ib yam kab mob loj heev uas tuaj yeem tua cov ntoo thuv hauv ob peb lub lis piam lossis ob peb lub hlis tom qab kis kab mob. Tsis tas li ntawd, kev kis kab mob nematode ntoo thuv yog ib qho uas tshwm sim ntau hauv ntau yam ecosystem, yog li ntawd cov kab mob sib kis tau tsim muaj1.
Kab mob Bursaphelenchus xylophilus yog ib hom kab mob uas nyob hauv cov nroj tsuag uas raug cais tawm uas yog los ntawm tsev neeg Aphelenchoidea thiab clade 102.5. Kab mob nematode noj cov nceb thiab tsim tawm hauv cov ntoo ntawm cov ntoo thuv, tsim ua plaub theem kab mob sib txawv: L1, L2, L3, L4 thiab ib tus neeg laus1,6. Nyob rau hauv cov xwm txheej ntawm kev tsis txaus zaub mov, kab mob nematode ntoo thuv dhau mus rau theem kab mob tshwj xeeb - dauer, uas ua rau nws tus kab mob - tus kab mob ntoo thuv (Monochamus alternatus) thiab raug xa mus rau cov ntoo thuv noj qab haus huv. Hauv cov tswv tsev noj qab haus huv, cov kab mob nematodes sai sai tsiv mus rau hauv cov ntaub so ntswg ntawm cov nroj tsuag thiab noj cov hlwb parenchymatous, uas ua rau muaj ntau yam kev tsis haum tshuaj, ntoo thuv lwj thiab tuag hauv ib xyoos tom qab kis kab mob1,7,8.
Kev tswj cov kab mob nematodes ntawm cov ntoo thuv tau ntev los ua ib qho kev sib tw, nrog rau kev ntsuas kev cais tawm txij li xyoo pua 20th. Cov tswv yim tam sim no rau kev tswj cov kab mob nematodes ntawm cov ntoo thuv feem ntau yog kev kho tshuaj, suav nrog kev tua ntoo thiab kev cog cov nematicides rau hauv cov ntoo. Cov nematicides uas siv ntau tshaj plaws yog avermectin thiab avermectin benzoate, uas yog tsev neeg avermectin. Cov tshuaj kim no muaj txiaj ntsig zoo rau ntau hom nematode thiab suav tias yog kev nyab xeeb rau ib puag ncig9. Txawm li cas los xij, kev siv cov nematicides no ntau zaus yuav tsim kev nyuaj siab xaiv uas yuav luag ua rau muaj cov nematodes ntawm cov ntoo thuv uas tiv taus, raws li tau pom rau ntau yam kab tsuag, xws li Leptinotarsa decemlineata, Plutella xylostella thiab cov nematodes Trichostrongylus colubriformis thiab Ostertagia circumcincta, uas tau maj mam tsim kev tiv taus rau avermectins10,11,12. Yog li ntawd, cov qauv tiv taus yuav tsum tau kawm tas li thiab cov nematicides raug tshuaj xyuas tas li kom nrhiav tau lwm txoj hauv kev, pheej yig thiab cov kev ntsuas zoo rau ib puag ncig los tswj PVD. Nyob rau xyoo caum dhau los no, ntau tus kws sau ntawv tau tawm tswv yim siv cov nroj tsuag rho tawm, cov roj tseem ceeb thiab cov volatiles ua cov tshuaj tswj nematode13,14,15,16.
Tsis ntev los no peb tau ua pov thawj txog kev ua haujlwm ntawm indole, ib qho molecule signaling intercellular thiab interkingdom, hauv Caenorhabditis elegans 17. Indole yog ib qho teeb liab intracellular dav dav hauv microbial ecology, tswj ntau yam haujlwm uas cuam tshuam rau microbial physiology, spore tsim, plasmid stability, tshuaj tiv thaiv, biofilm tsim, thiab virulence 18, 19. Kev ua haujlwm ntawm indole thiab nws cov derivatives tawm tsam lwm cov kab mob nematodes tsis tau kawm. Hauv kev tshawb fawb no, peb tau tshawb xyuas qhov kev ua haujlwm ntawm 34 indoles tawm tsam cov ntoo thuv nematodes thiab piav qhia txog cov txheej txheem ntawm kev ua haujlwm ntawm 5-iodoindole muaj zog tshaj plaws siv microscopy, time-lapse photography, thiab molecular docking experiments, thiab ntsuas nws cov teebmeem lom rau cov nroj tsuag siv cov noob germination assay.
Cov indole uas muaj ntau heev (>1.0 mM) tau tshaj tawm tias muaj cov nyhuv nematicidal rau nematodes17. Tom qab kev kho mob ntawm B. xylophilus (cov theem sib xyaw ua ke) nrog indole lossis 33 hom indole derivatives sib txawv ntawm 1 mM, kev tuag ntawm B. xylophilus tau ntsuas los ntawm kev suav cov nematodes uas muaj sia thiab tuag hauv cov pab pawg tswj thiab cov pab pawg kho. Tsib lub indoles tau ua rau muaj kev ua haujlwm nematicidal tseem ceeb; qhov muaj sia nyob ntawm pab pawg tswj tsis tau kho yog 95 ± 7% tom qab 24 teev. Ntawm 34 lub indoles uas tau sim, 5-iodoindole thiab 4-fluoroindole ntawm 1 mM ua rau muaj kev tuag 100%, thaum 5,6-difluoroindigo, methylindole-7-carboxylate, thiab 7-iodoindole ua rau muaj kev tuag kwv yees li 50% (Rooj 1).
Cov nyhuv ntawm 5-iodoindole rau kev tsim vacuole thiab kev ua haujlwm ntawm cov ntoo thuv nematode. (A) Cov nyhuv ntawm avermectin thiab 5-iodoindole rau cov txiv neej laus nematodes, (B) L1 theem nematode qe thiab (C) kev ua haujlwm ntawm B. xylophilus, (i) tsis pom cov vacuoles ntawm 0 teev, kev kho mob ua rau (ii) vacuoles, (iii) kev sib sau ua ke ntawm ntau lub vacuoles, (iv) o ntawm vacuoles, (v) kev sib xyaw ntawm vacuoles thiab (vi) kev tsim ntawm cov vacuoles loj. Cov xub liab qhia txog kev o ntawm vacuoles, cov xub xiav qhia txog kev sib xyaw ntawm vacuoles thiab cov xub dub qhia txog cov vacuoles loj. Scale bar = 50 μm.
Ntxiv rau, txoj kev tshawb fawb no kuj tau piav qhia txog cov txheej txheem sib law liag ntawm kev tuag los ntawm methane hauv cov nematodes ntoo thuv (Daim Duab 4C). Kev tuag los ntawm Methanogenic yog hom kev tuag ntawm tes uas tsis yog apoptotic uas cuam tshuam nrog kev sib sau ua ke ntawm cov cytoplasmic vacuoles27. Cov teeb meem morphological uas pom hauv cov nematodes ntoo thuv zoo li muaj feem cuam tshuam nrog cov txheej txheem ntawm kev tuag los ntawm methane. Kev kuaj xyuas microscopic ntawm ntau lub sijhawm sib txawv qhia tau tias cov vacuoles loj tau tsim tom qab 20 teev ntawm kev raug rau 5-iodoindole (0.1 mM). Cov vacuoles me me tau pom tom qab 8 teev ntawm kev kho mob, thiab lawv tus lej nce tom qab 12 teev. Ntau lub vacuoles loj tau pom tom qab 14 teev. Ntau lub vacuoles fused tau pom tseeb tom qab 12-16 teev ntawm kev kho mob, qhia tias vacuole fusion yog lub hauv paus ntawm cov txheej txheem tuag methanogenic. Tom qab 20 teev, ntau lub vacuoles loj tau pom thoob plaws hauv cov kab. Cov kev soj ntsuam no sawv cev rau thawj daim ntawv qhia ntawm metuosis hauv C. elegans.
Hauv cov kab mob uas kho nrog 5-iodoindole, kev sib sau ua ke ntawm vacuole thiab kev tawg kuj tau pom (Daim Duab 5), raws li pov thawj los ntawm kev khoov ntawm cov kab mob thiab kev tso tawm ntawm vacuole mus rau hauv ib puag ncig. Kev cuam tshuam ntawm vacuole kuj tau pom hauv daim nyias nyias ntawm lub plhaub qe, uas feem ntau khaws cia kom zoo los ntawm L2 thaum lub sijhawm hatching (Daim Duab Ntxiv S2). Cov kev soj ntsuam no txhawb nqa kev koom tes ntawm cov kua dej thiab kev ua tsis tiav ntawm osmoregulatory, nrog rau kev raug mob ntawm lub hlwb rov qab (RCI), hauv cov txheej txheem ntawm kev tsim vacuole thiab suppuration (Daim Duab 5).
Los ntawm kev xav txog lub luag haujlwm ntawm iodine hauv kev tsim vacuole uas tau pom, peb tau tshawb xyuas qhov kev ua haujlwm nematicidal ntawm sodium iodide (NaI) thiab potassium iodide (KI). Txawm li cas los xij, ntawm qhov concentration (0.1, 0.5 lossis 1 mM), lawv tsis cuam tshuam rau kev ciaj sia ntawm nematode lossis kev tsim vacuole (Supplementary Fig. S5), txawm hais tias 1 mM KI muaj qhov cuam tshuam me ntsis nematicidal. Ntawm qhov tod tes, 7-iodoindole (1 lossis 2 mM), zoo li 5-iodoindole, ua rau muaj ntau lub vacuoles thiab kev hloov pauv ntawm cov qauv (Supplementary Fig. S6). Ob lub iodoindoles tau qhia txog cov yam ntxwv zoo sib xws hauv cov ntoo thuv nematodes, thaum NaI thiab KI tsis tau. Qhov nthuav yog, indole tsis tau ua rau muaj vacuole hauv B. xylophilus ntawm qhov concentration sim (cov ntaub ntawv tsis tau qhia). Yog li, cov txiaj ntsig tau lees paub tias indole-iodine complex yog lub luag haujlwm rau vacuolization thiab metabolism ntawm B. xylophilus.
Ntawm cov indoles uas tau sim rau kev ua haujlwm nematicidal, 5-iodoindole muaj qhov ntsuas siab tshaj plaws ntawm -5.89 kcal / mol, ua raws li 7-iodoindole (-4.48 kcal / mol), 4-fluoroindole (-4.33), thiab indole (-4.03) (Daim Duab 6). Qhov kev sib txuas hydrogen muaj zog ntawm 5-iodoindole rau leucine 218 ua kom nws ruaj khov, thaum txhua lwm yam indole derivatives khi rau serine 260 ntawm cov saw hlau hydrogen. Ntawm lwm cov qauv iodoindoles, 2-iodoindole muaj tus nqi khi ntawm -5.248 kcal/mol, uas yog vim nws txoj kev sib txuas hydrogen tseem ceeb nrog leucine 218. Lwm cov kev khi paub suav nrog 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), thiab 6-fluoroindole (-2.6 kcal/mol) (Daim Duab Ntxiv S8). Feem ntau cov halogenated indoles thiab indole nws tus kheej, tshwj tsis yog 5-iodoindole thiab 2-iodoindole, tsim kev sib txuas nrog serine 260. Qhov tseeb tias hydrogen kev sib txuas nrog leucine 218 yog qhov qhia txog kev sib txuas ntawm receptor-ligand, raws li pom rau ivermectin (Supplementary Fig. S7), lees paub tias 5-iodoindole thiab 2-iodoindole, zoo li ivermectin, khi nruj rau qhov chaw nquag ntawm GluCL receptor ntawm leucine 218 (Daim Duab 6 thiab Supplementary Fig. S8). Peb xav tias qhov kev sib txuas no yog qhov yuav tsum tau ua kom tswj tau qhov qhib pore qauv ntawm GluCL complex thiab los ntawm kev khi nruj rau qhov chaw nquag ntawm GluCL receptor, 5-iodoindole, 2-iodoindole, avermectin thiab ivermectin yog li tswj tau qhov ion channel qhib thiab tso cai rau cov kua dej nqus.
Kev sib txuas ntawm indole thiab halogenated indole rau GluCL. Kev sib txuas ntawm (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, thiab (D) 5-iodoindole ligands rau qhov chaw nquag ntawm GluCL. Cov protein yog sawv cev los ntawm ib daim kab xev, thiab cov hydrogen bonds backbone tau qhia ua cov kab daj dotted. (A′), (B′), (C′), thiab (D′) qhia txog kev sib cuam tshuam ntawm cov ligands sib xws nrog cov amino acid residues nyob ib puag ncig, thiab cov hydrogen bonds sab-saw tau qhia los ntawm cov xub liab dotted.
Cov kev sim tau ua los ntsuam xyuas qhov cuam tshuam ntawm 5-iodoindole rau kev tawg paj ntawm cov noob zaub qhwv thiab radish. 5-iodoindole (0.05 lossis 0.1 mM) lossis avermectin (10 μg/mL) muaj qhov cuam tshuam me ntsis lossis tsis muaj rau kev tawg paj thaum pib thiab kev tawm ntawm cov nroj tsuag (Daim Duab 7). Tsis tas li ntawd, tsis pom muaj qhov sib txawv tseem ceeb ntawm qhov nrawm ntawm kev tawg paj ntawm cov noob tsis tau kho thiab cov noob kho nrog 5-iodoindole lossis avermectin. Qhov cuam tshuam rau kev ntev ntawm cov cag taproot thiab tus lej ntawm cov hauv paus sab nraud tsis tseem ceeb, txawm hais tias 1 mM (10 npaug ntawm nws qhov concentration) ntawm 5-iodoindole qeeb me ntsis rau kev loj hlob ntawm cov hauv paus sab nraud. Cov txiaj ntsig no qhia tau tias 5-iodoindole tsis muaj tshuaj lom rau cov hlwb cog thiab tsis cuam tshuam rau cov txheej txheem kev loj hlob ntawm cov nroj tsuag ntawm cov concentration uas tau kawm.
Cov nyhuv ntawm 5-iodoindole rau kev tawg noob. Kev tawg noob, kev tawg noob thiab kev tawm hauv paus ntawm B. oleracea thiab R. raphanistrum noob rau Murashige thiab Skoog agar medium nrog lossis tsis muaj avermectin lossis 5-iodoindole. Kev tawg noob tau sau tseg tom qab 3 hnub ntawm kev cog qoob loo ntawm 22 ° C.
Daim ntawv tshawb fawb no qhia txog ntau qhov xwm txheej ntawm kev tua nematode los ntawm indoles. Qhov tseem ceeb, qhov no yog thawj daim ntawv tshaj tawm ntawm iodoindole inducing methylation (ib qho txheej txheem tshwm sim los ntawm kev sib sau ua ke ntawm cov vacuoles me me uas maj mam sib koom ua ke rau hauv cov vacuoles loj, thaum kawg ua rau daim nyias nyias tawg thiab tuag) hauv cov koob ntoo thuv, nrog iodoindole qhia txog cov khoom nematicidal zoo ib yam li cov nematicide avermectin lag luam.
Yav dhau los, cov indoles tau tshaj tawm tias muaj ntau lub luag haujlwm signaling hauv prokaryotes thiab eukaryotes, suav nrog biofilm inhibition/formation, bacterial survival, thiab pathogenicity19,32,33,34. Tsis ntev los no, cov txiaj ntsig kho mob ntawm halogenated indoles, indole alkaloids, thiab semisynthetic indole derivatives tau nyiam kev tshawb fawb ntau yam35,36,37. Piv txwv li, halogenated indoles tau pom tias tua cov hlwb Escherichia coli thiab Staphylococcus aureus uas nyob mus ib txhis37. Tsis tas li ntawd, nws yog qhov txaus siab rau kev tshawb fawb los kawm txog kev ua haujlwm ntawm halogenated indoles tawm tsam lwm hom tsiaj, genera, thiab kingdoms, thiab kev tshawb fawb no yog ib kauj ruam mus rau kev ua tiav lub hom phiaj no.
Ntawm no, peb tawm tswv yim rau txoj hauv kev rau 5-iodoindole-induced lethality hauv C. elegans raws li kev raug mob ntawm tes rov qab (RCI) thiab methylation (Daim duab 4C thiab 5). Kev hloov pauv edematous xws li o thiab vacuolar degeneration yog cov cim qhia ntawm RCI thiab methylation, qhia ua cov vacuoles loj hauv cytoplasm48,49. RCI cuam tshuam nrog kev tsim hluav taws xob los ntawm kev txo qis ATP ntau lawm, ua rau lub twj tso kua mis ATPase tsis ua haujlwm, lossis cuam tshuam cov cell membranes thiab ua rau muaj kev nkag mus sai ntawm Na +, Ca2 +, thiab dej50,51,52. Intracytoplasmic vacuoles tshwm sim hauv cov tsiaj cov hlwb ua qhov tshwm sim ntawm cov kua dej hauv cytoplasm vim yog kev nkag mus ntawm Ca2 + thiab dej53. Qhov txaus nyiam yog tias txoj kev puas tsuaj ntawm cov cell no rov qab tau yog tias qhov kev puas tsuaj yog ib ntus xwb thiab cov cell pib tsim ATP rau ib lub sijhawm, tab sis yog tias qhov kev puas tsuaj tseem muaj lossis hnyav zuj zus, cov cell tuag.54 Peb cov kev soj ntsuam qhia tau tias cov nematodes kho nrog 5-iodoindole tsis tuaj yeem rov qab kho cov biosynthesis li qub tom qab raug kev ntxhov siab.
Qhov methylation phenotype uas 5-iodoindole ua rau hauv B. xylophilus tej zaum yuav yog vim muaj iodine thiab nws cov kev faib tawm molecular, vim 7-iodoindole muaj tsawg dua inhibitory cuam tshuam rau B. xylophilus dua li 5-iodoindole (Rooj 1 thiab Daim Duab Ntxiv S6). Cov txiaj ntsig no yog ib feem sib xws nrog kev tshawb fawb ntawm Maltese et al. (2014), uas tau tshaj tawm tias kev hloov pauv ntawm pyridyl nitrogen moiety hauv indole los ntawm para- mus rau meta-position tshem tawm vacuolization, kev loj hlob inhibition, thiab cytotoxicity hauv U251 hlwb, qhia tias kev sib cuam tshuam ntawm molecule nrog ib qho chaw tshwj xeeb hauv cov protein yog qhov tseem ceeb27,44,45. Kev sib cuam tshuam ntawm indole lossis halogenated indoles thiab GluCL receptors pom hauv kev tshawb fawb no kuj txhawb nqa lub tswv yim no, vim 5- thiab 2-iodoindole tau pom tias khi rau GluCL receptors muaj zog dua li lwm cov indoles kuaj xyuas (Daim Duab 6 thiab Daim Duab Ntxiv S8). Cov iodine ntawm qhov chaw thib ob lossis thib tsib ntawm indole tau pom tias khi rau leucine 218 ntawm GluCL receptor los ntawm cov hydrogen bonds backbone, thaum lwm cov halogenated indoles thiab indole nws tus kheej tsim cov hydrogen bonds tsis muaj zog nrog serine 260 (Daim Duab 6). Yog li ntawd, peb xav tias qhov chaw ntawm halogen ua lub luag haujlwm tseem ceeb hauv kev ua rau lub vacuolar degeneration, thaum qhov kev khi nruj ntawm 5-iodoindole ua rau lub ion channel qhib, yog li ua rau cov kua dej nkag sai thiab vacuole tawg. Txawm li cas los xij, cov txheej txheem ua haujlwm ntawm 5-iodoindole tseem yuav tsum tau txiav txim siab.
Ua ntej siv 5-iodoindole, nws cov teebmeem lom rau cov nroj tsuag yuav tsum tau soj ntsuam. Peb cov kev sim noob germination qhia tau hais tias 5-iodoindole tsis muaj kev cuam tshuam tsis zoo rau noob germination lossis cov txheej txheem kev loj hlob tom qab ntawm cov concentration kawm (Daim Duab 7). Yog li, txoj kev tshawb fawb no muab lub hauv paus rau kev siv 5-iodoindole hauv ib puag ncig ecological los tswj qhov kev phom sij ntawm cov ntoo thuv nematodes rau cov ntoo thuv.
Cov ntawv ceeb toom yav dhau los tau qhia tias kev kho mob raws li indole yog ib txoj hauv kev zoo los daws qhov teeb meem ntawm kev tiv thaiv tshuaj tua kab mob thiab kev loj hlob ntawm mob qog noj ntshav55. Tsis tas li ntawd, indoles muaj cov tshuaj tua kab mob, tiv thaiv mob qog noj ntshav, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative thiab antituberculosis thiab tej zaum yuav ua lub hauv paus zoo rau kev tsim tshuaj56,57. Kev tshawb fawb no qhia thawj zaug txog kev siv iodine ua tus neeg sawv cev tiv thaiv kab mob thiab anthelmintic.
Avermectin tau tshawb pom peb xyoo caum dhau los thiab yeej qhov khoom plig Nobel hauv xyoo 2015, thiab nws siv ua tshuaj anthelmintic tseem niaj hnub ua mus ntxiv. Txawm li cas los xij, vim muaj kev loj hlob sai ntawm kev tiv thaiv rau avermectins hauv nematodes thiab kab tsuag, lwm txoj hauv kev, pheej yig, thiab zoo rau ib puag ncig yog qhov xav tau los tswj kev kis tus kab mob PWN hauv cov ntoo thuv. Txoj kev tshawb fawb no kuj qhia txog txoj hauv kev uas 5-iodoindole tua cov nematodes ntoo thuv thiab tias 5-iodoindole muaj tshuaj lom tsawg rau cov hlwb cog, uas qhib kev cia siab zoo rau nws daim ntawv thov kev lag luam yav tom ntej.
Tag nrho cov kev sim tau pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Yeungnam University, Gyeongsan, Kauslim Teb, thiab cov txheej txheem tau ua raws li cov lus qhia ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Yeungnam University.
Cov kev sim qe qe tau ua tiav los ntawm kev siv cov txheej txheem uas tau tsim tsa43. Txhawm rau ntsuas qhov kev tawm ntawm qe (HR), cov nematodes laus hnub nyoog 1 hnub (kwv yees li 100 tus poj niam thiab 100 tus txiv neej) tau raug hloov mus rau hauv cov tais Petri uas muaj cov pwm thiab cia kom loj hlob rau 24 teev. Cov qe tom qab ntawd tau raug rho tawm thiab kho nrog 5-iodoindole (0.05 mM thiab 0.1 mM) lossis avermectin (10 μg / ml) ua cov dej ntws hauv cov dej ntshiab. Cov dej ntws no (500 μl; kwv yees li 100 lub qe) tau raug hloov mus rau qhov dej ntawm 24-qhov dej ntawm cov phaj kab lis kev cai thiab muab tso rau ntawm 22 ° C. L2 suav tau ua tom qab 24 teev ntawm kev muab tso rau hauv qe tab sis suav tias tuag yog tias cov hlwb tsis txav thaum raug txhawb nrog cov hlau platinum zoo. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua tus muaj rau qhov rov ua dua. Cov ntaub ntawv los ntawm ob qho kev sim tau muab tso ua ke thiab nthuav tawm. Feem pua ntawm HR tau suav raws li hauv qab no:
Kev tuag ntawm cov kab me me tau raug soj ntsuam siv cov txheej txheem uas tau tsim ua ntej. Cov qe nematode tau raug sau thiab cov embryos tau sib dhos los ntawm kev hatching hauv dej distilled sterile kom tsim cov kab me me theem L2. Cov kab me me sib dhos (kwv yees li 500 nematodes) tau kho nrog 5-iodoindole (0.05 mM thiab 0.1 mM) lossis avermectin (10 μg/ml) thiab yug rau ntawm B. cinerea Petri phaj. Tom qab 48 teev ntawm kev incubation ntawm 22 ° C, nematodes tau raug sau hauv dej distilled sterile thiab kuaj xyuas seb puas muaj theem L2, L3, thiab L4. Qhov muaj theem L3 thiab L4 qhia txog kev hloov pauv ntawm cov kab me me, thaum qhov muaj theem L2 qhia tias tsis muaj kev hloov pauv. Cov duab tau txais los ntawm kev siv iRiS ™ Digital Cell Imaging System. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua theem muaj rau qhov rov ua dua. Cov ntaub ntawv los ntawm ob qho kev sim tau muab tso ua ke thiab nthuav tawm.
Qhov tshuaj lom ntawm 5-iodoindole thiab avermectin rau cov noob tau raug soj ntsuam siv kev sim germination ntawm Murashige thiab Skoog agar phaj.62 B. oleracea thiab R. raphanistrum noob tau muab tso rau hauv dej ntshiab huv rau ib hnub, ntxuav nrog 1 ml 100% ethanol, sterilized nrog 1 ml 50% tshuaj dawb (3% sodium hypochlorite) rau 15 feeb, thiab ntxuav tsib zaug nrog 1 ml dej huv. Cov noob sterilized ces raug nias rau ntawm cov phaj germination agar uas muaj 0.86 g / l (0.2X) Murashige thiab Skoog nruab nrab thiab 0.7% bacteriological agar nrog lossis tsis muaj 5-iodoindole lossis avermectin. Cov phaj tau raug incubated ntawm 22 ° C, thiab cov duab tau raug coj tom qab 3 hnub ntawm incubation. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua tus muaj rau qhov rov ua dua.
Lub sijhawm tshaj tawm: Lub Ob Hlis-26-2025