Pine nematode yog ib qho quarantine migratory endoparasite paub tias ua rau muaj kev puas tsuaj loj hauv cov ntoo thuv hav zoov ecosystems. Txoj kev tshawb fawb tam sim no tshuaj xyuas cov haujlwm nematicidal ntawm halogenated indoles tawm tsam Pine nematodes thiab lawv cov txheej txheem ntawm kev ua. Kev ua haujlwm nematicidal ntawm 5-iodoindole thiab avermectin (zoo tswj) tawm tsam Pine nematodes tau zoo sib xws thiab siab ntawm qhov tsis tshua muaj siab (10 μg / mL). 5-iodoindole txo fecundity, kev ua me nyuam, embryonic thiab larval mortality, thiab locomotor cwj pwm. Molecular kev sib cuam tshuam ntawm ligands nrog invertebrate-specific glutamate-gated chloride channel receptors txhawb txoj kev xav tias 5-iodoindole, zoo li avermectin, khi nruj nreem rau lub chaw ua haujlwm receptor. 5-Iodoindole kuj induced ntau yam phenotypic deformations nyob rau hauv nematodes, nrog rau qhov txawv txav ntawm lub cev qhuav dej / shrinkage thiab nce vacuolization. Cov txiaj ntsig no qhia tias vacuoles tuaj yeem ua lub luag haujlwm hauv nematode methylation-mediated tuag. Qhov tseem ceeb, 5-iodoindole tsis muaj tshuaj lom rau ob hom nroj tsuag (cab thiab radish). Yog li, txoj kev tshawb no qhia tau hais tias iodoindole daim ntawv thov nyob rau hauv ib puag ncig tej yam kev mob tuaj yeem tswj cov ntoo thuv wilt raug mob.
Pine ntoo nematode (Bursaphelenchus xylophilus) belongs rau Pine ntoo nematodes (PWN), migratory endoparasitic nematodes paub ua rau kev puas tsuaj loj rau cov ntoo thuv hav zoov ecosystems1. Pine wilt disease (PWD) tshwm sim los ntawm ntoo thuv ntoo nematode tau dhau los ua teeb meem loj rau ntau lub tebchaws, suav nrog Asia thiab Europe, thiab hauv North America, cov nematode rhuav tshem cov ntoo thuv hom1,2. Pine ntoo poob yog ib qho teeb meem loj ntawm kev lag luam, thiab kev cia siab ntawm nws txoj kev sib kis thoob ntiaj teb yog kev txhawj xeeb3. Cov ntoo ntoo hauv qab no feem ntau raug tua los ntawm nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, thiab Pinus radiata4. Pine nematode yog ib yam kab mob loj uas tuaj yeem tua cov ntoo thuv hauv lub lis piam lossis ntau lub hlis ntawm kev kis kab mob. Tsis tas li ntawd, cov kab mob Pine nematode muaj ntau nyob rau hauv ntau yam ecosystems, yog li cov kab mob tsis tu ncua tau tsim muaj 1.
Bursaphelenchus xylophilus yog ib tsob nroj-parasitic nematode uas muaj nyob rau hauv superfamily Aphelenchoidea thiab clade 102.5. Cov nematode pub rau cov kab mob thiab rov tsim dua nyob rau hauv cov ntaub so ntswg ntawm cov ntoo thuv, tsim mus rau plaub theem sib txawv: L1, L2, L3, L4 thiab tus neeg laus 1,6. Nyob rau hauv tej yam kev mob ntawm cov zaub mov tsis txaus, cov ntoo thuv nematode kis mus rau ib tug tshwj xeeb larval theem - dauer, uas parasitizes nws vector - ntoo thuv bark kab (Monochamus alternatus) thiab pauv mus rau noj qab nyob zoo ntoo thuv. Hauv cov tswv tsev noj qab haus huv, nematodes sai sai tsiv los ntawm cov ntaub so ntswg thiab pub rau ntawm parenchymatous hlwb, uas ua rau muaj ntau yam kev xav tsis zoo, ntoo thuv wilting thiab tuag hauv ib xyoos tom qab kis kab mob 1,7,8.
Kev tswj kev lom neeg ntawm cov ntoo thuv nematodes tau ntev dhau los yog qhov nyuaj, nrog rau kev ntsuas cais tawm txij li xyoo pua 20th. Cov tswv yim tam sim no rau kev tswj cov ntoo thuv nematodes feem ntau koom nrog kev kho tshuaj, suav nrog kev cog ntoo thiab cog cov nematicides rau hauv tsob ntoo trunks. Feem ntau siv nematicides yog avermectin thiab avermectin benzoate, uas yog cov tsev neeg avermectin. Cov tshuaj kim kim no muaj txiaj ntsig zoo tiv thaiv ntau hom nematode thiab suav tias yog ib puag ncig zoo9. Txawm li cas los xij, rov siv cov nematicides yuav tsum tsim kom muaj kev xaiv siab uas yuav luag yuav ua rau muaj qhov tshwm sim ntawm cov ntoo thuv resistant nematodes, raws li tau qhia rau ntau yam kab kab, xws li Leptinotarsa decemlineata, Plutella xylostella thiab nematodes Trichostrongylus colubriformis thiab Ostertagia, uas maj mam tsim tawm. avermectins 10, 11, 12. Yog li ntawd, cov qauv kev tiv thaiv yuav tsum tau kawm tsis tu ncua thiab tshuaj xyuas cov tshuaj nematicides tsis tu ncua los nrhiav lwm txoj hauv kev, raug nqi thiab kev tiv thaiv ib puag ncig los tswj PVD. Nyob rau hauv xyoo tsis ntev los no, ib tug xov tooj ntawm cov kws sau ntawv tau npaj siv cov nroj tsuag rho tawm, cov roj yam tseem ceeb thiab volatiles li nematode tswj cov neeg sawv cev13,14,15,16.
Peb tsis ntev los no tau qhia txog kev ua haujlwm nematicidal ntawm indole, intercellular thiab interkingdom signaling molecule, hauv Caenorhabditis elegans 17 . Indole yog lub teeb liab intracellular thoob plaws hauv microbial ecology, tswj ntau lub zog uas cuam tshuam rau microbial physiology, tsim spore, plasmid stability, tshuaj tiv thaiv, biofilm tsim, thiab virulence 18, 19. Cov kev ua ntawm indole thiab nws cov derivatives tawm tsam lwm yam pathogenic nematodes tsis tau kawm. Hauv txoj kev tshawb no, peb tau tshawb xyuas cov haujlwm nematicidal ntawm 34 indoles tawm tsam Pine nematodes thiab elucidated lub mechanism ntawm kev txiav txim ntawm lub zog tshaj 5-iodoindole siv microscopy, time-lapse photography, thiab molecular docking thwmsim, thiab ntsuam xyuas nws cov tshuaj lom rau cov nroj tsuag siv cov noob germination assay.
Cov ntsiab lus siab (> 1.0 mM) ntawm indole tau tshaj tawm yav dhau los tias muaj cov nyhuv nematicidal ntawm nematodes17. Tom qab kev kho mob ntawm B. xylophilus (ib theem ntawm lub neej sib xyaw) nrog indole lossis 33 qhov sib txawv indole derivatives ntawm 1 mM, kev tuag ntawm B. xylophilus tau ntsuas los ntawm kev suav cov nematodes nyob thiab tuag hauv pawg tswj thiab kho. Tsib indoles nthuav tawm cov haujlwm tseem ceeb nematicidal; Kev ciaj sia ntawm pawg tswj tsis tau kho yog 95 ± 7% tom qab 24 teev. Ntawm 34 indoles kuaj, 5-iodoindole thiab 4-fluoroindole ntawm 1 mM ua rau 100% kev tuag, whereas 5,6-difluoroindigo, methylindole-7-carboxylate, thiab 7-iodoindole ua rau kwv yees li 50% kev tuag (Table 1).
Cov nyhuv ntawm 5-iodoindole ntawm vacuole tsim thiab metabolism ntawm ntoo thuv nematode. (A) Cov nyhuv ntawm avermectin thiab 5-iodoindole rau cov txiv neej laus nematodes, (B) L1 theem nematode qe thiab (C) metabolism ntawm B. xylophilus, (i) vacuoles tsis tau soj ntsuam ntawm 0 h, kev kho mob ua rau (ii) vacuoles, (iii) tsub zuj zuj ntawm ntau lub vacuoles, (iv) vacuoles ntawm swelling, (vi) tsim cov vacuoles loj heev. Cov xub liab qhia o ntawm vacuoles, xiav xub qhia fusion ntawm vacuoles thiab dub xub qhia loj vacuoles. Luas bar = 50 μm.
Tsis tas li ntawd, txoj kev tshawb no kuj tau piav qhia txog cov txheej txheem ua ntu zus ntawm methane-induced tuag hauv Pine nematodes (Daim duab 4C). Methanogenic tuag yog ib yam uas tsis yog-apoptotic cell tuag txuam nrog tsub zuj zuj ntawm cytoplasmic vacuoles tseem ceeb 27. Cov teeb meem morphological pom nyob rau hauv cov ntoo thuv nematodes zoo li muaj feem xyuam nrog cov txheej txheem ntawm kev tuag ntawm methane. Kev tshuaj xyuas microscopic ntawm lub sijhawm sib txawv tau pom tias cov vacuoles loj heev tau tsim tom qab 20 teev ntawm kev raug rau 5-iodoindole (0.1 mM). Microscopic vacuoles tau pom tom qab 8 teev ntawm kev kho mob, thiab lawv cov lej tau nce ntxiv tom qab 12 teev. Ob peb vacuoles loj tau pom tom qab 14 teev. Ntau lub fused vacuoles tau pom meej meej tom qab 12-16 teev ntawm kev kho mob, qhia tias vacuole fusion yog lub hauv paus ntawm cov txheej txheem methanogenic tuag. Tom qab 20 teev, ntau lub vacuoles loj heev tau pom thoob plaws hauv tus cab. Cov kev soj ntsuam no sawv cev rau thawj tsab ntawv ceeb toom ntawm metuosis hauv C. elegans .
Hauv 5-iodoindole-kho kab mob, vacuole aggregation thiab rupture kuj tau pom (Daim duab 5), raws li muaj pov thawj los ntawm cov kab mob bending thiab vacuole tso rau hauv ib puag ncig. Vacuole cuam tshuam kuj tau pom nyob rau hauv cov qe qe, uas ib txwm khaws cia tsis tu ncua los ntawm L2 thaum lub sij hawm hatching (Ntxiv Fig. S2). Cov kev soj ntsuam no txhawb kev koom tes ntawm cov kua dej sib xyaw thiab osmoregulatory tsis ua haujlwm, nrog rau kev raug mob ntawm tes rov qab (RCI), hauv cov txheej txheem ntawm vacuole tsim thiab suppuration (Fig. 5).
Kev xav txog lub luag haujlwm ntawm iodine hauv kev pom cov vacuole tsim, peb tau tshawb xyuas cov haujlwm nematicidal ntawm sodium iodide (NaI) thiab potassium iodide (KI). Txawm li cas los xij, ntawm cov concentrations (0.1, 0.5 lossis 1 mM), lawv tsis cuam tshuam rau nematode ciaj sia los yog vacuole tsim (Ntxiv daim duab S5), txawm hais tias 1 mM KI muaj cov nyhuv nematicidal me ntsis. Ntawm qhov tod tes, 7-iodoindole (1 lossis 2 mM), zoo li 5-iodoindole, induced ntau vacuoles thiab cov qauv deformations (Ntxiv daim duab S6). Ob lub iodoindoles tau pom cov yam ntxwv zoo sib xws hauv Pine nematodes, thaum NaI thiab KI tsis ua. Interestingly, indole tsis induce vacuole tsim nyob rau hauv B. xylophilus ntawm cov concentrations kuaj (cov ntaub ntawv tsis qhia). Yog li, cov txiaj ntsig tau lees paub tias indole-iodine complex yog lub luag haujlwm rau lub vacuolization thiab metabolism ntawm B. xylophilus.
Ntawm cov indoles kuaj rau nematicidal kev ua ub no, 5-iodoindole muaj qhov siab tshaj qhov ntsuas 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). Lub pob txha muaj zog hydrogen bonding ntawm 5-iodoindole rau leucine 218 stabilizes nws txoj kev khi, whereas tag nrho lwm yam indole derivatives khi rau serine 260 ntawm sab saw hydrogen bonds. Ntawm lwm cov qauv iodoindoles, 2-iodoindole muaj qhov sib txuam ntawm -5.248 kcal / mol, uas yog vim nws cov ntsiab lus hydrogen nrog leucine 218. Lwm cov kev sib txuas uas paub muaj xws li 3-iodoindole (-4.3 kcal / mol), 4-iodoindole (-6.2 kcal), thiab fluoro. kcal/mol) (Ntxiv daim duab S8). Feem ntau cov halogenated indoles thiab indole nws tus kheej, nrog rau kev zam ntawm 5-iodoindole thiab 2-iodoindole, tsim ib daim ntawv cog lus nrog serine 260. Qhov tseeb tias hydrogen bonding nrog leucine 218 yog qhov qhia tau hais tias muaj txiaj ntsig receptor-ligand binding, raws li pom rau ivermectin (Supplementary Fig). 2-iodoindole, zoo li ivermectin, khi nruj rau qhov chaw nquag ntawm GluCL receptor ntawm leucine 218 (Fig. 6 thiab Supplementary Fig. S8). Peb npaj siab tias qhov kev khi no yuav tsum tau ua kom muaj qhov qhib pore qauv ntawm GluCL complex thiab uas los ntawm kev khi nruj rau qhov chaw nquag ntawm GluCL receptor, 5-iodoindole, 2-iodoindole, avermectin thiab ivermectin yog li tswj cov ion channel qhib thiab tso cai rau cov kua dej.
Molecular docking ntawm indole thiab halogenated indole rau GluCL. Binding orientations ntawm (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, thiab (D) 5-iodoindole ligands mus rau qhov chaw nquag ntawm GluCL. Cov protein yog sawv cev los ntawm ib tug ribbon, thiab cov caj qaum hydrogen bonds yog pom raws li daj dotted kab. (A′), (B′), (C′), thiab (D′) qhia txog kev sib cuam tshuam ntawm cov ligands sib raug zoo nrog cov amino acid residues nyob ib puag ncig, thiab sab-chain hydrogen bonds yog qhia los ntawm liab dotted xub.
Kev sim tau ua los ntsuas cov tshuaj lom ntawm 5-iodoindole ntawm kev loj hlob ntawm cov zaub qhwv thiab radish noob. 5-iodoindole (0.05 lossis 0.1 mM) lossis avermectin (10 μg / mL) tsis muaj kev cuam tshuam me me los yog tsis muaj kev cuam tshuam rau thawj zaug germination thiab plantlet tshwm sim (Daim duab 7). Tsis tas li ntawd, tsis muaj qhov sib txawv tseem ceeb ntawm qhov germination tus nqi ntawm kev tswj tsis tau kho thiab cov noob kho nrog 5-iodoindole lossis avermectin. Cov nyhuv ntawm taproot elongation thiab tus naj npawb ntawm cov hauv paus hniav tsim yog qhov tsis tseem ceeb, txawm hais tias 1 mM (10 npaug ntawm nws cov active concentration) ntawm 5-iodoindole me ntsis ncua kev loj hlob ntawm cov hauv paus hniav. Cov txiaj ntsig no qhia tau hais tias 5-iodoindole tsis muaj tshuaj lom rau cov nroj tsuag thiab tsis cuam tshuam nrog cov txheej txheem kev loj hlob ntawm cov nroj tsuag hauv kev kawm.
Cov nyhuv ntawm 5-iodoindole ntawm noob germination. Germination, sprouting thiab lateral rooting ntawm B. oleracea thiab R. raphanistrum noob ntawm Murashige thiab Skoog agar nruab nrab nrog los yog tsis muaj avermectin los yog 5-iodoindole. Germination tau sau tseg tom qab 3 hnub ntawm incubation ntawm 22 ° C.
Txoj kev tshawb no qhia ntau qhov xwm txheej ntawm nematode tua los ntawm indoles. Qhov tseem ceeb, qhov no yog thawj daim ntawv tshaj tawm ntawm iodoindole inducing methylation (ib txheej txheem tshwm sim los ntawm kev sib sau ntawm cov vacuoles me me uas maj mam sib xyaw ua cov vacuoles loj, nws thiaj li ua rau daim nyias nyias tawg thiab tuag) hauv cov koob ntoo thuv, nrog iodoindole nthuav tawm cov khoom tseem ceeb nematicidal zoo ib yam li cov lag luam nematicide avermectin.
Indoles tau tshaj tawm yav dhau los kom siv ntau lub teeb liab ua haujlwm hauv prokaryotes thiab eukaryotes, suav nrog biofilm inhibition / tsim, kab mob ciaj sia, thiab pathogenicity19,32,33,34. Tsis ntev los no, cov teebmeem kev kho mob ntawm halogenated indoles, indole alkaloids, thiab semisynthetic indole derivatives tau nyiam kev tshawb fawb ntau yam txaus siab35,36,37. Piv txwv li, halogenated indoles tau pom tias tua kab mob Escherichia coli thiab Staphylococcus aureus hlwb37. Tsis tas li ntawd, nws yog qhov txaus siab rau kev tshawb fawb los kawm txog kev ua tau zoo ntawm halogenated indoles tawm tsam lwm hom, genera, thiab lub nceeg vaj, thiab txoj kev tshawb no yog ib kauj ruam mus rau kev ua tiav lub hom phiaj no.
Ntawm no, peb npaj ib lub tswv yim rau 5-iodoindole-induced lethality nyob rau hauv C. elegans raws li reversible cell raug mob (RCI) thiab methylation (Figures 4C thiab 5). Cov kev hloov pauv xws li puffiness thiab vacuolar degeneration yog cov cim ntawm RCI thiab methylation, tshwm sim raws li 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 dej 50,51,52. Intracytoplasmic vacuoles tshwm sim nyob rau hauv cov tsiaj hlwb los ntawm cov kua tsub zuj zuj nyob rau hauv lub cytoplasm vim influx ntawm Ca2+ thiab dej53. Interestingly, cov txheej txheem ntawm kev puas tsuaj ntawm tes yog thim rov qab yog tias qhov kev puas tsuaj yog ib ntus thiab cov hlwb pib tsim ATP rau qee lub sijhawm, tab sis yog tias qhov kev puas tsuaj txuas ntxiv mus lossis zuj zus, cov hlwb tuag.54 Peb cov kev soj ntsuam pom tau tias cov nematodes kho nrog 5-iodoindole tsis tuaj yeem rov ua dua li qub biosynthesis tom qab raug kev ntxhov siab.
Lub methylation phenotype induced los ntawm 5-iodoindole nyob rau hauv B. xylophilus tej zaum yuav yog vim muaj iodine thiab nws molecular tis, txij li thaum 7-iodoindole muaj tsawg inhibitory nyhuv ntawm B. xylophilus tshaj 5-iodoindole (Table 1 thiab ntxiv daim duab S6). Cov txiaj ntsig no yog ib feem nrog cov kev tshawb fawb ntawm Maltese li al. (2014), leej twg tau tshaj tawm tias kev hloov pauv ntawm pyridyl nitrogen moiety hauv indole los ntawm para- mus rau meta-txoj hauj lwm tshem tawm vacuolization, kev loj hlob inhibition, thiab cytotoxicity hauv U251 hlwb, qhia tias kev sib cuam tshuam ntawm cov molecule nrog ib qho chaw tshwj xeeb hauv cov protein yog qhov tseem ceeb27,44,45. Kev sib cuam tshuam ntawm indole los yog halogenated indoles thiab GluCL receptors tau pom nyob rau hauv txoj kev tshawb no kuj txhawb qhov kev xav no, raws li 5- thiab 2-iodoindole tau pom los khi rau GluCL receptors ntau dua li lwm cov indoles tshuaj xyuas (Daim duab 6 thiab ntxiv daim duab S8). Cov iodine ntawm qhov thib ob lossis thib tsib txoj haujlwm ntawm indole tau pom los khi rau leucine 218 ntawm GluCL receptor los ntawm cov pob txha hydrogen bonds, thaum lwm cov halogenated indoles thiab indole nws tus kheej tsim tsis muaj zog sab-chain hydrogen bonds nrog serine 260 (Daim duab 6). Yog li ntawd, peb xav tias qhov chaw ntawm lub halogen ua lub luag haujlwm tseem ceeb hauv kev cuam tshuam ntawm vacuolar degeneration, thaum lub kaw ntom nti ntawm 5-iodoindole ua rau lub ion channel qhib, yog li tso cai rau cov kua dej sai sai thiab vacuole rupture. Txawm li cas los xij, cov ncauj lus kom ntxaws ntawm kev ua ntawm 5-iodoindole tseem yuav txiav txim siab.
Ua ntej siv 5-iodoindole, nws yuav tsum tau soj ntsuam cov nyhuv toxic ntawm cov nroj tsuag. Peb cov noob germination thwmsim tau pom tias 5-iodoindole tsis muaj qhov tsis zoo rau cov noob germination los yog cov txheej txheem kev loj hlob tom ntej ntawm cov concentrations kawm (Daim duab 7). Yog li, txoj kev tshawb no muab lub hauv paus rau kev siv 5-iodoindole nyob rau hauv ecological ib puag ncig los tswj qhov teeb meem ntawm ntoo thuv nematodes rau ntoo thuv.
Cov ntawv ceeb toom dhau los tau pom tias kev kho mob indole-raws li sawv cev rau txoj hauv kev los daws qhov teeb meem ntawm cov tshuaj tiv thaiv kab mob thiab kev mob qog noj ntshav55. Tsis tas li ntawd, indoles muaj cov tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tua kab mob, tshuaj tiv thaiv kab mob thiab tshuaj tua kab mob thiab tuaj yeem ua lub hauv paus cog lus rau kev tsim tshuaj 56,57. Txoj kev tshawb no qhia thawj zaug qhov muaj peev xwm siv iodine ua ib qho tshuaj tiv thaiv kab mob thiab tshuaj tua kab mob.
Avermectin tau tshawb pom peb xyoo dhau los thiab yeej qhov khoom plig Nobel hauv 2015, thiab nws siv los ua tshuaj tua kab mob tseem ua haujlwm tsis tu ncua. Txawm li cas los xij, vim muaj kev loj hlob sai ntawm kev tiv thaiv avermectins hauv nematodes thiab kab kab, ib qho kev xaiv, tus nqi qis, thiab tus phooj ywg ib puag ncig yog xav tau los tswj PWN kab mob hauv cov ntoo thuv. Txoj kev tshawb no tseem qhia txog cov txheej txheem uas 5-iodoindole tua cov ntoo thuv nematodes thiab tias 5-iodoindole muaj toxicity tsawg rau cov nroj tsuag hlwb, uas qhib qhov kev cia siab zoo rau nws daim ntawv thov kev lag luam yav tom ntej.
Txhua qhov kev sim tau pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Yeungnam University, Gyeongsan, Kaus Lim Kauslim, thiab cov txheej txheem tau ua raws li cov lus qhia ntawm Pawg Saib Xyuas Kev Ncaj Ncees ntawm Yeungnam University.
Qe incubation thwmsim tau ua nrog cov txheej txheem tsim 43. Txhawm rau txheeb xyuas tus nqi hatching (HR), 1-hnub cov neeg laus nematodes (kwv yees li 100 tus poj niam thiab 100 txiv neej) raug xa mus rau Petri cov tais diav uas muaj cov kab mob thiab tso cai kom loj hlob rau 24 teev. Qe tau muab cais tawm thiab kho nrog 5-iodoindole (0.05 mM thiab 0.1 mM) lossis avermectin (10 μg / ml) raws li kev ncua hauv cov dej tsis huv. Cov kev ncua no (500 μl; kwv yees li 100 qe) tau pauv mus rau qhov dej ntawm 24-zoo cov ntaub so ntswg kab lis kev cai phaj thiab incubated ntawm 22 ° C. L2 suav tau ua tom qab 24 teev ntawm incubation tab sis raug suav hais tias tuag yog tias cov hlwb tsis txav thaum txhawb nqa nrog cov hlau platinum zoo. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua qhov muaj rau rov ua dua. Cov ntaub ntawv los ntawm ob qhov kev sim tau muab tso ua ke thiab nthuav tawm. Qhov feem pua ntawm HR yog xam raws li hauv qab no:
Larval mortality raug soj ntsuam siv cov txheej txheem tsim yav dhau los. Nematode qe tau sau thiab embryos tau synchronized los ntawm hatching hauv sterile distilled dej los tsim L2 theem larvae. Synchronized larvae (kwv yees li 500 nematodes) tau kho nrog 5-iodoindole (0.05 mM thiab 0.1 mM) los yog avermectin (10 μg / ml) thiab reared ntawm B. cinerea Petri daim hlau. Tom qab 48 teev ntawm incubation ntawm 22 ° C, nematodes tau sau nyob rau hauv sterile distilled dej thiab tshuaj xyuas rau lub xub ntiag ntawm L2, L3, thiab L4 theem. Lub xub ntiag ntawm L3 thiab L4 theem qhia tau hais tias larval transformation, whereas lub xub ntiag ntawm L2 theem qhia tsis muaj transformation. Cov duab tau txais los ntawm iRiS™ Digital Cell Imaging System. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua qhov muaj rau rov ua dua. Cov ntaub ntawv los ntawm ob qhov kev sim tau muab tso ua ke thiab nthuav tawm.
Cov tshuaj toxicity ntawm 5-iodoindole thiab avermectin rau cov noob tau raug soj ntsuam siv cov germination tests ntawm Murashige thiab Skoog agar daim hlau.62 B. oleracea thiab R. raphanistrum noob yog thawj zaug soaked nyob rau hauv sterile distilled dej rau ib hnub, ntxuav nrog 1 ml 100% ethanol, 1% sterilized coj mus muag. hypochlorite) rau 15 feeb, thiab ntxuav tsib zaug nrog 1 ml dej sterile. Cov noob sterilized ces nias mus rau germination agar daim hlau uas muaj 0.86 g / l (0.2X) Murashige thiab Skoog nruab nrab thiab 0.7% bacteriological agar nrog los yog tsis muaj 5-iodoindole los yog avermectin. Cov phiajcim tau muab tso rau ntawm qhov kub ntawm 22 ° C, thiab cov duab tau muab coj los ua tom qab 3 hnub ntawm incubation. Qhov kev sim no tau ua nyob rau hauv ob theem, txhua qhov muaj rau rov ua dua.
Post lub sij hawm: Feb-26-2025