Nematode pine bụ endoparasite na-akwaga iche iche mara na ọ na-akpata nnukwu mfu akụ na ụba na gburugburu oke ọhịa pine. Ọmụmụ ihe ugbu a na-enyocha ọrụ nematidal nke indoles halogenated megide nematodes pine na usoro ha si eme. Ihe omume nematidal nke 5-iodoindole na avermectin (nchịkwa dị mma) megide nematodes pine yiri ma dị elu na obere mkpokọta (10 μg/mL). 5-iodoindole belatara nwa ebu n'afọ, ọrụ ịmụ nwa, ọnwụ embrayo na nwa larval, na omume locomotor. Mmekọrịta molecular nke ligands na ndị na-anabata ọwa glutamate-gated chloride invertebrate na-akwado echiche ahụ na 5-iodoindole, dị ka avermectin, na-ejikọta ya na saịtị na-arụ ọrụ. 5-Iodoindole kpatakwara nrụrụ dị iche iche phenotypic na nematodes, gụnyere akụkụ ahụ na-adịghị mma daa / mbelata na ịba ụba. Nsonaazụ ndị a na-egosi na vacuoles nwere ike ịrụ ọrụ na ọnwụ mgbasa ozi nematode methylation. N'ụzọ dị mkpa, 5-iodoindole adịghị egbu egbu maka ụdị osisi abụọ ahụ (kabeeji na radish). Ya mere, ọmụmụ ihe a na-egosi na ngwa iodoindole n'okpuru ọnọdụ gburugburu ebe obibi nwere ike ịchịkwa pine wilt mmerụ ahụ.
nematode osisi pine (Bursaphelenchus xylophilus) bụ nke nematodes osisi pine (PWN), nematodes endoparasitic na-akwaga ebe a na-ama na ọ na-emebi oke gburugburu ebe obibi na gburugburu ebe obibi ọhịa pine1. Ọrịa pine wilt (PWD) nke nematode osisi pine kpatara na-aghọ nsogbu siri ike na kọntinent dị iche iche, gụnyere Asia na Europe, na North America, nematode na-ebibi ụdị pine ndị ewepụtara1,2. Mbelata osisi pine bụ nnukwu nsogbu akụ na ụba, na atụmanya nke mgbasa ya zuru ụwa ọnụ bụ ihe na-echegbu3. nematode na-awakpo ụdị pine ndị a: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, na Pinus radiata4. Pine nematode bụ ọrịa siri ike nke nwere ike igbu osisi pine n'ime izu ma ọ bụ ọnwa nke ọrịa. Na mgbakwunye, ntiwapụ nke nematode pine na-adịkarị n'ụdị dị iche iche nke gburugburu ebe obibi, ya mere ejirila ụdọ ọrịa na-adịgide adịgide1.
Bursaphelenchus xylophilus bụ nematode na-anapụ iche - parasitic nematode nke ezinụlọ Aphelenchoidea na clade 102.5. nematode na-eri nri fungi ma na-amụba n'ime anụ ahụ osisi nke osisi pine, na-etolite n'ụdị ahịrị anọ dị iche iche: L1, L2, L3, L4 na onye toro eto1,6. N'okpuru ọnọdụ ụkọ nri, pine nematode na-agafe n'ime oghere pụrụ iche - dauer, nke na-eme ka vector ya - osisi pine pine (Monochamus alternatus) na-ebufe ya na osisi pine dị mma. Na ndị ọbịa ahụike, nematodes na-akwaga ngwa ngwa site na anụ ahụ osisi ma na-eri nri na mkpụrụ ndụ parenchymatous, nke na-eduga n'ọtụtụ mmeghachi omume hypersensitivity, pine wilting na ọnwụ n'ime otu afọ mgbe ọrịa 1,7,8 gasịrị.
Ijikwa usoro ndu nke nematodes pine abụwo ihe ịma aka ogologo oge, yana usoro mgbochi iche malitere na narị afọ nke 20. Usoro dị ugbu a maka ịchịkwa nematodes pine na-agụnye ọgwụgwọ kemịkalụ, gụnyere nsị osisi na ịkụnye nematicides n'ime ogwe osisi. Nematides ndị a na-ejikarị eme ihe bụ avermectin na avermectin benzoate, nke sitere na ezinụlọ avermectin. Kemịkalụ ndị a dị oke ọnụ na-arụ ọrụ nke ukwuu megide ọtụtụ ụdị nematode ma na-ewere ya dị ka nchekwa gburugburu. Otú ọ dị, a na-atụ anya iji nematicides ndị a ugboro ugboro na-emepụta nrụgide nhọrọ nke ga-eme ka ọ bụrụ na ọ ga-eduga ná mmalite nke nematodes pine na-eguzogide ọgwụ, dị ka e gosipụtara maka ọtụtụ pests ụmụ ahụhụ, dị ka Leptinotarsa decemlineata, Plutella xylostella na nematodes Trichostrongylus colubriformis na Ostertagia ji nwayọọ nwayọọ na-eguzogide, 1. Ya mere, ọ dị mkpa ka a na-amụ usoro nguzogide oge niile ma na-enyocha nematicides mgbe niile ka ịchọta usoro ọzọ, ọnụ ahịa na gburugburu ebe obibi iji chịkwaa PVD. N'ime iri afọ ndị na-adịbeghị anya, ọtụtụ ndị na-ede akwụkwọ tụrụ aro ka a na-eji ihe ọkụkụ, mmanụ dị mkpa na ihe na-agbanwe agbanwe dị ka ndị ọrụ nchịkwa nematode13,14,15,16.
N'oge na-adịbeghị anya, anyị gosipụtara ọrụ nemacidal nke indole, ihe nrịbama intercellular na interkingdom, na Caenorhabditis elegans 17. Indole bụ mgbaàmà intracellular zuru ebe niile na microbial ecology, na-achịkwa ọtụtụ ọrụ na-emetụta physiology microbial, spore formation, plasmid nkwụsi ike, nkwụsị ọgwụ, mmepụta biofilm, na virulence 18, 19. A mụọbeghị ọrụ indole na ihe ndị sitere na ya megide nematodes pathogenic ndị ọzọ. N'ime ọmụmụ ihe a, anyị nyochara ọrụ nematicidal nke 34 indoles megide nematodes pine wee kọwaa usoro nke omume nke 5-iodoindole kachasị ike site na iji microscopy, foto oge na-adịghị anya, na nyocha nke molecular docking, ma nyochaa mmetụta ya na-egbu egbu na osisi site na iji nyocha mkpụrụ osisi.
Akọpụtala ọkwa dị elu (> 1.0 mM) nke indole nwere mmetụta nematidal na nematodes17. N'ịgbaso ọgwụgwọ nke B. xylophilus (ọgbakọ ndụ agwakọta) na indole ma ọ bụ 33 dị iche iche indole dị iche iche na 1 mM, a na-atụle ọnwụ nke B. xylophilus site na ịgụta ndị nematodes dị ndụ na ndị nwụrụ anwụ na njikwa na ndị otu a na-emeso ya. Indoles ise gosipụtara nnukwu ọrụ nematidal; nlanarị nke otu njikwa na-enweghị ọgwụgwọ bụ 95 ± 7% mgbe awa 24 gachara. N'ime 34 indoles nwalere, 5-iodoindole na 4-fluoroindole na 1 mM kpatara ọnwụ 100%, ebe 5,6-difluoroindigo, methylindole-7-carboxylate, na 7-iodoindole mere ihe dịka 50% ọnwụ (Table 1).
Mmetụta 5-iodoindole na nhazi vacuole na metabolism nke osisi pine nematode. (A) Mmetụta nke avermectin na 5-iodoindole na ndị okenye nematodes, (B) L1 ogbo nematode nsen na (C) metabolism nke B. xylophilus, (i) vacuoles na-adịghị ahụ anya na 0 h, ọgwụgwọ rụpụtara na (ii) vacuoles, (iii) nchịkọta nke vacuoles dị iche iche, (ivfusions), vacuoles (iii) vacuoles, vacuoles, vacuoles. (vi) nhazi nke nnukwu vacuoles. Àkụ́ na-acha uhie uhie na-egosi ọzịza nke vacuoles, akụ na-acha anụnụ anụnụ na-egosi njikọ nke vacuoles na akụ ojii na-egosi nnukwu vacuoles. Ogwe ọnụ ọgụgụ = 50 μm.
Tụkwasị na nke a, ọmụmụ ihe a kọwakwara usoro usoro nke ọnwụ nke methane kpatara na pine nematodes (Nyocha 4C). Methanogenic ọnwụ bụ ụdị ọnwụ cell na-abụghị apoptotic jikọtara ya na mkpokọta cytoplasmic vacuoles a ma ama27. Nrụrụ morphological a na-ahụ na nematodes pine yiri ka ọ nwere njikọ chiri anya na usoro nke methane kpatara ọnwụ. Nnyocha microscopic n'oge dị iche iche gosiri na e guzobere nnukwu vacuoles mgbe awa 20 nke ikpughe na 5-iodoindole (0.1 mM). A na-ahụ oghere microscopic mgbe awa 8 gasịrị, ọnụ ọgụgụ ha mụbara mgbe awa 12 gachara. Ahụrụ ọtụtụ nnukwu oghere mgbe awa 14 gachara. Ọtụtụ vacuoles fused pụtara nke ọma mgbe awa 12-16 gachara ọgwụgwọ, na-egosi na ngwakọta vacuole bụ ihe ndabere nke usoro ọnwụ methanogenic. Mgbe awa 20 gachara, a chọtara ọtụtụ nnukwu vacuoles n'ime irighiri ahụ. Ihe ngosi ndị a na-anọchite anya akụkọ mbụ nke metuosis na C. elegans.
N'ime ikpuru 5-iodoindole a na-agwọ, a na-ahụkwa nchịkọta vacuole na mgbawa (Fig 5), dị ka ọ na-egosi site na ikpuru ikpuru na ịhapụ oghere na gburugburu ebe obibi. A hụkwara nhụsianya Vacuole n'ime akpụkpọ akwa eggshell, bụ nke L2 na-echekwa na ọ naghị emebi emebi n'oge a na-amịpụta (Fig. S2). Nleba anya ndị a na-akwado ntinye aka nke nchịkọta mmiri na ọdịda osmoregulatory, yana mmerụ ahụ na-emegharị emegharị (RCI), na nhazi nke vacuole formation na suppuration (Fig 5).
N'ịkọwapụta ọrụ ayodiin na nhazi vacuole a hụrụ, anyị nyochara ọrụ nematidal nke sodium iodide (NaI) na potassium iodide (KI). Otú ọ dị, na mkpokọta (0.1, 0.5 ma ọ bụ 1 mM), ha emetụtaghị ma ọ bụ ndụ nematode ma ọ bụ nhazi vacuole (Mgbakwunye Fig. S5), ọ bụ ezie na 1 mM KI nwere ntakịrị mmetụta nematicidal. N'aka nke ọzọ, 7-iodoindole (1 ma ọ bụ 2 mM), dị ka 5-iodoindole, kpalitere ọtụtụ vacuoles na nrụrụ nhazi (Mgbakwunye Fig. S6). Iodoindoles abụọ ahụ gosipụtara njirimara phenotypic yiri ya na pine nematodes, ebe NaI na KI emeghị. N'ụzọ na-akpali mmasị, indole emeghị ka e guzobe vacuole na B. xylophilus na ntinye uche a nwalere (data egosighi). Ya mere, nsonaazụ ahụ gosipụtara na mgbagwoju anya indole-iodine bụ maka ikpochapụ na metabolism nke B. xylophilus.
N'ime indoles nke a nwalere maka ọrụ nematidal, 5-iodoindole nwere nchịkọta kachasị elu nke -5.89 kcal / mol, na-esote 7-iodoindole (-4.48 kcal / mol), 4-fluoroindole (-4.33), na indole (-4.03) (Nyocha 6). Njikọ dị n'azụ hydrogen dị ike nke 5-iodoindole na leucine 218 na-eme ka njide ya guzosie ike, ebe ihe mmepụta indole ndị ọzọ na-ejikọta na serine 260 site na agbụ hydrogen n'akụkụ. N'etiti iodoindoles ndị ọzọ a na-emepụta, 2-iodoindole nwere uru na-ejikọta -5.248 kcal / mol, nke bụ n'ihi njikọ hydrogen isi ya na leucine 218. Ihe ndị ọzọ a ma ama gụnyere 3-iodoindole (-4.3 kcal / mol), 4-iodoindole (-4.0 kcal/mol.6 kcal/mol). (Ihe atụ mgbakwunye S8). Ọtụtụ halogenated indoles na indole n'onwe ya, ma e wezụga 5-iodoindole na 2-iodoindole, na-emekọ njikọ na serine 260. Eziokwu ahụ bụ na hydrogen bonding na leucine 218 bụ ihe na-egosi nke ọma receptor-ligand bonding, dị ka a na-ahụ maka ivermectin (Mgbakwunye Fig. S5-iodoindoin) na-egosi na ọ dị mma. ivermectin, kechie nke ọma na saịtị GluCL na-arụ ọrụ site na leucine 218 (Fig. 6 na Mgbakwunye Fig. S8). Anyị na-atụ aro na a chọrọ njikọ a iji nọgide na-enwe oghere oghere nke mgbagwoju GluCL na site na ijikọ na saịtị na-arụ ọrụ nke GluCL receptor, 5-iodoindole, 2-iodoindole, avermectin na ivermectin si otú a na-eme ka ọwa ion meghee ma kwe ka mmiri nweta mmiri.
Docking molecular nke indole na halogenated indole na GluCL. Usoro njikọ nke (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, na (D) 5-iodoindole ligands na saịtị GluCL na-arụ ọrụ. A na-anọchi anya protein ahụ site na rịbọn, a na-egosikwa njikọ nke hydrogen azụ dị ka ahịrị nwere ntụpọ edo edo. (A′), (B′), (C′), na (D′) na-egosi mmekọrịta nke njikọ ndị kwekọrọ na ihe fọdụrụ na amino acid gbara ya gburugburu, na n'akụkụ-agbụ hydrogen ka ejiri akụ nwere ntụpọ pink egosi.
Emere nnwale iji nyochaa mmetụta nsi nke 5-iodoindole na mpụta nke kabeeji na mkpụrụ radish. 5-iodoindole (0.05 ma ọ bụ 0.1 mM) ma ọ bụ avermectin (10 μg/mL) nwere ntakịrị ma ọ bụ enweghị mmetụta na mbido mbụ na mpụta osisi (Nyocha 7). Na mgbakwunye, ọnweghị nnukwu ọdịiche dị n'etiti ọnụ ọgụgụ germination nke njikwa a na-agwọghị na mkpụrụ osisi 5-iodoindole ma ọ bụ avermectin na-agwọ. Mmetụta na taproot elongation na ọnụ ọgụgụ nke mpụta mgbọrọgwụ kpụrụ bụ ihe na-abaghị uru, ọ bụ ezie na 1 mM (ugboro 10 ya na-arụ ọrụ itinye uche) nke 5-iodoindole obere egbu oge mmepe nke mpụta mgbọrọgwụ. Nsonaazụ ndị a na-egosi na 5-iodoindole abụghị ihe na-egbu egbu na mkpụrụ ndụ ihe ọkụkụ na ọ dịghị egbochi usoro mmepe osisi na mkpokọta a mụrụ.
Mmetụta 5-iodoindole na mpụta mkpụrụ. Germination, na-epulite na mpụta mgbọrọgwụ nke B. oleracea na R. raphanistrum osisi na Murashige na Skoog agar ọkara nwere ma ọ bụ na-enweghị avermectin ma ọ bụ 5-iodoindole. A na-edekọ germination mgbe ụbọchị 3 nke incubation gasịrị na 22 Celsius.
Ọmụmụ ihe a na-akọ ọtụtụ ikpe nke ndị indoles gburu nematode. N'ụzọ dị mkpa, nke a bụ akụkọ mbụ nke iodoindole na-ebute methylation (usoro nke nchịkọta nke obere vacuoles na-eji nwayọọ nwayọọ na-abanye n'ime nnukwu vacuoles, na-eduga n'ịgbawa akpụkpọ ahụ na ọnwụ) na agịga pine, na iodoindole na-egosipụta njirimara nematicidal dị ịrịba ama yiri nke azụmahịa nematicide avermectin.
A kọwo na mbụ Indoles na-arụ ọtụtụ ọrụ mgbaàmà na prokaryotes na eukaryotes, gụnyere mgbochi biofilm / nhazi, nlanarị nje, na ọrịa nje19,32,33,34. N'oge na-adịbeghị anya, mmetụta ọgwụgwọ nwere ike ịnweta indoles halogenated, indole alkaloids, na semisynthetic indole derivatives adọtala mmasị nyocha dị ukwuu35,36,37. Dịka ọmụmaatụ, egosiri indoles halogenated igbu Escherichia coli na Staphylococcus aureus cell37. Na mgbakwunye, ọ bụ mmasị sayensị iji mụọ irè nke halogenated indoles megide ụdị ndị ọzọ, genera, na alaeze dị iche iche, na ọmụmụ ihe a bụ nzọụkwụ iji nweta ihe mgbaru ọsọ a.
N'ebe a, anyị na-atụ aro usoro maka 5-iodoindole na-akpata egbu egbu na C. elegans dabere na mmerụ ahụ na-emegharị ahụ (RCI) na methylation (Figures 4C na 5). Mgbanwe edematous dị ka nfụkasị ahụ na mmebi nke vacuolar bụ ihe na-egosi RCI na methylation, gosipụtara dị ka nnukwu vacuoles na cytoplasm48,49. RCI na-egbochi mmepụta ike site n'ibelata mmepụta ATP, na-akpata ọdịda nke mgbapụta ATPase, ma ọ bụ na-akpaghasị membranes cell na ime ka ngwa ngwa nke Na +, Ca2+, na mmiri50,51,52. Intracytoplasmic vacuoles na-ebilite na mkpụrụ ndụ anụmanụ n'ihi nchịkọta mmiri na cytoplasm n'ihi ntinye nke Ca2+ na mmiri53. N'ụzọ na-akpali mmasị, usoro a nke mmebi sel na-agbanwe ma ọ bụrụ na mmebi ahụ na-adịru nwa oge na mkpụrụ ndụ na-amalite ịmepụta ATP ruo oge ụfọdụ, ma ọ bụrụ na mmebi ahụ na-aga n'ihu ma ọ bụ na-akawanye njọ, mkpụrụ ndụ ahụ na-anwụ.54 Nleba anya anyị na-egosi na nematodes na-emeso ya na 5-iodoindole enweghị ike iweghachi biosynthesis nkịtị mgbe ekpughere ọnọdụ nrụgide.
Ụdị phenotype methylation nke 5-iodoindole na-eme na B. xylophilus nwere ike ịbụ n'ihi ọnụnọ nke ayodiin na nkesa molekụla ya, ebe ọ bụ na 7-iodoindole nwere mmetụta na-adịghị mma na B. xylophilus karịa 5-iodoindole (Table 1 na S6 na-agbakwụnye). Nsonaazụ ndị a dabara n'otu akụkụ na ọmụmụ nke Maltese et al. (2014), onye kọrọ na ntụgharị nke pyridyl nitrogen moiety indole site na para- na meta-ọnọdụ kagburu vacuolization, uto mgbochi, na cytotoxicity na U251 cell, na-atụ aro na mmekọrịta nke molekul na kpọmkwem na-arụ ọrụ saịtị na protein dị oké mkpa27,44,45. Mmekọrịta dị n'etiti indole ma ọ bụ halogenated indoles na ndị na-anabata GluCL hụrụ na ọmụmụ a na-akwadokwa echiche a, dịka 5- na 2-iodoindole na-ejikọta ya na ndị na-anabata GluCL karịa ndị ọzọ indoles nyochara (Nyocha 6 na S8 na-agbakwụnye). Achọpụtara iodine na ọnọdụ nke abụọ ma ọ bụ nke ise nke indole ka ọ na-ejikọta leucine 218 nke GluCL onye na-anabata ya site na agbụ hydrogen azụ, ebe indoles ndị ọzọ halogenated na indole n'onwe ya na-eme ka njikọ hydrogen na-adịghị ike na serine 260 (Nyocha 6). Ya mere, anyị na-eche na nhazi nke halogen na-arụ ọrụ dị mkpa na ntinye nke mmebi nke vacuolar degeneration, ebe njide siri ike nke 5-iodoindole na-eme ka ọwa ion ghe oghe, si otú ahụ na-ekwe ka mmiri na-asọba ngwa ngwa na nkwụsị nke vacuole. Otú ọ dị, a ka ga-ekpebi usoro ihe omume zuru ezu nke 5-iodoindole.
Tupu itinye 5-iodoindole n'ọrụ, a ga-enyocha mmetụta ya na-egbu egbu na osisi. Nnwale mịpụta mkpụrụ anyị gosiri na 5-iodoindole enweghị mmetụta na-adịghị mma na mpụta mkpụrụ ma ọ bụ usoro mmepe na-esote na mkpokọta a mụrụ (Nyocha 7). Ya mere, ọmụmụ ihe a na-enye ihe ndabere maka iji 5-iodoindole na gburugburu ebe obibi iji chịkwaa mmebi nke pine nematodes na osisi pine.
Akụkọ ndị gara aga egosila na ọgwụgwọ dabere na indole na-anọchite anya ụzọ nwere ike isi dozie nsogbu nke mgbochi ọgwụ nje na ọganihu ọrịa cancer55. Na mgbakwunye, indoles nwere antibacterial, anticancer, antioxidant, mgbochi mkpali, antidiabetic, antiviral, antiproliferative na antituberculosis omume na nwere ike bụrụ ihe ndabere na-ekwe nkwa mmepe ọgwụ56,57. Ọmụmụ ihe a na-atụ aro na nke mbụ enwere ike iji ayodiin mee ihe dị ka ihe mgbochi parasitic na anthelmintic.
Achọpụtara Avermectin afọ iri atọ gara aga wee nweta Nrite Nobel na 2015, iji ya dị ka anthelmintic ka na-aga n'ihu. Otú ọ dị, n'ihi ọganihu ngwa ngwa nke iguzogide avermectins na nematodes na ụmụ ahụhụ, a chọrọ ụzọ ọzọ, dị ọnụ ala, na atụmatụ enyi na gburugburu ebe obibi iji chịkwaa ọrịa PWN na osisi pine. Ọmụmụ ihe a na-akọkwa usoro nke 5-iodoindole na-egbu pine nematodes na 5-iodoindole nwere obere nsị na mkpụrụ ndụ ihe ọkụkụ, nke na-emepe atụmanya dị mma maka ngwa azụmahịa ya n'ọdịnihu.
Kọmitii ụkpụrụ omume nke Mahadum Yeungnam, Gyeongsan, Korea kwadoro nnwale niile, na usoro ndị a mere dịka ntuziaka nke Kọmitii Ethics nke Mahadum Yeungnam siri dị.
Emere nnwale nnabata akwa site na iji usoro guzosiri ike43. Iji chọpụta ọnụego hatching (HR), nematodes okenye dị otu ụbọchị (ihe dị ka ụmụ nwanyị 100 na ụmụ nwoke 100) ka ebufere n'ime efere Petri nwere ero ahụ ma hapụ ya ka ọ tolite maka awa 24. A na-ewepụ akwa dịpụrụ adịpụ ma jiri 5-iodoindole (0.05 mM na 0.1 mM) ma ọ bụ avermectin (10 μg/ml) dị ka nkwusioru n'ime mmiri na-adịghị ọcha. A na-ebufe nkwusioru ndị a (500 μl; ihe dị ka nsen 100) na olulu mmiri nke efere anụ ahụ nke ọma 24 ma tinye ya na 22 Celsius. A na-eme ọnụọgụ L2 ka awa 24 gachara mana a na-ewere ya dị ka onye nwụrụ anwụ ma ọ bụrụ na mkpụrụ ndụ ahụ emegharịghị mgbe ejiri ezigbo waya platinum kpalitere. Emere nnwale a na nkebi abụọ, nke ọ bụla nwere ugboro isii. Ejikọtara data sitere na nnwale abụọ ahụ wee gosipụta ya. A na-agbakọ pasentị HR dịka ndị a:
A na-enyocha onwu anwụ site na iji usoro emepụtara na mbụ. A na-achịkọta àkwá Nematode na embrayo na-ejikọta ya site n'ịkụnye n'ime mmiri a na-esighị ike iji mepụta L2 larvae. A na-emeso larvae mekọrịtara ọnụ (ihe dị ka nematodes 500) na 5-iodoindole (0.05 mM na 0.1 mM) ma ọ bụ avermectin (10 μg / ml) wee zụlite ya na efere B. cinerea Petri. Mgbe awa 48 gachara na 22 Celsius C, a na-achịkọta nematodes n'ime mmiri a na-esighị ike ma nyochaa ọnụnọ nke L2, L3, na L4. Ọnụnọ nke ọkwa L3 na L4 gosipụtara mgbanwe larval, ebe ọnụnọ nke L2 egosighi mgbanwe ọ bụla. E nwetara onyonyo site n'iji iRiS™ Digital Cell Imaging System. Emere nnwale a na nkebi abụọ, nke ọ bụla nwere ugboro isii. Ejikọtara data sitere na nnwale abụọ ahụ wee gosipụta ya.
A na-enyocha nsi nke 5-iodoindole na avermectin na mkpụrụ osisi site na iji ule germination na Murashage na Skoog agar plates.62 B. Oleracea na R. raphanistrum osisi a na-ebu ụzọ tinye ya na mmiri mmiri na-adịghị mma maka otu ụbọchị, jiri 1 ml 100% ethanol, nke 1 ml nke sodium chlorite, gbazere na 3% nke ọgwụ hypochloride. 15 min, wee saa ya ugboro ise na 1 ml mmiri na-adịghị mma. A na-apịnye mkpụrụ osisi a gbajiri agbaji na efere agar germination nwere 0.86 g/l (0.2X) Murasheji na Skoog media na 0.7% agar bacteriological nwere ma ọ bụ enweghị 5-iodoindole ma ọ bụ avermectin. A na-etinye efere ndị ahụ na 22 Celsius C, a na-esekwa foto mgbe ụbọchị 3 gachara. Emere nnwale a na nkebi abụọ, nke ọ bụla n'ime ha nwere ugboro isii.
Oge nzipu: Feb-26-2025