Nematode pine bụ endoparasite na-akwaga ebe a na-adọpụ iche nke a maara na ọ na-akpata nnukwu mfu akụ na ụba na gburugburu ebe obibi osisi pine. Ọmụmụ ihe a na-enyocha ọrụ nematicidal nke indoles halogenated megide pine nematodes na usoro ha si arụ ọrụ. Ọrụ nematicidal nke 5-iodoindole na avermectin (njikwa dị mma) megide pine nematodes yiri nke ahụ ma dị elu na obere njupụta (10 μg/mL). 5-iodoindole belatara ịmụ nwa, ọrụ ọmụmụ, ọnwụ nwa ebu n'afọ na larval, na omume locomotor. Mmekọrịta molekul nke ligands na ndị na-anabata ọwa glutamate-gated chloride nke invertebrate-specific na-akwado echiche ahụ na 5-iodoindole, dị ka avermectin, na-ejikọ nke ọma na ebe nnabata ahụ na-arụ ọrụ. 5-Iodoindole kpaliterekwa mgbanwe phenotypic dị iche iche na nematodes, gụnyere ọdịda akụkụ ahụ na-adịghị mma/mbelata na mmụba vacuolization. Nsonaazụ ndị a na-egosi na vacuoles nwere ike ịrụ ọrụ na ọnwụ nke nematode methylation. Ihe dị mkpa bụ na 5-iodoindole anaghị egbu egbu nye ụdị osisi abụọ ahụ (kabeeji na radish). Ya mere, ọmụmụ ihe a na-egosi na itinye iodoindole n'ọnọdụ gburugburu ebe obibi nwere ike ijikwa mmerụ ahụ pine wilt.
Nematode osisi pine (Bursaphelenchus xylophilus) bụ nke nematode osisi pine (PWN), nke na-agagharị agagharị nke endoparasitic nematodes a maara na ọ na-emebi gburugburu ebe obibi nke ọma na gburugburu ebe obibi nke osisi pine1. Ọrịa wilt nke osisi pine kpatara na-aghọ nnukwu nsogbu na kọntinent dị iche iche, gụnyere Eshia na Europe, na na North America, nematode na-ebibi ụdị pine ndị a webatara1,2. Ọdịda osisi pine bụ nnukwu nsogbu akụ na ụba, atụmanya nke mgbasa ya zuru ụwa ọnụ na-enye nsogbu3. Nematode na-awakpokarị ụdị pine ndị a: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, na Pinus radiata4. Nematode osisi pine bụ ọrịa dị oke njọ nke nwere ike igbu osisi pine n'ime izu ma ọ bụ ọnwa nke ọrịa. Na mgbakwunye, ọrịa pine nematode na-adịkarị na ọtụtụ gburugburu ebe obibi, yabụ e guzobela usoro ọrịa na-adịgide adịgide1.
Bursaphelenchus xylophilus bụ nematode osisi na-akpata ọrịa nke ezinụlọ Aphelenchoidea na clade 102.5. Nematode ahụ na-eri ero ma na-amụba n'ime anụ ahụ osisi pine, na-etolite n'ime ọkwa anọ dị iche iche nke larval: L1, L2, L3, L4 na onye toro eto1,6. N'ọnọdụ ụkọ nri, pine nematode na-agafe n'ọkwa larval pụrụ iche - dauer, nke na-emebi ihe na-akpata ya - pine bark beetle (Monochamus alternatus) ma na-ebufe ya na osisi pine dị mma. N'ime ndị ọbịa ahụike, nematodes na-agafe ngwa ngwa site na anụ ahụ osisi ma na-eri mkpụrụ ndụ parenchymatous, nke na-eduga na ọtụtụ mmeghachi omume hypersensitivity, pine wilt na ọnwụ n'ime otu afọ mgbe ọrịa gasịrị1,7,8.
Njikwa ihe ndị dị ndụ nke pine nematodes abụrụla ihe ịma aka ogologo oge, yana usoro mgbochi iche site na narị afọ nke 20. Usoro ugbu a maka ijikwa pine nematodes gụnyere ọgwụgwọ kemịkalụ, gụnyere ịfụ osisi na itinye nematicides n'ime ogwe osisi. Nematicides 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 ọma megide ọtụtụ ụdị nematodes ma a na-ewere ha dị ka ihe nchekwa gburugburu ebe obibi9. Agbanyeghị, a na-atụ anya na iji nematicides ndị a ugboro ugboro ga-emepụta nrụgide nhọrọ nke ga-eme ka ọ bụrụ na ha ga-ebute mpụta nke pine nematodes ndị na-eguzogide ọgwụ, dịka egosiri maka ọtụtụ nje ụmụ ahụhụ, dị ka Leptinotarsa decemlineata, Plutella xylostella na nematodes Trichostrongylus colubriformis na Ostertagia circumcincta, bụ ndị ejirila nwayọọ nwayọọ mepụta iguzogide avermectins10,11,12. Ya mere, ọ dị mkpa ka a na-amụ ụkpụrụ iguzogide mgbe niile ma na-enyocha nematicides mgbe niile iji chọta usoro ndị ọzọ, ndị dị ọnụ ala na ndị na-adịghị ize ndụ gburugburu ebe obibi iji chịkwaa PVD. N'ime iri afọ ndị na-adịbeghị anya, ọtụtụ ndị edemede atụọla aro ka e jiri ihe ndị sitere n'osisi, mmanụ dị mkpa na ihe ndị na-agbanwe agbanwe mee ihe dị ka ihe na-achịkwa nematode13,14,15,16.
N'oge na-adịbeghị anya, anyị gosipụtara ọrụ nematicidal nke indole, molekul signaling intercellular na interkingdom, na Caenorhabditis elegans 17. Indole bụ ihe mgbaàmà intracellular zuru oke na gburugburu ebe obibi microbial, na-achịkwa ọtụtụ ọrụ ndị na-emetụta physiology microbial, nhazi spore, nkwụsi ike plasmid, iguzogide ọgwụ, nhazi biofilm, na virulence 18, 19. Emebeghị ọmụmụ ihe gbasara ọrụ indole na ihe ndị sitere na ya megide nematodes ndị ọzọ na-akpata ọrịa. N'ọmụmụ ihe a, anyị nyochachara ọrụ nematicidal nke indoles 34 megide pine nematodes ma kọwaa usoro nke ọrụ nke 5-iodoindole kachasị ike site na iji microscopy, foto oge-lapse, na nnwale docking molecular, ma nyochaa mmetụta ya na osisi site na iji nnwale mkpụrụ.
Akọpụtala na oke indole (>1.0 mM) nwere mmetụta nematicidal na nematodes17. Mgbe ejiri indole ma ọ bụ 33 dị iche iche gwọọ B. xylophilus (oge ndụ agwakọtara) na 1 mM, a tụrụ ọnwụ nke B. xylophilus site na ịgụta nematodes dị ndụ na ndị nwụrụ anwụ n'ime otu njikwa na ndị a gwọrọ. Indoles ise gosipụtara ọrụ nematicidal dị ukwuu; ndụ nke otu njikwa a na-agwọghị bụ 95 ± 7% mgbe awa 24 gachara. N'ime indoles 34 a nwalere, 5-iodoindole na 4-fluoroindole na 1 mM kpatara ọnwụ 100%, ebe 5,6-difluoroindigo, methylindole-7-carboxylate, na 7-iodoindole kpatara ọnwụ ihe dị ka 50% (Tebụl 1).
Mmetụta nke 5-iodoindole na mmepụta vacuole na metabolism nke pine wood nematode. (A) Mmetụta nke avermectin na 5-iodoindole na nematodes ndị toro eto nke nwoke, (B) àkwá nematode nke ọkwa L1 na (C) metabolism nke B. xylophilus, (i) ahụghị vacuoles na awa 0, ọgwụgwọ butere (ii) vacuoles, (iii) mkpokọta nke ọtụtụ vacuoles, (iv) ọzịza nke vacuoles, (v) njikọta nke vacuoles na (vi) ịmepụta nnukwu vacuoles. Akụ uhie na-egosi ọzịza nke vacuoles, akụ anụnụ anụnụ na-egosi njikọta nke vacuoles na akụ ojii na-egosi nnukwu vacuoles. Ogwe nha = 50 μm.
Tinyere nke ahụ, ọmụmụ ihe a kọwakwara usoro nke ọnwụ methane kpatara n'ime pine nematodes (Foto 4C). Ọnwụ methanogenic bụ ụdị ọnwụ sel na-abụghị apoptotic nke jikọtara ya na mkpokọta nke cytoplasmic vacuoles a ma ama27. Nrụpụta ọdịdị nke a hụrụ na pine nematodes yiri ka ọ nwere njikọ chiri anya na usoro nke ọnwụ methane kpatara. Nnyocha microscopic n'oge dị iche iche gosiri na e mepụtara nnukwu vacuoles mgbe awa 20 nke ikpughe na 5-iodoindole (0.1 mM). A hụrụ nnukwu vacuoles mgbe awa 8 nke ọgwụgwọ gasịrị, ọnụọgụ ha mụbara mgbe awa 12 gasịrị. A hụrụ ọtụtụ nnukwu vacuoles mgbe awa 14 gachara. Ọtụtụ vacuoles a gwakọtara agwakọta pụtara ìhè nke ọma mgbe awa 12-16 nke ọgwụgwọ gasịrị, na-egosi na njikọ vacuole bụ ntọala nke usoro ọnwụ methanogenic. Mgbe awa 20 gachara, a hụrụ ọtụtụ nnukwu vacuoles n'ime ikpuru. Ihe ndị a na-egosi akụkọ mbụ nke metuosis na C. elegans.
N'ime ikpuru ndị e ji iodoindole gwọọ 5, a hụkwara nchịkọta vacuole na mgbawa (Foto 5), dịka ihe gosiri site na mgbagọ ikpuru na ntọhapụ vacuole n'ime gburugburu ebe obibi. A hụkwara mmebi vacuole n'ime akpụkpọ ahụ àkwá, nke L2 na-echekwa mgbe niile n'oge ọ na-amụ nwa (Foto mgbakwunye S2). Ihe ndị a na-egosi na-akwado itinye aka na mkpokọta mmiri na ọdịda osmoregulatory, yana mmerụ ahụ mkpụrụ ndụ a na-agbanwe agbanwe (RCI), n'usoro nhazi vacuole na suppuration (Foto 5).
N'ịkọwapụta ọrụ nke ayodiin na nhazi vacuole a hụrụ, anyị nyochara ọrụ nematicidal nke sodium iodide (NaI) na potassium iodide (KI). Agbanyeghị, na njupụta (0.1, 0.5 ma ọ bụ 1 mM), ha emetụtaghị ndụ nematode ma ọ bụ nhazi vacuole (Foto mgbakwunye S5), ọ bụ ezie na 1 mM KI nwere obere mmetụta nematicidal. N'aka nke ọzọ, 7-iodoindole (1 ma ọ bụ 2 mM), dị ka 5-iodoindole, butere ọtụtụ vacuoles na mgbanwe nhazi (Foto mgbakwunye S6). Iodoindoles abụọ ahụ gosipụtara njirimara phenotypic yiri nke ahụ na pine nematodes, ebe NaI na KI emeghị. N'ụzọ na-adọrọ mmasị, indole emeghị ka vacuole etolite na B. xylophilus na njupụta a nwalere (egosighi data). Ya mere, nsonaazụ ahụ kwadoro na mgbagwoju anya indole-iodine bụ maka vacuolization na metabolism nke B. xylophilus.
N'ime indoles a nwalere maka ọrụ nematicidal, 5-iodoindole nwere ndepụta slip kachasị elu nke -5.89 kcal/mol, nke sochiri ya bụ 7-iodoindole (-4.48 kcal/mol), 4-fluoroindole (-4.33), na indole (-4.03) (Foto nke 6). Njikọ hydrogen azụ siri ike nke 5-iodoindole na leucine 218 na-eme ka njikọ ya sie ike, ebe ihe ndị ọzọ niile indole na-ejikọ na serine 260 site na njikọ hydrogen dị n'akụkụ. N'etiti iodoindoles ndị ọzọ a na-eme ihe nlereanya, 2-iodoindole nwere uru njikọ nke -5.248 kcal/mol, nke bụ n'ihi njikọ hydrogen ya bụ isi na leucine 218. Njikọ ndị ọzọ a maara gụnyere 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), na 6-fluoroindole (-2.6 kcal/mol) (Ihe mgbakwunye S8). Ọtụtụ indoles na indole nke a na-akpọ halogenated n'onwe ya, ewezuga 5-iodoindole na 2-iodoindole, na-etolite njikọ na serine 260. Eziokwu ahụ bụ na njikọ hydrogen na leucine 218 na-egosi njikọ dị irè nke receptor-ligand, dịka a hụrụ maka ivermectin (Foto mgbakwunye S7), na-akwado na 5-iodoindole na 2-iodoindole, dịka ivermectin, na-ejikọ nke ọma na ebe na-arụ ọrụ nke receptor GluCL site na leucine 218 (Foto 6 na Foto mgbakwunye S8). Anyị na-atụ aro na njikọ a dị mkpa iji nọgide na-enwe nhazi oghere mepere emepe nke mgbagwoju anya GluCL na site na ijikọ nke ọma na ebe na-arụ ọrụ nke receptor GluCL, 5-iodoindole, 2-iodoindole, avermectin na ivermectin si otú a na-eme ka ọwa ion ahụ mepee ma kwe ka mmiri na-abata.
Njikọta nke indole na halogenated indole na GluCL. Nhazi njikọ nke (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, na (D) 5-iodoindole ligands na ebe GluCL na-arụ ọrụ. A na-egosi protein ahụ site na rịbọn, a na-egosikwa njikọ hydrogen azụ dị ka ahịrị ntụpọ odo odo. (A′), (B′), (C′), na (D′) na-egosi mmekọrịta nke ligands kwekọrọ na ihe fọdụrụ na amino acid gbara ya gburugburu, a na-egosikwa njikọ hydrogen n'akụkụ site na akụ pink ntụpọ.
E mere nnwale iji chọpụta mmetụta ọjọọ nke 5-iodoindole nwere n'ịmịpụta mkpụrụ kabeeji na radish. 5-iodoindole (0.05 ma ọ bụ 0.1 mM) ma ọ bụ avermectin (10 μg/mL) enweghị obere ma ọ bụ enweghị mmetụta ọ bụla na mmalite ịmịpụta na mpụta osisi (Foto 7). Na mgbakwunye, achọpụtaghị nnukwu ọdịiche dị n'etiti ọnụego ịmịpụta nke njikwa a na-agwọghị na mkpụrụ ejiri 5-iodoindole ma ọ bụ avermectin gwọọ. Mmetụta dị na ogologo taproot na ọnụọgụ mgbọrọgwụ dị n'akụkụ etolite adịchaghị mkpa, ọ bụ ezie na 1 mM (ugboro 10 nke mgbasawanye ya) nke 5-iodoindole belatara mmepe nke mgbọrọgwụ dị n'akụkụ. Nsonaazụ ndị a na-egosi na 5-iodoindole anaghị egbu mkpụrụ osisi ma ọ naghị egbochi usoro mmepe osisi na mgbasawanye a mụrụ.
Mmetụta nke 5-iodoindole na mkpụrụ osisi. Mmalite, ntolite na mgbọrọgwụ n'akụkụ nke mkpụrụ B. oleracea na R. raphanistrum na Murashige na Skoog agar medium nwere ma ọ bụ na-enweghị avermectin ma ọ bụ 5-iodoindole. E dekọrọ mmịpụta ahụ mgbe ụbọchị atọ nke ịmịnye ya na okpomọkụ 22°C gasịrị.
Nnyocha a na-akọ akụkọ gbasara ọtụtụ ikpe nke ogbugbu nematode site na indoles. Ihe dị mkpa bụ na nke a bụ akụkọ mbụ nke iodoindole na-akpata methylation (usoro sitere na mkpokọta obere vacuoles nke na-ejikọta nwayọ nwayọ gaa na nnukwu vacuoles, nke na-emesịa mee ka akpụkpọ ahụ gbawaa ma nwụọ) na agịga pain, yana iodoindole na-egosipụta nnukwu ihe ndị na-egbu egbu yiri nke nematicide avermectin azụmaahịa.
Akọpụtala na Indoles na-arụ ọrụ dị iche iche na prokaryotes na eukaryotes, gụnyere mgbochi/mmepụta biofilm, ndụ nje bacteria, na pathogenicity19,32,33,34. N'oge na-adịbeghị anya, mmetụta ọgwụgwọ nke indoles halogenated, indole alkaloids, na semisynthetic indole derivatives adọtala mmasị nyocha sara mbara35,36,37. Dịka ọmụmaatụ, egosila na indoles halogenated na-egbu mkpụrụ ndụ Escherichia coli na Staphylococcus aureus na-adịgide adịgide37. Na mgbakwunye, ọ dị mkpa ka sayensị mụọ irè nke indoles halogenated megide ụdị, genera, na alaeze ndị ọzọ, ọmụmụ ihe a bụkwa nzọụkwụ iji mezuo ebumnuche a.
N'ebe a, anyị na-atụ aro usoro maka ọnwụ 5-iodoindole kpatara na C. elegans dabere na mmerụ ahụ mkpụrụ ndụ a na-agbanwe agbanwe (RCI) na methylation (Foto 4C na 5). Mgbanwe edema dịka puffiness na vacuolar degeneration bụ ihe na-egosi RCI na methylation, nke a na-egosipụta dị ka nnukwu vacuoles na cytoplasm48,49. RCI na-egbochi mmepụta ike site na ibelata mmepụta ATP, na-akpata ọdịda nke ọkpọkọ ATPase, ma ọ bụ na-akpaghasị akpụkpọ ahụ mkpụrụ ndụ ma na-akpata mmụba ngwa ngwa nke Na+, Ca2+, na mmiri50,51,52. Vacuoles intracytoplasmic na-ebilite na mkpụrụ ndụ anụmanụ n'ihi mkpokọta mmiri na cytoplasm n'ihi mmụba nke Ca2+ na mmiri53. Ihe na-adọrọ mmasị bụ na usoro a nke mmebi mkpụrụ ndụ nwere ike ịgbanwe ma ọ bụrụ na mmebi ahụ adịru nwa oge ma mkpụrụ ndụ amalite imepụta ATP ruo oge ụfọdụ, mana ọ bụrụ na mmebi ahụ anọgide ma ọ bụ ka njọ, mkpụrụ ndụ ahụ na-anwụ.54 Ihe anyị hụrụ na-egosi na nematodes ejiri 5-iodoindole gwọọ enweghị ike iweghachi biosynthesis nkịtị mgbe ha kpughesịrị ọnọdụ nrụgide.
Ụdị methylation nke 5-iodoindole kpalitere na B. xylophilus nwere ike ịbụ n'ihi ọnụnọ nke iodine na nkesa molekul ya, ebe ọ bụ na 7-iodoindole nwere obere mmetụta mgbochi na B. xylophilus karịa 5-iodoindole (Tebụl 1 na Foto Mgbakwunye S6). Nsonaazụ ndị a kwekọrọ n'ọmụmụ ihe nke Maltese et al. (2014), bụ ndị kọrọ na ntụgharị nke pyridyl nitrogen moiety na indole site na para-to the meta-position kwụsịrị vacuolization, mgbochi uto, na cytotoxicity na mkpụrụ ndụ U251, na-atụ aro na mmekọrịta nke molekul na ebe a kapịrị ọnụ na protein dị oke mkpa27,44,45. Mmekọrịta dị n'etiti indole indole ma ọ bụ halogenated na ndị na-anabata GluCL hụrụ na ọmụmụ ihe a na-akwadokwa echiche a, ebe achọpụtara na 5- na 2-iodoindole na-ejikọ na ndị na-anabata GluCL karịa indoles ndị ọzọ enyochara (Foto 6 na Foto Mgbakwunye S8). A chọpụtara na ayodiin dị n'ọnọdụ nke abụọ ma ọ bụ nke ise nke indole na-ejikọ na leucine 218 nke onye na-anabata GluCL site na njikọ hydrogen azụ, ebe indoles halogenated ndị ọzọ na indole n'onwe ha na-etolite njikọ hydrogen n'akụkụ dị ala na serine 260 (Foto nke 6). Ya mere, anyị na-eche na ebe halogen dị na-arụ ọrụ dị mkpa na mkpali nke mmebi vacuolar, ebe njikọ siri ike nke 5-iodoindole na-eme ka ọwa ion mepee, si otú a na-ekwe ka mmiri mmiri ngwa ngwa na mgbawa vacuole. Agbanyeghị, a ka ga-ekpebi usoro zuru ezu nke 5-iodoindole na-arụ ọrụ.
Tupu e jiri 5-iodoindole mee ihe n'ụzọ bara uru, a ga-enyocha mmetụta ọjọọ ya n'ahụ osisi. Nnwale anyị mere iji 5-iodoindole amịpụta mkpụrụ gosiri na 5-iodoindole enweghị mmetụta ọjọọ ọ bụla n'ipụta mkpụrụ ma ọ bụ usoro mmepe na-esote na mkpokọta a mụrụ (Foto 7). Ya mere, ọmụmụ ihe a na-enye ntọala maka iji 5-iodoindole mịpụta gburugburu ebe obibi iji chịkwaa mmerụ ahụ 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 iguzogide ọgwụ nje na ọganihu kansa55. Na mgbakwunye, indoles nwere ọrụ nje bacteria, anticancer, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative na antituberculosis ma nwee ike ịbụ ihe ndabere maka mmepe ọgwụ56,57. Ọmụmụ ihe a na-atụ aro maka oge mbụ ojiji nke iodine dị ka ihe na-egbochi nje na ọgwụ mgbochi.
A chọpụtara Avermectin afọ iri atọ gara aga ma merie ihe nrite Nobel na 2015, ojiji ya dị ka ọgwụ mgbochi ọrịa ka na-aga n'ihu. Agbanyeghị, n'ihi mmepe ngwa ngwa nke iguzogide avermectins na nematodes na ụmụ ahụhụ, ọ dị mkpa atụmatụ ọzọ, dị ọnụ ala, na nke na-adịghị ize ndụ gburugburu ebe obibi iji chịkwaa ọrịa PWN na osisi pine. Ọmụmụ ihe a na-akọkwa usoro nke 5-iodoindole si egbu pine nematodes na 5-iodoindole nwere obere nsí na mkpụrụ ndụ osisi, nke na-emepe ohere dị mma maka ojiji azụmaahịa ya n'ọdịnihu.
Kọmitii Omume nke Mahadum Yeungnam, Gyeongsan, Korea kwadoro nnwale niile, e mekwara usoro ndị ahụ dịka ntuziaka nke Kọmitii Omume nke Mahadum Yeungnam si dị.
E jiri usoro ndị e guzobere mee nnwale mmalite àkwá43. Iji chọpụta ọnụego mmụba àkwá (HR), e zigara nematodes ndị toro eto dị otu ụbọchị (ihe dị ka ụmụ nwanyị 100 na ụmụ nwoke 100) na efere Petri nwere ero ahụ ma hapụ ha ka ha too ruo awa 24. E wee kewapụ àkwá ndị ahụ ma jiri 5-iodoindole (0.05 mM na 0.1 mM) ma ọ bụ avermectin (10 μg/ml) gwọọ ha dị ka ihe nkwusioru n'ime mmiri a gwakọtara agwakọta. E bufere ihe nkwusioru ndị a (500 μl; ihe dị ka àkwá 100) na olulu mmiri nke efere anụ ahụ nwere olulu 24 ma tinye ha na okpomọkụ 22 °C. E mere ọnụọgụ L2 mgbe awa 24 nke ịmịnye àkwá gasịrị mana a na-ewere ha dị ka ndị nwụrụ anwụ ma ọ bụrụ na mkpụrụ ndụ ahụ agagharịghị mgbe ejiri waya platinum dị mma kpalie ha. Emere nnwale a n'ọkwa abụọ, nke ọ bụla nwere ugboro isii. E jikọtara ma gosipụta data sitere na nnwale abụọ ahụ. A na-agbakọ pasentị nke HR dị ka ndị a:
A na-enyocha ọnwụ nke ụmụ ahụhụ site na iji usoro e mepụtara na mbụ. A na-anakọta àkwá Nematode ma jikọta embrayo site na ịgbapụta n'ime mmiri a gwakọtara agwakọta iji mepụta ụmụ ahụhụ nke ọkwa L2. A na-agwọ ụmụ ahụhụ ndị a gwakọtara agwakọta (ihe dị ka nematodes 500) na 5-iodoindole (0.05 mM na 0.1 mM) ma ọ bụ avermectin (10 μg/ml) ma zụlite ha na efere B. cinerea Petri. Mgbe awa 48 nke ịmịnye ha na okpomọkụ na 22 °C gasịrị, a na-anakọta ụmụ ahụhụ na mmiri a gwakọtara agwakọta ma lelee ma ha nwere ọkwa L2, L3, na L4. Ọnụnọ nke ọkwa L3 na L4 gosiri mgbanwe ụmụ ahụhụ, ebe ọnụnọ nke ọkwa L2 egosighi mgbanwe ọ bụla. E nwetara onyonyo site na iji Sistemụ Foto Cell Dijitalụ iRiS™. Emere nnwale a n'ọkwa abụọ, nke ọ bụla nwere ugboro isii. A jikọtara ma gosipụta data sitere na nnwale abụọ ahụ.
E jiri ule mkpụrụ osisi Murashige na efere agar nke Murashige na Skoog nyochaa nsí nke mkpụrụ osisi 5-iodoindole na avermectin na-akpata.62 B. oleracea na mkpụrụ R. raphanistrum buru ụzọ mikpuo na mmiri a gwakọtara agwakọta ruo otu ụbọchị, saa ya na 1 ml 100% ethanol, jiri 1 ml 50% commercial bleach (3% sodium hypochlorite) sachaa ya ruo nkeji iri na ise, ma jiri 1 ml mmiri a gwakọtara agwakọta saa ya ugboro ise. E mesịa pịa mkpụrụ osisi a gwakọtara agwakọta na efere agar germination nke nwere 0.86 g/l (0.2X) Murashige na Skoog medium na 0.7% bacteriological agar nwere ma ọ bụ na-enweghị 5-iodoindole ma ọ bụ avermectin. E mesịa tinye efere ndị ahụ na okpomọkụ 22 °C, e serekwa foto mgbe ụbọchị atọ nke mkpụrụ osisi ahụ gasịrị. E mere nnwale a n'ọkwa abụọ, nke ọ bụla nwere ugboro isii.
Oge ozi: Febụwarị-26-2025