Muti wepine nematode imhuka inotama-tama inozivikanwa nekukonzera kurasikirwa kukuru kwehupfumi mumasango epine. Chidzidzo chiripo chinoongorora mashandiro ekuuraya mahalogenated indoles kurwisa mapine nematodes uye mashandiro azvo. Mabasa ekuuraya mahalogenated indoles kurwisa mapine nematodes nemaitiro azvo. Mabasa ekuuraya mahalogenated e5-iodoindole neavermectin (kudzora kwakanaka) kurwisa mapine nematodes aive akafanana uye akakwira pahuwandu hwakaderera (10 μg/mL). 5-iodoindole yakaderedza kubereka, kubereka, kufa kwemwana ari mudumbu uye mabukira, uye maitiro ekufamba. Kudyidzana kwemamolecular kwema ligands ane invertebrate-specific glutamate-gated chloride channel receptors kunotsigira pfungwa yekuti 5-iodoindole, seavermectin, inosunga zvakasimba panzvimbo inoshanda yereceptor. 5-Iodoindole yakakonzerawo akasiyana-siyana ephenotypic deformations mumanematodes, kusanganisira kudonha/kuderera kwenhengo uye kuwedzera kwevacuolization. Izvi zvinoratidza kuti mavacuole anogona kuita basa mukufa kwenematode methylation-inokonzerwa nemetode. Zvakakosha, 5-iodoindole yakanga isina chepfu kumarudzi ese emiti (kabichi neradish). Saka, chidzidzo ichi chinoratidza kuti kushandiswa kweiodoindole mumamiriro ekunze kunogona kudzora kukuvara kwepine wilt.
Matanda emiti yepine wood (Bursaphelenchus xylophilus) ndeemiti yepine wood nematodes (PWN), inotama-tama endoparasitic nematodes inozivikanwa nekukonzera kukuvara kwakanyanya kwezvakatipoteredza kumasango epine wood ecosystems1. Chirwere chekuoma kwemiti yepine (PWD) chinokonzerwa nemiti yepine wood nematode chiri kuva dambudziko guru mumakondinendi akati wandei, kusanganisira Asia neEurope, uye muNorth America, matanda aya anoparadza marudzi emiti yepine akaunzwa1,2. Kuderera kwemiti yepine idambudziko guru rehupfumi, uye tarisiro yekupararira kwayo pasi rose iri kunetseka3. Mhando dzemiti yepine dzinotevera dzinowanzo rwiswa nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, uye Pinus radiata4. Matanda epine chirwere chakakomba chinogona kuuraya miti yepine mukati memavhiki kana mwedzi yehutachiona. Pamusoro pezvo, kubuda kwemiti yepine nematode kwakajairika munzvimbo dzakasiyana-siyana dzezvakatipoteredza, saka hutachiona hunoramba huripo hwavepo1.
Bursaphelenchus xylophilus imhuka inodya fungus uye inoberekana mumiti yemupaini, ichikura kuita zvikamu zvina zvakasiyana zvemakonye: L1, L2, L3, L4 uye munhu mukuru1,6. Mumamiriro ekushomeka kwechikafu, pine nematode inopinda muchikamu chemakonye - dauer, iyo inoparadzira hutachiona hwayo - pine bark beetle (Monochamus alternatus) uye inotamisirwa kumiti yemupaini ine hutano. Munzvimbo dzine hutano, makonye anokurumidza kufamba nepakati pemiti uye anodya masero ane parenchymatous, izvo zvinotungamira kune akawanda maitiro ehypersensitivity, pine wilting uye kufa mukati megore mushure mehutachiona1,7,8.
Kudzora zvipembenene zvepine nematodes kwagara kuri dambudziko, matanho ekuparadzanisa vanhu kubva muzana remakore rechi20. Nzira dziripo dzekudzivirira zvipembenene zvepine nematodes dzinosanganisira kurapwa nemakemikari, kusanganisira kufuridzira matanda uye kuisa mishonga inouraya zvipembenene mumatanda emiti. Mishonga inouraya zvipembenene inonyanya kushandiswa ndeye avermectin ne avermectin benzoate, iyo iri yemhuri ye avermectin. Mishonga iyi inodhura inoshanda zvikuru kurwisa mhando dzakawanda dzezvipembenene uye inoonekwa seyakachengeteka nharaunda9. Zvisinei, kushandiswa kwemishonga iyi kakawanda kunotarisirwa kugadzira kumanikidzwa kwekusarudza izvo zvichatungamira mukubuda kwezvipembenene zvinouraya zvipembenene zvepine, sezvakaratidzwa kune zvipembenene zvakawanda, zvakaita seLeptinotarsa decemlineata, Plutella xylostella uye mishonga inouraya zvipembenene Trichostrongylus colubriformis neOstertagia circumcincta, izvo zvishoma nezvishoma zvakatanga kuramba kurwisa avermectins10,11,12. Nokudaro, maitiro ekudzivirira anofanira kudzidzwa nguva dzose uye mishonga inouraya zvipembenene inoongororwa nguva dzose kuti iwane dzimwe nzira, dzinoshanda uye dzisingakuvadzi nharaunda dzekudzora PVD. Mumakumi emakore achangopfuura, vanyori vakati wandei vakakurudzira kushandiswa kwezvinomera zvakatorwa, mafuta anokosha uye zvinopisa semishonga yekudzivirira nematode13,14,15,16.
Munguva pfupi yapfuura takaratidza mashandiro ekuuraya zvipembenene zve indole, iyo molecule inopindirana nemasero uye inopindirana neumambo, muCaenorhabditis elegans 17. Indole chiratidzo chakapararira mukati memasero muzvinhu zvipenyu, ichidzora mabasa akawanda anokanganisa physiology yezvipembenene, kuumbwa kwespore, kugadzikana kweplasmid, kuramba mishonga, kuumbwa kwebiofilm, uye hutachiona 18, 19. Mabasa e indole nezvimwe zvinomera zvinorwisa zvipembenene haana kudzidzwa. Muchidzidzo ichi, takaongorora mashandiro ekuuraya zvipembenene e34 indole dzinorwisa pine nematodes uye takatsanangura mashandiro anoita 5-iodoindole ine simba zvikuru tichishandisa microscopy, time-lapse photography, uye molecular docking experiments, uye takaongorora migumisiro yayo ine chepfu pamiti tichishandisa nzira yekumera kwembeu.
Kuwanda kwakanyanya kweindole (>1.0 mM) kwakambotaurwa kuti kune simba rekuuraya nematodes17. Mushure mekurapwa kweB. xylophilus (zvikamu zvakasiyana zvehupenyu) ne indole kana 33 zvakasiyana zve indole derivatives pa 1 mM, kufa kweB. xylophilus kwakayerwa nekuverenga nematodes mhenyu neakafa mumapoka ekutonga nekurapwa. Indoles shanu dzakaratidza basa guru rekuuraya nematodes; kupona kweboka rekutonga risina kurapwa kwaive 95 ± 7% mushure memaawa makumi maviri nemana. Pama indoles 34 akaedzwa, 5-iodoindole ne 4-fluoroindole pa 1 mM zvakakonzera kufa kwe100%, nepo 5,6-difluoroindigo, methylindole-7-carboxylate, uye 7-iodoindole zvakakonzera kufa kweinenge 50% (Tafura 1).
Mhedzisiro ye5-iodoindole pakuumbwa kwevacuole uye kushandiswa kwemetabolism yepine wood nematode. (A) Mhedzisiro yeavermectin ne5-iodoindole pamhuka dzechirume dzekare, (B) mazai eL1 stage nematode uye (C) kushandiswa kwemetabolism yeB. xylophilus, (i) vacuoles hazvina kuonekwa pa0 awa, kurapwa kwakaguma ne (ii) vacuoles, (iii) kuunganidzwa kwevacuoles dzakawanda, (iv) kuzvimba kwevacuoles, (v) kusangana kwevacuoles uye (vi) kugadzirwa kwevacuoles hombe. Miseve mitsvuku inoratidza kuzvimba kwevacuoles, miseve yebhuruu inoratidza kusangana kwevacuoles uye miseve mitema inoratidza vacuoles hombe. Scale bar = 50 μm.
Pamusoro pezvo, chidzidzo ichi chakatsanangurawo maitiro ekufa kunokonzerwa nemethane mumapine nematodes (Mufananidzo 4C). Kufa kwemethanogenic rudzi rwekufa kwemasero kusingabvisi apoptotic kunobatanidzwa nekuunganidzwa kwema cytoplasmic vacuoles akakurumbira27. Kukanganisika kwechimiro kwakaonekwa mumapine nematodes kunoita sekunge kwakabatana zvakanyanya nemaitiro ekufa kunokonzerwa nemethane. Kuongorora kwe microscopic panguva dzakasiyana kwakaratidza kuti ma giant vacuoles akaumbwa mushure memaawa makumi maviri ekusangana ne 5-iodoindole (0.1 mM). Ma microscopic vacuoles akaonekwa mushure memaawa masere ekurapwa, uye huwandu hwawo hwakawedzera mushure memaawa gumi nemaviri. Mazhinji mahombe ma vacuoles akaonekwa mushure memaawa gumi nemana. Mazhinji ma fused vacuoles akaonekwa zvakajeka mushure memaawa gumi nemaviri kusvika gumi nematanhatu ekurapwa, zvichiratidza kuti vacuole fusion ndiyo hwaro hwe methanogenic death mechanism. Mushure memaawa makumi maviri, ma giant vacuoles akawanikwa muhonye yese. Izvi zvinomiririra mushumo wekutanga we metuosis muC. elegans.
Muhonye dzakarapwa ne5-iodoindole, kuungana kwevacuole nekuputika kwakaonekwawo (Mufananidzo 5), sezvinoratidzwa nekukombama kwehonye uye kuburitswa kwevacuole munzvimbo. Kukanganiswa kwevacuole kwakaonekwawo muganda remazai, iro rinowanzo chengetedzwa neL2 panguva yekuzvara (Mufananidzo S2). Izvi zvinotsigira kubatanidzwa kwehuwandu hwemvura uye kutadza kwe osmoregulatory, pamwe nekukuvara kwemasero anodzoreka (RCI), mukuumbwa kwevacuole uye suppuration (Mufananidzo 5).
Tichifunga nezvebasa reiodine mukuumbwa kwevacuole kwakaonekwa, takaongorora mashandiro e sodium iodide (NaI) uye potassium iodide (KI) ekuuraya utachiona. Zvisinei, pahuwandu (0.1, 0.5 kana 1 mM), hazvina kukanganisa kupona kwenematode kana kugadzirwa kwevacuole (Supplementary Fig. S5), kunyangwe 1 mM KI yaive nemhedzisiro yekuuraya utachiona zvishoma. Kune rumwe rutivi, 7-iodoindole (1 kana 2 mM), senge 5-iodoindole, yakakonzera vacuoles dzakawanda uye deformations yemaumbirwo (Supplementary Fig. S6). Ma iodoindole maviri aya akaratidza hunhu hwakafanana hwe phenotypic mumapine nematodes, nepo NaI neKI dzisina kudaro. Zvinofadza kuti, indole haina kukonzera kugadzirwa kwevacuole muB. xylophilus pahuwandu hwakaedzwa (data harina kuratidzwa). Saka, mhedzisiro yakasimbisa kuti indole-iodine complex ndiyo inokonzeresa vacuolization uye metabolism yeB. xylophilus.
Pakati pemaindole akaedzwa kuti aone kana paine basa rekuuraya zvipembenene, 5-iodoindole yaive ne-slip index yepamusoro ye -5.89 kcal/mol, ichiteverwa ne 7-iodoindole (-4.48 kcal/mol), 4-fluoroindole (-4.33), uye indole (-4.03) (Mufananidzo 6). Kubatana kwakasimba kwehydrogen yemusana kwe5-iodoindole ku leucine 218 kunoita kuti igare yakasimba, nepo zvimwe zvese zvinogadzirwa muindole zvinosunga ku serine 260 kuburikidza ne side chain hydrogen bonds. Pakati pemamwe ma iodoindoles akaumbwa, 2-iodoindole ine kukosha kwekusunga kwe -5.248 kcal/mol, izvo zvinokonzerwa nekusunga kwayo kukuru kwehydrogen ne leucine 218. Mamwe ma binding anozivikanwa anosanganisira 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), uye 6-fluoroindole (-2.6 kcal/mol) (Mufananidzo Wekuwedzera S8). Mazhinji ma indole ane halogenated ne indole pachayo, kunze kwe 5-iodoindole ne 2-iodoindole, anoumba chisungo ne serine 260. Chokwadi chekuti hydrogen bonding ne leucine 218 chinoratidza kubatana kwe receptor-ligand, sezvakaonekwa ne ivermectin (Supplementary Fig. S7), chinosimbisa kuti 5-iodoindole ne 2-iodoindole, se ivermectin, zvinosunga zvakasimba panzvimbo inoshanda ye GluCL receptor kuburikidza ne leucine 218 (Fig. 6 ne Supplementary Fig. S8). Tinokurudzira kuti kubatana uku kunodiwa kuchengetedza chimiro che open pore che GluCL complex uye kuti nekusunga zvakasimba panzvimbo inoshanda ye GluCL receptor, 5-iodoindole, 2-iodoindole, avermectin ne ivermectin nokudaro zvinochengetedza ion channel yakavhurika uye zvichibvumira kutorwa kwemvura.
Kuiswa kwemasero eindole ne halogenated indole kuGluCL. Kubatanidza marongero e (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, uye (D) 5-iodoindole ligands panzvimbo inoshanda yeGluCL. Puroteni iyi inomiririrwa neribhoni, uye backbone hydrogen bonds inoratidzwa semitsara yeyero ine madotsi. (A′), (B′), (C′), uye (D′) zvinoratidza kudyidzana kwema ligands anoenderana nema amino acid residues akapoteredza, uye side-chain hydrogen bonds inoratidzwa nemiseve ine madotsi epingi.
Kuedzwa kwakaitwa kuti kuongororwe chepfu ye5-iodoindole pakumera kwembeu dzekabheji neradish. 5-iodoindole (0.05 kana 0.1 mM) kana avermectin (10 μg/mL) haina kana mhedzisiro yakakura pakumera kwekutanga uye kubuda kwembeu dzezvirimwa (Mufananidzo 7). Pamusoro pezvo, hapana musiyano wakakosha wakawanikwa pakati pehuwandu hwekumera kwembeu dzisina kurapwa uye mbeu dzakarapwa ne5-iodoindole kana avermectin. Mhedzisiro pakureba kwemidzi yetaproot nehuwandu hwemidzi yemativi yakaumbwa yakanga isina kukosha, kunyangwe 1 mM (kagumi kakawanda huwandu hwayo hunoshanda) ye5-iodoindole yakanonotsa kukura kwemidzi yemativi. Mhedzisiro iyi inoratidza kuti 5-iodoindole haina chepfu kumasero emiti uye haikanganisi maitiro ekukura kwemiti pahuwandu hwakadzidzwa.
Mhedzisiro ye5-iodoindole pakumera kwembeu. Kumera, kumera uye midzi yembeu dzeB. oleracea neR. raphanistrum paMurashige neSkoog agar medium ine kana isina avermectin kana 5-iodoindole. Kumera kwakanyorwa mushure memazuva matatu ekuchengetwa kwembeu pa22°C.
Chidzidzo ichi chinoshuma nyaya dzinoverengeka dzekuurayiwa kwenematode ne indoles. Chinokosha ndechekuti, uyu ndiwo mushumo wekutanga we iodoindole inokonzera methylation (maitiro anokonzerwa nekuunganidzwa kwe vacuoles diki dzinobatana zvishoma nezvishoma kuita ma vacuoles makuru, zvichizopedzisira zvaita kuti membrane iparare uye ife) mupine tsono, iodoindole ichiratidza hunhu hwakakosha hwe nematicide hwakafanana nehwe commercial nematicide avermectin.
MaIndoles akambotaurwa kuti ane mabasa akawanda ekuratidza zviratidzo muprokaryotes ne eukaryotes, kusanganisira kudzivirira/kuumbwa kwebiofilm, kupona kwebhakitiriya, uye pathogenicity19,32,33,34. Munguva pfupi yapfuura, mhedzisiro inogona kuitika yekurapa kwe halogenated indoles, indole alkaloids, uye semisynthetic indole derivatives yakakwezva kufarira kukuru kwekutsvaga35,36,37. Semuenzaniso, halogenated indoles dzakaratidzwa kuti dzinouraya masero eEscherichia coli anoramba aripo uye Staphylococcus aureus37. Pamusoro pezvo, zvinonakidza zvesainzi kudzidza kushanda kwe halogenated indoles kurwisa mamwe marudzi, marudzi, uye humambo, uye chidzidzo ichi idanho rekuzadzisa chinangwa ichi.
Pano, tinokurudzira nzira yekuuraya masero ane iodoindole-induced lethality muC. elegans zvichibva pakukuvara kwemasero kunodzoserwa (RCI) uye methylation (Mifananidzo 4C ne5). Kuchinja kwemasero akadai sekuzvimba uye kuora kwemasero eropa zviratidzo zveRCI uye methylation, zvinoratidzwa sema vacuoles makuru mu cytoplasm48,49. RCI inokanganisa kugadzirwa kwesimba nekuderedza kugadzirwa kweATP, zvichikonzera kutadza kwepombi yeATPase, kana kukanganisa membrane yemasero uye zvichikonzera kupinda nekukurumidza kweNa+, Ca2+, uye water50,51,52. Ma vacuoles emukati meropa anomuka mumasero emhuka nekuda kwekuunganidzwa kwemvura mu cytoplasm nekuda kwekupinda kweCa2+ nemvura53. Zvinofadza kuti, nzira iyi yekukuvara kwemasero inogona kudzokororwa kana kukuvara kwacho kuri kwenguva pfupi uye masero akatanga kugadzira ATP kwenguva yakati, asi kana kukuvara kwacho kukaramba kuripo kana kuipa, masero acho anofa.54 Zvatakaona zvinoratidza kuti nematodes dzakarapwa ne5-iodoindole hadzikwanise kudzoreredza biosynthesis yakajairika mushure mekusangana nemamiriro ezvinhu ekushushikana.
Maitiro emethylation anokonzerwa ne5-iodoindole muB. xylophilus anogona kunge ari nekuda kwekuvapo kweiodine uye kupararira kwayo kwemamorekuru, sezvo 7-iodoindole isina simba rekudzivirira paB. xylophilus kupfuura 5-iodoindole (Tafura 1 neSupplementary Figure S6). Mhedzisiro iyi inoenderana zvishoma nezvidzidzo zveMaltese et al. (2014), avo vakataura kuti kutamiswa kwe pyridyl nitrogen moiety mu indole kubva papara- kuenda ku meta-position kwakabvisa vacuolization, kudziviswa kwekukura, uye cytotoxicity mumasero eU251, zvichiratidza kuti kudyidzana kwemorekuru nenzvimbo chaiyo inoshanda muprotein kwakakosha27,44,45. Kudyidzana pakati peindole kana halogenated indoles neGluCL receptors zvakaonekwa muchidzidzo ichi zvinotsigirawo pfungwa iyi, sezvo 5- ne2-iodoindole zvakawanikwa zvichibatana neGluCL receptors zvakasimba kupfuura mamwe ma indoles akaongororwa (Mufananidzo 6 neSupplementary Figure S8). Iodine iri panzvimbo yechipiri kana yechishanu yeindole yakawanikwa ichibatana neleucine 218 yeGluCL receptor kuburikidza nespine hydrogen bonds, nepo mamwe ma halogenated indoles ne indole pachayo zvichigadzira weak side-chain hydrogen bonds ne serine 260 (Mufananidzo 6). Saka tinofungidzira kuti nzvimbo ye halogen ine basa rakakosha mukutanga kwe vacuolar degeneration, nepo kubatana kwakasimba kwe 5-iodoindole kunochengetedza ion channel yakavhurika, nokudaro zvichibvumira kubuda kwemvura nekukurumidza uye vacuole kuputika. Zvisinei, mashandiro akadzama ekuita kwe 5-iodoindole achiri kuda kuzivikanwa.
Tisati tashandisa 5-iodoindole, mhedzisiro yayo ine chepfu pamiti inofanira kuongororwa. Kuedza kwedu kumera kwembeu kwakaratidza kuti 5-iodoindole haina mhedzisiro yakaipa pakumera kwembeu kana maitiro ekukura anotevera pahuwandu hwakadzidzwa (Mufananidzo 7). Saka, chidzidzo ichi chinopa hwaro hwekushandiswa kwe5-iodoindole munzvimbo yezvakatipoteredza kudzora kukuvadza kwepine nematodes kumiti yepine.
Mishumo yapfuura yakaratidza kuti kurapwa kwakavakirwa pa indole kunomiririra nzira inogona kugadzirisa dambudziko rekusagona kurwisa mabhakitiriya uye kufambira mberi kwekenza55. Pamusoro pezvo, indoles ine mabasa ekurwisa mabhakitiriya, ekurwisa kenza, antioxidant, anti-inflammatory, antidiabetic, antiviral, antiproliferative uye ekurwisa TB uye inogona kushanda sehwaro hunovimbisa hwekugadzirwa kwemishonga56,57. Chidzidzo ichi chinoratidza kekutanga kushandiswa kwe iodine semushonga wekurwisa tupukanana uye mushonga wekurwisa kenza.
Avermectin yakawanikwa makore makumi matatu apfuura uye yakahwina mubairo weNobel Prize muna 2015, uye kushandiswa kwayo semushonga wekurapa kuchiri kuenderera mberi. Zvisinei, nekuda kwekukura nekukurumidza kwekudzivirira maavermectin muzvipembenene nezvipembenene, imwe nzira, inodhura zvishoma, uye isingakuvadzi nharaunda inodiwa kudzora hutachiona hwePWN mumiti yemupaini. Chidzidzo ichi chinoshumawo nzira iyo 5-iodoindole inouraya zvipembenene zvemupaini uye kuti 5-iodoindole ine chepfu shoma kumasero emiti, izvo zvinovhura mikana yakanaka yekushandiswa kwayo mune ramangwana mukutengeserana.
Kuedza kwese kwakatenderwa neKomiti yeMaitiro eYeungnam University, Gyeongsan, Korea, uye nzira dzacho dzakaitwa zvichienderana nemirairo yeKomiti yeMaitiro eYeungnam University.
Kuyedza kwekuisa mazai mumazai kwakaitwa uchishandisa nzira dzakatarwa43. Kuti vaongorore huwandu hwekuchekwa kwemazai (HR), nematodes evakuru vane zuva rimwe chete (vakadzi vanenge zana nevarume zana) vakaendeswa kumadishi ePetri ane fungus ndokubvumirwa kukura kwemaawa makumi maviri nemana. Mazai akazobviswa ndokurapwa ne5-iodoindole (0.05 mM uye 0.1 mM) kana avermectin (10 μg/ml) semushonga mumvura yakacheneswa isina tsvina. Kuisa mazai aya (500 μl; mazai angangoita zana) akaendeswa kumatsime eplate ye24-well tissue culture ndokuiswa pa22 °C. Kuverengwa kweL2 kwakaitwa mushure memaawa makumi maviri nemana ekuisirwa mazai asi kwakanzi kwakafa kana masero asina kufamba paakakurudzirwa newaya yeplatinum yakanaka. Kuedza uku kwakaitwa muzvikamu zviviri, chimwe nechimwe nekudzokorora katanhatu. Data kubva mukuyedza kwese kwakasanganiswa uye kwakaratidzwa. Chikamu cheHR chinoverengerwa seinotevera:
Kufa kwembeva kwakaongororwa uchishandisa nzira dzakagadzirwa kare. Mazai eNematode akaunganidzwa uye maembryo akasanganiswa nekukandwa mumvura yakacheneswa isina tsvina kuti pave nembeva dzeL2 stage. Maembryo akasanganiswa (anenge 500 nematodes) akarapwa ne5-iodoindole (0.05 mM uye 0.1 mM) kana avermectin (10 μg/ml) ndokurerwa paB. cinerea Petri plates. Mushure memaawa makumi mana nemasere ekuiswa mudumbu pa22 °C, nematodes akaunganidzwa mumvura yakacheneswa isina tsvina uye akaongororwa kuti aone kana paine matanho eL2, L3, uye L4. Kuvapo kwematanho eL3 neL4 kwakaratidza kushanduka kwembeva, nepo kuvapo kwedanho reL2 kwakaratidza kuti hapana kushanduka. Mifananidzo yakawanikwa uchishandisa iRiS™ Digital Cell Imaging System. Kuedza uku kwakaitwa mumatanho maviri, imwe neimwe ichidzokororwa katanhatu. Data kubva mukuyedza kwese kwakasanganiswa uye kwakaratidzwa.
Chepfu ye5-iodoindole neavermectin pambeu yakaongororwa uchishandisa bvunzo dzekumera pamaplate eMurashige neSkoog agar.62 Mbeu dzeB. oleracea neR. raphanistrum dzakatanga kunyikwa mumvura yakacheneswa isina tsvina kwezuva rimwe chete, dzakagezwa ne1 ml ye100% ethanol, dzakacheneswa ne1 ml ye50% commercial bleach (3% sodium hypochlorite) kwemaminitsi gumi nemashanu, uye dzakagezwa kashanu nemvura yakacheneswa ne1 ml. Mbeu dzakacheneswa dzakabva dzadzvanywa pamaplate e germination agar ane 0.86 g/l (0.2X) Murashige neSkoog medium uye 0.7% bacteriological agar ine kana isina 5-iodoindole kana avermectin. Maplate acho akazoiswa muincubation pa22 °C, uye mifananidzo yakatorwa mushure memazuva matatu ekuicubation. Kuedza uku kwakaitwa muzvikamu zviviri, chimwe nechimwe chazvo chakaita katanhatu.
Nguva yekutumira: Kukadzi-26-2025