Nematode-ka geedku waa endoparasite socdaalka karantiil ee loo yaqaan inuu ku keeno khasaare dhaqaale oo ba'an nidaamyada deegaanka kaynta geedaha. Daraasadda hadda jirta waxay dib u eegis ku eegaysaa waxqabadka nematidal ee indoles halogenated ee lidka ku ah nematodes geed iyo habka ay wax u qabtaan. Dhaqdhaqaaqyada nematidal ee 5-iodoindole iyo avermectin (xakamaynta togan) ee ka dhanka ah nematodes-ka geedku waxay la mid ahaayeen oo aad u sarreeyay heerar hoose (10 μg/mL). 5-iodoindole waxay hoos u dhigtay uurka, dhaqdhaqaaqa taranka, dhimashada uurjiifka iyo dirxiga, iyo habdhaqanka lugaha. Isdhexgalka molecular ee ligands-ka-gaar ah ee glutamate-gated chloride channel reseptors ayaa taageeraya fikradda ah in 5-iodoindole, sida avermectin, ay si adag ugu xirto goobta firfircoon ee soo dhaweynta. 5-Iodoindole waxa kale oo ay keentay qallafsanaanta phenotypic ee kala duwan ee nematodes, oo ay ku jiraan xubno aan caadi ahayn oo burburay / yarayn iyo faaruqin kordhay. Natiijooyinkani waxay soo jeedinayaan in vacuoles laga yaabo inay door ka ciyaaraan nematode methylation dhimashada dhexdhexaadinta. Muhiimad ahaan, 5-iodoindole ma ahayn mid sun ah labada nooc ee dhirta (kaabajka iyo dabocase). Sidaa darteed, daraasaddan ayaa muujinaysa in codsiga iodoindole ee xaaladaha deegaanka ay xakameyn karto dhaawaca geedka.
nematode alwaax geed (Bursaphelenchus xylophilus) waxaa iska leh nematodes alwaax geed (PWN), nematodes endoparasitic guurguura oo loo yaqaan inay waxyeelo ba'an u geystaan deegaanka kaynta geedaha. Cudurka Pine wilt (PWD) ee uu keeno nematode alwaax geed ayaa noqonaya dhibaato halis ah oo ka dhacda qaaradaha dhowr ah, oo ay ku jiraan Aasiya iyo Yurub, iyo Waqooyiga Ameerika, nematode waxay burburisaa noocyada geedaha la soo bandhigay1,2. Hoos u dhaca geedka geedku waa dhibaato dhaqaale oo weyn, rajada faafitaankeeda adduunkuna waa welwel3. Noocyada geedaha soo socda ayaa inta badan ay weeraraan nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, iyo Pinus radiata4. Pine nematode waa cudur halis ah oo dili kara geedaha geedka toddobaadyo ama bilo gudahooda ee caabuqa. Intaa waxaa dheer, dillaaca nematode geedku waxay ku badan yihiin nidaamyada deegaanka ee kala duwan, sidaas darteed silsilado caabuq ah oo joogto ah ayaa la sameeyay1.
Bursaphelenchus xylophilus waa nematode-karantiil-karantiil ah oo karantiil ah oo ka tirsan qoyska superfamily Aphelenchoidea iyo clade 102.5. nematode waxay quudisaa fungi waxayna ku tarmeysaa unugyada alwaaxyada geedaha geedaha, waxayna u koraan afar marxaladood oo dirxiga ah: L1, L2, L3, L4 iyo qof weyn1,6. Marka ay jiraan xaalado cunto yaraan, nematode geedku waxa ay u gudubtaa marxalad dirxiga khaaska ah - dauer, kaas oo dulin ka dhigaya vector-keeda - jilif geedeedka (Monochamus alternatus) waxaana loo wareejiyaa geedo geed caafimaad qaba. Dadka martida loo yahay ee caafimaadka qaba, nematodes waxay si dhakhso ah ugu guuraan unugyada dhirta waxayna ku quudiyaan unugyada parenchymatous, taas oo keenta tiro dareen-celin xasaasiyadeed, geedo yaryar iyo dhimasho hal sano gudaheed ka dib caabuqa1,7,8.
Xakamaynta bayoolojiga ee nematodes-ka geedku muddo dheer ayay ahayd caqabad, iyada oo tillaabooyinka karantiilku socday ilaa qarnigii 20-aad. Xeeladaha hadda jira ee lagu xakameynayo nematodes-ka geedaha ayaa ugu horrayn ku lug leh daawaynta kiimikaad, oo ay ku jiraan qiiqa alwaaxyada iyo in nematicides lagu geliyo jirridda geedaha. Nemacides-ka ugu badan ee la isticmaalo waa avermectin iyo avermectin benzoate, kuwaas oo ka tirsan qoyska avermectin. Kiimikooyinkan qaaliga ah ayaa aad waxtar ugu leh noocyo badan oo nematode ah waxaana loo arkaa inay yihiin kuwo badbaado leh. Si kastaba ha ahaatee, isticmaalka soo noqnoqda ee nematicides ayaa la filayaa in ay abuurto cadaadis xulashada taas oo hubaal ah in ay horseedi doonto soo bixitaanka nematodes geed u adkaysta, sida lagu muujiyay dhowr cayayaan ah, sida Leptinotarsa decemlineata, Plutella xylostella iyo nematodes Trichostrongylus colubriformis iyo Ostertagia, si tartiib tartiib ah u soo baxay iska caabin ah. Sidaa darteed, qaababka iska caabbinta waxay u baahan yihiin in si joogto ah loo darso oo nematicides si joogto ah loo baadho si loo helo beddel, kharash-ku-ool iyo tallaabooyin deegaan u-saaxiib ah si loo xakameeyo PVD. Tobannaankii sano ee la soo dhaafay, tiro ka mid ah qorayaasha ayaa soo jeediyay isticmaalka soosaarka dhirta, saliidaha lagama maarmaanka ah iyo kuwa aan isbeddelin sida wakiilada kontoroolka nematode13,14,15,16.
Waxaan dhawaan soo bandhignay dhaqdhaqaaqa nematicidal ee indole, unug-dhexdhexaadiye ah iyo unugyo calaamadeeynaya interkingdom, gudaha Caenorhabditis elegans 17. Indole waa calaamad ballaaran oo gudaha gudaha ah oo ku jirta ecology microbial, xakamaynta hawlo badan oo saameeya physiology microbial, formation spore, xasiloonida plasmid, caabbinta daroogada, biofilm formation, iyo virulence 18, 19. Dhaqdhaqaaqa indole iyo waxyaabaha ka soo jeeda ee ka dhanka ah nematodes-ka cudur-sidaha kale lama baran. Daraasaddan, waxaan ku baarnay dhaqdhaqaaqa nematicidal ee 34 indoles oo ka dhan ah nematodes geed waxaanan qeexnay habka ficilka ee 5-iodoindole ee ugu awoodda badan iyadoo la adeegsanayo microscopy, sawir-qaadista waqti-dhaafka ah, tijaabooyinka docking molecular, waxaanan qiimeynay saameynta sunta ah ee dhirta iyadoo la adeegsanayo baaritaanka biqilka abuurka.
Heerarka sare (> 1.0 mM) ee indole ayaa hore loo soo sheegay inay saameyn nematicidal ah ku leeyihiin nematodes17. Ka dib daawaynta B. xylophilus (marxalado nololeed oo isku dhafan) oo leh indole ama 33 nooc oo kala duwan oo indole ah oo ah 1 mM, dhimashada B. xylophilus waxaa lagu qiyaasaa tirinta nematodes nool iyo kuwa dhintay ee kantaroolka iyo kooxaha la daweeyay. Shan indoles ayaa soo bandhigay waxqabad nematicidal oo muhiim ah; Badbaadinta kooxda xakamaynta aan la daaweynin waxay ahayd 95 ± 7% ka dib 24 saacadood. 34 ka mid ah indole-ka la tijaabiyay, 5-iodoindole iyo 4-fluoroindole ee 1 mM ayaa sababay 100% dhimasho, halka 5,6-difluoroindigo, methylindole-7-carboxylate, iyo 7-iodoindole ay sababeen ku dhawaad 50% dhimasho (Shaxda 1).
Saamaynta 5-iodoindole ee samaynta vacuole iyo dheef-shiid kiimikaadka nematode alwaax geed (A) Saamaynta avermectin iyo 5-iodoindole ee nematodes ragga qaangaarka ah, (B) L1 marxaladda nematode ukunta iyo (C) dheef-shiid kiimikaadka B. xylophilus, (i) vacuoles lama arkin 0 h, daaweyntu waxay keentay (ii) vacuoles, (iii) ururinta vacuoles badan, (ivfusions) vacuoles (vacuoles) oo barar ah. (vi) samaynta vacuoles giant. Fallaadhaha guduudan waxay muujinayaan bararka vacuoles, fallaadhaha buluuga ah waxay muujinayaan isku dhafka vacuoles iyo fallaadhaha madow waxay muujinayaan vacuoles weyn. Cabirka miisaanka = 50 μm.
Intaa waxaa dheer, daraasaddan ayaa sidoo kale sharraxday habka isku xigxiga ee dhimashada methane-ku-dhimashada ee nematodes-ka geedka (Jaantuska 4C). Dhimashada Methanogenic waa nooc aan apoptotic ahayn oo ah dhimashada unugyada ee la xidhiidha ururinta cytoplasmic vacuoles27 oo caan ah. Cilladaha morphological ee lagu arkay nematodes geed ayaa u muuqda inay si dhow ula xiriiraan habka dhimashada methane-ku keeno. Baaritaanka microscopic ee waqtiyo kala duwan ayaa muujiyay in vacuoles waaweyn la sameeyay ka dib 20 saacadood ee soo-gaadhista 5-iodoindole (0.1 mM). Vacuoles microscopic ayaa la arkay ka dib 8 saacadood oo daaweyn ah, tiradooduna waxay korodhay 12 saacadood ka dib. Dhowr faaruq oo waaweyn ayaa la arkay 14 saac kadib. Dhowr vacuoles oo isku dhafan ayaa si cad u muuqday 12-16 saacadood ka dib daaweynta, taas oo muujinaysa in fiyuuska vacuole uu yahay saldhigga habka dhimashada methanogenic. 20 saacadood ka dib, waxa laga helay gooryaanka oo dhan dhawr vacuoles waaweyn. Indho-indheyntani waxay ka dhigan tahay warbixintii ugu horreysay ee metuosis ee C. elegans.
Gooryaanka 5-iodoindole-lagu daweeyay, isugaynta vacuole iyo dillaac ayaa sidoo kale lagu arkay (Jaantus. 5), sida caddaynta foorarsiga gooryaanka iyo fakirka sii daynta deegaanka. Burburinta Vacuole ayaa sidoo kale lagu arkay xuubka ukunta, kaas oo sida caadiga ah u ilaalinaya L2 inta lagu jiro dillaacinta (Jaantuska Dheeraadka ah S2). Aragtiyadani waxay taageerayaan ku lug lahaanshaha ururinta dareeraha iyo fashilka osmoregulatory, iyo sidoo kale dhaawaca unugyada dib loo rogi karo (RCI), habka sameynta vacuole iyo suppuration (Jaantus. 5).
Qiyaasta doorka iodine ee samaynta vacuole ee la arkay, waxaanu baarnay dhaqdhaqaaqa nematidal ee sodium iodide (NaI) iyo potassium iodide (KI). Si kastaba ha ahaatee, marka la eego qiyaasta (0.1, 0.5 ama 1 mM), ma aysan saameynin badbaadada nematode ama samaynta vacuole (Jaantuska Dheeraadka ah. S5), inkastoo 1 mM KI uu lahaa saameyn yar oo nematicidal ah. Dhanka kale, 7-iodoindole (1 ama 2 mM), sida 5-iodoindole, waxay keentay vacuoles badan iyo qallafsanaan qaabdhismeed (Jaantuska Dheeraadka ah. S6). Labada iodoindoles waxay muujiyeen astaamo phenotypic la mid ah nematodes geed, halka NaI iyo KI aysan sameynin. Waxa xiisaha lihi leh, indole ma dhalin samaynta vacuole ee B. xylophilus marka la eego xadiga la tijaabiyey (xogta aan la muujin). Sidaa darteed, natiijadu waxay xaqiijisay in isku-dhafka indole-iodine uu mas'uul ka yahay faaruqinta iyo dheef-shiid kiimikaadka B. xylophilus.
Indoles-ka lagu tijaabiyay dhaqdhaqaaqa nematidal, 5-iodoindole wuxuu lahaa tusaha ugu sarreeya ee -5.89 kcal / mol, oo ay ku xigto 7-iodoindole (-4.48 kcal / mol), 4-fluoroindole (-4.33), iyo indole (-4.03) (Jaantus 6). Isku-xidhka hydrogen-ka ee adag ee 5-iodoindole ilaa leucine 218 waxay dejisaa xidhitaankeeda, halka dhammaan noocyada kale ee indole ay ku xidhan yihiin serine 260 iyada oo loo marayo silsiladaha hydrogen ee dhinaca. Iodoindoles kale oo la qaabeeyey, 2-iodoindole waxay leedahay qiime isku mid ah -5.248 kcal/mol, taas oo ay ugu wacan tahay dammaanadkeeda hydrogen ee ugu weyn ee leucine 218. Xirmooyinka kale ee la yaqaan waxaa ka mid ah 3-iodoindole (-4.3 kcal / mol), 4-iodoindole (-4.0 kcal/mol.6 kcal/mol). (Jaantuska Dheeraadka ah S8). Inta badan halogenated indoles iyo indole laftiisa, marka laga reebo 5-iodoindole iyo 2-iodoindole, waxay sameeyaan bond leh serine 260. Xaqiiqda ah in hydrogen bonding leucine 218 waa tilmaanta wax ku ool ah receptor-ligand xidhitaan, sida lagu arkay ivermectin (Supplementary Fig. 5-iodoindole. ivermectin, si adag ugu xidh goobta firfircoon ee GluCL receptor iyada oo loo sii marayo leucine 218 (Jaantus. 6 iyo Sawirka Dheeraadka ah S8). Waxaan soo jeedineynaa in xiritaankan loo baahan yahay si loo ilaaliyo qaab dhismeedka daloolka furan ee isku-dhafka GluCL iyo in si adag loogu xiro goobta firfircoon ee GluCL reseptor, 5-iodoindole, 2-iodoindole, avermectin iyo ivermectin si ay u ilaaliyaan kanaalka ion furan oo u oggolow qaadashada dareeraha.
Xiritaanka molecular ee indole iyo indole halogenated ilaa GluCL. Jihaynta isku xidhka ee (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, iyo (D) 5-iodoindole ligands ee goobta firfircoon ee GluCL. Borootiinku waxa uu u taagan yahay ribbon, iyo curaarta hydrogen ee laf dhabarta waxaa lagu muujiyey sida khadadka jaalaha ah. (A'), (B'), (C'), iyo (D) waxay muujinayaan isdhexgalka seedaha u dhigma hadhaaga amino acid ee ku hareeraysan, iyo silsiladaha silsilada ah ee hydrogen waxaa lagu muujiyay fallaadho casaan leh.
Tijaabooyin ayaa la sameeyay si loo qiimeeyo saameynta sunta ah ee 5-iodoindole ee biqilka kaabajka iyo miraha dabocase. 5-iodoindole (0.05 ama 0.1 mM) ama avermectin (10 μg/mL) wax yar ama wax saameyn ah kuma yeelan biqilka bilowga ah iyo soo bixitaanka dhirta (Jaantuska 7). Intaa waxaa dheer, wax farqi weyn ah looma helin inta u dhaxaysa heerka biqilka ee kontaroolada aan la daawayn iyo abuurka lagu daweeyay 5-iodoindole ama avermectin. Saamaynta dhererka taproot iyo tirada xididdada lateral ee la sameeyay ayaa ahaa mid aan micno lahayn, inkasta oo 1 mM (10 jeer ay diiradda saareyso firfircoonida) ee 5-iodoindole wax yar dib u dhigtay horumarinta xididdada lateral. Natiijooyinkani waxay muujinayaan in 5-iodoindole aanu sun ahayn unugyada dhirta oo aanu faragelinayn hababka horumarinta dhirta ee xoogga la bartay.
Saamaynta 5-iodoindole ee biqilka abuurka. Biqilka, biqilka iyo xididada lateral ee B. oleracea iyo R. raphanistrum iniinaha Murashiga iyo Skoog agar dhexdhexaad ah oo leh ama aan lahayn avermectin ama 5-iodoindole. Biqilka waxaa la duubay 3 maalmood ka dib markii la beeray 22 ° C.
Daraasaddan ayaa ka warbixisay dhowr xaaladood oo nematode ah oo ay dileen indoles. Muhiimad ahaan, tani waa warbixintii ugu horreysay ee iodoindole oo soo saarta methylation (habka ay keento ururinta vacuoles yar yar oo si tartiib tartiib ah ugu biira vacuoles weyn, ugu dambeyntiina horseedda dillaaca xuubka iyo dhimashada) cirbadaha geedaha, iyadoo iodoindole uu soo bandhigayo waxyaabo muhiim ah oo nematicidal ah oo la mid ah kuwa ganacsiga nematicide avermectin.
Indoles ayaa hore loo soo sheegay in ay sameeyaan hawlo badan oo calaamadeyn ah oo ku jira prokaryotes iyo eukaryotes, oo ay ku jiraan xakamaynta/samaynta biofilm, badbaadada bakteeriyada, iyo cudur-sidaha19,32,33,34. Dhowaan, saamaynta daawaynta ee suurtogalka ah ee indoles halogenated, alkaloids indole, iyo derivatives indole semisynthetic ayaa soo jiitay cilmi-baaris ballaaran35,36,37. Tusaale ahaan, indoles-ka halogenated ayaa la tusay inay dilaan Escherichia coli iyo Staphylococcus aureus unugyada37. Intaa waxaa dheer, waa dano cilmi ah in la barto waxtarka halogenated indoles ka dhanka ah noocyada kale, hiddaha iyo boqortooyooyinka, daraasaddan ayaa ah tallaabo loo qaaday yoolkan.
Halkan, waxaanu soo jeedinaynaa habka loogu talagalay dhimashada 5-iodoindole ee C. elegans oo ku salaysan dhaawaca unugyada dib loo celin karo (RCI) iyo methylation (Jaantus 4C iyo 5). Isbeddellada bararka sida bararka iyo hoos u dhaca vacuolar ayaa ah tilmaamayaasha RCI iyo methylation, oo u muuqda inay yihiin vacuoles weyn oo cytoplasm ah48,49. RCI waxay faragelisaa wax soo saarka tamarta iyadoo yaraynaysa wax soo saarka ATP, taasoo keenaysa fashilka bamka ATPase, ama carqaladeeya xuubabka unugyada oo keena qulqulka degdega ah ee Na +, Ca2+, iyo water50,51,52. Vacuoles intracytoplasmic waxay ka soo baxaan unugyada xayawaanka taas oo ay sabab u tahay ururinta dareeraha cytoplasmka sababtoo ah qulqulka Ca2+ iyo biyaha53. Waxa xiisaha lihi leh, habkan dhaawaca unuggu waa la beddeli karaa haddii dhaawacu yahay mid ku meel gaar ah oo unugyadu bilaabaan inay soo saaraan ATP muddo cayiman, laakiin haddii dhaawacu sii socdo ama ka sii daro, unugyadu way dhintaan.
Noocyada methylation phenotype ee ay keentay 5-iodoindole ee B. xylophilus waxaa laga yaabaa inay sabab u tahay joogitaanka iodine iyo qaybinteeda molecular, tan iyo markii 7-iodoindole uu saameyn yar oo xakameyn ah ku yeeshay B. xylophilus marka loo eego 5-iodoindole (Shaxda 1 iyo Sawirka Dheeraadka ah S6). Natiijooyinkani waxay qayb ahaan waafaqsan yihiin daraasadaha Maltese et al. (2014), kuwaas oo soo sheegay in tarjumaadda pyridyl nitrogen moiety in indole laga bilaabo para- ilaa meta-booska la tirtiray vacuolization, ka hortagga koritaanka, iyo cytotoxicity ee unugyada U251, oo soo jeedinaya in isdhexgalka molecule ee leh goob gaar ah oo firfircoon ee borotiinku waa muhiim27,44,45. Isdhexgalka ka dhexeeya indole ama halogenated indoles iyo GluCL reseptors ee lagu arkay daraasaddan ayaa sidoo kale taageeraya fikraddan, sida 5-iyo 2-iodoindole ayaa la ogaaday inay ku xidhaan GluCL reseptors si ka xoog badan kuwa kale ee indoles ee la baaray (Jaantus 6 iyo Jaantuska Dheeraadka ah S8). Iodine-ka booska labaad ama shanaad ee indole-ka ayaa la ogaaday inuu ku xidho leucine 218 ee GluCL reseptor iyada oo loo sii marayo curaarta hydrogen-ka laf-dhabarta, halka indoles kale oo halogenated ah iyo indole laftiisa ay sameeyaan curaarta silsiladda hydrogen ee daciifka ah oo leh serine 260 (Jaantus 6). Sidaa darteed waxaanu qiyaasnay in meelaynta halogen-ka ay door muhiim ah ka ciyaarto kicinta xumaanshaha vacuolar, halka isku-xidhka adag ee 5-iodoindole ay ilaalinayso kanaalka ion mid furan, taas oo u oggolaanaysa qulqulka dareeraha degdega ah iyo dillaaca vacuole. Si kastaba ha ahaatee, habka faahfaahsan ee ficilka 5-iodoindole ayaa weli ah in la go'aamiyo.
Kahor codsiga la taaban karo ee 5-iodoindole, saameynteeda sunta ah ee dhirta waa in la falanqeeyo. Tijaabooyinka biqilka abuurkayada waxay muujiyeen in 5-iodoindole uusan saameyn xun ku lahayn biqilka abuurka ama hababka korriinka ee xiga ee uruurinta la darsay (Jaantus 7). Sidaa darteed, daraasaddan ayaa saldhig u ah isticmaalka 5-iodoindole ee deegaanka deegaanka si loo xakameeyo waxyeelada nematodes-ka geedaha geedaha geedaha.
Warbixinadii hore waxay muujiyeen in daawaynta ku salaysan indole ay ka dhigan tahay hab suurtagal ah oo wax looga qabanayo dhibaatada iska caabbinta antibiyootiga iyo horumarka kansarka55. Intaa waxaa dheer, indoles waxay leeyihiin bakteeriyada bakteeriyada, anti-kansarka, antioxidant, anti-bararka, antidiabetic, antiviral, antiproliferative iyo hawlaha antituberculosis waxayna u adeegi kartaa aasaas rajo leh horumarinta daroogada56,57. Daraasadani waxay soo jeedinaysaa markii ugu horeysay suurtagalnimada isticmaalka iodine sida wakiilka antiparasitic iyo anthelmintic.
Avermectin waxa la helay soddon sano ka hor waxana uu ku guulaystay abaalmarinta Nobel Prize 2015, isticmaalkeeda sida anthelmintic-ka ayaa wali si firfircoon u socda. Si kastaba ha noqotee, iyadoo ay ugu wacan tahay horumarka degdega ah ee iska caabinta avermectins ee nematodes iyo cayayaanka cayayaanka, beddelka, kharash yar, iyo istiraatiijiyad deegaanka saaxiibtinimo ayaa loo baahan yahay si loo xakameeyo caabuqa PWN ee geedaha geedaha. Daraasaddan ayaa sidoo kale ka warbixisay habka ay 5-iodoindole u disho nematodes geed iyo in 5-iodoindole ay leedahay sunta hoose ee unugyada dhirta, taas oo u fureysa rajooyin wanaagsan oo loogu talagalay codsigeeda ganacsiga mustaqbalka.
Dhammaan tijaabooyinka waxaa ansixiyay Guddiga Anshaxa ee Jaamacadda Yeungnam, Gyeongsan, Kuuriya, iyo hababka waxaa loo sameeyay si waafaqsan tilmaamaha Guddiga Anshaxa ee Jaamacadda Yeungnam.
Tijaabooyinka ukunta ukunta ayaa la sameeyay iyadoo la adeegsanayo hababka la aasaasay43. Si loo qiimeeyo heerarka dillaaca (HR), nematodes qaangaar ah oo 1-maalmood jira (qiyaastii 100 dumar ah iyo 100 rag ah) ayaa loo wareejiyay suxuunta Petri oo ay ku jiraan fungus waxaana loo oggolaaday inay koraan 24 saacadood. Ukunta ayaa markaa la go'doomiyay waxaana lagu daaweeyay 5-iodoindole (0.05 mM iyo 0.1 mM) ama avermectin (10 μg/ml) sida ganaax biyo nadiif ah oo nadiif ah. Joojintan (500 μl; ku dhawaad 100 ukun) ayaa lagu wareejiyay ceelasha saxan dhaqameed 24-ceel ah waxaana lagu beeray 22 °C. Tirinta L2 ayaa la sameeyay 24 saac ka dib markii la tallaalay laakiin waxaa loo tixgeliyey inay dhinteen haddii unugyadu aysan dhaqaaqin markii lagu kiciyey silig platinum ah. Tijaabadan ayaa loo sameeyay laba marxaladood, mid walbana waxaa lagu soo celiyay lix jeer. Xogta labada tijaabo ayaa la isku daray oo la soo bandhigay. Boqolkiiba HR waxaa loo xisaabiyaa sida soo socota:
Dhimashada Larval waxaa lagu qiimeeyay iyadoo la adeegsanayo habab hore loo soo saaray. Ukunta Nematode waa la ururiyay embriyahana waa la isku daray iyadoo ku dillaacay biyo nadiif ah oo nadiif ah si loo dhaliyo dirxiga heerka L2. Dirxiga isku xiran (qiyaastii 500 nematodes) ayaa lagu daweeyay 5-iodoindole (0.05 mM iyo 0.1 mM) ama avermectin (10 μg/ml) waxaana lagu koray taarikada B. cinerea Petri. Ka dib 48 h ee ku-soo-baxa 22 ° C, nematodes waxaa lagu soo ururiyay biyo nadiif ah oo nadiif ah waxaana la baaray joogitaanka heerarka L2, L3, iyo L4. Joogitaanka marxaladaha L3 iyo L4 waxay muujiyeen isbeddel dirxiga, halka joogitaanka heerka L2 aanu muujin wax isbeddel ah. Sawirada waxaa lagu helay iRiS™ Nidaamka Sawirka Unug ee Dijital ah. Tijaabadan ayaa loo sameeyay laba marxaladood, mid walbana waxaa lagu soo celiyay lix jeer. Xogta labada tijaabo ayaa la isku daray oo la soo bandhigay.
Sunta 5-iodoindole iyo avermectin ee miraha ayaa lagu qiimeeyay iyada oo la adeegsanayo tijaabooyinka biqilka ee Murashige iyo Skoog agar plates.62 B. oleracea iyo abuurka R. raphanistrum ayaa markii hore lagu qooyay biyo nadiif ah oo nadiif ah hal maalin, lagu dhaqay 1 ml 100% ethanol sodium, 1 ml 50 lagu nadiifiyey 15 min, oo ku maydhay shan jeer 1 ml oo biyo nadiif ah. Abuurka jeermis-dilaaca ayaa markaas lagu riixay biqilka agar-ka taarikada ka kooban 0.86 g/l (0.2X) Murashige iyo Skoog dhexdhexaad ah iyo 0.7% agar bakteeriyada leh ama aan lahayn 5-iodoindole ama avermectin. Taarikada ayaa markaa lagu duray 22 °C, sawiradana waxaa la qaaday 3 maalmood ka dib markii la tallaalay. Tijaabadan ayaa loo sameeyay laba marxaladood, mid kasta oo ka mid ah waxay lahayd lix ku celcelin.
Waqtiga boostada: Febraayo-26-2025