Nematode na Pine wani nau'in endoparasite ne mai ƙaura wanda aka sani da ke haifar da asarar tattalin arziki mai tsanani a cikin yanayin halittu na dazuzzukan Pine. Wannan binciken ya sake duba ayyukan nematicidal na indoles masu halogenated akan pine nematodes da tsarin aikinsu. Ayyukan nematicidal na 5-iodoindole da avermectin (ikon sarrafawa mai kyau) akan pine nematodes sun kasance iri ɗaya kuma suna da yawa a ƙananan taro (10 μg/mL). 5-iodoindole ya rage yawan haihuwa, aikin haihuwa, mace-macen tayi da tsutsa, da halayen motsa jiki. Hulɗar kwayoyin halitta na ligands tare da masu karɓar tashar chloride mai gauraya ta musamman waɗanda ba su da ƙashin baya suna goyon bayan ra'ayin cewa 5-iodoindole, kamar avermectin, yana ɗaure sosai zuwa wurin aiki na mai karɓa. 5-Iodoindole kuma ya haifar da nakasu iri-iri a cikin nematodes, gami da rugujewar gabobi/raguwa mara kyau da ƙaruwar vacuolization. Waɗannan sakamakon sun nuna cewa vacuoles na iya taka rawa a cikin mutuwar nematode methylation. Abu mafi mahimmanci, 5-iodoindole ba shi da guba ga nau'ikan tsire-tsire (kabeji da radish). Don haka, wannan binciken ya nuna cewa amfani da iodoindole a ƙarƙashin yanayin muhalli na iya magance raunin wilt na pine.
Nematode na itacen Pine (Bursaphelenchus xylophilus) yana cikin ƙwayoyin nematode na itacen Pine (PWN), ƙwayoyin nematode na endoparasitic da aka sani da haifar da mummunan lalacewar muhalli ga yanayin halittu na gandun daji na Pine1. Cutar wilt ta Pine (PWD) da nematode na itacen Pine ke haifarwa yana zama babbar matsala a nahiyoyi da dama, ciki har da Asiya da Turai, kuma a Arewacin Amurka, ƙwayoyin nematode suna lalata nau'ikan pine da aka shigo da su1,2. Ragewar bishiyar Pine babbar matsala ce ta tattalin arziki, kuma yuwuwar yaɗuwarta a duniya abin damuwa ne3. Waɗannan nau'ikan pine galibi ana kai musu hari ta hanyar nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, da Pinus radiata4. Nematode na Pine cuta ce mai tsanani da za ta iya kashe bishiyoyin pine cikin makonni ko watanni bayan kamuwa da cuta. Bugu da ƙari, barkewar ƙwayoyin nematode na Pine ya zama ruwan dare a cikin nau'ikan halittu daban-daban, don haka an kafa sarƙoƙin kamuwa da cuta masu ɗorewa1.
Bursaphelenchus xylophilus wani nau'in nematode ne mai kama da na shuka wanda aka keɓe shi daga dangin Aphelenchoidea da clade 102.5. Nematode yana cin naman fungi kuma yana hayayyafa a cikin kyallen bishiyoyin pine, yana girma zuwa matakai huɗu daban-daban na tsutsotsi: L1, L2, L3, L4 da kuma mutum babba1,6. A ƙarƙashin yanayin ƙarancin abinci, pine nematode yana shiga wani matakin tsutsotsi na musamman - dauer, wanda ke lalata ƙwayoyin halittarsa - ƙwarƙwata na ɓawon pine (Monochamus alternatus) kuma ana canza shi zuwa bishiyoyin pine masu lafiya. A cikin masu masaukin lafiya, nematodes suna ƙaura cikin sauri ta cikin kyallen tsirrai kuma suna cin ƙwayoyin parenchymatous, wanda ke haifar da halayen rashin lafiyar jiki da yawa, bushewar pine da mutuwa cikin shekara guda bayan kamuwa da cuta1,7,8.
Kula da ƙwayoyin halittar pine nematodes ya daɗe yana da ƙalubale, inda matakan keɓewa suka fara tun ƙarni na 20. Dabaru na yanzu don magance ƙwayoyin pine nematodes sun haɗa da magungunan sinadarai, gami da feshin itace da dasa ƙwayoyin nematides a cikin gangar bishiyoyi. Kwayoyin nematides da aka fi amfani da su sune avermectin da avermectin benzoate, waɗanda ke cikin dangin avermectin. Waɗannan sinadarai masu tsada suna da tasiri sosai akan nau'ikan ƙwayoyin nematodes da yawa kuma ana ɗaukar su a matsayin masu aminci ga muhalli9. Duk da haka, ana sa ran amfani da waɗannan ƙwayoyin nematides akai-akai zai haifar da matsin lamba wanda kusan zai haifar da fitowar ƙwayoyin pine masu juriya, kamar yadda aka nuna ga kwari da yawa, kamar Leptinotarsa decemlineata, Plutella xylostella da ƙwayoyin nematodes Trichostrongylus colubriformis da Ostertagia circumcincta, waɗanda a hankali suka haɓaka juriya ga avermectins10,11,12. Saboda haka, ana buƙatar a yi nazari akai-akai game da yanayin juriya kuma a ci gaba da tantance ƙwayoyin nematides akai-akai don nemo wasu matakan madadin, masu inganci da aminci ga muhalli don sarrafa PVD. A cikin 'yan shekarun nan, wasu marubuta sun ba da shawarar amfani da abubuwan da aka samo daga tsire-tsire, mai mai mahimmanci da kuma abubuwan da ke haifar da gurɓatawa a matsayin magungunan hana ƙwayoyin cuta13,14,15,16.
Kwanan nan mun nuna aikin kashe ƙwayoyin cuta na indole, wani ƙwayar siginar intercellular da interkingdom, a cikin Caenorhabditis elegans 17. Indole siginar ƙwayoyin cuta ce da ta yaɗu a cikin ilimin halittu na ƙwayoyin cuta, tana sarrafa ayyuka da yawa waɗanda ke shafar ilimin halittar ƙwayoyin cuta, samuwar spore, kwanciyar hankali na plasmid, juriya ga magunguna, samuwar biofilm, da virulence 18, 19. Ba a yi nazarin ayyukan indole da abubuwan da suka samo asali daga gare shi ba akan sauran ƙwayoyin cuta masu cutarwa. A cikin wannan binciken, mun bincika ayyukan kashe ƙwayoyin cuta na indoles 34 akan pine nematodes kuma mun bayyana tsarin aikin mafi ƙarfi na 5-iodoindole ta amfani da na'urar microscopy, ɗaukar hoto na lokaci-lokaci, da gwaje-gwajen docking na ƙwayoyin cuta, kuma mun tantance tasirinsa mai guba akan tsire-tsire ta amfani da gwajin germination iri.
An riga an ruwaito cewa yawan indole mai yawa (>1.0 mM) yana da tasirin kashe ƙwayoyin cuta akan ƙwayoyin cuta17. Bayan an yi wa B. xylophilus (matakan rayuwa iri-iri) tare da indole ko nau'ikan indole daban-daban guda 33 a 1 mM, an auna mutuwar B. xylophilus ta hanyar ƙidaya ƙwayoyin cuta masu rai da matattu a cikin ƙungiyoyin da aka yi wa magani da waɗanda aka yi wa magani. Indoles guda biyar sun nuna babban aikin kashe ƙwayoyin cuta; rayuwar rukunin da ba a yi wa magani ba shine 95 ± 7% bayan sa'o'i 24. Daga cikin indoles guda 34 da aka gwada, 5-iodoindole da 4-fluoroindole a 1 mM sun haifar da mace-mace 100%, yayin da 5,6-difluoroindigo, methylindole-7-carboxylate, da 7-iodoindole suka haifar da kusan mace-mace 50% (Tebur 1).
Tasirin 5-iodoindole akan samuwar vacuole da metabolism na pine wood nematode. (A) Tasirin avermectin da 5-iodoindole akan manyan nematodes na maza, (B) ƙwai nematode na mataki na L1 da (C) metabolism na B. xylophilus, (i) ba a lura da vacuoles a cikin awanni 0 ba, magani ya haifar da (ii) vacuoles, (iii) tarin vacuoles da yawa, (iv) kumburin vacuoles, (v) haɗuwar vacuoles da (vi) samuwar manyan vacuoles. Kibiyoyi ja suna nuna kumburin vacuoles, kibiyoyi shuɗi suna nuna haɗuwar vacuoles da kibiyoyi baƙi suna nuna manyan vacuoles. Ma'aunin sikelin = 50 μm.
Bugu da ƙari, wannan binciken ya kuma bayyana tsarin da ke biye na mutuwar methane da ke haifar da shi a cikin ƙwayoyin pine (Hoto na 4C). Mutuwar methanogenic nau'in mutuwar ƙwayoyin halitta ne wanda ba apoptotic ba wanda ke da alaƙa da tarin manyan ƙwayoyin cytoplasmic27. Lalacewar siffar da aka gani a cikin ƙwayoyin pine suna da alaƙa da tsarin mutuwar da methane ke haifarwa. Binciken microscopic a lokuta daban-daban ya nuna cewa manyan ƙwayoyin halitta sun samo asali ne bayan sa'o'i 20 na fallasa ga 5-iodoindole (0.1 mM). An lura da ƙananan ƙwayoyin halitta bayan sa'o'i 8 na magani, kuma adadinsu ya ƙaru bayan sa'o'i 12. An lura da manyan ƙwayoyin halitta da yawa bayan sa'o'i 14. An ga wasu ƙwayoyin halitta da aka haɗa a sarari bayan sa'o'i 12-16 na magani, wanda ke nuna cewa haɗuwar ƙwayoyin halitta shine tushen tsarin mutuwar methanogenic. Bayan sa'o'i 20, an sami manyan ƙwayoyin halitta da yawa a cikin tsutsa. Waɗannan abubuwan da aka lura suna wakiltar rahoton farko na metuosis a cikin C. elegans.
A cikin tsutsotsi da aka yi wa magani da iodoindole guda 5, an kuma lura da tarin vacuole da fashewa (Hoto na 5), kamar yadda aka shaida ta hanyar lanƙwasa tsutsotsi da sakin vacuole cikin muhalli. An kuma lura da katsewar vacuole a cikin membrane na ƙwai, wanda yawanci L2 ke kiyaye shi ba tare da wata matsala ba yayin ƙyanƙyashewa (Hoto na Ƙarin S2). Waɗannan abubuwan da aka lura suna tallafawa shigar tarin ruwa da gazawar osmoregulatory, da kuma raunin ƙwayoyin halitta (RCI), a cikin tsarin samuwar vacuole da suppuration (Hoto na 5).
Da muka yi hasashen rawar da aidin ke takawa a cikin samuwar vacuole da aka lura, mun binciki aikin nematicidal na sodium iodide (NaI) da potassium iodide (KI). Duk da haka, a yawan (0.1, 0.5 ko 1 mM), ba su shafi rayuwar nematode ko samuwar vacuole ba (Fig. S5 na Ƙarin Bayani), kodayake 1 mM KI yana da ɗan tasirin nematicidal. A gefe guda kuma, 7-iodoindole (1 ko 2 mM), kamar 5-iodoindole, ya haifar da vacuoles da yawa da nakasar tsarin (Fig. S6 na Ƙarin Bayani). Iodoindoles guda biyu sun nuna irin waɗannan halaye na phenotypic a cikin pine nematodes, yayin da NaI da KI ba su yi ba. Abin sha'awa, indole bai haifar da samuwar vacuole a cikin B. xylophilus ba a yawan da aka gwada (bayanan da ba a nuna ba). Don haka, sakamakon ya tabbatar da cewa hadaddun indole-iodine shine ke da alhakin vacuolization da metabolism na B. xylophilus.
Daga cikin indoles da aka gwada don aikin kashe ƙwayoyin cuta, 5-iodoindole yana da mafi girman ma'aunin zamewa na -5.89 kcal/mol, sai kuma 7-iodoindole (-4.48 kcal/mol), 4-fluoroindole (-4.33), da indole (-4.03) (Hoto na 6). Ƙarfin haɗin hydrogen na baya na 5-iodoindole zuwa leucine 218 yana daidaita haɗinsa, yayin da duk sauran abubuwan da suka samo asali na indole suna ɗaure zuwa serine 260 ta hanyar haɗin hydrogen na sarkar sarkar. Daga cikin sauran iodoindoles masu ƙira, 2-iodoindole yana da ƙimar ɗaurewa ta -5.248 kcal/mol, wanda hakan ya faru ne saboda babban haɗin hydrogen ɗinsa da leucine 218. Sauran haɗin da aka sani sun haɗa da 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), da 6-fluoroindole (-2.6 kcal/mol) (Ƙarin Hoto na S8). Yawancin indoles masu halogenated da indole da kansu, ban da 5-iodoindole da 2-iodoindole, suna samar da haɗin gwiwa da serine 260. Gaskiyar cewa haɗin hydrogen da leucine 218 yana nuna ingantaccen haɗin receptor-ligand, kamar yadda aka lura da ivermectin (Ƙarin Hoto na S7), ya tabbatar da cewa 5-iodoindole da 2-iodoindole, kamar ivermectin, suna ɗaure sosai zuwa wurin aiki na mai karɓar GluCL ta hanyar leucine 218 (Hoto na 6 da Ƙarin Hoto na S8). Mun ba da shawarar cewa ana buƙatar wannan haɗin gwiwa don kula da tsarin buɗewar rami na mahaɗin GluCL kuma ta hanyar ɗaurewa sosai zuwa wurin aiki na mai karɓar GluCL, 5-iodoindole, 2-iodoindole, avermectin da ivermectin don haka suna kiyaye tashar ion a buɗe kuma suna ba da damar ɗaukar ruwa.
Haɗa indole da halogenated indole zuwa GluCL. Haɗa hanyoyin haɗin ligands na (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, da (D) 5-iodoindole zuwa wurin da GluCL ke aiki. Ana wakiltar furotin ta hanyar ribbon, kuma haɗin hydrogen na baya an nuna su azaman layuka masu digo-digo. (A′), (B′), (C′), da (D′) suna nuna hulɗar ligands masu dacewa da ragowar amino acid da ke kewaye, kuma haɗin hydrogen na gefe an nuna su da kibiyoyi masu digo-digo masu ruwan hoda.
An gudanar da gwaje-gwaje don tantance tasirin guba na 5-iodoindole akan tsiron tsaban kabeji da radish. 5-iodoindole (0.05 ko 0.1 mM) ko avermectin (10 μg/mL) ba su da wani tasiri ko kaɗan akan tsiron farko da fitowar shuke-shuke (Hoto na 7). Bugu da ƙari, babu wani bambanci mai mahimmanci da aka samu tsakanin ƙimar tsiron magungunan da ba a yi wa magani ba da kuma tsaba da aka yi wa magani da 5-iodoindole ko avermectin. Tasirin tsawaita taproot da adadin tushen gefe da aka samu bai yi yawa ba, kodayake 1 mM (sau 10 yawan aiki) na 5-iodoindole ya ɗan jinkirta ci gaban tushen gefe. Waɗannan sakamakon sun nuna cewa 5-iodoindole ba shi da guba ga ƙwayoyin shuka kuma baya tsoma baki ga tsarin ci gaban shuka a yawan da aka yi nazari a kai.
Tasirin iodoindole 5 akan tsiron iri. Tsirrai, tsiro da kuma tushen gefen tsaban B. oleracea da R. raphanistrum akan matsakaicin Murashige da Skoog agar tare da ko ba tare da avermectin ko 5-iodoindole ba. An yi rikodin tsiron bayan kwana 3 na shukawa a zafin jiki na 22°C.
Wannan binciken ya ba da rahoton wasu lamura da dama na kashe ƙwayoyin nematode ta hanyar indoles. Abin mahimmanci, wannan shine rahoto na farko na iodoindole da ke haifar da methylation (wani tsari da ke faruwa sakamakon tarin ƙananan ƙwayoyin da ke haɗuwa a hankali zuwa manyan ƙwayoyin vacuoles, wanda daga ƙarshe ke haifar da fashewar membrane da mutuwa) a cikin allurar pine, tare da iodoindole yana nuna manyan kaddarorin nematicide iri ɗaya da na nematicide avermectin na kasuwanci.
An ruwaito cewa Indoles suna yin ayyuka da yawa na sigina a cikin prokaryotes da eukaryotes, gami da hana/samfurin biofilm, tsira daga ƙwayoyin cuta, da kuma cututtuka19,32,33,34. Kwanan nan, tasirin warkewa na indoles masu halogenated, indole alkaloids, da abubuwan da suka samo asali daga indole masu semisynthetic sun jawo hankalin bincike mai zurfi35,36,37. Misali, an nuna cewa indoles masu halogenated suna kashe ƙwayoyin Escherichia coli masu ɗorewa da Staphylococcus aureus37. Bugu da ƙari, yana da sha'awar kimiyya a yi nazarin ingancin indoles masu halogenated akan wasu nau'ikan, jinsi, da masarautu, kuma wannan binciken mataki ne na cimma wannan burin.
A nan, mun gabatar da wata hanya ta kashe ƙwayoyin cuta masu ɗauke da iodoindole guda 5 a cikin ƙwayoyin C. elegans bisa ga raunin ƙwayoyin cuta masu juyawa (RCI) da methylation (Figures 4C da 5). Canje-canje a cikin ƙashi kamar kumburi da lalacewar ƙwayoyin cuta alamu ne na RCI da methylation, waɗanda aka bayyana a matsayin manyan ƙwayoyin cuta a cikin cytoplasm48,49. RCI yana tsoma baki ga samar da makamashi ta hanyar rage samar da ATP, yana haifar da gazawar famfon ATPase, ko kuma yana kawo cikas ga membranes na ƙwayoyin halitta da kuma haifar da kwararar Na+, Ca2+, da water50,51,52 cikin sauri. Kwayoyin cuta masu ɗauke da ƙwayoyin cuta a cikin ƙwayoyin halitta suna tasowa a cikin ƙwayoyin dabbobi sakamakon tarin ruwa a cikin ƙwayoyin halitta saboda kwararar Ca2+ da water53. Abin sha'awa, wannan hanyar lalacewar tantanin halitta za a iya mayar da ita idan lalacewar ta kasance ta ɗan lokaci kuma ƙwayoyin halittar suka fara samar da ATP na wani lokaci, amma idan lalacewar ta ci gaba ko ta tsananta, ƙwayoyin halittar suna mutuwa.54 Abubuwan da muka lura sun nuna cewa ƙwayoyin nematodes da aka yi wa magani da 5-iodoindole ba za su iya dawo da tsarin halittar halitta ba bayan sun fuskanci yanayin damuwa.
Tsarin methylation da 5-iodoindole ya haifar a cikin B. xylophilus na iya zama saboda kasancewar iodine da rarrabawar kwayoyin halittarsa, tunda 7-iodoindole yana da ƙarancin tasirin hana B. xylophilus fiye da 5-iodoindole (Tebur 1 da Ƙarin Hoto na S6). Waɗannan sakamakon sun yi daidai da nazarin Maltese et al. (2014), waɗanda suka ba da rahoton cewa canja wurin pyridyl nitrogen a cikin indole daga para-to the meta-position ya kawar da vacuolization, hana girma, da guba a cikin ƙwayoyin U251, yana nuna cewa hulɗar ƙwayar halitta tare da takamaiman wurin aiki a cikin furotin yana da mahimmanci27,44,45. Hulɗar tsakanin indole indole ko halogenated da masu karɓar GluCL da aka lura a cikin wannan binciken suma suna goyon bayan wannan ra'ayi, kamar yadda aka gano 5- da 2-iodoindole suna ɗaurewa da masu karɓar GluCL da ƙarfi fiye da sauran indoles da aka bincika (Hoto na 6 da Ƙarin Hoto na S8). An gano cewa aidin a matsayi na biyu ko na biyar na indole yana ɗaurewa da leucine 218 na mai karɓar GluCL ta hanyar haɗin hydrogen na baya, yayin da sauran indoles masu halogenated da indole da kansu suna samar da haɗin hydrogen mai rauni tare da serine 260 (Hoto na 6). Saboda haka, muna hasashen cewa wurin da halogen yake yana taka muhimmiyar rawa wajen haifar da lalacewar vacuolar, yayin da ɗaurewar 5-iodoindole mai tsauri ke sa tashar ion ta buɗe, ta haka ne ke ba da damar kwararar ruwa cikin sauri da fashewar vacuole. Duk da haka, har yanzu ba a tantance cikakken tsarin aikin 5-iodoindole ba.
Kafin a yi amfani da 5-iodoindole a aikace, ya kamata a yi nazarin tasirinsa mai guba ga shuke-shuke. Gwaje-gwajen da muka yi na 5-iodoindole sun nuna cewa 5-iodoindole ba shi da wani mummunan tasiri ga 5-iodoindole ko kuma ci gaba da aka samu a yawan da aka yi nazari a kai (Hoto na 7). Don haka, wannan binciken ya samar da tushen amfani da 5-iodoindole a cikin muhallin muhalli don sarrafa illolin pine nematodes ga bishiyoyin pine.
Rahotannin da suka gabata sun nuna cewa maganin da aka yi da indole yana wakiltar wata hanya mai yuwuwa don magance matsalar juriyar maganin rigakafi da ci gaban ciwon daji55. Bugu da ƙari, indoles suna da ayyukan kashe ƙwayoyin cuta, maganin ciwon daji, maganin antioxidant, maganin kumburi, maganin ciwon suga, maganin rigakafi, maganin yaduwa da kuma maganin tarin fuka kuma suna iya zama tushen ci gaban magunguna56,57. Wannan binciken ya nuna a karon farko cewa amfani da iodine a matsayin maganin kashe ƙwayoyin cuta da maganin kashe ƙwayoyin cuta.
An gano Avermectin shekaru talatin da suka gabata kuma ya lashe kyautar Nobel a shekarar 2015, kuma amfani da shi a matsayin maganin hana kumburi har yanzu yana ci gaba. Duk da haka, saboda saurin haɓakar juriya ga avermectins a cikin nematodes da kwari, ana buƙatar wata dabara ta daban, mai araha, kuma mai kyau ga muhalli don magance kamuwa da cutar PWN a cikin bishiyoyin pine. Wannan binciken ya kuma ba da rahoton hanyar da 5-iodoindole ke kashe pine nematodes da kuma cewa 5-iodoindole yana da ƙarancin guba ga ƙwayoyin shuka, wanda ke buɗe kyakkyawar dama ga amfani da shi a nan gaba a kasuwanci.
Kwamitin Da'a na Jami'ar Yeungnam, Gyeongsan, Koriya ta amince da dukkan gwaje-gwajen, kuma an gudanar da hanyoyin bisa ga ka'idojin Kwamitin Da'a na Jami'ar Yeungnam.
An gudanar da gwaje-gwajen ƙunƙun ƙwai ta amfani da hanyoyin da aka kafa43. Don tantance yawan ƙyanƙyashewa (HR), an mayar da ƙwayoyin nematodes manya na kwana 1 (kimanin mata 100 da maza 100) zuwa abincin Petri da ke ɗauke da naman gwari kuma aka bar su su girma na tsawon awanni 24. Daga nan aka ware ƙwai aka yi musu magani da iodoindole 5 (0.05 mM da 0.1 mM) ko avermectin (10 μg/ml) a matsayin dakatarwa a cikin ruwan da aka tace. An tura waɗannan dakatarwar (500 μl; kimanin ƙwai 100) zuwa rijiyoyin farantin nama mai rijiyoyi 24 kuma aka sanya su a zafin jiki na 22 °C. An yi ƙidayar L2 bayan awanni 24 na ƙunƙun amma an ɗauke su a matsayin matattu idan ƙwayoyin ba su motsa ba lokacin da aka motsa su da waya mai kyau ta platinum. An gudanar da wannan gwajin a matakai biyu, kowannensu da maimaitawa shida. An haɗa bayanan daga gwaje-gwajen biyu kuma an gabatar da su. An ƙididdige kashi na HR kamar haka:
An tantance mace-macen tsutsotsi ta amfani da hanyoyin da aka riga aka tsara. An tattara ƙwai na Nematode kuma an haɗa ƙwai ta hanyar ƙyanƙyashewa a cikin ruwan da aka tace don samar da tsutsotsi na matakin L2. An yi wa tsutsotsi masu daidaitawa (kimanin nematodes 500) magani da 5-iodoindole (0.05 mM da 0.1 mM) ko avermectin (10 μg/ml) kuma an haƙa su a kan faranti na B. cinerea Petri. Bayan awanni 48 na haɗuwa a zafin jiki na 22 °C, an tattara ƙwayoyin nematodes a cikin ruwan da aka tace kuma an duba ko akwai matakan L2, L3, da L4. Kasancewar matakan L3 da L4 ya nuna canjin tsutsotsi, yayin da kasancewar matakin L2 bai nuna wani canji ba. An samo hotuna ta amfani da Tsarin Hoton Kwayoyin Dijital na iRiS™. An gudanar da wannan gwajin a matakai biyu, kowannensu da maimaitawa shida. An haɗa bayanan daga gwaje-gwajen biyu kuma an gabatar da su.
An tantance gubar da ke tattare da 5-iodoindole da avermectin ga tsaba ta amfani da gwajin germination a kan faranti na Murashige da Skoog agar.62 B. oleracea da R. raphanistrum tsaba an fara jiƙa su a cikin ruwan da aka tace na kwana ɗaya, an wanke su da ethanol na 1 ml 100%, an yi musu maganin bleach na kasuwanci na 50% (3% sodium hypochlorite) na tsawon minti 15, sannan aka wanke su sau biyar da ruwan da aka tace na 1 ml. Sannan aka matse tsaban da aka tace a kan faranti na agar da aka shuka wanda ke ɗauke da 0.86 g/l (0.2X) na Murashige da Skoog matsakaici da 0.7% na agar bacteriological tare da ko ba tare da 5-iodoindole ko avermectin ba. Sannan aka sanya faranti a zafin jiki na 22 °C, kuma an ɗauki hotuna bayan kwana 3 na germination. An gudanar da wannan gwajin a matakai biyu, kowannensu yana da maimaitawa shida.
Lokacin Saƙo: Fabrairu-26-2025