Ko te nematode paina he endoparasite heke taratahi e mōhiotia ana he nui ngā mate ōhanga i roto i ngā pūnaha rauropi ngahere paina. E arotakehia ana e tēnei rangahau te mahi nematicidal o ngā indole halogenated ki ngā nematode paina me tā rātou tikanga mahi. He rite tonu, he teitei hoki ngā mahi nematicidal o te 5-iodoindole me te avermectin (mana pai) ki ngā nematode paina i ngā kukū iti (10 μg/mL). I whakaitihia e te 5-iodoindole te momonatanga, te mahi whakaputa uri, te mate o ngā kukune me ngā torongū, me te whanonga nekeneke. Ko ngā taunekeneke ngota o ngā ligands me ngā kaiwhakawhiwhi hongere chloride glutamate-gated motuhake ki ngā invertebrate e tautoko ana i te whakaaro ko te 5-iodoindole, pērā i te avermectin, ka piri pūmau ki te pae hohe o te kaiwhakawhiwhi. I puta anō hoki i te 5-Iodoindole ngā momo whakarerekētanga phenotypic i roto i ngā nematode, tae atu ki te hinga/te whakaiti okana rerekē me te pikinga o te vacuolization. E kī ana ēnei hua he wāhi pea ngā vacuole ki te mate o ngā nematode e whakahaerehia ana e te methylation. He mea nui, kāore te 5-iodoindole i pā kino ki ngā momo tipu e rua (kāpeti me te rātiti). Nō reira, e whakaatu ana tēnei rangahau ka taea e te whakamahinga o te iodoindole i raro i ngā āhuatanga taiao te whakahaere i te kino o te memenge o te paina.
Ko te nematode rakau paina (Bursaphelenchus xylophilus) nō roto i ngā nematode rakau paina (PWN), arā, ngā nematode endoparasitic heke e mōhiotia ana ka pā kino nui ki ngā pūnaha rauropi o te ngahere paina1. Ko te mate ngenge paina (PWD) i puta mai i te nematode rakau paina kei te noho hei raruraru nui i runga i ētahi whenua, tae atu ki Āhia me Ūropi, ā, i Amerika Te Raki, ka whakangaromia e te nematode ngā momo paina whakauru1,2. He raruraru ōhanga nui te hekenga o ngā rākau paina, ā, he mea āwangawanga te tūmanako ka horapa puta noa i te ao3. Ko ngā momo paina e whai ake nei ka tino whakaekea e te nematode: Pinus densiflora, Pinus sylvestris, Pinus thunbergii, Pinus koraiensis, Pinus thunbergii, Pinus thunbergii, me Pinus radiata4. He mate kino te nematode paina ka taea te patu i ngā rākau paina i roto i ngā wiki, i ngā marama rānei o te pangia. Hei tāpiri, he mea noa te pakarutanga o te nematode paina i roto i ngā momo pūnaha rauropi, nō reira kua whakatūria he mekameka pangia pumau1.
Ko te Bursaphelenchus xylophilus he nematode tipu-parasitic motuhake nō te whānau Aphelenchoidea me te clade 102.5. Ka kai te nematode i ngā harore, ka whakaputa uri i roto i ngā kiko rakau o ngā rākau paina, ka whanake hei whā ngā wāhanga torongū rerekē: L1, L2, L3, L4 me te tangata pakeke1,6. I raro i ngā āhuatanga o te kore kai, ka uru te nematode paina ki tētahi wāhanga torongū motuhake – te dauer, e parasitia ana tana kawe – te pītara kiri paina (Monochamus alternatus) ka whakawhitia ki ngā rākau paina hauora. I roto i ngā manaaki hauora, ka tere te heke o ngā nematode i roto i ngā kiko tipu ka kai i ngā pūtau parenchymatous, ka arahi ki te maha o ngā tauhohenga hypersensitivity, te memeha o te paina me te mate i roto i te tau kotahi i muri i te pangia1,7,8.
He wero te whakahaere koiora i ngā nematode paina, ā, mai anō i te rautau 20 ngā mahi karantini. Ko ngā rautaki o nāianei mō te whakahaere i ngā nematode paina ko ngā maimoatanga matū, tae atu ki te whakapūkara rakau me te whakato i ngā nematode ki roto i ngā pūtake rākau. Ko ngā nematode e whakamahia whānuitia ana ko te avermectin me te avermectin benzoate, nō te whānau avermectin. He tino whai hua ēnei matū utu nui ki te maha o ngā momo nematode, ā, e kiia ana he haumaru mō te taiao9. Heoi, ko te whakamahinga tonu o ēnei nematode e whakaarohia ana ka puta he pēhanga kōwhiringa ka tino arahi ki te putanga mai o ngā nematode paina ātete, e ai ki ngā whakaaturanga mō ētahi pepeke pests, pērā i a Leptinotarsa decemlineata, Plutella xylostella me ngā nematode Trichostrongylus colubriformis me Ostertagia circumcincta, kua āta whanake te ātete ki ngā avermectins10,11,12. Nō reira, me ako tonu ngā tauira ātete, ā, me tirotiro tonu i ngā nematode kia kitea ai he huarahi kē atu, he utu-whai hua, he pai hoki mō te taiao hei whakahaere i te PVD. I ngā tekau tau tata nei, he maha ngā kaituhi kua whakaaro ki te whakamahi i ngā tangohanga tipu, ngā hinu kakara me ngā matū whakakorikori hei whakahaere i ngā nematode13,14,15,16.
Nō nā tata nei mātou i whakaatu i te mahi nematicidal o te indole, he ngota tohu i waenga i ngā pūtau me ngā kingitanga, i roto i te Caenorhabditis elegans 17. Ko te indole he tohu pūtau whānui i roto i te rauropi moroiti, e whakahaere ana i ngā mahi maha e pā ana ki te āhua o te moroiti, te hanganga o ngā spore, te pumau o te plasmid, te ātete ki te rongoā, te hanganga biofilm, me te kaha 18, 19. Kāore anō kia rangahaua te mahi a te indole me ōna hua ki ētahi atu nematode pathogenic. I roto i tēnei rangahau, i tirotirohia e mātou te mahi nematicidal o te 34 indole ki ngā nematode paina, ā, i whakamāramahia te tikanga mahi a te 5-iodoindole tino kaha mā te whakamahi i te karuārai, te whakaahua wā-roa, me ngā whakamātautau hononga ngota, ā, i aromatawaihia ōna pānga paitini ki ngā tipu mā te whakamahi i te whakamātautau whakatipu purapura.
Kua pūrongohia i mua he pānga whakamate nematode ngā kukū teitei (>1.0 mM) o te indole ki ngā nematode17. Whai muri i te maimoatanga o te B. xylophilus (ngā wāhanga ora whakauru) ki te indole, ki te 33 rānei ngā momo indole rerekē i te 1 mM, i inehia te mate o te B. xylophilus mā te tatau i ngā nematode ora me ngā nematode mate i roto i ngā rōpū mana whakahaere me ngā rōpū i rongoatia. E rima ngā indole i whakaatu i te kaha whakamate nematode nui; ko te oranga o te rōpū mana whakahaere kāore i rongoatia he 95 ± 7% i muri i te 24 hāora. O ngā indole 34 i whakamatautauria, ko te 5-iodoindole me te 4-fluoroindole i te 1 mM i mate ai te 100%, ko te 5,6-difluoroindigo, te methylindole-7-carboxylate, me te 7-iodoindole i mate ai te tata ki te 50% (Ripanga 1).
Te pānga o te 5-iodoindole ki te hanganga o ngā vacuole me te pākia o te nematode rakau paina. (A) Te pānga o te avermectin me te 5-iodoindole ki ngā nematode tāne pakeke, (B) ngā hua nematode wāhanga L1 me te (C) pākia o te B. xylophilus, (i) kāore i kitea ngā vacuole i te 0 h, ko te hua o te maimoatanga he (ii) ngā vacuole, (iii) te kohikohinga o ngā vacuole maha, (iv) te pupuhi o ngā vacuole, (v) te hanumitanga o ngā vacuole me te (vi) te hanganga o ngā vacuole nunui. Ko ngā pere whero e tohu ana i te pupuhi o ngā vacuole, ko ngā pere kikorangi e tohu ana i te hanumitanga o ngā vacuole, ā, ko ngā pere mangu e tohu ana i ngā vacuole nunui. Pae tauine = 50 μm.
Hei tāpiritanga, i whakaahuahia hoki e tēnei rangahau te tukanga raupapa o te matenga i puta mai i te methane i roto i ngā nematode paina (Pikitia 4C). Ko te mate methanogenic he momo mate pūtau kore-apoptotic e pā ana ki te kohikohinga o ngā vacuoles cytoplasmic rongonui27. Ko ngā hapa āhua i kitea i roto i ngā nematode paina te āhua nei he hononga tata ki te tikanga o te matenga i puta mai i te methane. I whakaatuhia e te whakamātautau karuārai i ngā wā rerekē kua hangaia ngā vacuoles nunui i muri i te 20 hāora o te pānga ki te 5-iodoindole (0.1 mM). I kitea ngā vacuoles karuārai i muri i te 8 hāora o te maimoatanga, ā, kua piki ake tō rātou maha i muri i te 12 hāora. He maha ngā vacuoles nunui i kitea i muri i te 14 hāora. He maha ngā vacuoles kua honoa i kitea mārama i muri i te 12–16 hāora o te maimoatanga, e tohu ana ko te whakakotahitanga o ngā vacuole te pūtake o te tikanga mate methanogenic. I muri i te 20 hāora, he maha ngā vacuoles nunui i kitea puta noa i te kutukutu. Ko ēnei kitenga te pūrongo tuatahi o te metuosis i roto i te C. elegans.
I roto i ngā kutukutu i rongoatia ki te 5-iodoindole, i kitea anō te kohikohinga me te pakaru o ngā vacuole (Pikitia 5), e whakaatuhia ana e te piko o te kutukutu me te tukunga o ngā vacuole ki te taiao. I kitea anō hoki te pakaru o ngā vacuole i roto i te kiriuhi anga hua manu, e tiakina ana e te L2 i te wā e whānau ana (Pikitia Tāpiri S2). E tautoko ana ēnei kitenga i te whai wāhi o te kohikohinga wai me te korenga o te whakahaere osmore, tae atu ki te whara pūtau whakahurihuri (RCI), i roto i te tukanga hanganga me te pupuhi o ngā vacuole (Pikitia 5).
I te whakapae i te mahi a te iodine i roto i te hanganga o te vacuole i kitea, i tirotirohia e mātou te mahi nematicidal o te iodide konutai (NaI) me te iodide pāhare pākawa (KI). Heoi, i ngā kukū (0.1, 0.5, 1 mM rānei), kāore i pā ki te oranga o te nematode, ki te hanganga vacuole rānei (Pikitia Tāpiri S5), ahakoa he iti noa te pānga nematicidal o te 1 mM KI. I tētahi atu taha, i puta mai i te 7-iodoindole (1, 2 mM rānei), pērā i te 5-iodoindole, he maha ngā vacuole i puta mai me ngā whakarerekētanga hanganga (Pikitia Tāpiri S6). I whakaatuhia e ngā iodoindole e rua ngā āhuatanga phenotypic rite tonu i roto i ngā nematode paina, engari kāore i puta mai i te NaI me te KI. He mea whakamīharo, kāore te indole i puta mai te hanganga vacuole i roto i te B. xylophilus i ngā kukū i whakamatautauria (kāore i whakaaturia ngā raraunga). Nō reira, i whakaū ngā hua ko te matatini indole-iodine te kawenga mō te vacuolization me te metabolism o te B. xylophilus.
I roto i ngā indoles i whakamatautauria mō te mahi nematicidal, ko te 5-iodoindole te mea i nui rawa te taupū paheke o te -5.89 kcal/mol, ā, i muri mai ko te 7-iodoindole (-4.48 kcal/mol), 4-fluoroindole (-4.33), me te indole (-4.03) (Pikitia 6). Ko te hononga hauwai tuara kaha o te 5-iodoindole ki te leucine 218 ka whakau i tōna herenga, engari ko ngā atu indole katoa e herea ana ki te serine 260 mā roto i ngā here hauwai mekameka taha. I roto i ētahi atu iodoindole kua tauirahia, ko te uara herenga o te 2-iodoindole he -5.248 kcal/mol, nā tōna herenga hauwai matua ki te leucine 218. Ko ētahi atu herenga e mōhiotia ana ko te 3-iodoindole (-4.3 kcal/mol), 4-iodoindole (-4.0 kcal/mol), me te 6-fluoroindole (-2.6 kcal/mol) (Pikitia Tāpiri S8). Ko te nuinga o ngā indole halogenated me te indole tonu, haunga te 5-iodoindole me te 2-iodoindole, ka hanga hononga ki te serine 260. Ko te meka ko te hononga hauwai ki te leucine 218 he tohu mō te herenga pai o te kaiwhakawhiwhi-ligand, e kitea ana mō te ivermectin (Pikitia Tāpiri S7), e whakaū ana ko te 5-iodoindole me te 2-iodoindole, pērā i te ivermectin, ka herea piri ki te pae hohe o te kaiwhakawhiwhi GluCL mā te leucine 218 (Pikitia 6 me te Pikitia Tāpiri S8). E whakaaro ana mātou me whai hononga tēnei hei pupuri i te hanganga pore tuwhera o te matatini GluCL, ā, mā te herea piri ki te pae hohe o te kaiwhakawhiwhi GluCL, ka pupuri te 5-iodoindole, 2-iodoindole, avermectin me te ivermectin kia tuwhera te hongere katote, ā, ka taea te tango wai.
Te hononga ngota o te indole me te indole halogenated ki te GluCL. Ngā whakatakotoranga here o (A) indole, (B) 4-fluoroindole, (C) 7-iodoindole, me (D) 5-iodoindole ligands ki te pae hohe o GluCL. Ko te pūmua e tohuhia ana e te rīpene, ā, ko ngā here hauwai tuara e whakaatuhia ana hei rārangi iraira kōwhai. Ko (A′), (B′), (C′), me (D′) e whakaatu ana i ngā taunekeneke o ngā ligands e rite ana ki ngā toenga waikawa amino e karapoti ana, ā, ko ngā here hauwai mekameka taha e tohuhia ana e ngā pere iraira māwhero.
I whakahaerehia ngā whakamātautau hei aromatawai i te pānga paitini o te 5-iodoindole ki te whakatipuranga o ngā purapura kāpeti me te rātiti. He iti noa iho, he kore rānei he pānga o te 5-iodoindole (0.05, 0.1 mM rānei) te avermectin rānei (10 μg/mL) ki te whakatipuranga tuatahi me te putanga mai o te tipu iti (Pikitia 7). Hei tāpiri, kāore i kitea he rerekētanga nui i waenga i te tere whakatipuranga o ngā mana whakahaere kāore i rongoātia me ngā purapura i rongoātia ki te 5-iodoindole, ki te avermectin rānei. Kāore i te nui te pānga ki te roa o te pakiaka me te maha o ngā pakiaka taha i hangaia, ahakoa he iti noa te whakaroa o te whanaketanga o ngā pakiaka taha e te 1 mM (10 ngā wā o tōna kukū kaha) o te 5-iodoindole. E whakaatu ana ēnei hua kāore te 5-iodoindole i te paitini ki ngā pūtau tipu, ā, kāore e whakararuraru i ngā tukanga whanaketanga tipu i ngā kukū i rangahauhia.
Te pānga o te 5-iodoindole ki te whakatipu purapura. Te whakatipu, te pihi me te pakiaka taha o ngā purapura B. oleracea me R. raphanistrum ki runga i te ranunga agar Murashige me Skoog me te avermectin, te 5-iodoindole rānei, me te kore rānei. I tuhia te whakatipu i muri i ngā rā e 3 o te whakatipuranga i te 22°C.
E pūrongo ana tēnei rangahau i ētahi take o te matenga o ngā nematode e ngā indole. He mea nui, koinei te pūrongo tuatahi mō te methylation e puta mai ana i te iodoindole (he tukanga i puta mai i te kohikohinga o ngā vacuole iti e hanumi haere ana ki ngā vacuole nunui, ā, ka mutu ka pakaru te membrane me te mate) i roto i ngā ngira paina, me te whakaatu a te iodoindole i ngā āhuatanga nematicidal nui e rite ana ki ērā o te avermectin nematicide arumoni.
Kua pūrongohia i mua he maha ngā mahi tohu a ngā indole i roto i ngā prokaryote me ngā eukaryote, tae atu ki te aukati/te hanganga biofilm, te oranga o te huakita, me te pathogenicity19,32,33,34. Nō nā tata nei, kua kukumehia te hiahia nui ki ngā rangahau mō ngā pānga rongoā pea o ngā indole halogenated, ngā alkaloids indole, me ngā indole derivatives semisynthetic35,36,37. Hei tauira, kua whakaatuhia e ngā indole halogenated ka patu i ngā pūtau Escherichia coli me Staphylococcus aureus pumau37. Hei tāpiri, he mea nui ki te pūtaiao te ako i te whai huatanga o ngā indole halogenated ki ētahi atu momo, puninga, me ngā kingitanga, ā, he taahiraa tēnei rangahau ki te whakatutuki i tēnei whāinga.
I konei, ka whakaarohia e mātou he tikanga mō te mate i puta mai i te 5-iodoindole i roto i te C. elegans i runga i te whara pūtau whakahurihuri (RCI) me te methylation (Pikitia 4C me te 5). Ko ngā huringa edematous pēnei i te pupuhi me te hekenga o te vacuolar he tohu o te RCI me te methylation, e whakaatuhia ana hei vacuoles nunui i roto i te cytoplasm48,49. Ka whakararu te RCI i te whakaputanga pūngao mā te whakaiti i te whakaputanga ATP, ka kore e taea te papu ATPase, ka whakararuraru rānei i ngā kiriuhi pūtau ka puta he rerenga tere o Na+, Ca2+, me te wai50,51,52. Ka puta ake ngā vacuoles intracytoplasmic i roto i ngā pūtau kararehe hei hua o te kohikohinga wai i roto i te cytoplasm nā te rerenga mai o Ca2+ me te wai53. He mea whakamīharo, ka taea te whakahurihuri i tēnei tikanga o te kino o te pūtau mena he wā poto te kino, ā, ka tīmata ngā pūtau ki te whakaputa ATP mō tētahi wā, engari ki te mau tonu te kino, ki te kino rānei, ka mate ngā pūtau.54 E whakaatu ana ā mātou kitenga kāore e taea e ngā nematodes i rongoatia ki te 5-iodoindole te whakahoki mai i te biosynthesis noa i muri i te pānga ki ngā āhuatanga ahotea.
Ko te āhua methylation i puta mai i te 5-iodoindole i roto i te B. xylophilus, tērā pea nā te aroaro o te iodine me tōna tohatoha ngota, nā te mea he iti ake te pānga aukati o te 7-iodoindole ki te B. xylophilus i te 5-iodoindole (Ripanga 1 me te Pikitia Tāpiri S6). He rite tonu ēnei hua ki ngā rangahau a Maltese et al. (2014), nāna i kōrero ko te nekehanga o te wāhanga hauota pyridyl i roto i te indole mai i te para- ki te meta-tūnga i whakakore i te vacuolization, te aukati tipu, me te cytotoxicity i roto i ngā pūtau U251, e kī ana he mea nui te taunekeneke o te ngota me tētahi pae hohe motuhake i roto i te pūmua27,44,45. Ko ngā taunekeneke i waenga i ngā indole indole, i ngā indole halogenated rānei me ngā kaiwhakawhiwhi GluCL i kitea i roto i tēnei rangahau e tautoko ana hoki i tēnei whakaaro, nā te mea i kitea he kaha ake te here o te 5- me te 2-iodoindole ki ngā kaiwhakawhiwhi GluCL i ngā indole kē atu i tirotirohia (Pikitia 6 me te Pikitia Tāpiri S8). I kitea te iodine i te tuarua, i te tuarima rānei o te indole e here ana ki te leucine 218 o te kaiwhakawhiwhi GluCL mā roto i ngā here hauwai tuara, ko ētahi atu indole halogenated me te indole tonu e hanga ana i ngā here hauwai taha-taha ngoikore ki te serine 260 (Pikitia 6). Nō reira, e whakapae ana mātou he mea nui te wāhi o te halogen ki te whakaoho i te pirau vacuolar, ko te here pūmau o te 5-iodoindole e pupuri ana i te hongere ion kia tuwhera, kia tere ai te rere o te wai me te pakaru o te vacuole. Heoi, kāore anō kia whakatauhia te tikanga mahi taipitopito o te 5-iodoindole.
I mua i te whakamahinga o te 5-iodoindole, me tātarihia tōna pānga paitini ki ngā tipu. I whakaatuhia e ā mātou whakamātautau whakatipu purapura kāore he pānga kino o te 5-iodoindole ki te whakatipu purapura, ki ngā tukanga whanaketanga rānei i muri mai i ngā kukū i akohia (Pikitia 7). Nō reira, ka whakaratohia e tēnei rangahau he pūtake mō te whakamahinga o te 5-iodoindole i roto i te taiao taiao hei whakahaere i te kino o ngā nematode paina ki ngā rākau paina.
Kua whakaatuhia e ngā pūrongo o mua ko te rongoā indole he huarahi pea hei whakatika i te raruraru o te ātete ki ngā paturopi me te whanaketanga o te mate pukupuku55. Hei tāpiri, he mahi patu huakita, patu mate pukupuku, ārai matū, ārai mumura, ārai mate huka, ārai huaketo, ārai tipu me te ārai mate kohi kei roto i ngā indole, ā, ka taea te whakamahi hei pūtake whai hua mō te whakawhanaketanga rongoā56,57. E kī ana tēnei rangahau mō te wā tuatahi i te whakamahinga pea o te iodine hei rongoā patu parataiao me te rongoā anthelmintic.
I kitea te Avermectin i ngā tekau tau e toru kua pahure ake nei, ā, i toa i te Tohu Nobel i te tau 2015, ā, kei te haere tonu te whakamahinga hei rongoā ārai i ngā pepeke. Heoi, nā te tere o te whanaketanga o te ātete ki ngā avermectin i roto i ngā nematode me ngā pepeke riha, me whai rautaki rerekē, iti te utu, me te pai ki te taiao hei whakahaere i te mate PWN i roto i ngā rākau paina. E pūrongo ana hoki tēnei rangahau i te tikanga e patu ai te 5-iodoindole i ngā nematode paina, ā, he iti te paitini o te 5-iodoindole ki ngā pūtau tipu, e whakatuwhera ana i ngā tūmanako pai mō tana tono arumoni ā muri ake nei.
I whakaaetia ngā whakamātautau katoa e te Komiti Matatika o Te Whare Wānanga o Yeungnam, Gyeongsan, Korea, ā, i mahia ngā tikanga i runga anō i ngā aratohu a te Komiti Matatika o Te Whare Wānanga o Yeungnam.
I whakahaerehia ngā whakamātautau whakatipu hua manu mā te whakamahi i ngā tikanga kua whakatūria43. Hei aromatawai i ngā tere pao (HR), i whakawhitia ngā nematode pakeke 1-rā te pakeke (tata ki te 100 uwha me te 100 tāne) ki ngā rihi Petri kei roto te harore, ā, i tukuna kia tupu mō te 24 hāora. Kātahi ka wehea ngā hua manu, ā, ka rongoatia ki te 5-iodoindole (0.05 mM me te 0.1 mM) te avermectin rānei (10 μg/ml) hei wai whakatārewatanga i roto i te wai purehurehu parakore. I whakawhitia ēnei wai whakatārewatanga (500 μl; tata ki te 100 hua manu) ki ngā poka o tētahi pereti ahurea kiko 24-poka, ā, i whakatārewatia ki te 22 °C. I tatauhia ngā L2 i muri i te 24 hāora o te whakatārewatanga engari i kiia kua mate mēnā kāore ngā pūtau i neke i te wā i whakaohokia ki te waea konukawata pai. I whakahaerehia tēnei whakamātautau i roto i ngā wāhanga e rua, ia wāhanga me ngā tukuruatanga e ono. I whakakotahihia ngā raraunga mai i ngā whakamātautau e rua, ā, i whakaaturia. Ka tatauhia te ōrau o te HR penei:
I aromatawaihia te mate o ngā torongū mā te whakamahi i ngā tikanga kua whakawhanakehia i mua. I kohia ngā hua nematode, ā, i tukuna ngā kukune mā te whakatō ki roto i te wai whakapūmau hei whakaputa i ngā torongū wāhanga L2. I rongoatia ngā torongū tukutahi (tata ki te 500 nematode) ki te 5-iodoindole (0.05 mM me te 0.1 mM) te avermectin rānei (10 μg/ml) ā, i whakatipuria ki runga i ngā pereti Petri B. cinerea. I muri i te 48 hāora o te whakatipu i te 22 °C, i kohia ngā nematode ki roto i te wai whakapūmau, ā, i tirotirohia mō te aroaro o ngā wāhanga L2, L3, me te L4. Ko te aroaro o ngā wāhanga L3 me te L4 i tohu i te huringa torongū, engari ko te aroaro o te wāhanga L2 i tohu kāore he huringa. I kohia ngā whakaahua mā te whakamahi i te Pūnaha Whakaahua Pūtau Mamati iRiS™. I whakahaerehia tēnei whakamātautau i roto i ngā wāhanga e rua, ia wāhanga me ngā tukuruatanga e ono. I whakakotahihia ngā raraunga mai i ngā whakamātautau e rua, ā, i whakaaturia.
I aromatawaihia te paitini o te 5-iodoindole me te avermectin ki ngā purapura mā te whakamahi i ngā whakamātautau whakatipu i runga i ngā pereti agar Murashige me Skoog.62 I rumakina tuatahitia ngā purapura B. oleracea me R. raphanistrum ki te wai whakapūmau mō te rā kotahi, i horoia ki te 1 ml 100% etanoru, i whakapūmau ki te 1 ml 50% wai māori (3% konutai hypochlorite) mō te 15 meneti, ā, i horoia kia rima ngā wā ki te 1 ml wai whakapūmau. Kātahi ka pēhia ngā purapura kua whakapūmau ki runga i ngā pereti agar whakatipu kei roto ko te 0.86 g/l (0.2X) te reo Murashige me Skoog me te 0.7% agar bacteriological me te 5-iodoindole, avermectin rānei, me te kore rānei. Kātahi ka whakatōngia ngā pereti ki te 22 °C, ā, i tangohia ngā whakaahua i muri i te 3 rā o te whakatōngia. I whakahaerehia tēnei whakamātautau i roto i ngā wāhanga e rua, ia wāhanga e ono ngā tukuruatanga.
Te wā tuku: 26-Pepuere-2025