I roto i tēnei rangahau, ko ngā pānga whakaohooho o te maimoatanga whakakotahi ongā kaiwhakahaere tipu tipu(2,4-D me te kinetin) me ngā matūriki rino waikura (Fe₃O₄-NPs) mō te morphogenesis in vitro me te whakaputanga metabolite tuarua i roto i te *Hypericum perforatum* L. I tirotirohia te maimoatanga kua arotauhia [2,4-D (0.5 mg/L) + kinetin (2 mg/L) + Fe₃O₄-NPs (4 mg/L)] i tino whakapai ake i ngā tawhā tipu tipu: i piki ake te teitei o te tipu mā te 59.6%, te roa o te pakiaka mā te 114.0%, te maha o ngā puku mā te 180.0%, me te taumaha hou o te karu mā te 198.3% ki te whakataurite ki te rōpū whakahaere. I whakarei ake hoki tēnei maimoatanga whakakotahi i te whai huatanga o te whakaoranga (50.85%) me te whakanui ake i te ihirangi hypericin mā te 66.6%. I whakaatuhia e te tātaritanga GC-MS he nui te ihirangi o te hyperoside, te β-patholene, me te cetyl alcohol, e 93.36% o te horahanga tihi katoa, ā, ko te ihirangi o ngā phenolics me ngā flavonoids katoa i piki ake tae atu ki te 80.1%. E whakaatu ana ēnei hua he pānga tahi ngā kaiwhakahaere tipu tipu (PGR) me ngā nanoparticles Fe₃O₄ (Fe₃O₄-NP) mā te whakaoho i te organogenesis me te kohikohi o ngā pūhui bioactive, e tohu ana i tētahi rautaki whai hua mō te whakapai ake i te hangarau koiora o ngā tipu rongoā.
Ko te wort o Hato Hoani (Hypericum perforatum L.), e mōhiotia ana ko te wort o Hato Hoani, he tipu otaota ā-tau o te whānau Hypericaceae he uara ōhanga.[1] Ko ōna wāhanga koiora pea ko ngā tannins taiao, ngā xanthones, te phloroglucinol, te naphthalenedianthrone (hyperin me te pseudohyperin), ngā flavonoids, ngā waikawa phenolic, me ngā hinu kakara.[2,3,4] Ka taea te whakatipu i te wort o Hato Hoani mā ngā tikanga tuku iho; heoi, ko te ā-wā o ngā tikanga tuku iho, te iti o te tipu o te purapura, me te ngāwari ki ngā mate ka whakawhāiti i tōna kaha ki te whakatipu nui me te hanga tonu o ngā matū tuarua.[1,5,6]
Nō reira, e kiia ana te whakatipu kiko in vitro he tikanga whai hua mō te whakatipu tere i ngā tipu, te tiaki i ngā rauemi germplasm, me te whakapiki ake i te hua o ngā matū rongoā [7, 8]. He mahi nui tā ngā kaiwhakahaere tipu tipu (PGR) ki te whakahaere i te morphogenesis, ā, he mea tika mō te whakatipu in vitro o te callus me ngā rauropi katoa. He mea nui te arotau i ō rātou kukū me ngā huinga mō te otinga angitu o ēnei tukanga whanaketanga [9]. Nō reira, he mea nui te mārama ki te hanganga tika me te kukū o ngā kaiwhakahaere hei whakapai ake i te tipu me te kaha whakaora o te wort St. John (H. perforatum) [10].
Ko ngā matūriki rino waikura (Fe₃O₄) he momo matūriki kua whakawhanakehia, kei te whakawhanakehia rānei mō te whakatipu kiko. He nui ngā āhuatanga aukume o te Fe₃O₄, he pai te hototahitanga ki te koiora, me te kaha ki te whakatairanga i te tipu o te tipu me te whakaiti i te ahotea taiao, nō reira kua aro nui ki ngā hoahoa whakatipu kiko. Ko ngā tono pea o ēnei matūriki ko te arotau i te ahurea in vitro hei whakatairanga i te wehenga pūtau, te whakapai ake i te mimiti matūkai, me te whakahohe i ngā whākōkī ārai mate [11].
Ahakoa kua whakaatuhia e ngā matūriki nano he pānga whakatairanga pai ki te tipu o ngā tipu, kei te onge tonu ngā rangahau mō te whakamahinga ngātahi o ngā matūriki nano Fe₃O₄ me ngā kaiwhakahaere tipu tipu kua arotauhia i roto i te *H. perforatum*. Hei whakakī i tēnei āputa mōhiotanga, i arotakehia e tēnei rangahau ngā pānga o ō rātou pānga ngātahi ki te morphogenesis in vitro me te whakaputanga metabolite tuarua hei whakarato i ngā māramatanga hou mō te whakapai ake i ngā āhuatanga o ngā tipu rongoā. Nō reira, e rua ngā whāinga o tēnei rangahau: (1) te arotau i te kukū o ngā kaiwhakahaere tipu tipu hei whakatairanga i te hanganga callus, te whakaoranga wana, me te pakiaka i roto i te vitro; me te (2) te aromatawai i ngā pānga o ngā matūriki nano Fe₃O₄ ki ngā tawhā tipu i roto i te vitro. Ko ngā mahere ā muri ake nei ko te aromatawai i te tere ora o ngā tipu kua whakahoutia i te wā o te whakaōrite (in vitro). E matapaetia ana ka whakapai ake ngā hua o tēnei rangahau i te whai huatanga o te horapa iti o *H. perforatum*, ā, ka whai wāhi ki te whakamahinga pumau me ngā tono hangarau koiora o tēnei tipu rongoā nui.
I roto i tēnei rangahau, i tangohia mai i a mātou ngā tipu rau i whakatipuria i te mara, arā, te wort St. John's wort (ngā tipu whaea). I whakamahia ēnei tipu hei whakapai ake i ngā āhuatanga whakatipu in vitro. I mua i te whakatipu, i horoia pai ngā rau i raro i te wai whakapūmau e rere ana mō ētahi meneti. Kātahi ka horoia ngā mata o ngā tipu mā te rumaki ki roto i te 70% etanoru mō te 30 hēkona, ā, i muri mai ka rumaki ki roto i te otinga 1.5% konutai hypochlorite (NaOCl) kei roto ētahi pata o Tween 20 mō te 10 meneti. Hei whakamutunga, i horoia ngā tipu e toru ngā wā ki te wai whakapūmau parakore i mua i te whakawhiti ki te ipu whakatipu e whai ake nei.
I roto i ngā wiki e whā e whai ake nei, i inehia ngā tawhā whakaora wana, tae atu ki te tere whakaora, te maha o ngā wana mō ia tipu whakatō, me te roa o ngā wana. Ina tae ngā wana kua whakatipuria ki te roa o te iti rawa o te 2 cm, ka whakawhitia ki tētahi ranunga pakiaka kei roto ko te ranunga MS haurua-kaha, 0.5 mg/L waikawa indolebutyric (IBA), me te 0.3% kapia guar. I haere tonu te whakatipu pakiaka mō ngā wiki e toru, ā, i roto i taua wā ka inehia te tere pakiaka, te maha o ngā pakiaka, me te roa o te pakiaka. E toru ngā wā i whakahokia ai ia maimoatanga, me te 10 ngā tipu whakatō i whakatipuria mō ia tāruarua, ā, tata ki te 30 ngā tipu whakatō i puta mō ia maimoatanga.
I inehia te teitei o te tipu i roto i ngā henimita (cm) mā te whakamahi i te rūrū, mai i te pūtake o te tipu ki te pito o te rau teitei rawa. I inehia te roa o te pakiaka i roto i ngā mirimita (mm) i muri tonu i te tangohanga āta i ngā tipu me te tango i te oneone tipu. I tatauhia tika te maha o ngā puku mō ia tipu i runga i ia tipu. I inehia ā-kanohi te maha o ngā ira pango i runga i ngā rau, e mōhiotia ana ko ngā nodules. E whakaponohia ana ko ēnei nodules pango he repe kei roto te hypericin, ngā ira oxidative rānei, ā, e whakamahia ana hei tohu ā-tinana mō te urupare a te tipu ki te maimoatanga. I muri i te tangohanga o te oneone tipu katoa, i inehia te taumaha hou o ngā tipu mā te whakamahi i te tauine hiko me te tika o ngā mirikaramu (mg).
Ko te tikanga mō te tatau i te tere o te hanganga o te karu: i muri i te whakatipu i ngā tipu i roto i tētahi reo kei roto ngā momo kaiwhakahaere tipu (kinase, 2,4-D, me Fe3O4) mō ngā wiki e whā, ka tatauhia te maha o ngā tipu ka taea te hanga karu. Ko te tātai mō te tatau i te tere o te hanganga o te karu:
E toru ngā tukuruatanga o ia maimoatanga, me te iti rawa kia 10 ngā explant i tirohia i ia tukuruatanga.
Ko te tere o te whakaoranga e whakaata ana i te ōwehenga o te kiko karu e whakaoti angitu ana i te tukanga whakaputa puku i muri i te wāhanga hanganga karu. E whakaatu ana tēnei tohu i te kaha o te kiko karu ki te huri hei kiko kua rerekē, ā, ka tipu hei okana tipu hou.
Ko te tauwehenga pakiaka te ōwehenga o te maha o ngā manga ka taea te pakiaka ki te maha katoa o ngā manga. E whakaatu ana tēnei tohu i te angitu o te wāhanga pakiaka, he mea nui tēnei i roto i te whakatipu moroiti me te whakatipu tipu, nā te mea ka āwhina te pakiaka pai i ngā tipu kia ora ake i roto i ngā āhuatanga tipu.
I tangohia ngā matū hypericin ki te 90% methanol. E rima tekau mg o ngā tipu maroke i tāpirihia ki te 1 ml o te methanol, ā, i whakamātauria mō te 20 meneti i te 30 kHz i roto i te horoi ultrasonic (tauira A5120-3YJ) i te pāmahana rūma i te pōuri. I muri i te whakamātauria, i whakatōngia te tauira i te 6000 rpm mō te 15 meneti. I kohia te wai i roto i te ipu, ā, i inehia te mimiti o te hypericin i te 592 nm mā te whakamahi i te inenga Plus-3000 S e ai ki te tikanga i whakaahuahia e Conceiçao et al. [14].
Ko te nuinga o ngā maimoatanga me ngā kaiwhakahaere tipu tipu (PGR) me ngā matūriki waikura rino (Fe₃O₄-NP) kāore i puta he pona pango ki ngā rau wana kua whakahoutia. Kāore i kitea he pona i roto i tētahi o ngā maimoatanga me te 0.5, 1 mg/L rānei o te 2,4-D, 0.5, 1 mg/L rānei o te kinetin, 1, 2, 4 mg/L rānei o ngā matūriki waikura rino. I whakaatuhia e ētahi huinga he pikinga paku o te whanaketanga o ngā pona (engari kāore i te tino hira i te tatauranga) i ngā kukū teitei ake o te kinetin me/ranei ngā matūriki waikura rino, pērā i te huinga o te 2,4-D (0.5–2 mg/L) me te kinetin (1–1.5 mg/L) me ngā matūriki waikura rino (2–4 mg/L). Kei te whakaatuhia ēnei hua i te Pikitia 2. Ko ngā pona pango he repe momona-hypericin, he mea hanga noa, he mea whai hua hoki. I roto i tēnei rangahau, ko ngā pona pango te nuinga o te wā e pā ana ki te parauri o ngā kiko, e tohu ana i tētahi taiao pai mō te kohikohinga hypericin. Ko te maimoatanga ki ngā matūriki nano 2,4-D, kinetin, me Fe₃O₄ i whakatairanga i te tipu o te callus, te whakaiti i te parauri, me te whakanui ake i te ihirangi chlorophyll, e kī ana he pai ake te mahi pākia me te whakaiti pea i te kino oxidative [37]. I arotakehia e tēnei rangahau ngā pānga o te kinetin i te taha o ngā matūriki nano 2,4-D me Fe₃O₄ ki te tipu me te whanaketanga o te callus wort a St. John (Pikitia 3a–g). Kua whakaatuhia e ngā rangahau o mua he mahi antifungal me antimicrobial ngā matūriki nano Fe₃O₄ [38, 39] ā, ina whakamahia ngātahi me ngā kaiwhakahaere tipu tipu, ka taea te whakaoho i ngā tikanga tiaki tipu me te whakaiti i ngā tohu ahotea pūtau [18]. Ahakoa he mea whakahaere ira te biosynthesis o ngā metabolites tuarua, he tino whakawhirinaki tō rātou hua tuturu ki ngā āhuatanga taiao. Ka taea e ngā huringa pākia me te āhua te awe i ngā taumata metabolite tuarua mā te whakahaere i te whakaaturanga o ngā ira tipu motuhake me te urupare ki ngā āhuatanga taiao. Hei tāpiri atu, ka taea e ngā mea whakaoho te whakaoho i te whakahohenga o ngā ira hou, ka whakaoho i te mahi whākōkī, ā, ka mutu, ka whakahohe i ngā ara biosynthetic maha, ka arahi ki te hanganga o ngā metabolites tuarua. Hei tāpiri atu, i whakaatuhia e tētahi atu rangahau ko te whakaiti i te atarangi ka whakanui ake i te pānga o te rā, ā, ka piki ake te pāmahana o te awatea i te kāinga tūturu o *Hypericum perforatum*, e whai wāhi ana hoki ki te pikinga o te hua hypericin. I runga i ēnei raraunga, i rangahauhia e tēnei rangahau te tūranga o ngā matūriki rino hei mea whakaoho pea i roto i te ahurea kiko. I whakaatuhia e ngā hua ka taea e ēnei matūriki nano te whakahohe i ngā ira e uru ana ki te biosynthesis hesperidin mā te whakaoho enzymatic, ka arahi ki te pikinga o te kohikohinga o tēnei matū (Pikitia 2). Nō reira, ki te whakatauritea ki ngā tipu e tipu ana i raro i ngā āhuatanga tūturu, ka taea te tautohe ka taea hoki te whakanui ake i te hanga o aua matū i roto i te tinana ina honoa te ahotea ngawari me te whakahohenga o ngā ira e uru ana ki te biosynthesis o ngā metabolites tuarua. Ko ngā maimoatanga whakakotahi he pānga pai ki te tere whakaora, engari i ētahi wā, ka ngoikore tēnei pānga. He mea nui, mā te maimoatanga ki te 1 mg/L 2,4-D, 1.5 mg/L kinase, me ngā kukū rerekē ka taea te whakanui ake i te tere whakaora mā te 50.85% ki te whakataurite ki te rōpū whakahaere (Pikitia 4c). E whakaatu ana ēnei hua ka taea e ngā huinga motuhake o ngā nanohormone te mahi tahi hei whakatairanga i te tipu tipu me te whakaputa metabolite, he mea nui tēnei mō te ahurea kiko o ngā tipu rongoā. I whakaatu a Palmer rāua ko Keller [50] ka taea e te maimoatanga 2,4-D te whakaoho motuhake i te hanganga callus i roto i te St. perforatum, i te mea ko te tāpiri o te kinase i whakarei ake i te hanganga callus me te whakaora. Nā te whakapainga o te taurite homoni me te whakaoho i te wehenga pūtau i puta ai tēnei pānga. I kitea e Bal et al. [51] ka taea e te maimoatanga Fe₃O₄-NP te whakarei ake i te mahi a ngā whākōkī antioxidant, ā, mā reira ka whakatairanga i te tipu pakiaka i roto i te St. perforatum. Nā te whakamahinga o te kinetin, 2,4-dichlorobenzothiazolinone, me te 1.5 mg/L i whakapai ake te tere o te whakaoranga o ngā tipu harakeke [52]. Nā te whakamahinga o te kinetin, 2,4-dichlorobenzothiazolinone, me ngā nanoparticles Fe₃O₄ i tino whakapai ake i te tere o te hanganga o te callus me te pakiaka, heoi, me whai whakaaro ki ngā pānga taha pea o te whakamahi i ēnei homoni mō te whakaoranga in vitro. Hei tauira, ko te whakamahinga roa, te whakamahinga nui rānei o te 2,4-dichlorobenzothiazolinone, kinetin rānei, ka hua ake pea he rerekētanga o te somatic clonal, he ahotea oxidative, he āhua callus rerekē, he vitrification rānei. Nō reira, kāore te tere whakaoranga teitei e tohu i te pumau ira. Me aromatawai ngā tipu katoa kua whakahoutia mā te whakamahi i ngā tohu ngota (hei tauira, RAPD, ISSR, AFLP) me te tātari cytogenetic rānei hei whakatau i tō rātou ōritetanga me te rite ki ngā tipu in vivo [53,54,55].
I whakaatuhia e tēnei rangahau mō te wā tuatahi ko te whakamahinga ngātahi o ngā kaiwhakahaere tipu tipu (2,4-D me te kinetin) me ngā matūriki nano Fe₃O₄ ka taea te whakarei ake i te morphogenesis me te kohikohinga o ngā metabolites bioactive matua (tae atu ki te hypericin me te hyperoside) i roto i te *Hypericum perforatum*. Ko te tikanga maimoatanga kua arotauhia (1 mg/L 2,4-D + 1 mg/L kinetin + 4 mg/L Fe₃O₄-NPs) ehara i te mea i whakanui noa i te hanganga callus, te organogenesis, me te hua metabolite tuarua engari i whakaatu hoki i te pānga whakaoho ngawari, ka whakapai ake pea i te manawanui o te tipu ki te ahotea me te uara rongoā. Ko te whakakotahitanga o te nanotechnology me te ahurea kiko tipu ka whakarato i tētahi tūāpapa pumau me te whai hua mō te whakaputa nui i roto i te vitro o ngā matū rongoā. Mā ēnei hua ka whakatuwhera i te ara mō ngā tono ahumahi me ngā rangahau ā-muri ki ngā tikanga ngota, te arotautanga horopeta me te tika o te ira, ā, ka hono i te rangahau taketake mō ngā tipu rongoā ki te hangarau koiora mahi.
Wā tuku: Hakihea-12-2025