Muchidzidzo ichi, mhedzisiro yekurapa pamwe chete kwevanodzora kukura kwezvirimwa(2,4-D uye kinetin) uye iron oxide nanoparticles (Fe₃O₄-NPs) pa in vitro morphogenesis uye kugadzirwa kwemetabolite yechipiri mu *Hypericum perforatum* L. zvakaongororwa. Kurapa kwakagadziridzwa [2,4-D (0.5 mg/L) + kinetin (2 mg/L) + Fe₃O₄-NPs (4 mg/L)] kwakavandudza zvakanyanya kukura kwemiti: kureba kwemiti kwakawedzera ne59.6%, kureba kwemidzi ne114.0%, nhamba yemabukira ne180.0%, uye huremu hutsva hwe callus ne198.3% zvichienzaniswa neboka rekutonga. Kurapa kwakabatana uku kwakasimbisawo kushanda kwekuvandudzwa (50.85%) uye kwakawedzera huwandu hwe hypericin ne66.6%. Ongororo yeGC-MS yakaratidza huwandu hwakawanda hwe hyperoside, β-patholene, uye cetyl alcohol, zvichiita 93.36% yenzvimbo yese yepamusoro, nepo huwandu hwe phenolics ne flavonoids hwakawedzera kusvika 80.1%. Izvi zvinoratidza kuti zvinodzora kukura kwezvirimwa (PGRs) uye Fe₃O₄ nanoparticles (Fe₃O₄-NPs) zvine simba rekubatana nekukurudzira organogenesis uye kuunganidzwa kwemakemikari anoshanda, izvo zvinomiririra nzira inovimbisa yekuvandudza biotechnological yemiti yemishonga.
Muti weSt. John's wort (Hypericum perforatum L.), unozivikanwawo seSt. John's wort, muti unogara kwenguva refu wemhuri yeHypericaceae une kukosha kwehupfumi.[1] Zvinhu zvawo zvinogona kuita kuti uve nehupenyu zvinosanganisira tannins dzechisikigo, xanthones, phloroglucinol, naphthalenedianthrone (hyperin uye pseudohyperin), flavonoids, phenolic acids, uye mafuta anokosha.[2,3,4] Muti weSt. John's wort unogona kuparadzirwa nenzira dzechinyakare; zvisinei, mwaka wenzira dzechinyakare, kumera kwembeu shoma, uye kubatwa nezvirwere zvinoderedza mukana wayo wekurima kwakakura uye kuumbwa kwemetabolites dzechipiri nguva dzose.[1,5,6]
Saka, kumera kwezvinyama mu vitro kunoonekwa senzira inoshanda yekukurumidzira kupararira kwezvirimwa, kuchengetedza zviwanikwa zve germplasm, uye kuwedzera goho remishonga [7, 8]. Zvinodzora kukura kwezvirimwa (PGRs) zvinoita basa rakakosha mukutonga morphogenesis uye zvinodiwa pakurima callus nezvisikwa zvese mu vitro. Kugadzirisa kuwanda kwavo nekusanganiswa kwavo kwakakosha kuti maitiro aya ekukura abudirire [9]. Nokudaro, kunzwisisa kuumbwa kwakakodzera uye kuwanda kwezvinodzora kwakakosha pakuvandudza kukura nekugona kudzorerazve kweSt. John's wort (H. perforatum) [10].
Zvidimbu zvidiki zvesimbi oxide (Fe₃O₄) imhando yezvikamu zvidiki zvakambogadzirwa kana kuti zviri kugadzirwa kuti zvikure nyama. Fe₃O₄ ine simba guru remagineti, inokwanisa kugara zvakanaka mumuviri, uye inokwanisa kukurudzira kukura kwezvirimwa nekuderedza kushushikana kwezvakatipoteredza, saka yakakwezva kutariswa kukuru mumagadzirirwo ekugadzira nyama. Mashandisirwo angangoitwa nezvikamu izvi zvinogona kusanganisira kugadzirisa tsika ye in vitro kukurudzira kupatsanurwa kwemasero, kuvandudza kutorwa kwezvinovaka muviri, uye kumutsa ma enzymes e antioxidant [11].
Kunyangwe hazvo nanoparticles dzakaratidza mhedzisiro yakanaka pakukura kwezvirimwa, zvidzidzo pakushandiswa pamwe chete kweFe₃O₄ nanoparticles uye optimized plant growth controllers mu *H. perforatum* zvichiri kushomeka. Kuti zvizadze musiyano uyu weruzivo, chidzidzo ichi chakaongorora mhedzisiro yemigumisiro yazvo yakabatana pa in vitro morphogenesis uye kugadzirwa kwechipiri kwemetabolite kuti zvipe ruzivo rutsva rwekuvandudza hunhu hwemiti yemishonga. Nokudaro, chidzidzo ichi chine zvinangwa zviviri: (1) kugadzirisa huwandu hwezvinodzora kukura kwemiti kuti zvikurudzire zvinobudirira kuumbwa kwe callus, kudzoreredza nhunzi, uye kudzika midzi mu vitro; uye (2) kuongorora mhedzisiro yeFe₃O₄ nanoparticles pamatanho ekukura mu vitro. Zvirongwa zveramangwana zvinosanganisira kuongorora mwero wekurarama kwemiti yakadzokororwa panguva yekujairana (in vitro). Zvinotarisirwa kuti mhedzisiro yechidzidzo ichi ichavandudza zvakanyanya kushanda kwe *H. perforatum* micropropagation, nokudaro zvichibatsira mukushandiswa kwakasimba uye kushandiswa kwebiotechnology kwemuti uyu unokosha wemishonga.
Muchidzidzo ichi, takawana zvirimwa zvemashizha zvakabva kumiti yeSt. John's wort (zvirimwa zvepagore zveSt. John's wort). Izvi zvakashandiswa kugadzirisa mamiriro ekurima muvhu. Tisati tagadzira, mashizha acho akasukurudzwa zvakanaka nemvura yakasanganiswa nemvura inoyerera kwemaminitsi akati wandei. Nzvimbo dzakasanganiswa dzakazobviswa mushonga wekuuraya utachiona nekunyudzwa mu70% ethanol kwemasekonzi makumi matatu, zvichiteverwa nekunyudzwa mu1.5% sodium hypochlorite (NaOCl) solution ine madonhwe mashoma eTween 20 kwemaminitsi gumi. Pakupedzisira, zvirimwa zvakasanganiswa nemvura yakasanganiswa nemvura yakasanganiswa nemvura isina tsvina zvisati zvaendeswa kune imwe nzira yekukudziridza.
Mumavhiki mana akatevera, zviyero zvekudzorerwa kwemuti zvakayerwa, kusanganisira mwero wekudzorerwa kwemuti, huwandu hwemuti pamuti wega wega, uye urefu hwemuti. Kana muti wega wega wasvika pakureba kwe2 cm, wakaendeswa kunzvimbo ine hafu yesimba reMS medium, 0.5 mg/L indolebutyric acid (IBA), uye 0.3% guar gum. Kukura kwemuti kwakaramba kuripo kwemavhiki matatu, panguva iyo mwero wemidzi, huwandu hwemuti, uye urefu hwemuti zvakayerwa. Kurapa kwega kwega kwakadzokororwa katatu, nemiti gumi yakakuriswa pamuti wega wega, zvichibereka miti inosvika makumi matatu pamuti wega wega.
Kureba kwemuti kwakayerwa nemasendimita (cm) uchishandisa ruler, kubva pasi pemuti kusvika pamucheto weshizha refu. Kureba kwemidzi kwakayerwa nemamilimita (mm) mushure mekubvisa mbesa nokungwarira uye kubvisa nzvimbo inokura. Huwandu hwemabukira pamuti wega wega hwakaverengwa zvakananga pamuti wega wega. Huwandu hwemapundu matema pamashizha, anozivikanwa semapundu, hwakayerwa nemaziso. Mapundu matema aya anofungidzirwa kuti matumbu ane hypericin, kana kuti mapundu e oxidative, uye anoshandiswa sechiratidzo chemuviri chemhinduro yemuti pakurapwa. Mushure mekubvisa nzvimbo yese inokura, huremu hutsva hwembesa hwakayerwa uchishandisa chikero chemagetsi chine kururama kwe milligrams (mg).
Nzira yekuverenga mwero wekuumbwa kwe callus ndeiyi inotevera: mushure mekurimisa explants mune imwe nzvimbo ine akasiyana-siyana ekukura (kinases, 2,4-D, uye Fe3O4) kwemavhiki mana, huwandu hwe explants dzinokwanisa kugadzira callus hunoverengwa. Fomura yekuverenga mwero wekuumbwa kwe callus ndeiyi inotevera:
Kurapa kwega kwega kwakadzokororwa katatu, uye kanenge ka10 kakaongororwa pakudzokorora kwega kwega.
Mwero wekugadzurwa kwembeu unoratidza huwandu hwenyama yeganda inopedzisa zvinobudirira maitiro ekusiyana kwemabukira mushure mekuumbwa kweganda. Ichi chiratidzo chinoratidza kugona kwenyama yeganda kushanduka kuita nyama yakasiyana uye kukura kuita nhengo itsva dzemiti.
Kuwanda kwemidzi kunoratidza huwandu hwematavi anokwanisa kudzika midzi kusvika kuhuwandu hwematavi ese. Chiratidzo ichi chinoratidza kubudirira kwedanho remidzi, iro rakakosha pakupararira kwemiti midiki uye kukura kwemiti, sezvo midzi yakanaka ichibatsira mbesa kurarama zviri nani mumamiriro ekukura.
Mishonga yeHypericin yakabviswa ne90% methanol. Makumi mashanu mg emiti yakaomeswa yakawedzerwa ku1 ml yemethanol uye yakasanganiswa kwemaminitsi makumi maviri pa30 kHz mumuchina wekuchenesa we ultrasonic (model A5120-3YJ) patembiricha yemukamuri murima. Mushure mekushandiswa kwe sonication, sampuro yakatenderedzwa pa6000 rpm kwemaminitsi gumi nemashanu. Supernatant yakaunganidzwa, uye kunyudzwa kwe hypericin kwakayerwa pa592 nm uchishandisa Plus-3000 S spectrophotometer zvichienderana nenzira yakatsanangurwa naConceiçao et al. [14].
Kurapa kwakawanda nezvinogadzirisa kukura kwezvirimwa (PGRs) uye iron oxide nanoparticles (Fe₃O₄-NPs) hakuna kukonzera kuumbwa kwemapundu matema pamashizha emuti akadzorerwa. Hapana mapundu akaonekwa mune chero kurapwa ne 0.5 kana 1 mg/L 2,4-D, 0.5 kana 1 mg/L kinetin, kana 1, 2, kana 4 mg/L iron oxide nanoparticles. Kusanganiswa kushoma kwakaratidza kuwedzera kudiki mukukura kwemapundu (asi kwete kukosha kwehuwandu) pahuwandu hwakanyanya hwekinetin uye/kana iron oxide nanoparticles, senge kusanganiswa kwe 2,4-D (0.5–2 mg/L) nekinetin (1–1.5 mg/L) uye iron oxide nanoparticles (2–4 mg/L). Mhedzisiro iyi inoratidzwa muMufananidzo 2. Mapundu matema anomiririra matumbu ane hypericin yakawanda, ese ari echisikigo uye anobatsira. Muchidzidzo ichi, mapundu matema ainyanya kusanganiswa nekusviba kwematishu, zvichiratidza nzvimbo yakanaka yekuunganidzwa kwehypericin. Kurapa ne2,4-D, kinetin, uye Fe₃O₄ nanoparticles kwakasimudzira kukura kwe callus, kuderedza browning, uye kuwedzera kwe chlorophyll content, zvichiratidza kuvandudzika kwebasa re metabolic uye kuderedzwa kwekukuvadzwa kwe oxidative [37]. Chidzidzo ichi chakaongorora mhedzisiro ye kinetin pamwe chete ne 2,4-D uye Fe₃O₄ nanoparticles pakukura nekukura kwe St. John's wort callus (Mufananidzo 3a–g). Zvidzidzo zvekare zvakaratidza kuti Fe₃O₄ nanoparticles dzine mabasa e fungal uye antimicrobial [38, 39] uye, kana ikashandiswa pamwe chete ne plant growth regulators, inogona kukurudzira nzira dzekudzivirira zvirimwa uye kuderedza cellular stress indices [18]. Kunyangwe biosynthesis ye secondary metabolites ichidzorwa nemajini, goho rayo chairo rinoenderana zvikuru nemamiriro ekunze. Kuchinja kwe metabolic uye morphological kunogona kukanganisa secondary metabolite levels nekudzora kuratidzwa kwemajini chaiwo emiti uye kupindura kune zvakatipoteredza. Uyezve, zvinhu zvinokurudzira majini matsva zvinogona kukonzera kushanda kwemajini matsva, izvo zvinozokurudzira kushanda kwema enzyme, zvichizopedzisira zvaita kuti nzira dzakawanda dze biosynthetic dzishande uye zvichitungamira pakuumbwa kwema secondary metabolites. Uyezve, imwe ongororo yakaratidza kuti kuderedza mumvuri kunowedzera kuratidzwa kwezuva, nokudaro zvichikwidza tembiricha yemasikati munzvimbo yechisikigo ye *Hypericum perforatum*, iyo inobatsirawo mukuwedzera kwe hypericin goho. Zvichibva padata iri, ongororo iyi yakaongorora basa re iron nanoparticles sezvingangoita zvinokurudzira mukugadzirwa kwetishu. Mhedzisiro yacho yakaratidza kuti idzi nanoparticles dzinogona kumutsa majini anobatanidzwa mu hesperidin biosynthesis kuburikidza ne enzymatic stimulation, zvichikonzera kuwedzera kuunganidzwa kweiyi compound (Mufananidzo 2). Naizvozvo, zvichienzaniswa nemiti inokura mumamiriro echisikigo, zvinogona kutaurwa kuti kugadzirwa kwema composites akadaro mumuviri kunogonawo kuwedzerwa kana kushushikana kuri pakati nepakati kuchibatanidzwa nekushanda kwemajini anobatanidzwa mu biosynthesis yema secondary metabolites. Kurapa kwakabatana kunowanzova nemhedzisiro yakanaka pakukura kwekuvandudzwa, asi mune dzimwe nguva, mhedzisiro iyi inodzikira. Zvinonyanya kukosha, kurapwa ne1 mg/L 2,4-D, 1.5 mg/L kinase, uye huwandu hwakasiyana hwemasero zvinogona kuwedzera mwero wekuvandudzwa kwemasero ne50.85% zvichienzaniswa neboka rekutonga (Mufananidzo 4c). Izvi zvinoratidza kuti kusanganiswa kwemahormone akasiyana kunogona kushanda pamwe chete kukurudzira kukura kwezvirimwa uye kugadzirwa kwemetabolite, izvo zvakakosha zvikuru pakuumbwa kwematishu emiti yemishonga. Palmer naKeller [50] vakaratidza kuti kurapwa kwe2,4-D kunogona kukonzera kuumbwa kwecallus muSt. perforatum, ukuwo kuwedzerwa kwekinase kuchivandudza zvikuru kuumbwa kwecallus. Izvi zvakakonzerwa nekuvandudzwa kwemazinga emahormone uye kukurudzira kupatsanurwa kwemasero. Bal et al. [51] vakaona kuti kurapwa kweFe₃O₄-NP kunogona kuwedzera kushanda kwema enzymes e antioxidant, nokudaro kuchikurudzira kukura kwemidzi muSt. perforatum. Midziyo yekurima ine Fe₃O₄ nanoparticles pahuwandu hwe0.5 mg/L, 1 mg/L, uye 1.5 mg/L yakavandudza huwandu hwekugadziriswazve kwemiti yeflax [52]. Kushandiswa kwekinetin, 2,4-dichlorobenzothiazolinone, uye Fe₃O₄ nanoparticles kwakavandudza zvakanyanya huwandu hwekuumbwa kwecallus nemidzi, zvisinei, mhedzisiro inogona kuitika yekushandisa mahormone aya pakuvandudzwa kwe in vitro inofanirwa kufungwa nezvayo. Semuenzaniso, kushandiswa kwenguva refu kana kwakanyanya kwe2,4-dichlorobenzothiazolinone kana kinetin kunogona kukonzera shanduko ye somatic clonal, oxidative stress, abnormal callus morphology, kana vitrification. Nokudaro, huwandu hwakawanda hwekugadziriswazve hahurevi kugadzikana kwemajini. Miti yese yakagadziriswazve inofanira kuongororwa uchishandisa mamorekuru (semuenzaniso RAPD, ISSR, AFLP) kana cytogenetic analysis kuti ione homogeneity uye kufanana kwayo nemiti ye in vivo [53,54,55].
Chidzidzo ichi chakaratidza kekutanga kuti kushandiswa pamwe chete kwezvinodzora kukura kwezvirimwa (2,4-D uye kinetin) neFe₃O₄ nanoparticles kunogona kuwedzera morphogenesis uye kuunganidzwa kwe key bioactive metabolites (kusanganisira hypericin uye hyperoside) mu *Hypericum perforatum*. Nzira yekurapa yakagadziriswa (1 mg/L 2,4-D + 1 mg/L kinetin + 4 mg/L Fe₃O₄-NPs) haina kungowedzera kuumbwa kwe callus, organogenesis, uye secondary metabolite goho asi yakaratidzawo mhedzisiro inokurudzira zvishoma, zvichigona kuvandudza kushivirira kwechirimwa kushushikana uye kukosha kwemishonga. Kubatanidzwa kwe nanotechnology ne plant tissue culture kunopa puratifomu inogara kwenguva refu uye inoshanda yekugadzira mishonga yakawanda mu vitro. Mhedzisiro iyi inovhura nzira yekushandiswa kwemaindasitiri uye kutsvagurudza mune ramangwana muma molecular mechanisms, kugadzirisa dosage uye genetic usahihi, nokudaro ichibatanidza kutsvagurudza kwakakosha pamiti yemishonga ne biotechnology inoshanda.
Nguva yekutumira: Zvita-12-2025