N'ọmụmụ ihe a, mmetụta mkpali nke ọgwụgwọ ejikọtara ọnụ nkendị na-achịkwa uto osisiA nyochachara (2,4-D na kinetin) na iron oxide nanoparticles (Fe₃O₄-NPs) na in vitro morphogenesis na mmepụta metabolite nke abụọ na *Hypericum perforatum* L.. Ọgwụgwọ kachasị mma [2,4-D (0.5 mg/L) + kinetin (2 mg/L) + Fe₃O₄-NPs (4 mg/L)] mere ka paramita uto osisi dịkwuo mma nke ukwuu: elu osisi mụbara site na 59.6%, ogologo mgbọrọgwụ site na 114.0%, nọmba osisi site na 180.0%, na ibu ọhụrụ callus site na 198.3% ma e jiri ya tụnyere otu njikwa. Ọgwụgwọ a jikọtara ọnụ mekwara ka arụmọrụ mmeghari ohuru (50.85%) dịkwuo mma ma mụbaa ọdịnaya hypericin site na 66.6%. Nnyocha GC-MS gosiri na hyperoside, β-patholene, na cetyl alcohol dị elu, nke ruru 93.36% nke oke kachasị elu, ebe ọdịnaya nke phenolics na flavonoids mụbara ruo 80.1%. Nsonaazụ ndị a na-egosi na ndị na-achịkwa uto osisi (PGRs) na Fe₃O₄ nanoparticles (Fe₃O₄-NPs) na-enwe mmetụta synergistic site na ịkpalite organogenesis na mkpokọta nke ihe ndị na-arụ ọrụ bioactive, nke na-anọchite anya atụmatụ na-ekwe nkwa maka mmezi biotechnology nke osisi ọgwụ.
St. John's wort (Hypericum perforatum L.), nke a makwaara dị ka St. John's wort, bụ osisi ahịhịa na-eto eto nke ezinụlọ Hypericaceae nke nwere uru akụ na ụba.[1] Ihe ndị nwere ike ime ka ihe ndị dị ndụ pụta ìhè gụnyere tannins eke, xanthones, phloroglucinol, naphthalenedianthrone (hyperin na pseudohyperin), flavonoids, phenolic acids, na mmanụ dị mkpa.[2,3,4] Enwere ike ịgbasa wort St. John site na usoro ọdịnala; agbanyeghị, oge nke usoro ọdịnala, obere mkpụrụ na-eto, na nnabata ọrịa na-ebelata ikike ya maka ịkọ nnukwu ihe na ịmepụta metabolites nke abụọ na-aga n'ihu.[1,5,6]
Ya mere, a na-ewere ọdịbendị anụ ahụ n'ime vitro dị ka ụzọ dị irè maka mgbasa ngwa ngwa nke osisi, nchekwa akụrụngwa germplasm, na mmụba nke mkpụrụ ọgwụ [7, 8]. Ndị na-achịkwa uto osisi (PGRs) na-arụ ọrụ dị oke mkpa n'ịchịkwa morphogenesis ma dị mkpa maka ịkọ callus na ihe ndị dị ndụ n'ime vitro. Imeziwanye mkpokọta ha na njikọta ha dị oke mkpa maka mmezu nke usoro mmepe ndị a nke ọma [9]. Ya mere, ịghọta nhazi na ntinye nke ndị na-achịkwa dị mkpa maka imeziwanye uto na ikike mmeghari nke St. John's wort (H. perforatum) [10].
Nanoparticles Iron oxide (Fe₃O₄) bụ otu n'ime nanoparticles ndị e mepụtara ma ọ bụ na-emepụta maka ọdịbendị anụ ahụ. Fe₃O₄ nwere nnukwu ihe ndọta, ezigbo ndakọrịta bio, na ikike ịkwalite uto osisi na ibelata nrụgide gburugburu ebe obibi, ya mere ọ dọtara uche dị ukwuu na nhazi omenala anụ ahụ. Ojiji nke nanoparticles ndị a nwere ike ịgụnye imeziwanye ọdịbendị in vitro iji kwalite nkewa sel, melite nnabata nri, na ime ka enzymes antioxidant rụọ ọrụ [11].
Ọ bụ ezie na nanoparticles egosila mmetụta dị mma n'ịbawanye uto osisi, ọmụmụ ihe gbasara ojiji nke Fe₃O₄ na ndị na-achịkwa uto osisi ka dị ụkọ. Iji mejupụta oghere ihe ọmụma a, ọmụmụ ihe a tụlere mmetụta nke mmetụta ha jikọtara ọnụ na in vitro morphogenesis na mmepụta metabolite nke abụọ iji nye nghọta ọhụrụ maka imeziwanye njirimara nke osisi ọgwụ. Ya mere, ọmụmụ ihe a nwere ebumnuche abụọ: (1) imeziwanye ntinye nke ndị na-achịkwa uto osisi iji kwalite nhazi callus nke ọma, imeghari oge, na ịgba mgbọrọgwụ na in vitro; na (2) nyochaa mmetụta nke nanoparticles Fe₃O₄ na paramita uto na in vitro. Atụmatụ ndị ga-eme n'ọdịnihu gụnyere inyocha ọnụego ndụ nke osisi ndị a na-emegharị n'oge nnabata (in vitro). A na-atụ anya na nsonaazụ nke ọmụmụ ihe a ga-eme ka arụmọrụ micropropagation nke *H. perforatum* ka mma nke ukwuu, si otú a na-enye aka na ojiji na-adịgide adịgide na ojiji biotechnological nke osisi ọgwụ dị mkpa a.
N'ọmụmụ ihe a, anyị nwetara ihe ọkụkụ akwụkwọ sitere na osisi wort St. John's kwa afọ (osisi nne) a na-akọ n'ubi. E jiri ihe ọkụkụ ndị a mee ka ọnọdụ ọdịbendị in vitro dịkwuo mma. Tupu a kụọ ha, a na-asachapụ akwụkwọ ndị ahụ nke ọma n'ime mmiri a na-agba agba ruo ọtụtụ nkeji. E mesịa, e tinyere ihe ọkụkụ ahụ n'ime ethanol 70% ruo sekọnd 30, wee mikpuo ha n'ime mmiri sodium hypochlorite (NaOCl) 1.5% nke nwere ntakịrị ntapụ Tween 20 maka nkeji 10. N'ikpeazụ, e ji mmiri a na-agba agba sachaa ihe ọkụkụ ndị ahụ ugboro atọ tupu e bufee ha na ihe ọkụkụ ọzọ.
N'ime izu anọ sochirinụ, a tụrụ paramita mmụba nke ịgba égbè, gụnyere ọnụego mmụba, ọnụọgụ nke ịgba égbè kwa mkpụrụ osisi, na ogologo oge ịgba égbè. Mgbe mkpụrụ osisi ndị a gbanyere ọhụrụ ruru ogologo ma ọ dịkarịa ala 2 cm, a na-ebufe ha na usoro mgbọrọgwụ nke nwere ọkara ike MS, 0.5 mg/L indolebutyric acid (IBA), na 0.3% guar gum. Usoro mgbọrọgwụ gara n'ihu ruo izu atọ, n'oge ahụ a tụrụ ọnụego mgbọrọgwụ, nọmba mgbọrọgwụ, na ogologo mgbọrọgwụ. A na-emegharị ọgwụgwọ ọ bụla ugboro atọ, na-akụ mkpụrụ osisi iri kwa mkpụrụ osisi, na-enye ihe dị ka mkpụrụ osisi 30 kwa ọgwụgwọ.
A tụrụ ogologo osisi na sentimita (cm) site na iji ihe na-achịkwa osisi ahụ, site na ntọala osisi ahụ ruo na nsọtụ akwụkwọ kachasị elu. A tụrụ ogologo mgbọrọgwụ na milimita (mm) ozugbo ewepụchara mkpụrụ osisi ahụ nke ọma ma wepụ ihe na-eto eto. A gụrụ ọnụọgụ nke mkpụrụ osisi ahụ kpọmkwem na osisi ọ bụla. A tụrụ ọnụọgụ ntụpọ ojii na akwụkwọ ahụ, nke a maara dị ka nodules, anya. A kwenyere na nodules ojii ndị a bụ gland nwere hypericin, ma ọ bụ ntụpọ oxidative, a na-ejikwa ha dị ka ihe ngosi nke mmeghachi omume osisi ahụ na ọgwụgwọ. Mgbe ewepụsịrị ihe niile na-eto eto, a tụrụ ibu ọhụrụ nke mkpụrụ osisi ahụ site na iji nha eletrọniki na izi ezi nke milligrams (mg).
Usoro e si agbakọ ọnụego mmepụta callus bụ dịka ndị a: mgbe e mechara kụọ ihe ndị e ji emepụta callus n'ime ihe nwere ọtụtụ ihe na-achịkwa uto (kinases, 2,4-D, na Fe3O4) ruo izu anọ, a na-agụ ọnụọgụgụ ihe ndị e ji emepụta callus. Usoro e ji agbakọ ọnụego mmepụta callus bụ ndị a:
A na-emeghachi ọgwụgwọ ọ bụla ugboro atọ, a na-enyocha ma ọ dịkarịa ala ihe iri e ji mee ihe ugboro ugboro ọ bụla.
Ọnụọgụ mmụgharị ahụ na-egosi oke nke anụ ahụ callus nke na-emecha usoro nhazi nke mkpụrụ osisi mgbe usoro nhazi callus gasịrị. Ihe ngosi a na-egosi ikike nke anụ ahụ callus ịgbanwe ka ọ bụrụ anụ ahụ dị iche iche ma too ka ọ bụrụ akụkụ osisi ọhụrụ.
Ọnụọgụ mgbọrọgwụ bụ oke nke ọnụọgụ alaka nwere ike ịgba mgbọrọgwụ na ọnụọgụ alaka niile. Ihe ngosi a na-egosi ihe ịga nke ọma nke ọkwa mgbọrọgwụ, nke dị oke mkpa na obere mgbasa na mgbasa osisi, ebe ọ bụ na ezigbo mgbọrọgwụ na-enyere mkpụrụ osisi aka ịlanarị nke ọma n'ọnọdụ uto.
E jiri 90% methanol wepụta ngwakọta Hypericin. E tinyere 50 mg nke ihe osisi kpọrọ nkụ na 1 ml nke methanol ma tinye sonicates ruo nkeji 20 na 30 kHz na ihe nhicha ultrasonic (ụdị A5120-3YJ) na okpomọkụ ụlọ n'ọchịchịrị. Mgbe sonication gasịrị, e tinyere centrifuge na 6000 rpm maka nkeji 15. E chịkọtara ihe na-asọfe ahụ, a tụọkwa mmịpụta nke hypericin na 592 nm site na iji Plus-3000 S spectrophotometer dịka usoro Conceiçao et al. [14] si kọwaa.
Ọtụtụ ọgwụgwọ e ji ihe ndị na-achịkwa uto osisi (PGRs) na ígwè oxide nanoparticles (Fe₃O₄-NPs) emeghị ka e nwee oghere ojii n'akwụkwọ osisi ndị a gbanyere ọhụrụ. Ahụghị oghere ọ bụla n'ime ọgwụgwọ ọ bụla e ji 0.5 ma ọ bụ 1 mg/L 2,4-D, 0.5 ma ọ bụ 1 mg/L kinetin, ma ọ bụ 1, 2, ma ọ bụ 4 mg/L iron oxide nanoparticles. Nchikọta ole na ole gosiri mmụba dị nta na mmepe nodule (mana ọ bụghị nke dị mkpa n'ọnụ ọgụgụ) na oke mkpokọta kinetin na/ma ọ bụ iron oxide nanoparticles, dị ka njikọta nke 2,4-D (0.5-2 mg/L) na kinetin (1-1.5 mg/L) na iron oxide nanoparticles (2-4 mg/L). E gosiri nsonaazụ ndị a na Foto nke 2. Oghere ojii na-anọchite anya glands bara ụba na hypericin, ma na-eme n'ụzọ ebumpụta ụwa ma baa uru. N'ọmụmụ ihe a, eriri ojii na-ejikọtakarị na agba aja aja nke anụ ahụ, na-egosi gburugburu ebe dị mma maka mkpokọta hypericin. Ọgwụgwọ na 2,4-D, kinetin, na Fe₃O₄ nanoparticles kwalitere uto callus, belata agba aja aja, na mmụba ọdịnaya chlorophyll, na-egosi na ọrụ metabolic ka mma na mbelata mmebi oxidative nwere ike [37]. Ọmụmụ ihe a tụlere mmetụta nke kinetin na njikọta nanoparticles 2,4-D na Fe₃O₄ na uto na mmepe nke St. John's wort callus (Fig. 3a–g). Ọmụmụ ihe ndị gara aga egosila na nanoparticles Fe₃O₄ nwere ọrụ antifungal na antimicrobial [38, 39], ma, mgbe ejiri ya na ndị na-achịkwa uto osisi, nwere ike ịkpalite usoro nchekwa osisi ma belata ihe ngosi nrụgide sel [18]. Ọ bụ ezie na a na-achịkwa biosynthesis nke metabolites nke abụọ, mmepụta ha n'ezie dabere na ọnọdụ gburugburu ebe obibi. Mgbanwe metabolic na morphological nwere ike imetụta ọkwa metabolite nke abụọ site na ịchịkwa ngosipụta nke mkpụrụ ndụ ihe nketa osisi kpọmkwem na ịzaghachi ihe ndị metụtara gburugburu ebe obibi. Ọzọkwa, ihe ndị na-akpali akpali nwere ike ịkpalite mkpali nke mkpụrụ ndụ ihe nketa ọhụrụ, nke na-akpali ọrụ enzymatic, na-eme ka ọtụtụ ụzọ biosynthesis rụọ ọrụ ma na-eduga n'ịmepụta metabolites nke abụọ. Ọzọkwa, nnyocha ọzọ gosiri na ibelata onyinyo na-eme ka ìhè anyanwụ dịkwuo elu, si otú a na-eme ka okpomọkụ ehihie dịkwuo elu na ebe obibi eke nke *Hypericum perforatum*, nke na-enyekwa aka na mmụba nke mkpụrụ hypericin. Dabere na data ndị a, ọmụmụ ihe a nyochachara ọrụ nke nanoparticles ígwè dị ka ihe ndị nwere ike ịkpalite ọdịbendị anụ ahụ. Nsonaazụ gosiri na nanoparticles ndị a nwere ike ime ka mkpụrụ ndụ ihe nketa ndị metụtara na hesperidin biosynthesis site na mkpali enzymatic, na-eduga na mmụba nke mkpokọta nke ihe a (Foto 2). Ya mere, ma e jiri ya tụnyere osisi na-eto n'okpuru ọnọdụ eke, enwere ike ikwu na mmepụta nke ihe ndị dị otú ahụ na vivo nwekwara ike ime ka ọ dịkwuo mma mgbe ejikọtara nrụgide dị ala na mkpali nke mkpụrụ ndụ ihe nketa metụtara na biosynthesis nke metabolites nke abụọ. Ọgwụgwọ njikọta na-enwekarị mmetụta dị mma na ọnụego mmụgharị, mana n'ọnọdụ ụfọdụ, mmetụta a na-ebelata. N'ụzọ dị mkpa, ọgwụgwọ na 1 mg/L 2,4-D, 1.5 mg/L kinase, na njupụta dị iche iche nwere ike ime ka ọnụego mmụgharị dịkwuo elu site na 50.85% n'adabereghị onwe ha ma e jiri ya tụnyere otu njikwa (Foto 4c). Nsonaazụ ndị a na-egosi na njikọta kpọmkwem nke nanohormones nwere ike ịrụ ọrụ ọnụ iji kwalite uto osisi na mmepụta metabolite, nke dị oke mkpa maka ọdịbendị anụ ahụ nke osisi ọgwụ. Palmer na Keller [50] gosiri na ọgwụgwọ 2,4-D nwere ike ịkpalite nhazi callus n'onwe ya na St. perforatum, ebe mgbakwunye nke kinase mere ka nhazi callus na mmụgharị dịkwuo mma. Mmetụta a bụ n'ihi mmụba nke nguzozi homonụ na mkpali nke nkewa sel. Bal et al. [51] chọpụtara na ọgwụgwọ Fe₃O₄-NP nwere ike ime ka ọrụ nke enzymes antioxidant dịkwuo mma n'adabereghị onwe ya, si otú a na-akwalite uto mgbọrọgwụ na St. perforatum. Usoro ihe ọkụkụ nwere Fe₃O₄ nanoparticles na mkpokọta nke 0.5 mg/L, 1 mg/L, na 1.5 mg/L mere ka ọnụego mmụgharị nke osisi flax dịkwuo mma [52]. Ojiji nke kinetin, 2,4-dichlorobenzothiazolinone, na Fe₃O₄ nanoparticles mere ka ọnụego mmụgharị nke callus na mgbọrọgwụ dịkwuo mma nke ukwuu, agbanyeghị, a ga-atụle mmetụta ndị nwere ike ịnweta site na iji homonụ ndị a maka mmụgharị in vitro. Dịka ọmụmaatụ, ojiji ogologo oge ma ọ bụ nke dị elu nke 2,4-dichlorobenzothiazolinone ma ọ bụ kinetin nwere ike ibute mgbanwe clonal somatic, nrụgide oxidative, ọdịdị callus na-adịghị mma, ma ọ bụ vitrification. Ya mere, ọnụego mmụgharị dị elu anaghị ebu amụma nkwụsi ike nke mkpụrụ ndụ ihe nketa. A ga-enyocha osisi niile e megharịrị ọhụrụ site na iji akara molekul (dịka ọmụmaatụ RAPD, ISSR, AFLP) ma ọ bụ nyocha cytogenetic iji chọpụta nha ha na myirịta ha na osisi in vivo [53,54,55].
Ọmụmụ ihe a gosiri nke mbụ na ojiji nke ndị na-achịkwa uto osisi (2,4-D na kinetin) na Fe₃O₄ nanoparticles nwere ike ime ka morphogenesis dịkwuo mma na mkpokọta nke metabolites bioactive dị mkpa (gụnyere hypericin na hyperoside) na *Hypericum perforatum*. Usoro ọgwụgwọ kachasị mma (1 mg/L 2,4-D + 1 mg/L kinetin + 4 mg/L Fe₃O₄-NPs) abụghị naanị na ọ mere ka nhazi callus, organogenesis, na mmepụta metabolite nke abụọ dịkwuo elu kamakwa ọ gosipụtara mmetụta dị nro na-akpali akpali, nke nwere ike ime ka ntachi obi osisi ahụ na uru ọgwụ dịkwuo mma. Nchikota nke nanotechnology na ọdịbendị anụ ahụ osisi na-enye ikpo okwu na-adịgide adịgide ma dị irè maka mmepụta ọgwụ na vitro buru ibu. Nsonaazụ ndị a na-emeghe ụzọ maka ngwa ụlọ ọrụ mmepụta ihe na nyocha n'ọdịnihu na usoro molekul, nhazi dose na nkenke mkpụrụ ndụ ihe nketa, si otú a na-ejikọ nyocha bụ isi na osisi ọgwụ na biotechnology bara uru.
Oge ozi: Disemba-12-2025