Muri ubu bushakashatsi, ingaruka zo gukangura ubuvuzi buhuriweho bwaabashinzwe kugenzura imikurire y'ibimera(2,4-D na kinetin) na nanoparticles za oxide y'icyuma (Fe₃O₄-NPs) ku miterere ya in vitro no ku musaruro wa metabolite ya kabiri muri *Hypericum perforatum* L. byarasuzumwe. Uburyo bwo kuvura [2,4-D (0.5 mg/L) + kinetin (2 mg/L) + Fe₃O₄-NPs (4 mg/L)] bwazamuye cyane ibipimo by'imikurire y'ibimera: uburebure bw'ibimera bwiyongereyeho 59.6%, uburebure bw'imiziho kuri 114.0%, umubare w'amashu ku 180.0%, n'uburemere bushya ku 198.3% ugereranije n'itsinda ryagenzuwe. Ubu buryo bwo kuvura hamwe bwanongereye ubushobozi bwo kuvugurura (50.85%) kandi bwongera igipimo cya hypericin ku kigero cya 66.6%. Isesengura rya GC-MS ryagaragaje ko ibikubiye mu bimera birimo hyperoside, β-patholene, na cetyl alcohol byinshi, bingana na 93.36% by'ubuso bwose bw'agasongero, mu gihe ibikubiye mu bimera byose bya phenolics na flavonoids byiyongereyeho 80.1%. Ibi bisubizo bigaragaza ko ibintu bigenzura imikurire y'ibimera (PGRs) na Fe₃O₄ nanoparticles (Fe₃O₄-NPs) bigira uruhare mu guhuza ibimera binyuze mu gukurura organogenesis no kwibumbira mu bimera, ibi bikaba ari ingamba nziza yo kunoza ikoranabuhanga ry'ibimera bivura.
Umutobe wa Mutagatifu Yohani (Hypericum perforatum L.), uzwi kandi nka Umutobe wa Mutagatifu Yohani, ni ikimera cy’ibimera kiramba cyo mu muryango wa Hypericaceae gifite agaciro k’ubukungu.[1] Ibice byacyo bishobora kugira ingaruka ku buzima birimo tannins karemano, xanthones, phloroglucinol, naphthalenedianthrone (hyperin na pseudohyperin), flavonoids, aside phenolic, n’amavuta y’ingenzi.[2,3,4] Umutobe wa Mutagatifu Yohani ushobora gukwirakwizwa hakoreshejwe uburyo gakondo; icyakora, uburyo gakondo bukoreshwa mu bihe bitandukanye, imbuto nke zimera, no kwandura indwara bigabanya ubushobozi bwabyo bwo guhinga ku bwinshi no gukomeza gukora metabolites za kabiri.[1,5,6]
Bityo, gukwirakwiza imiterere y’uturemangingo mu buryo bwa vitro bifatwa nk'uburyo bwiza bwo gukwirakwiza vuba ibimera, kubungabunga umutungo kamere w’uturemangingo, no kongera umusaruro w’imiti [7, 8]. Ibipimo bigenzura imikurire y’ibimera (PGRs) bigira uruhare runini mu kugenzura imiterere y’uturemangingo kandi ni ngombwa mu guhinga callus n’ibinyabuzima byose mu buryo bwa vitro. Kunoza ingano yabyo n’uburyo bivangavanze ni ingenzi kugira ngo iyi mikorere irangire neza [9]. Kubwibyo, gusobanukirwa imiterere ikwiye n’uburyo ibipimo biyigenzura ni ingenzi mu kunoza imikurire n’ubushobozi bwo kuvugurura bwa St. John's wort (H. perforatum) [10].
Uduce duto twa okiside y'icyuma (Fe₃O₄) ni ubwoko bw'uduce duto twakozwe cyangwa turimo gutegurwa kugira ngo tuvumbure ingingo. Fe₃O₄ ifite ubushobozi bukomeye bwo gukura kw'ibimera, ihuza neza n'ibinyabuzima, kandi ifite ubushobozi bwo guteza imbere imikurire y'ibimera no kugabanya stress ku bidukikije, bityo yakuruye ibitekerezo byinshi mu miterere y'uduce duto. Ikoreshwa ry'utu duce duto rishobora kuba rikubiyemo kunoza ubuhinzi bwakozwe muri vitro kugira ngo buteze imbere igabanyuka ry'uturemangingo, kunoza uburyo intungamubiri zinjira, no gukora enzymes za antioxidant [11].
Nubwo uduce duto tw’ibimera twagaragaje ingaruka nziza ku mikurire y’ibimera, ubushakashatsi ku ikoreshwa rya Fe₃O₄ nanoparticles hamwe n’ibikoresho bigenzura imikurire y’ibimera muri *H. perforatum* biracyari bike cyane. Kugira ngo zuzuze iki cyuho cy’ubumenyi, ubu bushakashatsi bwasuzumye ingaruka z’ingaruka zabyo ku miterere ya in vitro no mu musaruro wa metabolite ya kabiri kugira ngo butange ibitekerezo bishya byo kunoza imiterere y’ibimera bivura. Kubwibyo, ubu bushakashatsi bufite intego ebyiri: (1) kunoza ubwinshi bw’ibikoresho bigenzura imikurire y’ibimera kugira ngo biteze imbere kurema neza callus, kongera imikurire, no gushinga imizi muri vitro; na (2) gusuzuma ingaruka za Fe₃O₄ nanoparticles ku bipimo by’imikurire muri vitro. Gahunda z’ejo hazaza zirimo gusuzuma igipimo cyo kubaho kw’ibimera bishya mu gihe cyo kumenyera (in vitro). Biteganijwe ko ibisubizo by’ubu bushakashatsi bizamura cyane imikorere ya *H. perforatum* mu mikorobe, bityo bigatanga umusanzu mu ikoreshwa rirambye n’ikoranabuhanga ry’ibinyabuzima muri iki gihingwa cy’ingenzi mu miti.
Muri ubu bushakashatsi, twabonye imbuto z’amababi zikomoka ku bimera bya St. John's wort (ibimera nyiramuhinzi) byahinzwe mu murima. Izi mbuto zakoreshejwe mu kunoza imiterere y’ubuhinzi mu buryo bwa vitro. Mbere yo guhinga, amababi yogejwe neza mu mazi atemba mu gihe cy’iminota myinshi. Hanyuma ubuso bw’imbuto bwasukwaga hifashishijwe imiti yica udukoko mu kwinjizwa muri ethanoli ya 70% mu gihe cy’amasegonda 30, hagakurikiraho kwinjizwa mu gisubizo cya sodium hypochlorite (NaOCl) cya 1.5% kirimo udutonyanga duke twa Tween 20 mu gihe cy’iminota 10. Amaherezo, imbuto zasukwaga gatatu n’amazi yasukuwe mbere yo kwimurirwa mu bundi buryo bwo guhinga.
Mu byumweru bine byakurikiyeho, ibipimo byo kuvugurura amashami byapimwe, harimo igipimo cyo kuvugurura amashami, umubare w’amashu kuri buri gihingwa, n’uburebure bw’amashu. Iyo amashami yavuguruwe yageraga ku burebure bwa cm 2, yimurirwa mu buryo bwo gushora imizi bugizwe n’igice cy’imbaraga za MS, aside indolebutyric (IBA) ya 0.5 mg/L, na guar gum ya 0.3%. Guteza imbere imizi byakomeje mu gihe cy’ibyumweru bitatu, muri icyo gihe igipimo cyo gushinga imizi, umubare w’amashu, n’uburebure bw’amashu. Buri buryo bwo kuvura bwasubiwemo inshuro eshatu, hahingwa amashami 10 kuri buri gihingwa, hagatangwa amashami agera kuri 30 kuri buri gihingwa.
Uburebure bw'ikimera bwapimwe muri santimetero (cm) hakoreshejwe agakoresho, kuva ku gice cyo hasi cy'ikimera kugeza ku isonga ry'ikibabi kirekire cyane. Uburebure bw'umuzi bwapimwe muri milimetero (mm) ako kanya nyuma yo gukuraho neza ingemwe no gukuraho igice cyo gukuraho. Umubare w'utubuto kuri buri gihingwa wabazwe neza kuri buri kimera. Umubare w'utubuto tw'umukara ku mababi, tuzwi nka nodules, wapimwe mu buryo bugaragara. Utu tubuto tw'umukara twitwa gland turimo hypericin, cyangwa utubuto twa oxidative, kandi dukoreshwa nk'ikimenyetso cy'imiterere y'ikimera ku buryo kibyakira umuti. Nyuma yo gukuraho igice cyo gukuraho, uburemere bushya bw'ingemwe bwapimwe hakoreshejwe icyuma gipima neza miligarama (mg).
Uburyo bwo kubara igipimo cyo kurema kwa callus ni ubu bukurikira: nyuma yo guhingwa kwa explants mu buryo burimo ibintu bitandukanye bigenzura imikurire (kinases, 2,4-D, na Fe3O4) mu gihe cy'ibyumweru bine, umubare w'explants zishobora kurema callus urabarirwa. Formula yo kubara igipimo cyo kurema kwa callus ni iyi ikurikira:
Buri buryo bwo kuvura bwasubiwemo gatatu, nibura hasuzumwa 10 muri buri buryo bwo kuvura.
Igipimo cyo kuvugurura kigaragaza ingano y'uturemangingo tw'inyama z'inka (callus) turangiza neza igikorwa cyo gutandukanya amashu nyuma yo kurema callus. Iki kimenyetso kigaragaza ubushobozi bw'uturemangingo tw'inyama z'inka bwo guhinduka uturemangingo dutandukanye no gukura tukavamo ingingo nshya z'ibimera.
Igipimo cy’imizi ni igipimo cy’umubare w’amashami ashobora gushinga imizi n’umubare wose w’amashami. Iki kimenyetso kigaragaza ko intambwe yo gushinga imizi yagenze neza, ari ingenzi mu gukwirakwiza no gukwirakwiza ibimera, kuko gushinga imizi neza bifasha ibimera gukomeza kubaho neza mu bihe bikura.
Ibinyabutabire bya Hypericin byakuwemo 90% bya methanol. Miligarama mirongo itanu y’ibimera byumye byongewe muri ml 1 ya methanol maze bishyirwa muri sonic mu minota 20 kuri 30 kHz mu cyuma gisukura hakoreshejwe ultrasonic (model A5120-3YJ) ku bushyuhe bw’icyumba mu mwijima. Nyuma yo gukaraba, icyitegererezo cyashyizwe muri centrifuge kuri 6000 rpm mu minota 15. Supernatant yakusanyijwe, maze uburyo hypericin yafatanywe bupimwa kuri 592 nm hakoreshejwe spectrophotometer ya Plus-3000 S hakurikijwe uburyo bwasobanuwe na Conceiçao et al. [14].
Uburyo bwinshi bwo kuvura hakoreshejwe uburyo bwo kugenzura imikurire y'ibimera (PGRs) na nanoparticles za oxide y'icyuma (Fe₃O₄-NPs) ntibwatumye habaho utudomo tw'umukara ku mababi y'imishibu yavuguruwe. Nta tudomo twagaragaye mu buryo ubwo aribwo bwose bwo kuvura hakoreshejwe kinetin ya 0.5 cyangwa 1 mg/L 2,4-D, 0.5 cyangwa 1 mg/L, cyangwa nanoparticles za oxide y'icyuma ya 1, 2, cyangwa 4 mg/L. Uruvange ruto rwagaragaje ukwiyongera gato mu mikurire y'utudomo (ariko ntabwo ari ingenzi mu mibare) ku gipimo cyo hejuru cya kinetin na/cyangwa nanoparticles za oxide y'icyuma, nko guhuza 2,4-D (0.5–2 mg/L) na kinetin (1–1.5 mg/L) na nanoparticles za oxide y'icyuma (2–4 mg/L). Ibi bisubizo bigaragara ku Ishusho ya 2. Utudomo tw'umukara tugaragaza imiterere y'utudomo twinshi, tubaho mu buryo busanzwe kandi dufite akamaro. Muri ubu bushakashatsi, utudomo tw'umukara twagaragaye ahanini mu gutuma ingirabuzima fatizo zihinduka umukara, bigaragaza ahantu heza ho kwibumbira hypericin. Ubuvuzi hakoreshejwe uduce duto twa 2,4-D, kinetin, na Fe₃O₄ bwateje imbere gukura kwa callus, kugabanya ibara ry'umukara, no kwiyongera kwa chlorophyll, bigaragaza ko imikorere ya metabolike irushaho kuba myiza no kugabanya kwangirika kwa oxidative [37]. Ubu bushakashatsi bwasuzumye ingaruka za kinetin hamwe na duce duto twa 2,4-D na Fe₃O₄ ku mikurire n'iterambere rya callus ya St. John's wort (Ishusho ya 3a–g). Ubushakashatsi bwabanje bwagaragaje ko uduce duto twa Fe₃O₄ dufite imikorere yo kurwanya no kurwanya mikorobe [38, 39] kandi, iyo bikoreshejwe hamwe n'ibikoresho bigenzura imikurire y'ibimera, bishobora gukangura uburyo bwo kwirinda ibimera no kugabanya ibimenyetso by'imihangayiko y'uturemangingo [18]. Nubwo imikorere ya metabolite ya kabiri igenzurwa na gene, umusaruro wayo nyakuri ushingira cyane ku miterere y'ibidukikije. Impinduka mu mikorere ya metabolite n'imiterere yazo zishobora kugira ingaruka ku rugero rwa metabolite ya kabiri binyuze mu kugenzura igaragara ry'uturemangingo tw'ibimera runaka no gusubiza ibintu bifitanye isano n'ibidukikije. Byongeye kandi, ibintu bitera imbaraga bishobora gutera imbaraga za jeni nshya, ibyo bigatuma habaho imikorere ya enzyme, amaherezo bigatera imbaraga inzira nyinshi za biosynthetic biganisha ku iremwa rya metabolite ya kabiri. Byongeye kandi, ubundi bushakashatsi bwagaragaje ko kugabanya igicucu byongera urumuri rw'izuba, bityo bigatera ubushyuhe bw'amanywa mu gace karemano ka *Hypericum perforatum*, kandi bigatera imbaraga zo kongera umusaruro wa hypericin. Hashingiwe kuri aya makuru, ubu bushakashatsi bwasuzumye uruhare rwa nanoparticles z'icyuma nk'ibintu bishobora gutera imbaraga mu mikurire y'uturemangingo. Ibisubizo byagaragaje ko izi nanoparticles zishobora gutera imbaraga za jeni zigize hesperidin biosynthesis binyuze mu gukangura enzymes, bigatuma iyi enzyme yiyongera (Ishusho ya 2). Kubwibyo, ugereranije n'ibimera bikura mu bihe bisanzwe, bishobora kuvugwa ko umusaruro w'ibyo bintu mu mubiri ushobora kwiyongera iyo stress iri hagati ihujwe no gukora kwa jeni zigize uruhare mu gukora kwa jeni za kabiri. Uburyo bwo kuvura buvanze muri rusange bugira ingaruka nziza ku muvuduko wo kuvugurura, ariko mu bihe bimwe na bimwe, iyi ngaruka iragabanuka. Ikigaragara ni uko kuvura hakoreshejwe 1 mg/L 2,4-D, 1.5 mg/L kinase, hamwe n’ibipimo bitandukanye bishobora kongera igipimo cyo kuvugurura ku kigero cya 50.85% ku giti cyabyo ugereranije n’itsinda ry’abagenzuye (Ishusho ya 4c). Ibi byavuye mu bushakashatsi bigaragaza ko imvange yihariye ya nanohormones ishobora gukorana mu buryo buhuriweho kugira ngo iteze imbere gukura kw’ibimera no gukora metabolite, ibyo bikaba bifite akamaro kanini mu guteza imbere imiterere y’ibimera bivura. Palmer na Keller [50] bagaragaje ko kuvura 2,4-D bishobora gutuma habaho callus muri St. perforatum ku giti cyabyo, mu gihe kongeramo kinase byongereye cyane callus mu buryo bufatika no kongera kuvugurura. Ibi byatewe no kunoza uburinganire bw’imisemburo no gushishikariza igabanyuka ry’uturemangingo. Bal et al. [51] basanze kuvura Fe₃O₄-NP bishobora kongera imikorere ya enzymes zirwanya antioxidant ku giti cyabyo, bityo bigatera imbere gukura kw’imizi muri St. perforatum. Imbuto zirimo uduce duto twa Fe₃O₄ ku gipimo cya 0.5 mg/L, 1 mg/L, na 1.5 mg/L byazamuye igipimo cyo kuvugurura ibimera bya flax [52]. Gukoresha kinetin, 2,4-dichlorobenzothiazolinone, na Fe₃O₄ nanoparticles byazamuye cyane igipimo cyo gushinga imizi n'imizi, icyakora, ingaruka mbi zishobora guterwa no gukoresha iyi misemburo mu kuvugurura imizi muri vitro zigomba kwitabwaho. Urugero, gukoresha 2,4-dichlorobenzothiazolinone cyangwa kinetin mu gihe kirekire cyangwa mu buryo bwinshi bishobora gutera impinduka mu miterere y'uturemangingo, stress ya oxidative, imiterere idasanzwe ya callus, cyangwa vitrification. Kubwibyo, igipimo cyo kuvugurura kinini ntigisobanura ko imiterere y'uturemangingo ihamye. Ibimera byose byavuguruwe bigomba gusuzumwa hakoreshejwe ibimenyetso bya molekile (urugero: RAPD, ISSR, AFLP) cyangwa isesengura rya cytogenetiki kugira ngo hamenyekane imiterere n'uko bisa n'ibimera muri vivo [53,54,55].
Ubu bushakashatsi bwagaragaje ku nshuro ya mbere ko gukoresha hamwe ibikoresho bigenzura imikurire y'ibimera (2,4-D na kinetin) hamwe na nanoparticles za Fe₃O₄ bishobora kongera imiterere n'ubwiyongere bw'ibintu by'ingenzi bikora (harimo hypericin na hyperoside) muri *Hypericum perforatum*. Uburyo bwo kuvura bunoze (1 mg/L 2,4-D + 1 mg/L kinetin + 4 mg/L Fe₃O₄-NPs) ntibwatumye gusa habaho callus nyinshi, organogenesis, n'umusaruro wa kabiri wa metabolite ariko kandi bwagaragaje ingaruka nziza, bishobora kongera ubushobozi bw'ikimera bwo kwihanganira imihangayiko n'agaciro k'ubuvuzi. Guhuza nanotechnology n'imiti y'ibimera bitanga urubuga rurambye kandi rwiza rwo gukora imiti myinshi mu buryo bwa vitro. Ibi bisubizo bitanga inzira yo gukoresha mu nganda no gukora ubushakashatsi bw'ejo hazaza ku mikorere ya molekile, kunoza ikoreshwa ry'ibipimo n'ubuziranenge bw'uturemangingo, bityo bigahuza ubushakashatsi bw'ibanze ku bimera bivura n'ikoranabuhanga ry'ibinyabuzima.
Igihe cyo kohereza: Ukuboza 12-2025