20.500.12556/DKUM-83214 Biosinteza fenolnih spojin v ozkolistnem koščcu (berula erecta) in vitro pod vplivom svetlobe in temperature ter njihovo določanje s hplc-uv magistrsko delo Biosynthesis of phenolic compounds in berula erecta in vitro under different light and temperaturconditions and their determination by hplc-uv method V okviru magistrske naloge smo najprej v tkivni kulturi s tehniko mikropropagacije namnožili rastlinski material vrste ozkolistni koščec (Berula erecta) in ga in vitro izpostavili različnim temperaturnim in svetlobnim razmeram. V nadaljevanju raziskave smo ugotavljali kako različne temperaturne in svetlobne razmere vplivajo na vsebnost celokupnih fenolnih spojin v vzorcih in tudi antioksidativno aktivnost ekstraktov rastlin. Razmnoženi rastlinski material smo po 14 dneh aklimatizacije za 28 dni izpostavili trem različnim temperaturno-svetlobnim pogojem: 23 °C in beli svetlobi, 12 °C in modri svetlobi in 12 °C in rdeči svetlobi. Material smo vzorčili na 14 dni in opazovali spreminjanje sveže mase in suhe mase glede na gojitvene pogoje. V drugi fazi raziskave smo določili najprimernejše ekstrakcijsko topilo za ekstrahiranje fenolnih spojin iz predhodno liofiliziranih rastlinskih vzorcev izpostavljenih različnim temperaturno-svetlobnimi razmeram, izbrali najprimernejšo metodo za določevaje fenolnih spojin s HPLC-UV, z metodo HPLC-UV določali vsebnost izbranih fenolnih kislin in flavonoidov v vzorcih in določili celokupno vsebnost fenolnih spojin v vzorcih, ter določili antioksidativno aktivnost etanolnih ekstraktov vzorcev. Ozkolistni koščec je pridobil največ sveže in suhe biomase 28 dni po končani aklimatizaciji pri 23 ̊C pod belo svetlobo. Kot najprimernejše ekstrakcijsko topilo se je izkazal 70 % etanol. Vseboval je kvercetin, kvercetin-3-beta-D-glukozid, kavno kislino in klorogensko kislino. Etanolni ekstrakti vzorcev ozkolistnega koščca so vsebovali največ klorogenske kisline. Najvišje koncentracije klorogenske kisline so vsebovale rastline, ki so bile gojene 28 dni po aklimatizaciji pri 12 ̊C in pod modro svetlobo. Prav tako so rastline gojene pod temi gojitvenimi pogoji dosegle najvišjo koncentracijo celokupnih fenolnih spojin in njihovi ekstrakti so izkazali najvišjo antioksidativno aktivnost. Ugotovili smo tudi, da bi glede na vsebnost klorogenske kisline v primerjavi z nekaterimi drugimi rastlinskimi vrstami ozkolistni koščec potencialno lahko gojili v namene pridobivanja klorogenske kisline, vendar bi morali gojitvene pogoje s podrobnejšimi raziskavami še dodatno optimizirati. As a first part of the master's thesis, we propagated the plant material of Berula erecta in tissue culture using the micropropagation technique and exposed it to different temperature and light conditions. In the second part of the research, we determined how different temperature and light conditions affect the content of total phenolic compounds in the samples and also the antioxidant activity of plant extracts. After 14 days of acclimatization, the propagated plant material was exposed to three different temperature and light conditions for 28 days: 23 °C and white light, 12 °C and blue light, and 12 °C and red light. We sampled the material every 14 days and observed changes in fresh weight and dry weight. In the second phase of the research, we determined the most suitable extraction solvent for extracting phenolic compounds from previously lyophilized plant samples exposed to different temperature and light conditions, selected the most suitable method for determining phenolic compounds with HPLC-UV, determined the content of selected phenolic acids and flavonoids in the HPLC-UV method samples and determined the total content of phenolic compounds in the samples, and determined the antioxidant activity of the ethanol extracts. Berula erecta obtained the most fresh and dry biomass 28 days after the end of acclimatization at 23 ̊C under white light. 70% ethanol proved to be the most suitable extraction solvent. Extracts contained quercetin, quercetin-3-beta-D-glucoside, caffeic acid and chlorogenic acid. Ethanol extracts of Berula erecta contained the most chlorogenic acid. Plants grown 28 days after acclimatization at 12 ̊C and under blue light contained the highest concentrations of chlorogenic acid. Also, plants grown under these growing conditions reached the highest concentration of total phenolic compounds and their extracts showed the highest antioxidant activity. We also found that, considering the content of chlorogenic acid, compared to some other plant species, Berula erecta could potentially be cultivated for the purpose of obtaining chlorogenic acid, but the cultivation conditions should be further optimized with more detailed research. in vitro Berula erecta HPLC sekundarni metaboliti fenolne spojine antioksidativnost fenolne kisline flavonoidi rastlinska tkivna kultura in vitro Berula erecta HPLC secondary metabolites phenolic compounds antioxidant activity phenolic acids flavonoids plant tissue culture true true false [J. Senekovič] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2022-09-28 16:53:51 2022-10-28 11:20:53 2022-10-29 03:08:59 0000-00-00 00:00:00 2022 0 Maribor 0 Maribor XVI, 124 f. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 581.5:582.795(043.2) 127486979 EMAG_Senekovic_Jan_2022.pdf EMAG_Senekovic_Jan_2022.pdf 1 93E2049A65888E21DC0A877C71F1AF9B 8eb67a888464236b79e40177224db4305407d162bc4c41402a019b25ded07cf7 0da912c7-3f3f-11ed-8c93-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=163579 Fakulteta za naravoslovje in matematiko 0 0 0