20.500.12556/RUL-185915 Analiza globalne metilacije citozina v DNA pri različnih genotipih konoplje (Cannabis sativa) gojenih v pogojih in vivo in in vitro z metodo HPLC Analysis of global cytosine methylation in the DNA of different Cannabis sativa genotypes grown under in vivo and in vitro conditions using the HPLC method Konoplja (Cannabis sativa L.) je gospodarsko vse pomembnejša rastlinska vrsta, pri kateri mikropropagacija omogoča hitro pridobivanje enotnega rastlinskega materiala. Daljše gojenje in vitro lahko sproži epigenetske spremembe, med katerimi je osrednjega pomena globalna metilacija DNA. Namen naloge je bil z metodo HPLC-UV določiti delež 5-mC v genomski DNA štirih genotipov konoplje ter primerjati matične rastline, poganjke po prvi in četrti subkultivaciji na gojišču DKW brez dodatka in z dodatkom TDZ, poganjke na gojišču za izdolževanje, aklimatizirane rastline in vegetativne klone. Analiza deleža 5-mC v genomu rastlin je pokazala, da so razlike že prisotne na ravni matičnih rastlin, pri čemer se je skozi subkultivacije pri polovici vseh genotipov pojavil trend hipometilacije. Učinek TDZ ni bil enoznačen ne v smeri metilacije ne v velikosti in je izkazoval veliko odvisnost od genotipa rastline ter njene razvojne stopnje. Učinek gojenja v tkivnih kulturah je ostal zaznaven tudi po aklimatizaciji in se je kazal kot višja raven 5-mC pri večini genotipov po prehodu v odrasle rastline v primerjavi z vegetativno pridobljenimi kloni. Rezultati potrjujejo, da genotip ključno uravnava tako izhodiščno raven kot dinamiko globalne metilacije DNA med mikropropagacijo konoplje, metoda HPLC pa se je izkazala za dovolj občutljivo za zaznavo teh razlik. Hemp (Cannabis sativa L.) is a plant of growing economic importance, in which micropropagation enables rapid production of uniform plant material. However, prolonged in vitro cultivation can trigger epigenetic changes, among which global DNA methylation is of central importance. The aim of the thesis was to use the HPLC-UV method to determine the proportion of 5-mC in the genomic DNA of four hemp genotypes, and to compare mother plants, shoots after the first and fourth subculture on DKW medium with and without added TDZ, shoots on elongation medium, acclimatized plants, and vegetative clones. Analysis of the 5-mC proportion in the plant genome showed that differences were already present at the level of the mother plants, while a trend of hypomethylation appeared across subcultures in half of all genotypes. The effect of TDZ was not consistent, neither in the direction nor in the magnitude of methylation and showed strong dependence on both plant genotype and developmental stage. The effect of growth in tissue culture remained detectable after acclimatization and manifested as a higher level of 5-mC in most genotypes after transition to adult plants, compared with vegetatively obtained clones. The results confirm that genotype is a key factor governing both the baseline level and the dynamics of global DNA methylation during hemp micropropagation, and that the HPLC method proved sensitive enough to detect these differences. globalna DNA metilacija HPLC konoplja epigenetske spremembe 5-metilcitozin global DNA methylation HPLC cannabis epigenetic changes 5-methylcytosine true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-08-23 07:16:53 2026-08-23 07:17:03 2026-08-25 14:09:31 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 2027-08-20 285496 288835843 60451.pdf 60451.pdf 1 55C5D4BA59B5CAB283AA19E7B3FC81F7 573e980c7c2a60a5fcf0102076c671bb87502032469965f357b0592d05c47319 7ac3af5a-9eb1-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=244922 Biotehniška fakulteta 0 0 0