20.500.12556/RUL-149080 Sodobne metode indukcije haploidov po poti mikrosporne embriogeneze Modern methods of haploid induction by microspore embryogenesis Mikrosporna embriogeneza je eden glavnih postopkov, ki jih uporabljajo za pridobivanje podvojenih haploidov, ki jih nato uporabljajo za žlahtnjenje rastlin in razvoj superiornih F1 hibridnih sort. Pri mnogih vrstah in genotipih je uspeh uveljavljenih pristopov androgeneze nezadovoljiv, zato razvijajo vedno nove metode indukcije haploidov. V tem diplomskem delu so opisani različni pristopi izboljšanja mikrosporne embriogeneze, kot je dodajanje TDZD-8, inhibitorja sesalčjega GSK-3, ki dokazano povečuje uspešnost mikrosporne embriogeneze pri oljni ogrščici. Poleg tega obetavne rezultate kažejo tudi eksperimenti s 5-azacitidinom, ki vpliva na reprogramiranje celic prek demetilacije DNA in povzroči veliko povečanje regeneriranih embrijev pri oljni ogrščici in ječmenu. BIX-01294 s svojo inhibicijo metiltransferaz histonskih lizinov zmanjša stopnjo metilacije histonov in je tako povzročil reprogramiranje mikrospor ječmena in oljne ogrščice. Trihostatin A inhibira histonske deacetilaze rastlin in tako deregulira dostop transkripcijskih faktorjev do molekule DNA, kar povzroča reprogramiranje mikrospor. Dodatek trihostatina A v kulture mikrospor oljne ogrščice je pri neodzivnih genotipih povečal število nastalih embrijev za več kot 600 %. Fitosulfokin alfa je še ena kemikalija, ki deluje kot peptidni rastni faktor. Pri kulturah mikrospor pšenice in tritikale je bila najvišja uporabljena koncentracija najuspešnejša pri regeneraciji embrijev iz mikrospor. Vse te metode zaenkrat kažejo obetavne rezultate pri preizkušenih vrstah in bodo v prihodnosti morda širše uporabljene pri vrstah in genotipih, ki so neodzivni na dosedanje metode indukcije mikrosporne embriogeneze. Microspore embryogenesis is one of the main procedures utilized for obtaining doubled haploids, which are used for breeding and producing superior F1 hybrid cultivars. The success of established androgenesis approaches is unsatisfactory in many species and genotypes, prompting the development of new methods for inducing haploids. This thesis discusses the addition of chemical substances such as the TDZD-8 inhibitor of mammalian GSK-3, which enhances the efficiency of microspore embryogenesis in oilseed rape. Additionally, promising results were shown in experiments with 5-azacytidine, influencing cell reprogramming through DNA demethylation, resulting in a significant increase in regenerated embryos in oilseed rape and barley. BIX-01294, with its inhibitory effect on histone lysin methyltransferases, reduces the amount of histone methylation and thus induced reprogramming of barley and rapeseed microsporse. Triostatin A inhibits plant histone deacetylases, thus deregulating transcription factor access to DNA molecules, causing microspore reprogramming. In oilseed rape this represents a key step in microspore embryogenesis of recalcitrant genotypes, yielding over 600% better results compared to controls. Phytosulfokine alpha, a chemical functioning as a peptide growth factor, was successful in wheat and triticale microspore cultures, with the highest applied concentration yielding the most successful embryo regeneration. These methods currently show promising results in specific species, and in the future, some may become widely adopted in breeding and research programs of genotypes recalcitrant to current inducing techniques. mikrosporna embriogeneza haploid podvojeni haploid TDZD-8 5-azacitidin BIX-1294 trihostatin A fitosulfokin alfa oljna ogrščica ječmen microspore embryogenesis haploid doubled haploid TDZD-8 5-azacytidine BIX-01294 trichostatin A phytosulfokine alpha rapeseed barley true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2023-09-03 07:15:57 2023-09-03 07:16:01 2023-10-04 10:36:15 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 228224 163269379 34776.pdf 34776.pdf 1 0DB04C485CF5E04E51086002AA78C340 29ac99822eeea85846db9ef521bc2fd5ddc63bb77d94f7281b41aa21a7a27a59 f72cd5b8-4a18-11ee-af86-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=173467 Biotehniška fakulteta 0 0 0