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Sodobne metode indukcije haploidov po poti mikrosporne embriogeneze
ID Hauptman Pepelnjak, Maj (Author), ID Murovec, Jana (Mentor) More about this mentor... This link opens in a new window

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Abstract
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.

Language:Slovenian
Keywords:mikrosporna embriogeneza, haploid, podvojeni haploid, TDZD-8, 5-azacitidin, BIX-1294, trihostatin A, fitosulfokin alfa, oljna ogrščica, ječmen
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:BF - Biotechnical Faculty
Year:2023
PID:20.500.12556/RUL-149080 This link opens in a new window
COBISS.SI-ID:163269379 This link opens in a new window
Publication date in RUL:03.09.2023
Views:552
Downloads:20
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Secondary language

Language:English
Title:Modern methods of haploid induction by microspore embryogenesis
Abstract:
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.

Keywords:microspore embryogenesis, haploid, doubled haploid, TDZD-8, 5-azacytidine, BIX-01294, trichostatin A, phytosulfokine alpha, rapeseed, barley

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