Oxylipins in automictic yeast life cycles

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Date
2007-11
Authors
Swart, Chantel Wendy
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University of the Free State
Abstract
English: 3-OH oxylipins are saturated and unsaturated oxidized fatty acids which are produced in mitochondria via incomplete b-oxidation or fatty acid synthesis type II. These compounds possibly play an important role in the sexual cycle of yeasts by assisting with ascospore liberation. Literature suggests that 3-OH oxylipins act as a lubricant during ascospore liberation, thereby ensuring efficient release of ascospores. Since the discovery of oxylipins i.e. 3-hydroxy (OH) oxylipins in the early 1990’s, these compounds were found to be distributed in various species of the fungal domain. Studies performed thus far, however, focused mainly on non-fermenting yeast species such as Ascoidea, Dipodascopsis and Eremothecium. According to various studies performed so far, 3-OH oxylipins were found to accumulate specifically in the sexual structures (asci and surrounding ascospores) of various nonfermenting yeasts. These studies also revealed that the sexual stages of nonfermenting yeasts are most susceptible to acetylsalicylic acid (ASA), a known mitochondrial (respiration and 3-OH oxylipin production) inhibitor. No information regarding oxylipin accumulation in asci or ASA-sensitivity of fermentative yeasts, however, has so far been reported. Using confocal laser scanning microscopy in combination with an oxylipin probe for 3-OH oxylipins and coupled to a fluorescing secondary antibody, the accumulation of these oxylipins was discovered in the asci of the following fermentative yeasts i.e. Pichia anomala, Pichia farinosa and Schizosaccharomyces octosporus. Interestingly, no 3-OH oxylipin accumulation was observed in the asci of the fermenting yeast Zygosaccharomyces bailii. This could be ascribed to the fact that this yeast depends more on a fermentative pathway for growth and sexual reproduction, than on mitochondrial respiration. Since 3-OH oxylipins are produced in the mitochondria, it is expected that there should be an increase in mitochondrial activity associated with these sexual structures. Using confocal laser scanning microscopy and a mitochondrial fluorescing probe (Rhodamine 123), an increase in mitochondrial activity was also observed in the asci of the fermenting yeasts tested, again with the exception of Z. bailii. Furthermore, during this study links between yeast sexual reproduction, 3-OH oxylipin accumulation/production, mitochondrial activity and oxygen requirement were established. This study revealed that fermenting yeasts are more resistant to ASA than non-fermenting yeasts when grown in liquid media. This is probably due to the fact that these yeasts can use either aerobic respiration or a fermentative pathway for growth and reproduction. This research prompted the development of a bio-assay that may find application in screening for effective antimitochondrial antifungals.
Afrikaans: 3-OH oksilipiene is versadigde en onversadigde geoksideerde vetsure wat in die mitochondria geproduseer word via ß-oksidasie of vetsuur sintese tipe II. Hierdie komponente speel moontlik ‘n belangrike rol in die seksuele fase van giste deur te help met die vrystelling van askospore. Literatuur stel voor dat 3-OH oksilipiene dien as ‘n smeermiddel tydens askospoorvrystelling en verseker dus effektiewe verspreiding. Sedert die ontdekking van oksilipiene nl. 3-hidroksie (OH) oksilipiene in die vroeë 1990’s, is hierdie verbindings in verskeie fungus species gevind. Verskeie studies alreeds uitgevoer, fokus meestal op nie-fermenterende gisspecies soos byvoorbeeld Ascoidea, Dipodascopsis en Eremothecium. Na aanleiding van verskeie studies is gevind dat 3-OH oksilipiene in spesifiek die seksuele strukture (askus en askospore) van verskeie nie-fermenterende giste akkumuleer. Hierdie studies het ook onthul dat die seksuele fase van nie-fermenterende giste meer vatbaar is vir asetielsalisiensuur (ASA), ‘n bekende mitochondriale (respirasie en 3-OH oksilipien produksie) inhibeerder. Geen inligting rakende oksilipienakkumulasie in aski of ASA-sensitiwiteit in fermenterende giste is tot dusver beskikbaar nie. Deur die gebruik van konfokale laser skanderingsmikroskopie in kombinasie met ‘n oksilipienmerker vir 3-OH oksilipiene (gebind aan ‘n fluoresserende sekondêre teenliggaam) is die akkumulasie van hierdie oksilipiene ontdek in die aski van die volgende fermenterende giste: Pichia anomala, Pichia farinosa en Schizosaccharomyces octosporus. Geen 3-OH oksilipien akkumulasie is egter in die aski van die fermenterende Zygosaccharomyces bailii gevind nie. Dit kan toegeskryf word aan die feit dat hierdie organisme moontlik meer op fermentasie staatmaak vir groei en reproduksie as mitochondriale respirasie. Aangesien 3-OH oksilipiene in die mitochondria geproduseer word, verwag ons ‘n toename in mitochondriale aktiwiteit in die seksuele strukture. Deur die gebruik van konfokale laser skanderingsmikroskopie en ‘n fluoresserende merker (Rhodamine 123) is ‘n toename in mitochondriale aktiwiteit in die aski van al die bogenoemde giste behalwe weereens Z. bailii waargeneem. Tydens hierdie studie is daar ‘n verband tussen seksuele reproduksie van giste, 3-OH oksilipien akkumulasie/produksie, mitochondriale aktiwiteit en suurstof benodigdheid gevind. Ook het hierdie studie onthul dat fermenterende giste meer weerstandbiedend is teen ASA as nie-fermenterende giste wanneer dit in vloeibare media gekweek word. Dit kan moontlik toegeskryf word daaraan dat fermenterende giste aerobiese respirasie of fermentasie kan gebruik vir groei en voortplanting. Hierdie navorsing het die weg gebaan vir die ontwikkeling van ‘n bio-essaëring wat gebruik kan word vir die vinnige uitkenning van nuwe effektiewe antimitochondriale antifungale middels.
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Keywords
Confocal laser scanning microscopy, Fermentation, Mitochondrial activity, Mitochondrial respiration, Rhodamine 123, 3-hydroxy oxylipins, Yeast, Ascus, Ascospore release, Acetylsalicylic acid, Lipids, Yeast fungi -- Reproduction, Yeast fungi -- Genetics, Dissertation (M.Sc. (Microbial, Biochemical and Food Biotechnology))--University of the Free State, 2007
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