Several new pterins have been discovered in halophilic and thermoacidophilic archaebacteria. Two of these were identified in the extreme halophiles and were thus called halopterins. One of these halopterins is produced by Halobacterium salinarium, Halobacterium halobium, and Halococcus morrhuae and is called phosphohalopterin-1. It was given this name because it was the first halopterin discovered and it has four monophosphate esters per dimeric pterin. The proposed structure of phosphohalopterin-1 is as follows. [see document for diagram of chemical structure] The other halopterin, which is produced by Halobacterium marismortui, Halobacterium volcanii, and Halobacterial strain GN-1, is called sulfohalopterin-2 because it has two sulfate esters per dimeric pterin and it was isolated and recognized after the isolation of phosphohalopterin-1. The proposed structure of sulfohalopterin-2 is as follows. [see document for diagram of chemical structure] As shown above, both pterins are dimers with an ether linkage connecting the polyol side chains. Both of the halopterins are negatively charged because of the phosphate and sulfate esters on the side chains. In addition to the halopterins, a positively charged pterin has been isolated from Sulfolobus solfataricus. This pterin is very special since no positively charged unconjugated pterin had ever been found in nature before. This pterin is named solfapterin after the species name of the bacteria from which it was obtained. The structure of this pterin is still unknown but the preliminary data indicate that it is an unconjugated pterin with a polyol containing an amine on the side chain. Another positively charged pterin which is different from solfapterin was found in Thermoplasma. All of the above pterins are different from any previously described pterins and thus represent new pterins in the archaebacterial kingdom. The discovery of these new pterins is important both to pterin biochemistry and to archaebacterial taxonomy. These discoveries also open up a new field, that is, the exploration of the function of these new pterins in norunethanogenic archaebacteria. / Ph. D.

Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: In southern Africa, the ticks Amblyomma hebraeum and Hyalomma rufipes are the main vectors of animal Heartwater and human Crimean Congo Haemorragic Fever, respectively. In order to investigate the phylogeographic structure of these ectoparasites, population genetic studies were conducted throughout southern and partly eastern Africa. Knowing the levels of genetic diversities and population connectivities throughout the species’ native ranges, may be useful for implementing control measures (cross country borders for example), evaluate the potential for drug resistance in introduced locations as well as for elucidating its vectorial capacities in the region. Mitochondrial cytochrome oxidase I (COI) and the internal transcriber spacer 2 (ITS-2) sequences were obtained from eleven sampled localities. In general, haplotype networks and molecular diversity values showed significant genetic structure between most of the sampled populations. Fu’s Fs and mismatch distributions suggested recent population expansions following a bottleneck. A Mantel regression analysis also revealed a significant isolation by distance for H. rufipes. Comparing the two species, H. rufipes showed much stronger signals of geographic genetic population structure in relation to A. hebraeum. A single taxonomic unit is suggested for A. hebraeum while H. rufipes is characterized by the existence of at least three distinct genetic assemblages that may point to incipient speciation. The difference between the two species may be explained by gene flow restriction within the region, by the number of hosts required to complete the life cycle (2 and 3 hosts, respectively), type of host species (domestic and wild animals; small and big mammals, birds) and its availability, and host species overlap. Also, a geographic break between possible host species during the Pleistocene, domestication of animals and climatic conditions in the region played a role on the genetic structure of H. rufipes. / AFRIKAANSE OPSOMMING: Die suider Afrikaanse bosluise, Amblyomma hebraeum en Hyalomma rufipes is die hoof vektore van Hartwater in diere en Crimean Kongo Haemorargiese koors in mense. Om die filogeografiese struktuur van hierdie ektoparasiete te ondersoek, is bevolking-genetiese studies uitgevoer in suidelike Afrika asook gedeeltelik in oos Afrika. Deur die vlakke van genetiese diversiteit en bevolkingsverbindings regdeur die spesies se natuurlike verspreiding te weet, sal dit dalk moontlik wees om: beheermaatreëls (byvoorbeeld oorgrens) in te stel, die waarskynlikheid van weerstandbiedendheid in nuut losgelate gebiede te verstaan, en meer insigte in die vektoriale kapasiteit te verkry. Mitochondriale sitochroom oksidae (COI) en die interne transkriberende spasieerder 2 (ITS- 2) basisnukleotiedopeenvolgings van elf bevolkings is versamel. Haplotiepe netwerke en molekulêre diversiteitwaardes het in die algemeen, betekenisvolle genetiese verkille tussen meeste van die versamelde bevolkings aangetoon. Fu’s Fs en gemiste-paringverspreidings het voorgestel dat die bevolkings onlangs uitgebrei het nadat hulle 'n bottelnek ondergaan het. 'n Mantel regressie analise het ook betekenisvolle isolasie met afstand aangetoon vir H. rufipes. As die twee spesies vergelyk word het H. rufipes baie sterker seine van genetiese geografiese bevolkingstruktuur gewys as A. hebraeum. 'n Enkele taksonomiese eenheid word voorgestel vir A. hebraeum terwyl H. rufipes gekarakteriseer word deur ten minste drie taksonomiese eenhede. Die verskil tussen die twee spesies kan moontlik verklaar word deur geen-vloeibeperkings in die gebied, die aantal gasheer spesies wat benodig word om die lewenssiklus te voltooi (2 of 3 gashere onderskeidelik), tipe gasheer (domestikeerd- of wildlewend; klein of groot soogdiere, voëls) en hulle beskikbaarheid, en die gasheerspesies se oorvleueling. Dit is ook moontlik dat 'n geografiese breek tussen moontlike gasheerspesies gedurende die Pleistoseen, domestikering van diere en klimaatstoestande in die streek 'n rol kon speel by die genetiese struktuur van H. rufipes. / The NRF and the University of Stellenbosch for funding and bursary

Diversos estudos filogenéticos moleculares em Iridaceae confirmam o monofiletismo de Trimezieae. Entretanto, a tribo é pobremente caracterizada e carece de sinapomorfias morfológicas que a sustentem. Por outro lado, o gênero Pseudotrimezia, apesar de morfologicamente bem caracterizado, nunca teve seu monofiletismo investigado. Estudos envolvendo a tribo Trimezieae demonstram que caracteres anatômicos podem contribuir para a taxonomia do grupo além de auxiliar na sua reconstrução filogenética. No presente estudo foram realizados estudos filogenéticos envolvendo caracteres morfológicos e moleculares, separadamente e em análise de evidência total. Foram levantados 50 caracteres morfológicos, incluindo anatomia foliar, em Pseudotrimezia e nos gêneros relacionados, Neomarica e Trimezia. Os caracteres moleculares foram obtidos de três regiões plastidias (trnG, trnH-psbA e trnK) e uma nuclear (ITS). A análise de parcimônia com os dados morfológicos resultou em uma filogenia com Pseudotrimezia monofilético, sem sustentação, enquanto Neomarica e Trimezia emergem como polifiléticos. Assim, os caracteres morfológicos mostraram-se insuficientes para a compreensão da história evolutiva do grupo. A análise de evidência total apresenta Pseudotrimezia monofilético em um clado bem caracterizado morfologicamente sustentado por 10 sinapomorfias homoplásticas. Não obstante, Trimezia e Neomarica emergem agrupados em quatro clados distintos, a maioria sem sustentação, revelando que os gêneros reconhecidos na tribo Trimezieae não correspondem a grupos monofiléticos. As análises filogenéticas fornecem um arcabouço para discutir a sistemática de Pseudotrimezia, gênero endêmico dos campos rupestres da porção mineira da Cadeia do Espinhaço. Apesar de sua distribuição restrita e morfologia floral relativamente uniforme, 24 nomes surgiram desde sua criação em 1945, e muitas das espécies são endêmicas de localidades restritas. É apresentada a revisão de 16 espécies reconhecidas, respectivos mapas de distribuição, ilustrações e chave de identificação / Several molecular phylogenetic studies focusing on Iridaceae contribute to confirm Trimezieae as a monophyletic group. Despite that, the tribe is poorly characterized and lacks morphological synapomorphies as support. On the other hand, Pseudotrimezia is morphologically well characterized, but its monophyly has never been investigated. Anatomical studies carried out with tribe Trimezieae indicates that this kind of data can be a good source for taxonomic and phylogenetic analyses. This study performed phylogenetic analysis using morphological and molecular data, separately and combined in a total evidence approach. Molecular data were obtained from three plastid regions (trnG, trnH-psbA and trnK), and one nuclear (ITS). Fifty morphological characters, including leaf anatomy, were selected within Pseudotrimezia and the related genera Neomarica and Trimezia. Parsimony morphological analysis results in a monophyletic Pseudotrimezia with no support, and, Neomarica and Trimezia aspolyphyletic; therefore, morphological characters were insufficient to help understand the evolutionary history of this group.The total evidence analysis shows Pseudotrimezia monophyletic in a morphologically well-characterized clade, suppported by 10 homoplastic synapomorfies. Nevertheless, Trimezia and Neomarica arise mixed together in four distinct clades, most of them with no support, suggesting that tradicionally recognized genera in the tribe do not correspond to monophyletic groups. Neverthless, phylogenetic analyses provide an important framework for systematic studies of Pseudotrimezia (Iridaceae), an endemic genus from \"campos rupestres\" in Espinhaço Range, Minas Gerais State, Brazil. Despite its restricted distribution and quite uniform morphology, 24 names have been created since the genus was first described in 1945, and many species are endemic of particular localities. The revision of 16 recognized species, their distribution maps, illustrations, and identification key are presented.

Chironius, um dos maiores gêneros da subfamília Colubrinae na América do Sul, inclui 14 espécies, diagnosticadas pela presença de 10 ou 12 fileiras de escamas dorsais no meio do corpo. O gênero apresenta uma distribuição Neotropical, que se estende desde a costa norte de Honduras até o Uruguai e o nordeste da Argentina. Neste trabalho foi realizada uma análise filogenética combinada de 75 caracteres morfológicos e 1397 caracteres moleculares (provenientes das seqüências de dois genes mitocondriais, 12S e 16S, e um gene nuclear, c-mos) pelos métodos de máxima parcimônia e inferência bayesiana. A monofilia do gênero Chironius foi corroborada e os resultados obtidos sugerem que o gênero é dividido em 10 componentes monofiléticos. Os gêneros Drymobius, Leptophis e Dendrophidion são apontados como os mais relacionados à Chironius, corroborando trabalhos anteriores. Chironius laevicollis e C. scurrulus formam o clado mais basal entre as espécies, e sua distribuição restrita a América do Sul sugere que Chironius tenha tido origem neste continente e que posteriormente tenha invadido a América Central através de eventos de dispersão. / Chironius, one of the greatest genera of the Colubrinae subfamily in South America, consists of 14 species, diagnosed by the presence of 10 or 12 rows of dorsal scales in the mid-body. The genus presents a Neotropical distribution extending from the northern coast of Honduras to Uruguay and northeastern Argentina. This study presents a combined phylogenetic analysis of 75 morphological characters and 1397 molecular characters (from the sequences of two mitochondrial genes, 12S and 16S, as well as a nuclear gene, c-mos) by means of Maximum Parsimony and Bayesian Inference. The monophylety of the genus Chironius was corroborated and the obtained results suggest that the genus is subdivided in ten monophyletic components. The genera Drymobius, Leptophis and Dendrophidion come out as the most closely related to Chironius, corroborating previous studies. Chironius laevicollis and C. scurrulus composed the most basal clade among the species. Their distribution is restricted to South America, which suggests that Chironius originated in South America and later invaded Central America through dispersion.

Borya mirabilis is one of the world's most critically endangered plants. The research in this thesis has illuminated key aspects of: its reproductive biology; interspecies and intraspecies molecular relationships, mycorrhizal status, tissue culture potential and disease threats. Each of these aspects has fundamental management implications for the active management of B. mirabilis. Floral observations of B. mirabilis and related species affirmed the uniqueness of the Boryaceae amongst the Asparagales. B. mirabilis had an unusually high number of floral abnormalities compared with other species of Borya observed. B. mirabilis is fly-pollinated. Pollen of Borya species showed little difference in the characteristics of mature pollen between species, with viable pollen being prolate and unicolpate with a single colpa-style aperture and a unique patterning of the pila. The structural immaturity of B. mirabilis pollen correlated with evidence from pollen growth experiments, where B. mirabilis pollen had extremely low germination rates, with those grains that did germinate being slow to do so and with slow-growing pollen tubes compared to those of fertile Borya species. Examination of the ovules of B. mirabilis showed that morphologically they were viable compared to viable Borya species. The field population of B. mirabilis was crossed, with one seed produced (the first recorded seed for th is species). Cross-pollination using the pollen of the closely related B. constricta and B. sphaerocephala with B. mirabilis ovules proved unsuccessful. Examination of the chromosome number of B. mirabilis showed that it had approximately 66 chromosomes and is probably hexaploid, relative to the diploid number of 26 in B. constricta. This may explain its low fertility. Interspecies and intraspecies relationships of the Boryaceae and Borya mirabilis were investigated using sequences of chloroplast and nuclear DNA. The closest similarities to B. mirabilis were B. constricta and B. sphaerocephala. B. mirabilis may have emerged from alloploidy of these species in the past. Because of the consistent similarities of B. mirabilis and B. constricta chloroplast sequences, it is proposed that both shared a common ancestor with a chromosome number of 2n=22. A malfunction n meiosis may have resulted in ovules with 2n=44. The high similarity of the nuclear ribosomal ITS region DNA suggests that the nuclear DNA was derived from B. sphaerocephela. B. mirabilis may be an allopolyploid, from fertilisation of a diploid ovule of B. constricta with haploid pollen of B. sphaerocephala, resulting in a reproductively isolated polyploidy of low fertility. The wild population of B. mirabilis was determined to have a small amount of genetic variation. The genetic variation in the field population w as not fully reflected in the ex-situ population. An effective means of micro-propagation of B. nitida for use in B. mirabilis has been established, providing an effective means of mass production of the species. The research has determined: a suitable explant (shoot tips) for regeneration; an effective means of reducing contamination in tissue culture (PPM); what medium is required to micro-propagate the species (LMHM); an appropriate gelling agent (Phytagel); and a practical method for inducing roots on the shoots grown in tissue culture. B. mirabilis has been established as mycorrhizal. The predominant mycorrhizal association is a nodular arbuscular mycorrhiza, present in the form of coils in root nodules over wetter months and as spores in these nodules over dryer months. A significant increase in the health of the ex-situ population of B. mirabilis was recorded after addition of soil containing fine roots of the wild population. Of the plants associated with the wild population, Callitris rhomboidea had the most morphologically similar vesicular arbuscular mycorrhizal relationship. But molecular identification was not achieved due to recalcitrance of DNA in PCR attempts. Potential translocation sites for some of the ex-situ population of B. mirabilis were examined for Phytophthora infestation. Reid's Lookout and Mackey's Peak were infected with P. cinnamomi. Vegetation at Mackey's Peak displayed characteristic infection symptoms, resulted in isolates of P. cinnamomi from baiting and would directly receive runoff from both the walking track and the existing infested B .mirabilis site. At the Reid's Lookout site, both walking track and proposed translocation site were infested with P. cinnamomi, yet did not display the associated symptoms in the vegetation. The Pine Plantation translocation site was uninfected at the level of sampling undertaken. Its vegetation did not display any characteristic infection symptoms and was not isolated when soil samples were baited. It was therefore chosen for translocation and so far the plants are healthy and actively growing. This research has provided critical knowledge to aid the recovery team in its current and future endeavours to manage this species and bring it back from the brink of extinction.

The taxonomy and biogeography of the southern African pheasant shell fauna are poorly known. Thirty–one nominal taxa referable to Phasianelloidea have been described or recorded in this region, but no systematic revision of these has ever been undertaken. Morphological evidence suggests that 16 taxa represent valid species, 13 are synonyms and two represent incorrect identifications. DNA sequence data from mitochondrial COI and 16S markers are used to assess the validity of the described nominal southern African Tricolia species. Phylogenetic analyses recovered seven distinct clades. Tricolia adusta, T. elongata, T. formosa, T. kochii, T. saxatilis and T. neritina were recovered as distinct species. Tricolia africana and T. capensis are genetically indistinguishable. However, morphological characters of the shell are clearly diagnosable. This could be due to incomplete sorting (ancestral polymorphism) reflecting recent speciation with rapid morphological and ecological divergence co–incident with geographical separation. Similarly, there is little genetic differentiation between T. bicarinata, T. insignis and T. kraussi. In this case the similarity is also supported by morphological data as the three species are conchologically close with intergrading shell characters, and might even be one species exhibiting ecogeographic variation in shell form. Monophyly of the southern African Tricolia species is not supported as well as the relationship between these and the European Tricolia pullus. In the last chapter a molecular phylogeny based on sequence data from mtDNA (COI and 16S), nuclear (18S and 28S) and the combined data (COI, 16S, 18S and 28S) is presented for the Phasianelloidea. Bayesian inference analyses performed on the combined data support the monophyly of Tricolia sensu stricto, Eulithidium and Phasianella. Tricolia sensu lato is not monophyletic, as its southern Australian and Indo–West Pacific species do not cluster with its southern African and Eastern Atlantic representatives. The position of Hiloa and Gabrielona within the Phasianelloidea is unresolved. Phylogenetic reconstructions using bayesian inference support monophyly of the Phasianelloidea. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Chironius, um dos maiores gêneros da subfamília Colubrinae na América do Sul, inclui 14 espécies, diagnosticadas pela presença de 10 ou 12 fileiras de escamas dorsais no meio do corpo. O gênero apresenta uma distribuição Neotropical, que se estende desde a costa norte de Honduras até o Uruguai e o nordeste da Argentina. Neste trabalho foi realizada uma análise filogenética combinada de 75 caracteres morfológicos e 1397 caracteres moleculares (provenientes das seqüências de dois genes mitocondriais, 12S e 16S, e um gene nuclear, c-mos) pelos métodos de máxima parcimônia e inferência bayesiana. A monofilia do gênero Chironius foi corroborada e os resultados obtidos sugerem que o gênero é dividido em 10 componentes monofiléticos. Os gêneros Drymobius, Leptophis e Dendrophidion são apontados como os mais relacionados à Chironius, corroborando trabalhos anteriores. Chironius laevicollis e C. scurrulus formam o clado mais basal entre as espécies, e sua distribuição restrita a América do Sul sugere que Chironius tenha tido origem neste continente e que posteriormente tenha invadido a América Central através de eventos de dispersão. / Chironius, one of the greatest genera of the Colubrinae subfamily in South America, consists of 14 species, diagnosed by the presence of 10 or 12 rows of dorsal scales in the mid-body. The genus presents a Neotropical distribution extending from the northern coast of Honduras to Uruguay and northeastern Argentina. This study presents a combined phylogenetic analysis of 75 morphological characters and 1397 molecular characters (from the sequences of two mitochondrial genes, 12S and 16S, as well as a nuclear gene, c-mos) by means of Maximum Parsimony and Bayesian Inference. The monophylety of the genus Chironius was corroborated and the obtained results suggest that the genus is subdivided in ten monophyletic components. The genera Drymobius, Leptophis and Dendrophidion come out as the most closely related to Chironius, corroborating previous studies. Chironius laevicollis and C. scurrulus composed the most basal clade among the species. Their distribution is restricted to South America, which suggests that Chironius originated in South America and later invaded Central America through dispersion.

Diversos estudos filogenéticos moleculares em Iridaceae confirmam o monofiletismo de Trimezieae. Entretanto, a tribo é pobremente caracterizada e carece de sinapomorfias morfológicas que a sustentem. Por outro lado, o gênero Pseudotrimezia, apesar de morfologicamente bem caracterizado, nunca teve seu monofiletismo investigado. Estudos envolvendo a tribo Trimezieae demonstram que caracteres anatômicos podem contribuir para a taxonomia do grupo além de auxiliar na sua reconstrução filogenética. No presente estudo foram realizados estudos filogenéticos envolvendo caracteres morfológicos e moleculares, separadamente e em análise de evidência total. Foram levantados 50 caracteres morfológicos, incluindo anatomia foliar, em Pseudotrimezia e nos gêneros relacionados, Neomarica e Trimezia. Os caracteres moleculares foram obtidos de três regiões plastidias (trnG, trnH-psbA e trnK) e uma nuclear (ITS). A análise de parcimônia com os dados morfológicos resultou em uma filogenia com Pseudotrimezia monofilético, sem sustentação, enquanto Neomarica e Trimezia emergem como polifiléticos. Assim, os caracteres morfológicos mostraram-se insuficientes para a compreensão da história evolutiva do grupo. A análise de evidência total apresenta Pseudotrimezia monofilético em um clado bem caracterizado morfologicamente sustentado por 10 sinapomorfias homoplásticas. Não obstante, Trimezia e Neomarica emergem agrupados em quatro clados distintos, a maioria sem sustentação, revelando que os gêneros reconhecidos na tribo Trimezieae não correspondem a grupos monofiléticos. As análises filogenéticas fornecem um arcabouço para discutir a sistemática de Pseudotrimezia, gênero endêmico dos campos rupestres da porção mineira da Cadeia do Espinhaço. Apesar de sua distribuição restrita e morfologia floral relativamente uniforme, 24 nomes surgiram desde sua criação em 1945, e muitas das espécies são endêmicas de localidades restritas. É apresentada a revisão de 16 espécies reconhecidas, respectivos mapas de distribuição, ilustrações e chave de identificação / Several molecular phylogenetic studies focusing on Iridaceae contribute to confirm Trimezieae as a monophyletic group. Despite that, the tribe is poorly characterized and lacks morphological synapomorphies as support. On the other hand, Pseudotrimezia is morphologically well characterized, but its monophyly has never been investigated. Anatomical studies carried out with tribe Trimezieae indicates that this kind of data can be a good source for taxonomic and phylogenetic analyses. This study performed phylogenetic analysis using morphological and molecular data, separately and combined in a total evidence approach. Molecular data were obtained from three plastid regions (trnG, trnH-psbA and trnK), and one nuclear (ITS). Fifty morphological characters, including leaf anatomy, were selected within Pseudotrimezia and the related genera Neomarica and Trimezia. Parsimony morphological analysis results in a monophyletic Pseudotrimezia with no support, and, Neomarica and Trimezia aspolyphyletic; therefore, morphological characters were insufficient to help understand the evolutionary history of this group.The total evidence analysis shows Pseudotrimezia monophyletic in a morphologically well-characterized clade, suppported by 10 homoplastic synapomorfies. Nevertheless, Trimezia and Neomarica arise mixed together in four distinct clades, most of them with no support, suggesting that tradicionally recognized genera in the tribe do not correspond to monophyletic groups. Neverthless, phylogenetic analyses provide an important framework for systematic studies of Pseudotrimezia (Iridaceae), an endemic genus from \"campos rupestres\" in Espinhaço Range, Minas Gerais State, Brazil. Despite its restricted distribution and quite uniform morphology, 24 names have been created since the genus was first described in 1945, and many species are endemic of particular localities. The revision of 16 recognized species, their distribution maps, illustrations, and identification key are presented.

The phylogenetics of eight species of the Peromyscus truei and P. boylii species groups from 15 populations were analyzed based on mitochondrial DNA sequence differentiation, using 13 hexanucleotide specific restriction enzymes. P. difficilis, P. nasutus, and P. attwateri were found to be members of the same clade. P. leucopus was not found to be closely related to any of the species of the boylii or truei species groups. Phylogenetic interpretations for the remaining species differed based on Wagner and Dollo parsimony analyses. P. true appears to be most closely related to P. gratus based on Wagner parsimony and the phenetic analysis, while the relationship of P. gratus to other species could not be resolved based on Dollo parsimony.

In eukaryotes, the defined loci on each chromosome, the centromeres, accomplish the critical task of correct cell division. In some organisms, centromeres are composed of a euchromatic central core region embedded in a stretch of heterochromatin and the inheritance and maintenance of centromeres are controlled by dynamic epigenetic phenomena. Although the size of centromeres differs between organisms, its organization, and the placement of euchromatic and heterochromatic regions is conserved from the fission yeast, Schizosaccharomyces pombe, to humans, Homo sapiens. However, relatively little is known about centromeres in the filamentous fungi from the Ascomycota, representing the largest group of fungi and fungal pathogens. Further, studies from humans, flies, yeast and plants have shown that the inheritance of centromeres is not strictly guided by centromeric DNA content, which is highly AT-rich, repetitive and constantly evolving. Therefore, it is difficult to align ans assemble the sequenced contigs of centromeric regions of higher eukaryotes, including most filamentous fungi. A genetic technique, tetrad (or octad) analysis has helped to map the centromeres of the filamentous fungus Neurospora crassa early on. The research presented in this dissertation used N. crassa as a model to focus on characterizing different features of centromeres with an emphasis on the centromere-specific histone H3 (CenH3) protein. Data included here represent the first study on centromere-specific proteins in Neurospora, and demonstrate that the central core of the centromeres are heterochromatic, showing enrichment of silent histone marks, which is in contrast to the centromere arrangement in fission yeast. The CenH3 protein, whose deposition on the genome licenses formation or maintenance of centromeres, shows highly divergent N-terminal regions and a conserved histone fold domain (HFD) in all eukaryotes. This bipartite nature of CenH3 is also observed in the Ascomycota, which provides an opportunity for functional complementation assays by replacing Neurospora CenH3 (NcCenH3) with CenH3 genes from other species within the Ascomycota. The results from this experimental approach provide good measures for (1) determining the specific regions of CenH3 required for the assembly of centromeres during meiotic and mitotic cell divisions and (2) analyzing the resistance to changes in the organization of centromeres in N. crassa. The genetic analysis showed that the divergent N-terminal region is essential for the proper assembly of centromeres, and that the conserved carboxy-terminus of CenH3 is important for the process of meiosis but not mitotic cell division. ChIP-seq analyses suggest that the observed loss of Podospora anserina CenH3 (PaCenH3- GFP) from certain N. crassa centromeres does not result in obvious phenotypic defects, e.g. diminished growth or evidence for aneuploidy. Further, the low enrichment of PaCenH3-GFP at certain centromeres is possibly predetermined during meiosis, which results in irreversible and progressive decreases in enrichment. It remains to be determined if this process is random as far as selection of centromeres is concerned. Together the results presented here suggest that during meiosis more stringent structural requirements for centromere assembly apply and that these are dependent on CenH3, and that depletion of CenH3 from centromeres does not critically affect mitosis in the asynchronously dividing nuclei of Neurospora hyphae. / Graduation date: 2013