Made available in DSpace on 2014-12-17T14:10:26Z (GMT). No. of bitstreams: 1 TiaraSC_DISSERT_PARCIAL.pdf: 3904486 bytes, checksum: c7dbd69e3fa57b6310c7eaf6781c6f83 (MD5) Previous issue date: 2011-07-15 / Gasteroids fungi are characterized by the basidiospores maturation inside the basidioma, from which spores liberation occurs in a passive manner. These fungi were once seen as a well definite class of Basidiomycota, but nowadays they are considered an artificial assemblage, because the organisms have independent evolutionary histories forming a polyphyletic group with a vast morphological variety. Despite their diversity, studies with this group in the tropics are incipient, and the phylogenetic relationships of the species from temperate climate remain unknown. Thus, this work aimed to elucidate the phylogenetic relationships of gasteroids fungi from the Geastrales and Phallales orders, with the inclusion of tropical and temperate species, and with these analyses suggest a systematic position of species like Asero? floriformis and Phallus roseus, as well as to verify if the lignicolous habit can indicate parental relationship in the Geastrum genus. For this, basidiomata were collected at Atlantic rain forest areas, during the rainy season, and the specimen identification followed specific literature for gasteroid fungi. The phylogenetic analyses were performed with Maximum Parsimony and Bayesian Analysis, making use of RPB2 and 28S nuclear genes and atp6 mitochondrial gene. It could be observed on the Phallales dendogram, that Asero? floriformis did not cluster with A. rubra, and that it has an anterior divergence from all others species of the family Clathraceae used in this analysis, assuming a basal position in the clade. Phallus roseus, which once was recognized as Itajahya, has previous divergence from the group formed by Phallus species. At the Geastrales dendogram, in the group corresponding to Geastrum genus, it could be observed that species with lignicolous habitat clustered in a clade with high support values. So, the results suggest the creation of a new genus to accommodate A. floriformis, and the revalidation of Itajahya, as well as it can be affirmed that the lignicolous habitat on the Geastrum genus in fact indicates parental relationships, and that it has arised only once at the evolutionary history of the genus / Os fungos gasteroides s?o reconhecidos pela matura??o dos basidi?sporos dentro do basidioma, cuja libera??o ocorre de forma passiva. Esses fungos j? foram vistos como uma classe bem definida de Basidiomycota, mas atualmente s?o considerados um grupo artificial, por tratar-se de organismos com hist?rias evolutivas independentes formando um grupo polifil?tico com grande diversidade morfol?gica. Apesar da grande diversidade, estudos com esse grupo nos tr?picos ainda ? incipiente, e as rela??es filogen?ticas com esp?cies de regi?es temperadas permanecem desconhecidas. Dessa forma, objetivou-se neste trabalho elucidar as rela??es filogen?ticas de fungos gasteroides das ordens Geastrales e Phallales, com a inclus?o de esp?cies tropicais e de regi?es temperadas, e atrav?s dessas an?lises conhecer a rela??o de Asero? floriformis e Phallus roseus com outros membros do grupo, assim como verificar se o h?bito lign?cola em Geastrum pode indicar rela??o de parentesco. Para isso, coletas de basidiomas foram realizadas em ?reas de Mata Atl?ntica em per?odo chuvoso, com a identifica??o dos esp?cimes seguindo bibliografia espec?fica para fungos gasteroides. Para as an?lises filogen?ticas foram utilizados os m?todos de M?xima Parcim?nia e An?lise Bayesiana, utilizando-se os genes nucleares RPB2 e 28S, e o gene mitocondrial atp6. Observou-se no dendograma obtido para a ordem Phallales, que Asero? floriformis n?o se agrupou com A. rubra, apresentando diverg?ncia anterior a todas as esp?cies da fam?lia Clathraceae utilizadas na an?lise, assumindo uma posi??o basal no clado. J? Phallus roseus, antes reconhecido como Itajahya, possui diverg?ncia anterior ao grupo formado pelas esp?cies de Phallus. No dendograma da ordem Geastrales, no grupamento correspondente ao g?nero Geastrum, p?de-se observar esp?cies que possuem h?bito lign?cola se agrupando com alto valor de suporte. Assim, os resultados sugerem a cria??o de um novo g?nero para acomodar A. floriformis, e a revalida??o do g?nero Itajahya, assim como se pode afirmar que o h?bito lign?cola em Geastrum de fato indica rela??es de parentesco, aparentemente tendo surgindo apenas uma vez na hist?ria evolutiva do grupo

I studied evolutionary history in the angiosperm order Apiales, with a special emphasis on interactions between form, time, and space. Four broad categories of problems were addressed: interfamilial relationships in Apiales, the assignment of genera traditionally assigned to the Apiaceae subfamily Hydrocotyloideae, the estimation of divergence times of the major clades, and the reconstruction of the biogeographic history of Apiales. We used molecular markers with different evolutionary properties and rates derived from the plastid (trnD-trnT and rpl16), nuclear (RPB2), and mitochondrial (nad1 intron 2) genomes, from more than 250 species representing all major clades in the order. The nuclear RPB2 region exhibited evidence of at least six duplication events in Apiales and provided a rich source of information for understanding the origins of polyploid lineages, especially in Araliaceae. Sequence comparisons among the copies show that exon regions are highly conserved. All copies appear to be functional but may have undergone subfunctionalization. Phylogenetic analyses of the three genomes suggest that Hydrocotyloideae should be divided into as many as six evolutionary lineages, but that most taxa should be included in subfamilies Azorelloideae and Mackinlayoideae. Relationships among and within the major clades of Azorelloideae need further analyses since many genera appeared non-monophyletic (e.g., Azorella, Schizeilema, and Eremocharis). Mackinlayoideae appeared as the earliest diverging lineage of Apiaceae, but the plastid and nuclear trees were incongruent in the placement of the Platysace clade relative to Mackinlayoideae and the rest of Apiaceae. Among the remaining clades of suborder Apiineae, Myodocapaceae appeared sister to Apiaceae in both plastid and nuclear trees, preceded by the divergence of Araliaceae and then Pittosporaceae. At the base of the gene trees in Apiales, Griseliniaceae and Torricelliaceae formed successive sisters to Apiineae. The placement of Pennantiaceae as sister to the rest of Apiales was confirmed by plastid data, but was not found in the nuclear trees. The order appears to have originated in the Cretaceous, with Apiineae having an age of c. 100 Mya. Australasia appears to be the most likely center of origin for Apiineae and most of its major clades, except Azorelloideae (South America) and Apioideae-Saniculoideae (sub-Saharan Africa).

<p>The genus <i>Arnica</i> (Asteraceae) was investigated for phylogenetic relationships and sesquiterpene lactone (STL) content with the aims to trace the evolutionary history of the genus and to investigate possible congruencies between DNA sequence data, secondary chemistry, and biological activity. </p><p>Complex evolutionary patterns in <i>Arnica</i> are evident from phylogenetic analyses of chloroplast regions (the <i>rpl16</i> and <i>rps16</i> introns and the <i>psbA–trnH</i>, <i>ycf4–cemA</i>, and <i>trnT–L</i> spacers), nuclear ribosomal regions (the internal and external transcribed spacers) and the nuclear low-copy DNA region coding for the second largest subunit of RNA polymerase II (<i>RPB2</i>) between exons 17 and 23. Polymorphism was detected in nuclear ribosomal and low-copy regions<i>,</i> likely caused by polyploidy and agamospermy. Lineage sorting and/or hybridization is a possible explanation for incongruencies between topologies of the different DNA regions. None of the five subgenera in <i>Arnica</i> constitute a monophyletic group according to any of our analyses. </p><p>Sesquiterpene lactone profiles were compared to nuclear ribosomal DNA data using phylogenetic inference and principal component analysis for 33 accessions of 16 species. Clusters supported by both STL chemistry and ribosomal DNA sequence data consist of multiple accessions of the same species (e.g.<i> A montana </i>and<i> A. longifolia</i>), indicating that these species are well defined both genetically and chemically, based on our sampling. Support for subspecies classification of <i>A. chamissonis</i> and <i>A. parryi</i> was found in chemical data. For the first time STLs are reported from subtribe Madiinae, sister to Arniciinae.</p><p>Anti-inflammatory properties, as measured by inhibition of human neutrophil elastase release from neutrophils and inhibition of the binding of transcription factor NF-κB to DNA, were investigated for extracts of 12 <i>Arnica</i> species. <i>Arnica montana</i>, <i>A. chamissonis</i> and <i>A. longifolia</i> accessions show high inhibitory effects in both bioassays. Generally, species with a more diverse STL chemistry also possess the strongest inhibitory activity in the bioassays.</p>

The genus Arnica (Asteraceae) was investigated for phylogenetic relationships and sesquiterpene lactone (STL) content with the aims to trace the evolutionary history of the genus and to investigate possible congruencies between DNA sequence data, secondary chemistry, and biological activity. Complex evolutionary patterns in Arnica are evident from phylogenetic analyses of chloroplast regions (the rpl16 and rps16 introns and the psbA–trnH, ycf4–cemA, and trnT–L spacers), nuclear ribosomal regions (the internal and external transcribed spacers) and the nuclear low-copy DNA region coding for the second largest subunit of RNA polymerase II (RPB2) between exons 17 and 23. Polymorphism was detected in nuclear ribosomal and low-copy regions, likely caused by polyploidy and agamospermy. Lineage sorting and/or hybridization is a possible explanation for incongruencies between topologies of the different DNA regions. None of the five subgenera in Arnica constitute a monophyletic group according to any of our analyses. Sesquiterpene lactone profiles were compared to nuclear ribosomal DNA data using phylogenetic inference and principal component analysis for 33 accessions of 16 species. Clusters supported by both STL chemistry and ribosomal DNA sequence data consist of multiple accessions of the same species (e.g. A montana and A. longifolia), indicating that these species are well defined both genetically and chemically, based on our sampling. Support for subspecies classification of A. chamissonis and A. parryi was found in chemical data. For the first time STLs are reported from subtribe Madiinae, sister to Arniciinae. Anti-inflammatory properties, as measured by inhibition of human neutrophil elastase release from neutrophils and inhibition of the binding of transcription factor NF-κB to DNA, were investigated for extracts of 12 Arnica species. Arnica montana, A. chamissonis and A. longifolia accessions show high inhibitory effects in both bioassays. Generally, species with a more diverse STL chemistry also possess the strongest inhibitory activity in the bioassays.

This thesis focuses on phylogenetic, taxonomic, ecological, and conservation aspects of the crustose lichen genus Lecania (Ramalinaceae, lichenized Ascomycota). Lecania has previously been defined on basis of relatively few morphological characters, and the genus had never been treated in molecular phylogenies. The molecular phylogeny of the genus is inferred from DNA sequences. Twenty-five species traditionally placed in Lecania are included in the study along with 21 species from closely related genera. Lecania is a polyphyletic genus. A well-supported monophyletic group containing 16 Lecania species, including the type species L. fuscella is discovered, i.e. Lecania s. str. Nine species formerly included in Lecania do not belong in the genus. A new species, L. belgica, is described. The relationships of a group of morphologically similar Lecania species, i.e. the L. cyrtella group are investigated using morphological and molecular methods. Haplotype network and phylogenetic analyses indicate that the included species, as conceived in the morphological examinations, all are monophyletic. Two new species, L. leprosa and L. madida, are described, L. proteiformis is resurrected from synonymy, and the known range of L. prasinoides is greatly expanded. The type species Lecania fuscella has become endangered in many countries. Twelve localities in Sweden where the species had been found historically are investigated, but L. fuscella is only recovered in one locality. The species composition in these 12 localities, 58 old and 5 new collections with L. fuscella is determined and analyzed. The vegetation community differs between the old and the new collections, and between the locality where the species is recovered and those where it is not. Lecania fuscella has not been able to adapt to environmental changes and now only appears in a specific type of vegetation community. The phylogenetic diversity of the species is calculated, but does not reflect the species’ evolutionary potential.

<p>DNA sequences from the <i>rps16</i> intron and the <i>psbE-petL</i> spacer from the chloroplast genome, the ribosomal nuclear ITS region, and introns from the low copy nuclear genes <i>RPA2</i>, <i>RPB2</i>, <i>RPD2a</i> and <i>RPD2b</i>, are in different combinations used to infer phylogenetic relationships in <i>Sileneae</i> (<i>Caryophyllaceae</i>). Used in concert, the biparentally inherited nuclear regions are useful to distinguish between paralogy due to allopolyploidy and single gene duplications, respectively, because the latter are not expected to give rise to repeated phylogenetic patterns in potentially unlinked sequence regions. In addition, the sequences resolve previously poorly known relationships in the tribe <i>Sileneae</i>. Several independent losses and incomplete concerted evolution are inferred between the two <i>RPD2</i> paralogues in a subgroup of <i>Silene</i>.</p><p>An allopolyploid origin is suggested for the tetraploid <i>S. aegaea</i>, with the maternal ancestor from the diploid <i>S. pentelica</i> lineage, and the paternal contributor from the diploid <i>S. sedoides</i> lineage.</p><p><i>Silene involucrata</i> originated as an allotetraploid with the diploid lineage of Arctic <i>S. uralensis</i> as cytoplasmic donor and the diploid Siberian/Northeast Asian <i>S. ajanensis</i> lineage as pollen donor. A subsequent allopolyploidization with the <i>S. ajanensis</i> lineage as pollen donor and the tetraploid <i>S. involucrata</i> lineage as cytoplasmic donor resulted in the hexaploid lineage of <i>S. sorensenis sensu lato</i>.</p><p>A monophyletic origin of the North American polyploids is rejected. One lineage consists of tetraploid <i>S. menziesii</i> and its diploid allies. A separate lineage leads to a clade consisting of both diploid and polyploid Arctic, European and Asian taxa in addition to the majority of the North American polyploids. The tetraploid <i>S. californica</i> and the hexaploid <i>S. hookeri</i> are derived from separate allopolyploidization events between these two lineages.</p>

DNA sequences from the rps16 intron and the psbE-petL spacer from the chloroplast genome, the ribosomal nuclear ITS region, and introns from the low copy nuclear genes RPA2, RPB2, RPD2a and RPD2b, are in different combinations used to infer phylogenetic relationships in Sileneae (Caryophyllaceae). Used in concert, the biparentally inherited nuclear regions are useful to distinguish between paralogy due to allopolyploidy and single gene duplications, respectively, because the latter are not expected to give rise to repeated phylogenetic patterns in potentially unlinked sequence regions. In addition, the sequences resolve previously poorly known relationships in the tribe Sileneae. Several independent losses and incomplete concerted evolution are inferred between the two RPD2 paralogues in a subgroup of Silene. An allopolyploid origin is suggested for the tetraploid S. aegaea, with the maternal ancestor from the diploid S. pentelica lineage, and the paternal contributor from the diploid S. sedoides lineage. Silene involucrata originated as an allotetraploid with the diploid lineage of Arctic S. uralensis as cytoplasmic donor and the diploid Siberian/Northeast Asian S. ajanensis lineage as pollen donor. A subsequent allopolyploidization with the S. ajanensis lineage as pollen donor and the tetraploid S. involucrata lineage as cytoplasmic donor resulted in the hexaploid lineage of S. sorensenis sensu lato. A monophyletic origin of the North American polyploids is rejected. One lineage consists of tetraploid S. menziesii and its diploid allies. A separate lineage leads to a clade consisting of both diploid and polyploid Arctic, European and Asian taxa in addition to the majority of the North American polyploids. The tetraploid S. californica and the hexaploid S. hookeri are derived from separate allopolyploidization events between these two lineages.

The study of fungal diversity may lead to many fundamental discoveries and conclusions. Molecular genetics, and particularly high throughput sequencing methods using short DNA fragments as barcodes, has recently experienced a boom. The most frequently used marker for fungal research is the partial region of nuclear ribosomal DNA called ITS (Internal Transcribed Spacer). It occurs in the form of tandem repetitions of up to 200 copies. This fact greatly simplifies its amplification from the environment but also introduces some negatives. One of them can be an existence of intragenomic and intraspecific variability which confounds diversity estimates by exaggerating the real number of species. Using alternative low-copy markers can easily prevent these problems. In this study EF-1α and RPB2 protein- coding genes were compared with traditionally used ITS1 and ITS2 markers. An artificial mock community was created by blending genomic DNA of different fungal lineages. The community was sequenced for all markers and the data were processed according to guidelines commonly used in environmental studies. The results show that ITS2 is unequivocally a more suitable marker for environmental studies than other compared markers. The average coefficient of overestimation was deemed to be approximately two for ITS1, ITS2,...

This thesis focuses on phylogenetic relationships among some of the major lineages in Silene subgenus Behenantha (Caryophyllaceae) using DNA sequences from multiple, potentially unlinked gene regions from a large taxonomic and geographic sample. Both traditional phylogenetic analyses and a strategy to infer species trees and gene trees in a joint approach are used. A new strategy to optimize species classifications, based on the likelihoods of the observed gene trees, is presented. Silene latifolia, S. dioica and the other dioecious species previously classified in section Elisanthe are not closely related to the type of the section (S. noctiflora). The correct name for the group of dioecious species is section Melandrium. The chloroplast DNA data presented indicate a geographic, rather than a taxonomic, structure in section Melandrium. The nuclear genes investigated correlate more to the current taxonomy, although hybridization has likely been influencing the relationships within section Melandrium. Incongruence between different parts of the gene SlXY1 in two Silene lineages is investigated, using phylogenetic methods and a novel probabilistic, multiple primer-pair PCR approach. The incongruence is best explained by ancient hybridization and recombination events. A survey of mitochondrial substitution rate variation in Sileneae is presented. Silene section Conoimorpha, S. noctiflora and the closely related S. turkestanica have elevated synonymous substitution rates in the mitochondrial genes investigated. Morphological and phylogenetic data reject that the Californian S. multinervia should be treated as a synonym to the Asian S. coniflora, as has previously been suggested. Furthermore, none of the genes investigated, or a chromosome count, support the inclusion of S. multinervia in section Conoimorpha. Data from multiple genes suggest that S. noctiflora and S. turkestanica form a sister group to section Conoimorpha. The calyx nervature, which is a potential synapomorphy for S. multinervia and section Conoimorpha, may be explained either by parallelism or by sorting effects.

Heliosperma (nom. cons prop.) comprises 15—20 taxa, most of them endemic to the Balkan Peninsula. DNA sequences from the chloroplast (rps16 intron, psbE-petG spacer) and the nuclear genome (ITS and four putatively unlinked RNA polymerase genes) are used to elucidate phylogenetic relationships within Heliosperma, and its position within Sileneae. Three main lineages are found within Heliosperma: Heliosperma alpestre, H. macranthum and the H. pusillum-clade. The relationships among the lineages differ between the plastid and the nuclear trees. Relative dates are used to discriminate among inter- and intralineage processes causing such incongruences, and ancient homoploid hybridisation is the most likely explanation. The chloroplast data strongly support two, geographically correlated clades in the H. pusillum-group, whereas the relationships appear poorly resolved by the ITS data, when analysed under a phylogenetic tree model. However, a network analysis finds a geographic structuring similar to that in the chloroplast data. Ancient vicariant divergence followed by hybridisation events best explains the observed pattern. The morphological and taxonomical diversity in the H. pusillum-group is possibly ecology-induced, and is not correlated with the molecular data. Phylogenetic patterns regarding the origin of Heliosperma are complicated, probably influenced by reticulate and sorting events. At least two ancient lineages have been involved in its evolution, one most closely related to Viscaria/Atocion and the other to Eudianthe/Petrocoptis. Atocion and Viscaria are sister genera, most species-rich on the Balkans, and including six/three species. Phylogenies do not support their traditional classification, and provide a framework for a taxonomic revision. Atocion compactum is found in three different positions in the chloroplast tree, and in a single clade in the nuclear gene trees. Using relative dates we demonstrate that hybridisation with subsequent chloroplast capture is a feasible explanation for the pattern observed. This, and other observed reticulate patterns, highlights the importance of hybridisation in plant evolution.