The existence and widespread distribution of sexual reproduction despite its obvious disadvantages is an evolutionary enigma. According to the theory of frozen plasticity, the evolution of sexual and asexual species is fundamentally different. One of the implications of this theory are different ecological preferences of these species. Sexual species should prefer biotically and abiotically variable environments because of their ability to quickly and reversibly respond to unpredictable changes. On the other hand, asexual species should prosper in biotically and abiotically stable environments because of their ability to perfectly adapt even to extreme environmental factors in the long term. I decided to test these predictions in the metastudy comparing ancient asexual clades (in which we can be sure about their obligate asexuality and evolutionary longevity) with their sexual sister or closely related ecologically comparable clades. The hypothesis of the preference of asexual species to biotically and abiotically homogenous environments was supported by the results of this metastudy. On the other hand, the hypothesis of supposed ability of asexual species to perfectly adapt to a broader range of environmental conditions, tested on the comparison of temperature ranges of activity of previously...

Aspergillus fumigatus é um fungo saprófita e um dos principais fungos patogênicos humanos. Os seus conídios são inalados e chegam ao pulmão, onde se tornam ativos e germinam, dando origem a uma doença conhecida como aspergilose. A aspergilose é um conjunto de formas clínicas que apresenta grande risco, principalmente, para indivíduos imunossuprimidos. A sinalização de cálcio desempenha importante papel na regulação de diversas respostas fisiológicas, já que o Ca2+ corresponde a um mensageiro secundário que participa de diferentes processos biológicos, como ciclo celular, diferenciação, ou homeostase. Além disso, as equinocandinas, como a caspofungina, são antifúngicos cujo alvo é a parede celular do fungo. Esses agentes reduzem a concentração total de 1,3-?-D-glucanos na parede da célula, através da inibição da atividade da 1,3-?-glucano sintase. A inibição da síntese de componentes da parede celular através de drogas antifúngicas pode ser um alvo importante contra infecções causadas por A. fumigatus. Através de um screening de fatores de transcrição, foram identificados genes pertinentes a esses processos. A deleção do gene nsdC resultou em maior sensibilidade ao CaCl2, EGTA, Congo Red, Nikkomycin Z, Calcoflúor White, Caspofungina (efeito paradoxical reduzido), 44oC, Menadiona e T-butil hidroperóxido. Além disso, o mutante nulo ?nsdC mostrou-se mais resistente ao Sorbitol e à Ciclosporina. O fator de transcrição NsdC parece estar envolvido na via de sinalização de cálcio, por ter sua expressão alterada na ausência do fator de transcrição CrzA. Além disso, NsdC mostrou-se importante em processos relacionados à parede celular, pois sua deleção leva à formação de hifas mais ramificadas e desorganizadas, e parede celular mais espessa. Esse fator de transcrição aparenta, ainda, ser repressor da reprodução assexual. Isso se deve ao fato de o mutante nulo ?nsdC apresentar conídios em meio líquido e aumento signficativo da expressão de brlA. Ademais, a proteína NsdC encontra-se majoritariamente presente no núcleo, após tratamento com CaCl2. Outros mutantes de fatores de transcrição, que apresentam sensibilidade à Caspofungina, foram identificados: ?cbfA, ?nctA, ?nctB e ?nctC. Algumas evidências apontam que NctA e NctB fazem parte de um mesmo complexo. Já NctC parece estar relacionado a estresses de temperatura e osmótico, sendo importante para a virulência em A. fumigatus. Portanto, mais estudos são necessários a fim de melhor compreender os papéis desses genes, podendo ser importantes para trazer maiores informações acerca de potenciais alvos terapêuticos no tratamento de aspergilose. / Aspergillus fumigatus is a saprophytic fungus and one of the main human pathogenic fungi. Their conidia are inhaled and reach the lungs, where they activate and germinate, and can trigger aspergillosis. Aspergillosis clinical forms presents great risk, mainly for the immunosuppressed patients. Calcium signaling has important role in physiological responses. Calcium corresponds to a secondary messenger that participates in different biological processes, such as cell cycle, differentiation, or homeostasis. In addition, echinocandins, such as caspofungin, are antifungal drugs that target the fungal cell wall. These agents reduce the total concentration of 1,3-?- D-glucans on the cell wall, by inhibiting the activity of 1,3-?-glucan synthase. Inhibition of the synthesis of cell wall components through antifungal drugs may be an important target against infections caused by A. fumigatus. Screening of transcription factors genes have been identified. The deletion of the nsdC gene results from increased sensitivity to CaCl2, EGTA, Congo Red, Nikkomycin Z, Calcofluor White, Caspofungin (reduced paradoxical effect), 44C, Menadione and Tbutyl hydroperoxide. In addition, the ?nsdC null mutant is more resistant to Sorbitol and Cyclosporin. The transcription factor NsdC seems to be involved in the signaling of calcium, and its expression is altered in the absence of the CrzA transcription factor. In addition, NsdC has been shown to be important in processes related to the cell wall, since its deletion leads to the formation of more branched and disorganized hyphae and thicker cell wall. This transcription factor also appears to be a repressor of asexual reproduction. This is due to the fact that the null mutant ?nsdC produces conidia in liquid medium and leads to a significant increase in the expression of brlA. In addition, the NsdC protein is mostly present in the nucleus, after treatment with CaCl2. Other mutants of transcription factors sensitivity to Caspofungin have been identified: ?cbfA, ?nctA, ?nctB and ?nctC. Some evidence indicates that NctA and NctB are part of the same complex. Besides that, NctC seems to be related to temperature and osmotic stresses, being important for virulence in A. fumigatus. Therefore, more studies are needed in order to better understand the roles of these genes and may be important in bringing greater information about potential therapeutic targets in the treatment of aspergillosis.

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The traditional view of a lifelong monogamy between a king and a queen has recently been challenged in termites. In several species, multiple parthenogenetically-produced secondary queens replace the primary queen and mate with the primary king; this strategy is referred to as “Asexual Queen Succession” (AQS). The aim of my thesis was to investigate the modalities of reproduction and the ecology of neotropical soil-feeding termites from the Termitinae, with a focus on the inquiline termite Cavitermes tuberosus in the Termes group.In the first axis, we investigated the modalities of reproduction of C. tuberosus. (i) AQS is the main reproductive strategy of this species. (ii) The evolution of AQS requires the propensity of parthenogens to develop into neotenic queens. In C. tuberosus, secondary queens develop from a developmental stage of “aspirants” which participate to the social tasks usually undertaken by workers, as long as the primary queen is alive. (iii) In AQS species, a female-biased sex ratio is expected in the dispersing reproductives. In C. tuberosus, sex ratio varies among years and according to the type of reproductives, and the population sex ratio is balanced. These results raise hints on queen-king conflict over the sex ratio.In the second axis, we described the ecology and symbioses of C. tuberosus. (iv) Wolbachia, an endosymbiotic bacterium mainly known for manipulating the reproduction of arthropods in order to enhance its own transmission, infects all individuals in societies. This bacterium, particularly abundant in a gut-associated bacteriome, may play a role in the nutrition of C. tuberosus; both partners would have evolved a mutualistic symbiosis. (v) Inquiline termites live in a nest built by other termite species and do not forage outside. Physico-chemical measures and microbiota sequencing revealed that C. tuberosus is a generalist nest-feeder.Finally, we expanded our study of the breeding systems in the phylogenetic proximity of C. tuberosus. (vi) We described Palmitermes impostor, a new genus and species as a sister-group to the genus Cavitermes. (vii) AQS is the main reproductive strategy in P. impostor, and queens of Spinitermes trispinosus and Inquilinitermes inquilinus are able to reproduce parthenogenetically. Therefore, it appears likely that the conditional use of sexual and asexual reproductions is a preadaptation common to the whole Termes group, and that it evolved into a stable element of their breeding system at least in some species.Overall, our results open new perspectives in the understanding of reproductive strategies in termites and their relationships with their bacterial symbionts. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Les Porifères et les Cténophores sont probablement les deux lignées animales les plus anciennes. Leur étude permet de retracer l’histoire précoce des métazoaires, et d’aborder l'origine de la distinction entre les lignées somatique et germinale. En effet, chez les éponges cette distinction n’existe pas: les archéocytes et choanocytes pouvant donner les cellules somatiques et les gamètes.Après avoir établi la liste des gènes considérés comme impliqués dans la gamétogenèse chez les métazoaires, j’ai caractérisé leurs séquences et retracé leur histoire grâce à des analyses comparatives intégrant les principales lignées animales. Enfin, un suivi in situ d'Oscarella lobularis m’a permis d’affiner son cycle de vie et d’accéder à toutes les étapes du développement pour étudier l’expression de ces gènes et de tester leur implication dans la gamétogenèse.Ainsi, j'ai montré que 18 gènes du GMP (Germline Multipotency Program) sont présents ancestralement chez les animaux. Parmi ceux-ci 11 s’expriment pendant la gamétogenèse, au cours de l’embryogenèse, de la reproduction asexuée et de la régénération. Enfin, le suivi in situ a montré l’influence de la variation de la température et de la matière organique sur le déclenchement de la gamétogenèse.Mon travail suggère d’une part, que la spécification des cellules germinales est régie par des mécanismes génétiques communs à l’échelle de métazoaires, et d’autre part que ces gènes pourraient être impliqués dans la multipotence. Ces résultats renforcent l’hypothèse proposant une origine commune de la lignée germinale et des cellules souches somatiques. / Porifera and Ctenophora are probably the two most ancient animal lineages. Their study therefore allows to trace back the early history of metazoans and to address the origin of the distinction between somatic and germ lines. Indeed, in sponges this distinction does not exist: archeocytes and choanocytes can give rise to both somatic cells and gametes.After establishing the list of genes considered to be involved in gametogenesis in metazoans, I searched for these candidate genes (by local blast) in the transcriptomes of two sponge species (Oscarella lobularis and Oopsacas minuta). I thereby managed to characterize their sequences (phylogenetic and protein domain analyzes) and to trace their evolution through comparative analyzes including all main animal phyla. Finally, the in situ monitoring of O. lobularis enabled me to refine its life cycle and access all key developmental stages in order to study the expression of candidate genes in order to test their possible involvement in gametogenesis in this species.I have shown that 18 GMP (Germline Multipotency Program) genes are present ancestrally in animals. Among them, at least 11 are expressed not only during gametogenesis but also during embryogenesis, asexual reproduction and regeneration. Finally, in situ monitoring showed the influence of temperature variations and organic matter availability on gametogenesis.My work suggests, firstly that the specification of germ cells is controlled by common genetic mechanisms across metazoans, and secondly that these ancestral genes might be involved in pluripotency. These results reinforce the hypothesis suggesting a common origin of the germline and somatic stem cells.

<p>The brackish Baltic Sea is a marginal environment for both marine and freshwater species. The rate of ecological differentiation is presumably high due to strong selection pressure from a gradient of decreasing salinity that has been present in its current state for only about 3 000 years. Even more recently, increased nutrient loading due to human activities has affected the growth rate of species, with potential effects on their competitive interactions and responses to other regulating factors. I have investigated the potential effects of low salinity and nutrient enrichment on the distributional ranges of two marine macroalgae with a wide distribution in the Baltic Sea, the red alga <i>Ceramium tenuicorne</i> (Kütz.) Wærn and the brown alga <i>Fucus vesiculosus</i> L. </p><p>A field study in the northern Baltic Sea indicated a strong relationship between the community structure of macroalgae and abiotic factors even on a small, local scale. The abiotic factors are potentially modulated by eutrophication, which may have a strong effect on the depth distribution and abundance of macroalgae. On a regional scale, laboratory experiments suggested that nutrient enrichment is unlikely to affect the distribution of <i>Ceramium</i> and <i>Fucus</i> along the salinity gradient. Growth in <i>Ceramium</i> from the Baltic Proper was enhanced by nitrate and phosphate, but the response did not override growth constraints due to low salinity. <i>Ceramium</i> from the Gulf of Bothnia had an inherently lower growth rate that was not positively affected by nitrate and phosphate increase. In <i>Fucus vesiculosus</i>, reproductive performance was impaired by nitrate and phosphate levels corresponding to ambient levels in eutrophicated areas of the Baltic Sea, when measured by their effect on zygote attachment, germination, and rhizoid development.</p><p>The wide distribution of <i>Ceramium</i> in the inner Baltic Sea is probably related to local adaptation, rather than a generalized tolerance of different salinity levels. Ecotypic differences were observed when comparing strains from the Baltic Proper (salinity 7 psu) and the Gulf of Bothnia (4 psu). A high rate of vegetative reproduction was evident, although sexual reproduction was occasionally observed in salinity 4. In <i>Fucus vesiculosus</i>, genetic and morphological analyses of sympatric and allopatric populations of the common, vesicular, morphotype and a dwarf morphotype, characteristic for the Gulf of Bothnia, showed that the dwarf morphotype represents a separate evolutionary lineage. Also, vegetative reproduction was observed in <i>Fucus</i> for the first time, as supported by genetic and experimental data. </p><p>The results show that the biota of the inner Baltic Sea may have unique adaptive and genetic properties, and that it is highly relevant to consider subspecies diversity in Baltic Sea management.</p>

The brackish Baltic Sea is a marginal environment for both marine and freshwater species. The rate of ecological differentiation is presumably high due to strong selection pressure from a gradient of decreasing salinity that has been present in its current state for only about 3 000 years. Even more recently, increased nutrient loading due to human activities has affected the growth rate of species, with potential effects on their competitive interactions and responses to other regulating factors. I have investigated the potential effects of low salinity and nutrient enrichment on the distributional ranges of two marine macroalgae with a wide distribution in the Baltic Sea, the red alga Ceramium tenuicorne (Kütz.) Wærn and the brown alga Fucus vesiculosus L. A field study in the northern Baltic Sea indicated a strong relationship between the community structure of macroalgae and abiotic factors even on a small, local scale. The abiotic factors are potentially modulated by eutrophication, which may have a strong effect on the depth distribution and abundance of macroalgae. On a regional scale, laboratory experiments suggested that nutrient enrichment is unlikely to affect the distribution of Ceramium and Fucus along the salinity gradient. Growth in Ceramium from the Baltic Proper was enhanced by nitrate and phosphate, but the response did not override growth constraints due to low salinity. Ceramium from the Gulf of Bothnia had an inherently lower growth rate that was not positively affected by nitrate and phosphate increase. In Fucus vesiculosus, reproductive performance was impaired by nitrate and phosphate levels corresponding to ambient levels in eutrophicated areas of the Baltic Sea, when measured by their effect on zygote attachment, germination, and rhizoid development. The wide distribution of Ceramium in the inner Baltic Sea is probably related to local adaptation, rather than a generalized tolerance of different salinity levels. Ecotypic differences were observed when comparing strains from the Baltic Proper (salinity 7 psu) and the Gulf of Bothnia (4 psu). A high rate of vegetative reproduction was evident, although sexual reproduction was occasionally observed in salinity 4. In Fucus vesiculosus, genetic and morphological analyses of sympatric and allopatric populations of the common, vesicular, morphotype and a dwarf morphotype, characteristic for the Gulf of Bothnia, showed that the dwarf morphotype represents a separate evolutionary lineage. Also, vegetative reproduction was observed in Fucus for the first time, as supported by genetic and experimental data. The results show that the biota of the inner Baltic Sea may have unique adaptive and genetic properties, and that it is highly relevant to consider subspecies diversity in Baltic Sea management.

The Pezizomycotina, commonly known as the filamentous fungi, are a diverse group of organisms that have a major impact on human life. The filamentous fungi diverged from a common ancestor approximately 200 – 700 million years ago. Because of the diversity and the wealth of biological and genomic tools for the filamentous fungi it is possible to track the evolutionary history of genes and gene networks in these organisms. In this dissertation I focus on the evolution of two genes (lolC and lolD) in the LOL secondary metabolite gene cluster in Epichloë and Neotyphodium genera, the evolution of the MAP kinase-signaling cascade in the filamentous fungi, the regulation of the gene networks involved in asexual development in Neurospora crassa, and the identification of two genes in the N. crassa asexual development gene network, acon-2 and acon-3. I find that lolC and lolD originated as an ancient duplication in the ancestor of the filamentous fungi, which were later recruited in the LOL gene cluster in the fungal endophyte lineage. In the MAP kinase-signaling cascade, I find that the MAPK component is the most central gene in the gene network. I also find that the MAPK signaling cascade originated as three copies in the ancestor to eukaryotes, an arrangement that is maintained in filamentous fungi. My observations of gene expression profiling during N. crassa asexual development show tissue specific expression of genes. Both the vegetative mycelium and the aerial hyphae contribute to the formation of macroconidiophores. Also, with the help of genomic tools recently developed by researchers in the filamentous fungal community, I identified NCU00478 and NCU07617 as the genes with mutations responsible for two aconidial strains of N. crassa, acon-2 and acon-3 respectively.

The effects of energy quantity to the reproductive strategies of the sea anemone Aiptasia pulchella Carlgren 1943 Chien-Hsun Chen (Advisors: Drs. Keryea Soong, Chao-Lun Chen) Institute of Marine Biology, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan, R.O.C. Thesis abstract The influences of energy quantity on the sexual and asexual reproduction and possible trade-offs between them in the sea anemone, Aiptasia pulchella were investigated. Different feeding frequencies, light intensities and presence of zooxanthellae were the treatments of energy quantity. Lacerate numbers, lacerate sizes and total weight of lacerates were used to quantify investment in asexual reproduction; oocyte sizes and gonad weights were used as the measures of sexual reproduction. The correlation between total weight of lacerates and that of gonads provided an opportunity for evidence of trade-offs between sexual and asexual reproduction. Under higher feeding frequencies, A. pulchella produced more and larger lacerates as well as larger oocytes and heavier gonads than under lower feeding frequencies. Under lower light intensities, A. pulchella produced more and heavier lacerates with than without zooxanthellae. Lacerate sizes were influenced by light intensities, presence of zooxanthellae and the interaction of the two factors. At intermediate light intensity (2000 lux), Aiptasia pulchella produced heavier gonads than at 0 lux or 10000 lux. A. pulchella produced heavier gonads with than without zooxanthellae. The oocyte sizes of A. pulchella were also influenced by light intensities, zooxanthellae and the interaction of the two. In both sexual or asexual modes of reproduction, A. pulchella performed better under higher than lower feeding frequencies. There were higher level of asexual reproduction for A. pulchella under lower light intensities with zooxanthellae. A. pulchella achieved the highest levels of fecundity, as an indication of sexual reproduction, under a light intensity of 2000 lux in individuals containing zooxanthellae. There is a positive correlation between sexual and asexual investment under different energy regimes. This is not compatible with the prediction of the trade-off hypothesis.

Thesis (Ph.D.)--University of Wollongong, 2006. / Typescript. Includes bibliographical references: p. 161-192.