Thesis (Ph. D.)--University of Washington. / Bibliography: l. 358-361.

Thesis (M.S.) in Ecology and Environmental Science--University of Maine, 2008. / Includes vita. Includes bibliographical references (leaves 80-86).

In this study, inter- and intraspecific matings were made between mycelia of different species in the genus Coprlnus. The species studied were: C. comatus (Schum.) Fries, C. micaceus (Bull.) Fries, C. domesticus (Pers.) Fries, C. macrorrhizus (Pers.) Rea, C. myceliocephalus Lange, C. narcoticus, Fries, C. Patouillardii Quel., C. lagopus Fries, and C. picaceua (Bull.) Fries. Matings were carried out using both heterothallic and homothallic species. The cultural characteristics of all species were studied and described following growth on semisynthetic solid culture media. The results of the inter- and intraspecific crosses were assessed by microscopic observations of the area of contact between paired species and by further growth and examination of the mycelia and fruit bodies arising from inocula taken from the contact area. Mycelial macerates as well as plug inocula were employed in the study. Chemical monokaryotization techniques were applied to the homodikaryotic mycelium of C. myceliocephalus. Hyphal fusions or anastomoses were not observed in the confrontation of non-compatible mycelia of the same species. This observation was true in pairings of monokaryophase x monokaryophase, dikaryophase x dikaryophase, and monokaryophase x dikaryophase, involving mycelia of the same species. Close contact, taking the form of hyphal coils or hyphal "knots," was observed in pairings of compatible monokaryophases. Similar contact was obtained between hyphae of two different isolates of the homothallic species, C. Patouillardii, but no cytoplasmic continuity could be seen between the paired mycelia. Apparent cytoplasmic continuity and hyphal contact was established in crosses between the dikaryophases of C. micaceus and C. comatus and C. macrorrhizus and C. narcoticus. In addition, similar relationships including hyphal contact and cytoplasmic continuity were observed between mycelia of the homothallic species. C. myceliocephalus and C. Patouillardii. No nuclear transfer was observed between these different species. The cultural characteristics of mycelium subsequently isolated from the cross of C. micaceus x C. comatus and C. myceliocephalus x C. Patouillardii were significantly altered from those of the original species. The linear growth rate was increased in both cases. In addition, changes in coloration and mycelial form were observed. The mycelium arising from the cross of C. narcoticus x C. macrorrhizus was identical in every respect to that of C. macrorrhizus alone. No evidence of the growth of C. narcoticus could be seen in mycelium arising from this cross. All other species crosses studied revealed no anastomoses or formation of clamp connections following mating. Three monokaryotic isolates were obtained from the treatment of the homodikaryophase of C. myceliocephalus. Two of these isolates were obtained following sixteen weeks growth on medium containing .002 M sodium arsenate, while the third was isolated after three weeks growth on liquid medium containing 0.5% cholic acid. Cytological examination of the mycelia of the three isolates showed them to be monokaryotic and all three isolates lacked clamp connections. The clampless isolates produced no fruit bodies in culture on any medium, while the original isolate readily fruited on supplemented malt agar or on oat agar. Pairings of the isolates in all possible combinations resulted neither in mating nor in clamp connection formation. The stability and distinctiveness of cultural characteristics recommended their use in any future monographic studies of this genus. The presence of hyphal interactions, including hyphal continuity between different species, suggested the need for the examination of interspecific crosses for evidence of mating other than clamp connection formation. Homothallic and heterothallic species should be considered in studies of interspecific mating. Putative hybrid mycelia were produced from matings between different species. The absence of fruit body formation by these mycelia prevented demonstration of their hybridity in this study. The recovery of clampless, monokaryotic mycelia from the homodikaryon of C. myceliocephalus suggested that this species was secondarily homothallic. / Science, Faculty of / Botany, Department of / Graduate

Thesis (Ph. D.)--University of Wisconsin--Madison, 1955. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 65-72).

No description available.

Techn. Universiẗat, Diss.--Braunschweig, 1995.

C. cinereus, any two of which are sufficient to promote B-regulated development following cell fusion. The isolation of the B42 locus is described along with the DNA sequence analysis that identified nine B mating type genes within a 27kb B42 -specific DNA sequence. Six of the genes, with small transcripts of 800-900nt, encode the mating pheromone precursors and the other three, with 1.9 to 2-5kb transcripts, encode the transmembrane pheromone receptors. The genes are arranged in three groups, designated group 1, 2 and 3, each consisting of one receptor gene and two pheromone genes. B42 and B6 share the same alleles of the group 1 genes, but not those of groups 2 and 3. This was demonstrated by DNAsequence analysis and Southern blot analysis. None of the group 1 genes from B42 were able to activate B -regulated development in a B6 host when introduced by transformation but with one exception, all genes from group 2 and group 3 were able to do so. This analysis led to the recognition that the three genes in any one group are held together in an allele-specific DNA sequence and that Southern blot analysis and transformation can be used to identify shared alleles in uncloned loci. Extensive Southern analyses using cloned genes to probe genomic DNAs from strains having other B mating specificites showed that different B loci may share identical alleles of two groups of genes. Mating partners thus require different alleles of only one group of genes to generate a compatible B mating interaction. Transformation analyses with the same cloned genes confirmed the conclusions derived from the hybridisation data. Multiple B mating specificities thus appear to be derived from three groups of multiallelic and functionally redundant genes. A tenth gene located within the B42- specific DNA sequence encodes a putative transporter protein belonging to the major facilitator superfamily (MFS). In other genomic backgrounds this gene lies in homologous flanking sequences and its presence within the B42 locus is unlikely to be related to mating type function.

<p>Mushrooms are believed to have a great potential for production of bioactive metabolites e. g. terpenes, a group of interesting compounds with diverse chemical properties such as antitumour and antibacterial activity. Cop6 is a terpene cyclase isolated from the mushroom <em>Coprinus cinereus</em> that catalyzes the cyclization of farnesyl diphosphate (FPP) to mainly α-cuprenene. In this study gas chromatography combined with mass spectroscopy (GC-MS) is used to analyze the product profile of Cop6 mutants created by PCR based site directed mutagenesis. The goal is to produce trichodiene, the parent hydrocarbon in the biosynthesis of trichothecene antibiotics and mycotoxins. Valine instead of tyrosine in amino acid position 195 resulted in cyclisation of (E)-β-Farnesene and (3Z,6E)-α-Farnesene besides the products of the wild type enzyme. Another mutant with aspartic acid instead of asparagine in position 224 resulted in the synthesis of β-Bisabolene except for α-cuprenene and methionine in position 74 instead of isoleucine killed the activity of the cyclase. Furthermore, an attempt to saturation of position 98 was made, resulting in four mutants. Two of them essentially killed the activity of the cyclase whereas two had minor effect of the product profile compared to the wild type. </p>

Mushrooms are believed to have a great potential for production of bioactive metabolites e. g. terpenes, a group of interesting compounds with diverse chemical properties such as antitumour and antibacterial activity. Cop6 is a terpene cyclase isolated from the mushroom Coprinus cinereus that catalyzes the cyclization of farnesyl diphosphate (FPP) to mainly α-cuprenene. In this study gas chromatography combined with mass spectroscopy (GC-MS) is used to analyze the product profile of Cop6 mutants created by PCR based site directed mutagenesis. The goal is to produce trichodiene, the parent hydrocarbon in the biosynthesis of trichothecene antibiotics and mycotoxins. Valine instead of tyrosine in amino acid position 195 resulted in cyclisation of (E)-β-Farnesene and (3Z,6E)-α-Farnesene besides the products of the wild type enzyme. Another mutant with aspartic acid instead of asparagine in position 224 resulted in the synthesis of β-Bisabolene except for α-cuprenene and methionine in position 74 instead of isoleucine killed the activity of the cyclase. Furthermore, an attempt to saturation of position 98 was made, resulting in four mutants. Two of them essentially killed the activity of the cyclase whereas two had minor effect of the product profile compared to the wild type.

Fungi are integral parts of terrestrial ecosystems. Sporocarps propagate by spreading their spores from fruiting bodies. Their fruiting bodies are looked for both by foragers and conservationists. Potential changes to spore production and the timing of fruit body production could be useful information for both foragers and conservationists. This report uses e-DNA from air filters gathered from Kiruna in northern Sweden to measure shifts in the spore-producing period of the fungal genera Boletus, Morchella, Coprinus, Cantharellus, Hydnellum, and Ramaria, both in terms of amount of spores released and timing of fruit body formation. The start week, peak week of sporulation, estimation of seasonal end, and cumulative sum of spores released for each genus were calculated and then correlated against each other. The relationship between these variables and the climatic variables of temperature and precipitation, both for the current year and previous year were also calculated.    The report found no significant trends in the timing of the start of sporulation, but spore production for both Morchella and Ramaria increased along the timeseries. The report found an overall positive response to increases in precipitation and temperature for the studied genera, with the exception of Boletus, indicating that climate change is unlikely to negatively impact these genera in northern Sweden.