Lichens produce a diverse array of bioactive secondary metabolites, many of which are unique to the organisms. Their potential applications, however, are limited by their finite sources and the slow-growing nature of the organisms in both laboratory and environmental conditions. This thesis set out to investigate polyketide synthase genes in lichens, with the ultimate goal of providing a sustainable source of lichen natural products to support these applications. To expand the diversity of PKS genes that could be detected in lichens, new degenerate primers targeting ketoacylsynthase (KS) domains of specific clades of PKS genes have been developed and tested on various lichen samples. Using these primers, 19 KS domains from various lichens were obtained. Phylogenetic analysis of the KS domains was used to infer the function of the PKS genes based on the predicted PKS domain architecture and chemical analysis by TLC and/or HPLC. KS domains from PKS clades not previously known in lichens were identified; this included the clade III NR (non-reducing)-PKSs, PR (partially reducing)-PKSs and HR (highly reducing)-PKSs. The discovery of clade III NR-PKSs with C-methyltransferase (CMeT) domain and their wide occurrence in lichens was especially significant. Based on the KS domain phylogenetic analysis and compounds detected in the individual lichens, the clade III NR-PKSs were hypothesized to be involved in the biosynthesis of β-orsellinic acid and methylphloroacetopheno ne - the monoaromatic precursors for many lichen coupled phenolic compounds, such as β-orcinol depsides/depsidones and usnic acid. A strategy has been developed to isolate clade III NR-PKSs directly from environmental lichen DNA using clade III NR-type KS amplified from the degenerate primers (NR3KS-F/R) as homologous probes. Another pair of degenerate primers specific to the CMeT domain of NR-PKSs has also been developed to facilitate the cloning and probing of new clade III NR-PKS genes in lichens. A clade III NR-PKS gene (xsepks1) from X. semiviridis was cloned successfully. This is the first report of the isolation of a full-length PKS gene from environmental lichen DNA. The domain architecture of xsepks1 is KS-AT-ACP-CMeT, as expected for a clade III NR-PKS, suggesting that the newly developed clade-specific primers are useful for cloning new clade III NR-PKS genes and that KS domain phylogenetic analysis can predict the functional domains in PKSs. Attempts were made to characterize the function of xsepks1 by heterologous expression in Aspergillus species. Both A. nidulans (transformed with 5´partial xsepks1 including native promoter) and A. oryzae (transformed with full-length xsepks1 under the regulation of starch-inducible amyB promoter) were tested as potential hosts for the expression of lichen PKS genes. Transcriptional analysis showed that A. nidulans could potentially utilize the lichen PKS gene promoter and both fungal hosts could splice the introns of a lichen PKS gene. Several compounds unique to the A. oryzae transformants carrying xsepks1 were detected, but they could not be reproduced in subsequent fermentations even though the gene was transcribed into mRNA. None of the expected products (β-orsellinic acid, methylphloroacetophenone or similar methylated monoaromatic compounds) was detected in A. oryzae transformants, and the function of xsepks1 remains to be determined. The other clade III NR-PKS genes detected in X. semiviridis cou ld also be responsible for the biosynthesis of β-orsellinic acid or methylphloroacetophenone, as precursors of the major secondary metabolites detected in X. semiviridis (i.e. fumarprotocetraric acid, succinprotocetraric acid and usnic acid). Overall, the work in this thesis demonstrated the prospect of using a molecular approach to access the lichen biosynthetic potential without going through the cumbersome culturing stage.

This thesis consists of three manuscripts describing ecophysiological research on the cyanolichen Lobaria oregana. The first manuscript includes a re-evaluation of the assumptions underlying past estimates of N fixation by this species and provides an estimate of annual N fixation at the Wind River Canopy Crane (WRCC). Based upon litterfall data, canopy biomass data, N content of lichen tissue, and published growth rates, L. oregana fixes 0.4-1.6 kg N₂ ha⁻¹ yr⁻¹. The second manuscript presents a series of physiological response curves and a model of N fixation by L. oregana. Temperature is the most important parameter controlling nitrogenase activity in hydrated thalli. The model is used to predict annual N fixation at the WRCC and at the H.J. Andrews (HJA) Experimental Forest. Lobaria oregana fixes 1.4-1.8 kg N₂ ha⁻¹ yr⁻¹ at the WRCC, and low winter temperatures often inhibit nitrogenase activity. Temperatures at the HJA are slightly warmer during the winter, and L. oregana fixes 2.6-16.5 kg N₂ ha⁻¹ yr⁻¹ depending on its stand-level biomass. The third manuscript investigates the effects of thallus water content, light, and temperature on CO₂ exchange in L. oregana. This species shows a typical photosynthetic response upon rehydration, and like other lichens it becomes light-saturated at low PAR levels. Positive net photosynthesis in L. oregana occurs only between 1-12°C. High respiration rates prevent carbon gain at warmer temperatures. The temperature constraints on carbon gain and nitrogen fixation may explain some of the landscape distribution patterns of L. oregana. / Graduation date: 2002

Riparian forests in the Idaho Panhandle, north of Whitebird, were surveyed for rare riparian lichen species. The region was stratified into nine geographic units and by stream size. Eighty-one plots were surveyed for lichen community, stand and river characteristics. Variables important to lichen community composition included regional differences, elevation, climatic affinity, floodplain cross-section type, and the amount of basal area in hardwoods. Seventeen species were reported to be rare or uncommon in northern Idaho by lichenologists familiar with eastern Washington, northern Idaho and northwestern Montana. Of the seventeen target species, Cetraria sepincola, Pseudocyphellaria anomala, Ramalina pollinaria and Ramalina subleptocarpha were determined to be rare, with less than 25 occurrences each in northern Idaho. Lobaria hallii, Physconia americana, and Ramalina thrausta were determined to be locally abundant when found, but should continue to be species of concern in northern Idaho because of their limited distribution and narrow habitat requirements. Collema curtisporum is more common in the riparian forests of northern Idaho than previously thought, but appears to be restricted to old Populus balsamfera ssp. trichocarpa (black cottonwood) stands that receive seasonal inundation. Collema occultatum, Nephroma laevigatum, Leptogium cellulosum, Phaeophyscia hirtella, and P. ciliata are all new records for northern Idaho. Management recommendations include maintaining or restoring natural flood cycles in riparian forests and protecting mature black cottonwood stands on floodplains. / Graduation date: 2002

Thesis (doctoral)--Universität Göttingen, 2004. / Includes bibliographical references (leaves 106-117). Also available in PDF format.

Graduation date: 1991

The objective of the current study was to analyse the effect of landscape structure (habitat size and exposure to farmland) on the occurrence and abundance of Lobaria pulmonaria, a foliose cyanolichen. Since the agrarian revolution during the 19th century the agricultural landscape has become increasingly fragmented resulting in isolated meadows and wood-pastures surrounded by farmland. Lobaria pulmonaria is one of the species being affected by this habitat change, much due to their dispersal limitations, specific habitat demands and susceptibility to air pollution. 36 localities of two different size classes (< 1.5 ha and > 4.5 ha) and two different exposure classes (exposed or unexposed to farmland) were studied. The occurrence, size of lichen thallus and height of lichen patches on tree trunks were significantly positively affected by habitat size and negatively affected by habitat exposure. The implications of these findings for strategies to manage and conserve L. pulmonaria in a fragmented landscape are discussed.

Inaug.-Dissert. Kgl. Bayer. Julius-Maximilians-Universität Würzburg. 17 Feb. 1904.

Inaug.-Dissert. Kgl. Bayer. Julius-Maximilians-Universität Würzburg. 17 Feb. 1904.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1962. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M. Sc.)--Acadia University, 1998. / Includes bibliographical references (leaves 75-82). Also available on the Internet via the World Wide Web.