ADVANCES IN MICROBIAL TOXIN RESEARCH AND ITS BIOTECHNOLOGICAL EXPLOITATION Edited by Rajeev K. Upadhyay Directorate of Plant Protection, Quarantine and Storage Faridabad, India Kluwer Academic / ee ee New York, Boston, Dordre nd 2002STRUCTURAL DIVERSITY OF LICHEN METABOLITES AND THEIR POTENTIAL USE Joanne G. Romagni’ and Franck E, Dayan" *University of St. Thomas, Houston, TX; “USDA-ARS, Natural Products Utilization Research Unit, University, MS, 38677 USA 1, INTRODUCTION Lichens are symbiotic organisms composed of a fungal partner, the mycobiont, usually in association with one or more photosynthetic partners, the photobiont(s). The photobionts can be green alga, cyanobacteria or both. There are approximately 17,000 species of mycobionts whereas there are only ca. 40 species of photobionts. Therefore, the taxonomicname of the lichen is traditionally determined by the species of the fungal partner. The mycobiont, being the host partner for the photobiont, generally determines the morphology of the lichen species. Most lichen mycobionts are not known to exist naturally without the photobiont, whereas many of the photobionts, like the cyanobacteria Nostoc, Scytonema, and the green algae, Trentepohlia, are knownto flourish alone in nature. However, because the symbiosis is so complex, lichens are normally referred to as individual organisms rather than separate organisms. ‘The majority of lichen secondary metabolites originate from the fungal partner and are normally produced when the organisms are in their symbiotic association. These compounds are produced in the cells, but are usually excreted and deposited on the external surface of the hyphae rather than inside the cell. Most of the compounds are insoluble in water and must be extracted with organic solvents, Jtis rather interesting, however, that of the 630 known metabolites, only about 50—60 occur in other fungi or higher plants (Culberson, 1969; Elix, 1996). Forexample, the anthraquinone Parietin is an orange pigment that is common in the lichen family Teloschistales, and it also occurs in several fungal genera such as Aspergillus and Penicillium, as well as in the vascular plant species Rheum, Rumex and Ventilago. : Secondary metabolites were originally used by scientists as markers for the chemotaxonomic, classification of lichens. The first chemical analyses of lichen thalli were done by Nylander in the 1860°s (Nylander, 1866). Asahina and his co-workers in the 1930s (Asahina and Shibata, 1954) established one of the most comprehensive elucidations of the chemical strictures oflichens. Although numerous new lichen compounds have been reported since then, these early studies have laid the foundation forall modem workinthisarea. Whilemodem molecular analyses such as PCR amplification nd restriction fragment length polymorphism (RFLP) are becoming more widespread for the Classification of lichens according to their genetic traits (Guzow-Krzeminska and Wegrzyn, 2000), ‘Advanées in Microbial Torin Research and its Biotechnological Exploitation Edled by Rajeev K. Upadhyay, Kluwer Academic/Plenum Publishers, New York, 2002 151traditional chemical profilingis still useful for the chemotaxonomic classification of some lichen species (Harris e al., 2000; Trinkaus and Mayrhofer, 2000; Rosato and Scutari, 2000). ‘The morphological and chemical analyses done by Nylander indicated that the distribution of pigments was not uniformly distributed throughout the thallus. Some pigments were restricted to the upper cortex, while others were in the medullary layer (Figure 1). <— Upper cortex Algal layer +— Medulla <— Lower cortex — Rhizines Figure 1, Cross section of a lichen thallus Scientists found a 10-fold higher concentration of the metabolite usnic acid in the outer regions ofthe thallus than in the inner region in the lichen Usnia diffracta (Taguchi et al., 1969). Current electron microscopy techniques, mass spectrometry and nuclear magnetic resonance have helped to identify the crystals located on or in the thallus (Culberson and Elix, 1989). There appears to bea concentration of these compounds on vegetative and reproductive portions of the thallus (Lawrey, 1995). 2. LICHEN DIVERSITY As previously mentioned, lichens have been traditionally characterized and categorized atcording to their lichenized fungal partners, Lichen mycobionts are an ancient group of fungi with slow rates of evolution (Kamefelt, 1990). According to Taylor(1987), they probably arose during the evolutionary radiations of the ascomycetes approximately 400 million years ago. The diversity of lichens suggests that lichenization as a character has occurred more than once (Tehler, 1996). The estimated number of species varies between ca. 14,000 (Hawksworth and Hill, 1984) and 17,000 (Hale, 1983), depending on the stringency of the classification scheme used. Most lichens have an Ascomycete species as their mycobiont and almost half the described Ascomycetes are lichenized (Tehler, 1994). Lichenized Basiodiomycetes and Deuteromycetes are much less common, but can also be found, although the latter group is sterile (Nash, 1996). Lichen morphology is extremely diverse, and these organisms come in a fantastic array of colors, including orange, yellow, red, green, grey, brown and black (Wirth, 1987). They can vary in size from very small individuals (< 1 mm?) to drooping structures over a meter long (Nash, 1996), They are found inall ecosystems. Lichens can colonize bare rock surfaces, occur on soil, grow endolithically onrocks, grow as epiphytes on trees, and in some tropical species, develop on the surface of leaves. Some species grow in freshwater streams and others in intertidal zones (Nash, 1996). Lichen morphology and thallus structures are generally determined by the mycobiont and the few exceptions will not be mentioned in this paper. The general morphology can be subdivided into three main categories : crustose, foliose and fruticose, with a fourth subgroup, the gelatinous lichens, which isnot covered here. Crustose lichens have a thallus tightly adhered to the substrate at the lower surface, and they cannot be removed from the substrate without seriously damaging the organism. They often are primary successional species surviving in inhospitable areas and, in many cases, the weak lichen acids produced slowly break down rocks for later successional species. Foliose lichens are two-dimensional leafy structures, typically with an upper and lower side, and, are divided into lobes. This group displays a large range of thallus diversity. The lower surface of these lichens usually has some sort of attachment structure, such as rhizines or cilia. 152