Protocols in Lichenology

As an intricate association between a fungus and one or more green algae or cyanobacteria, lichens are one of the most successful examples of symbiosis. These fascinating organisms survive extreme desiccation and temperatures. They are adapted to a great variety of habitats, from coastal fog zones of deserts to intertidal zones, from plant leaves in tropical rain forests to the glacial moraines of the Himalayas, and they are dominant components of communities in circumpolar ecosystems. Possibly, because of their tendency to grow in nutrient-poor habitats, lichens are extremely efficient accumulators of atmospherically deposited pollutants, and are therefore widely used to monitor environmental pollution. The wide range of secondary products only found in lichens show pharmaceutically interesting fungicidal, antibacterial and antiviral properties. Lichens are extremely difficult to culture, grow very slowly, and their secondary metabolites very often complicate the analyses of other compounds. As a result, they require special techniques. This manual provides well-tested protocols, including tissue culture protocols and methods for studying lichen ultrastructure, (eco)physiology, primary and secondary compounds, and nucleic acids. Protocols for using lichens to monitor environmental pollution and to document lichen biodiversity are also provided. Special terms used in lichenology are explained in a glossary.

Contenu

Culture and Cultivation.- 1 Isolation and Culture of Lichen Photobionts and Mycobionts.- 2 Culture of Thallus Fragments and Redifferentiation of Lichens.- 3 Resynthesis of Photosymbiodemes.- 4 Protoplast Isolation from Lichen Mycobionts.- 5 Differentiation Processes in Lichens-in Vivo Cultivation and Transplantation Methods.- 6 Isolation and Culture of Lichenicolous Fungi.- Ultrastructure.- 7 Preparative Techniques for Transmission Electron Microscopy and Confocal Laser Scanning Microscopy of Lichens.- 8 Preparative Techniques for Low Temperature Scanning Electron Microscopy of Lichens.- Physiology and Ecophysiology.- 9 Measurement of Chlorophyll Fluorescence in Lichens.- 10 Characterising Photosynthesis and Respiration in Freshly Isolated or Cultured Lichen Photobionts.- 11 Analysis of Ethylene and ACC in Lichens.- 12 Measuring Activities of the Enzymes Superoxide Dismutase and Glutathione Reductase in Lichens.- 13 Studying the Effects of Elevated Concentrations of Carbon Dioxide on Lichens Using Open Top Chambers.- 14 In situ Measurements of the Water Content of Lichens.- 15 Determination of the Parameters of Liehen Water Relations.- 16 Measurement of Lichen Growth.- Lichen Compounds.- 17 Analysis of Phenolic Products in Lichens for Identification and Taxonomy.- 18 Investigating the Production of Secondary Compounds in Cultured Lichen Mycobionts.- 19 Analysing Lichen Enzymes by Isoelectricfocussing.- 20 Analysis of Lipids in Lichens.- 21 Measuring Ergosterol and Chitin in Lichens.- 22 Analysis of Chlorophylls, Carotenoids, and Tocopherols in Lichens.- Nucleic acids.- 23 Isolation of Nucleic Acids from Lichens.- 24 PCR Techniques and Automated Sequencing in Lichens.- 25 RAPD-PCR of Lichens.- Bioindication and Biomonitoring.- 26 Biomonitoring Radionuclide Deposition with Lichens.- 27 Biomonitoring Heavy Metal Pollution with Lichens.- 28 Bioindication of Sulphur Dioxide Pollution with Lichens.- Biodiversity and Information Systems.- 29 Management of a Lichen Herbarium.- 30 Documentation of Lichens by Macrophotography.- 31 Computer-Aided Identification Systems for Biology, with Particular Reference to Lichens.- 32 On-line Documentation of Lichen Biodiversity.