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Microbes are key drivers of the world's ecosystems. The vast majority of the world's diversity and metabolic potential lies within micro-organisms, yet we are just beginning to understand and utilize this ultimate resource of biological diversity. Critical to our exploration of the microbial world are methods that allow for the analysis of organisms that are invisible to our eyes, difficult to distinguish from each other, and often impossible to grow using available culture methods. The field of microbial ecology has been revolutionized in the past two decades by the introduction of molecular methods into the toolbox of the microbial ecologist. This molecular arsenal has helped to unveil the enormity of microbial diversity across the breadth of the earth's ecosystems, and has revealed that we are only familiar with a very small minority of the organisms that carry out key microbial functions in diverse habitats. The Molecular Microbial Ecology Manual, Second Edition (MMEM-II) provides a detailed and user-friendly description of the methods that have made this revolution in microbial ecology possible.However, what is perhaps most exciting about MMEM-II is that it contains a large number of new chapters, highlighting the newest trends in microbial ecology research, which seek to provide more quantitative and statistically robust data, and means of coupling microbial identity and function. In addition, the majority of the proven methods described in MMEM's first version have undergone significant revisions to provide the most up-to-date applications available. The state-of-the-art methods described in MMEM-II have not only been provided by experts in the field, but in most cases by the laboratories that actually first developed and applied the methods, thus providing the MMEM-II user with unique first-hand tips and insight. The new on-line format available for MMEM-II should also add to the utility of MMEM-II by allowing users to search for key topics throughout the manual, skip between interrelated chapters at the push of a button, and by providing immediate availability to protocol updates and new chapters dedicated to future technical developments.
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Five years have now passed since the first symposium on frankiae was held at Harvard Forest, Petersham, Massachusetts, USA and the inauguration of the term actinorhiza. Many advances have been made during these five years in our understanding of the actinorhizal symbioses. Evidence for this was provided by the papers presented at the Wageningen Workshop on Frankia symbioses, held in Wageningen at the Department of Microbiology of the Agricultural University. Most of these papers are now published in this volume of PLANT AND SOIL. We kindly acknowledge the assistance of Anton Houwers, editor of the journal, in planning, reviewing and publishing these studies. Although the papers presented at Wageningen described the active research areas, they also illuminated those aspects of these symbioses which remain beyond our understanding. Primary among the areas of our ignorance is the concept of species within the bacterial symbiont, Frankia. At present groupings of bacterial strains are based on cell chemistry, physiology, serology, DNA homology and symbiotic capa bilities (cross-inoculation). When these classification schemes are merged no clear species framework is obtained. Undoubtedly part of the difficulty is due to a lack of strains for analysis. Currently bacterial strains from only half of the actinorhizal symbioses known to exist, have been isolated and studied in pure culture. We must postpone there fore any comprehensive taxonomic classification until a larger majority of the symbioses are represented. Another research area wherein our understanding is insufficient is host-symbiont interaction.
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For a long time microbial ecology has been developed as a distinct field with in Ecology. In spite of the important role of microorganisms in the environ ment, this group of 'invisible' organisms remained unaccessable to other ecologists. Detection and identification of microorganisms remain largely dependent on isolation techniques and characterisation of pure cultures. We now realise that only a minor fraction of the microbial community can be cultivated. As a result of the introduction of molecular methods, microbes can now be detected and identified at the DNA/RNA level in their natural environment. This has opened a new field in ecology: Molecular Microbial Ecology. In the present manual we aim to introduce the microbial ecologist to a selected number of current molecular techniques that are relevant in micro bial ecology. The first edition of the manual contains 33 chapters and an equal number of additional chapters will be added this year. Since the field of molecular ecology is in a continuous progress, we aim to update and extend the Manual regularly and will invite anyone to deposit their new protocols in full detail in the next edition of this Manual.