Preface for PHYSCOmanual


The moss Physcomitrella patens is an emerging model organism for various fields in science, and it will be more extensively used in near future. There should be several advantages to use this moss, in which the most prominent is its feasibility in gene targeting based on the high homologous recombination rate similar to that of fission yeast. P. patens nuclear genome will be fully sequenced and annotated in a couple of years, which will facilitate the reverse genetic approaches. Forward genetics are also applicable for this moss and other new techniques especially on imaging at cellular level are enormously improved recently. We intend this laboratory manual to be useful for new in P. patens research. Furthermore, we hope that this laboratory manual will be useful also for more established researches to improve their experimental techniques. Actually, contents of this manual are extensively changed in the last few years.

This manual starts from the cultivation and the observation of wild type moss. Then, necessary techniques for reverse genetics, such as gene isolation, southern hybridization, northern hybridization, and gene targeting are described in detail. These are mostly similar to regular techniques in molecular biology, but there are some minor but critical tips for P. patens, which are stressed in the text. Useful techniques to characterize gene functions, such as the observation of disruptant and overexpressor phenotypes and the detection of protein localization, were fully described. The last chapter focuses how to use PHYSCObase. Many plasmids listed at the end of this manual are all available.

Japanese version of this manual was originally made to share laboratory techniques in our laboratory, and then opened for public in our web site since 1998. All protocols were written by former and present members in our laboratory, and I deeply appreciate for their enthusiasm to Physcomitrella patens. Rumiko Kofuji (Faculty of Science, Kanazawa University), Keiko Sakakibara (Graduate School of Science, Hiroshima University), Tomoaki Nishiyama (Advanced Science Research Center, Kanazawa University), and Tomomichi Fujita (Graduate School of Sciences, Hokkaido University) left my laboratory, and are now expanding moss biology intheir new laboratories. All authors of this manual appreciate the open-minded Physco community for sharing knowledge and many techniques, especially David Cove, Didier Schaefer, Jean-Pierre Zryd, Andy Cumming, Celia Knight, Ralf Reski, Ralph Quatrano and more moss friends. The old version was expanded to cover a broader range of techniques for the laboratory course in National Institute for Basic Biology, and revised year by year.

We hope this manual will be useful for moss researchers, and any comments and suggestions on the improvement of this manual are very welcome.



                                    Mitsuyasu Hasebe