Protocols for plant cell biologists

Abstract

Plant cell biology (2nd edition) Edited by C. R. Hawes & B. Satiat-Jeunemaitre. 338 pages. Oxford, UK: Oxford University Press, 2001. £70.00 hours/b. ISBN 0 19 963866 7 In Plant cell biology (2nd edition) a practical approach, an attempt has been made to do the impossible. Editors Chris Hawes and Béatrice Satiat-Jeunemaitre have attempted to collect into a single, reasonably priced volume descriptions of, and protocols for, most of the techniques used by plant cell biologists. A wide range of methodologies are presented, including light and electron microscopy, immunocytochemistry and in situ hybridization, transient expression of genes in plant cells, and use of the green fluorescent protein (GFP) as a reporter molecule. Each chapter is written by researchers who share with the reader insights that have come from their own experience, and protocols from their own laboratories. The editors, perhaps deliberately, have done little to impose a uniform writing style. Although this is slightly off putting when liberties are taken with standard English, it reinforces the impression that what is presented is based on the experience of individuals and on the success they have had with particular cell types, tissues and plant species. Of note are excellent, thorough chapters on optical microscopy, fluorescent probes for live cells, in situ hybridization, and organelle isolation. The chapter ‘Plant histology’ provides a brief introduction to the subject, and will be useful to those needing a set of standard protocols. There is a wealth of information in the chapter ‘Electron microscopy’, though at times the inclusion of so much material obscures the organization of the chapter and is confusing. Likewise ‘Immunocytochemistry for light microscopy’ lacks the focus that would make it most useful to novices. Coverage of transient expression seems, at first glance, to be extremely limited since only electroporation is described in any detail. Fortunately, protocols for plant transformation using polyethylene glycol, Agrobacterium, Potato Virus X and biolistics are included in the chapter ‘The green fluorescent protein (GFP) as reporter in plant cells’. The chapter on microinjection scarcely mentions transformation via microinjection, but does discuss in detail the unique challenges posed by various cell types. Perhaps least useful to most readers, except as a description of the methodology, is ‘Micromanipulation by laser microbeams and optical tweezers’. The chapter on electrophysiological methods, surprisingly, is restricted to capacitance measurements as an assay for exo- and endocytosis. More basic voltage clamp and patch clamp techniques have been extremely useful tools for plant cell biologists during the past two decades. It is unfortunate that explicit protocols were not included for making measurements of, for example, voltage difference across the plasma membrane or whole cell currents. Flow cytometry is a powerful technique that is rarely used by plant biologists, but commonly used by other cell biologists. It is disappointing therefore that the chapter on flow cytometry covers only the quantification of DNA in nuclei in any depth. Illustrating a wider range of applications would have been beneficial to the plant science community. This book will be an excellent resource for anyone who needs more than a cursory introduction to an unfamiliar methodology. Those that will find it useful are scientists who want to use a new technique for their own research, and instructors trying to develop contemporary laboratory exercises. Plant cell biology (2nd edition) a practical approach, does not accomplish the impossible. Yet it is a remarkably good attempt, and exceptionally good value. I highly recommend it.