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Microscopy techniques

Methods in plant electron microscopy and cytochemistry

Ed. by William V. Dashek. xi + 300 pages. Totawa, NJ, USA: Humana Press, 2000. US$ 89.50; £67.50 p/b. ISBN 089603 589 1

There is certainly scope for morebooks on microscopic techniquesfor studying plants, because mostof the ‘standard’ methods weredeveloped for animal tissues andoften give poor results with planttissues. A new book on planttechniques could be useful toeveryone from schoolteachersplanning plant biology classes tosenior researchers needing toacquire specific new skills. Doesthis book fulfil this potential? The

answer, as so often, is yes and no. Yes, because of the severalexcellent chapters which give all the information requiredfor using the methods described. But on balance, no. It appearsthat the hard-pressed editor, valiantly trying to meet publica-tion deadlines but failing to extract the promised chaptersfrom a number of colleagues, was forced to step in late tomake up the shortfall: he is author or coauthor of almosthalf the main chapters and coverage of the subject is veryuneven.

There are some startling omissions. Open up any issue ofa plant journal which has a molecular slant, and every sec-ond or third paper features illustrations of

in situ

hybridiza-tions of riboprobe to transcript, a key technique in the studyof tissue specificity of gene expression. Both hybridization totissue sections followed by light microscopy, and tissue printanalysis, have been used extensively. But except for fleetingmentions, RNA detection isn’t covered at all in the lightmicroscopy section, and is given only a brief note in the finalchapter, ‘EM and molecular biology’, insufficient to enableanyone to use the method. Moreover, there’s more on detect-ing bacterial surface antigens than on plant RNA.

Another microscopy technique which has revolutionizedplant cytology is the application of fluorescence

in situ

hybridization (FISH) to plant chromosomes, using eithertotal genomic DNA (GISH) or specific probes. Again, noth-ing in this book: studies on chromatin are limited to a brief

description of 4,6-diamidino-2-phenylindole dihydrochloride(DAPI) staining for nuclei, and a couple of pages outlining SEMobservation of chromosomes, visualization of DNA helix andnucleosomes, and determination of chromatin loop size. Whilethe first method is described for

Vicia faba

, the latter two aremethods developed for chicken erythrocytes and humanleukaemia cells, respectively. Will they work for plant (orfungal) material? We aren’t told. There’s nothing at all oncytological/microscopic techniques for looking at meiosis.

Now I come to my biggest criticism. Three of my col-leagues (plant biochemist, physiologist and molecularbiologist) independently considered the defining features ofa plant (as opposed to other eukaryote) cell to be, in differ-ent orders, chloroplasts /plastids, cell walls, and vacuoles/tonoplasts. Except in the introductory chapter, there ishardly a mention of any of these compartments. Thechapters on topics such as fluorescence microscopy of anilineblue stained pistils, dark-field microscopy and its applica-tion to pollen tube culture, and isolation and characteriza-tion of endoplasmic reticulum from mulberry corticalparenchyma cells are more suited to a methodological jour-nal or the proceedings of a methods workshop: someonewanting to use dark-field microscopy to study pollen tubeswouldn’t buy this book just for the relevant chapter, andthose of us who don’t work on pollen tubes aren’t going togain much (except a couple of attractive pictures). I imaginethat some people who assumed that they were investing in awork devoted to higher plants might feel similarly about‘Scanning electron microscopy – preparations for diatoms’,although diatoms are so breathtakingly beautiful that I can’tsee enough of them. Some equivalents from higher plantswould have been welcome too – how about SEM studies ofmeristem/floral development, a technique which is oftenvery informative? More to the point, I expected chapters onthe study of plant cell walls, vacuoles/tonoplasts, and, mostimportantly, plastids. In the otherwise useful ‘Identificationof isolated organelles’, a table of marker enzymes for identi-fying particular organelles includes Golgi apparatus, plasmamembranes, ER, secretory vesicles, intact vacuoles and tono-plast, but omits mitochondria and plastids.

A few other gripes before I come to the positive points.Several errors have slipped through at the proof stage. Someof the diagrams, which have been reproduced from otherpublications, are confusing because the text doesn’t tie inwith them very well. But more worrying are safety issues,especially important in a book which might be used inschools and in the design of undergraduate practicals. Inseveral chapters coverage was reasonable, but I was some-what taken aback by a figure showing the method for

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preparation of stripping-film autoradiographs, in which amicroscope slide is manoeuvred under water to pick up apiece of stripping film. Although this slide would carryspecimens radiolabelled with tritium, 14C or even a higher-energy isotope, the worker’s hand is ungloved. The radiationhazard apart, the ribonucleases on the worker’s skin woulddamage the specimen if RNA were the macromolecule ofinterest. Still on safety, I was surprised to read that radioactivewaste must be ‘further broken down into solid waste, liquidwaste and animal carcasses to aid in its proper disposal’.How many radioactive animal carcasses does the averageplant or fungal research lab produce? Flies which have falleninto the stock isotope solution, maybe? Or graduate studentswho have failed to observe proper safety precautions?

So what’s good about this book? Many of the chapters areexcellent. The first gives a good general introduction toplant cells and tissues – useful to an animal biologist or hard-line molecular biologist needing to get up to speed on plantstructure and composition. Chapters 2–4 contain a greatdeal of information about localization of chemicals in planttissues, radioautography, and fluorescence methods. ‘A shortintroduction to immunocytochemistry and a protocol forimmunovisualization of proteins with alkaline phosphatase’could certainly have been longer, but would be extremelyhelpful to those wanting to implement the methods in theirown labs. It is well illustrated and gives sensible tips aboutchoice of controls and how to get started (choose a simplemethod; consult your colleagues; ideally try the favouritemethod of a nearby lab or Hospital ). My only criticism isthat monoclonal antibodies are recommended for over-coming the problems presented by polyclonals. Often, becauseeach monoclonal by definition recognizes only one epitope,the epitope in question, exposed in the denatured proteinused to raise the antibody, is inaccessible in the tissue sec-tion being challenged. Hence a monoclonal will sometimeswork well with western blots but not in immunolocaliza-tion. The reader could do with a warning about this. Sev-eral of the electron microscopy sections are also useful: theone about immunogold localization for EM is particularlygood on controls to avoid artefacts, although protocolsshould be more detailed. Since I didn’t know much aboutmicroanalysis, I found the coverage of the principles andapplications of the different types of analytical instrument(electron probe microanalysis, particle induced X-ray emission

(PIXE), laser microprobe mass analyzer (LAMMA), electronenergy loss spectroscopy (EELS), secondary ion mass spec-trometry (SIMS) ) informative. Although the title of Chapter15 is ‘Methods for atomic force and scanning tunnelingmicroscopies’ it is actually an introduction to the principlesand a summary of some recent applications, which directthe reader to further information.

Finally, my favourite chapter, on computer-aided micro-photometry (CAM), is a systematic guide to everything youneed to know about using a photometer attached to a lightmicroscope for computer-aided image analysis and quantifi-cation of cell components, whether you’ve just acquired aready-made commercial kit or want to construct your own.I loved the thought that a teenage nerd might be inspired toturn her, or his, school’s ancient microscope and a mid-range PC into a fairly sophisticated analytical facility. Thecomponents of a generic CAM system, with or withoutepifluorescence and polarized-light capability on the camera;illumination, shutters (real or virtual), photometers andmonochromators; choice of interface and software are allcovered. Programming is described using BASIC, andalthough this language might not be many people’s choice,the concepts transfer well enough. The section on imageanalysis is excellent. I thought the description and illustra-tions of image matrix smoothing using a kernel should berequired reading for anyone working with computer images.At the end of the chapter is a troubleshooting guide, some-thing which would have been useful elsewhere in the book.Throughout, the style is clear, readable and witty, and I likethe author’s attitude, summed up in his remark, ‘For thosewith minimal funding, this is a way to have some fun anddo good science’.

Do I recommend this book? The value of individualchapters would make it a useful addition to an Institutionallibrary, but I can’t recommend it for individual purchasebecause it doesn’t cover all the main areas of plant electronmicroscopy and cytochemistry by a long way.

Helen Ougham

Cell Biology Department, IGER, Plas Gogerddan,Aberystwyth SY23 3 EB, UK

(tel +44 1970823 094; fax +44 1970823 242;email [email protected])


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The facts of photosynthesis – an accessible resource

Photosynthesis: a comprehensive treatise, 2nd edn

Edited by A. S. Raghavendra. xviii + 376 pages. Cambridge, UK: Cambridge University Press, 2000. £29.95 (US$47.95) p/b. ISBN 0 51 78444 1

Photosynthesis research is theone area of plant science thatwins Nobel prizes. Having receivedthe attention of some of the big-gest brains, and deepest pockets,the subject has matured over thecourse of the last century to thestage where its fundamental pro-cesses can be taken out of thehands of biologists and left to thechemists and quantum physicists.

This is good news for students and teachers because it meansthat the essentials of the subject do not change from week to weekand the Z scheme or the Calvin-Benson cycle are descriptionsof light and dark reactions as accurate today as they were ageneration ago. It also means that textbooks dealing with themajor principles of photosynthesis do not go out of style quiteas quickly as those dealing with other aspects of plant biology.

Thus it stands with

Photosynthesis

, the corrected paper-back edition of a book first published in 1998. The liter-ature cited includes a few 1998 papers and one or two from1999, but essentially it represents a view of the field in themid-nineties. Even so, I would guess that it has a few moreyears of usefulness left and is a handy source of the factsof photosynthesis presented at a level accessible to bothresearchers and advanced students.

In 26 chapters the 44 authors, drawn from nine countriesin Europe, North America and Australasia, cover the whole scopeof the subject from cell and molecular biology to agronomy,evolution and biotechnological applications. The editor hasmade a good job of blending multiauthor and multidiscipli-nary contributions into a consistent style. This generouslyand, for the most part, clearly illustrated book lives up to itssubtitle: it is indeed a comprehensive treatise on the processthat defines plants and sustains the whole of life on earth.

Howard Thomas

Cell Biology Department, IGER, Plas Goderddan,Aberystwyth SY23 3EB, UK

(tel +44 1970828255; fax +44 1970828357;email [email protected])

150no issue no.2001163BooksBooksBooks000000Graphicraft Limited, Hong Kong

Surveying the plant kingdom

Green plants: their origin and diversity, 2nd edn

By P. R. Bell and A. R. Hemsley. x + 349 pages. Cambridge, UK: Cambridge University Press, 2000. £19.95 (US$31.95) p/b. ISBN 0521 64673 1. £55.00 (US$90.00) h/b. ISBN 0521 64109 8

Form and function, classifica-tion, and evolution are at theheart of the classical discipline ofbotany, a discipline that has hadmore than its share of ups anddowns. Renamed and riding themolecular wave, plant biology hasbeen refreshed and reinvigorated.Not so long ago hand sections,morphological specimens andtaxonomic trees had a dusty,

yesterday’s-science image; now we are beginning to look atthem with new understanding and they are becoming onceagain the indispensable tools of the botanical trade.

All the more reason therefore to welcome the secondedition of Bell and Hemsley’s well organized and clearlywritten survey of the plant kingdom published in 1992.Starting with an admirably terse and to-the-point overviewof plant origins, adaptations and life cycles, the book worksits way systematically from cyanobacteria through to themost advanced angiosperms, covering en route the morpho-logy, reproductive development and evolutionary signific-ance of representatives of the major phyla.

Structures are illustrated by well executed line drawingsand detailed photographs, and a useful glossary of botanicalterms is included

. Green plants

would serve as a basic text forstudents of plant or general biological sciences and also arefresher and reference source for researchers.

Howard Thomas

Cell Biology Department, IGER, Plas Goderddan,Aberystwyth SY23 3EB, UK

(tel +44 1970828255;fax +44 1970828357;

email [email protected])

150no issue no.2001164BooksBooksBooks000000Graphicraft Limited, Hong Kong


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Biopesticides

Biopesticides – use and delivery

Ed. by F. R. Hall and J. J. Menn. 626 pages. Totowa, NJ, USA: Humana Press, 1999. h/b. ISBN 089603 515 8

This fifth volume in the series

Methods in biotechnology

addressesthe basic biology of key organisms,examples of their use, the develop-ment of recombinant biopesti-cides and the potential for the

joint action of different groups.The full range of biopesticides is considered, from

biologically derived forms of pest control, such as neem,pheromones and transgenic plants, to the more traditionalviruses, bacteria, fungi and nematodes.

Particularly useful information on the registration ofbiopesticides is given, although much relates to the USA,and there are general overviews of opportunities for thedevelopment of biopesticides in Europe, North America and

the Developing World. The chapters on delivery systemsand strategies for resistance management are particularlygood and thought-provoking.

If I have any criticism it is that there is no chapter dealingsolely with potential environmental impact. Nontarget impactof biological control agents in the classical sense is attractingincreasing scrutiny. This aspect of microbial pesticides isoften only briefly covered, however, is undoubtedly an issuethat will eventually need to be addressed. As is often the casein a book of wide scope and short extent, there is a tendencyto deal rather briefly with issues and data, and to rely onother reviews, rather than primary source material.

Biopesticides – use and delivery

provides a good startingpoint for anyone interested in biopesticides in the broadestsense and the wide range of urgent issues that they raise.

Jenny Cory

Ecological and Biocontrol Centre, NERC CEH Oxford,Mansfield Road, Oxford OX1 3SR, UK

(tel +44 1865281 643; fax +44 1865281 696;email [email protected])


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