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Tissue-culture guide
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TRENDS in Plant Science Vol.6 No.2 February 2001
85
interested in how experimentation in this field was being taught by others and, in particular, how they were teaching experimental design and statistical analyses of data – plant tissue culturists have, for decades, been notoriously cavalier in their treatment of these matters. Imagine my disappointment on finding no emphasis on the selection of appropriate experimental designs, no discussion on statistical analyses and, few experiments. Discussing the selection of culture media for a plant, Roberta Smith states that ‘the development of a suitable medium is based on trial and error’ – why not on experimentation? The subtitle to this book should read ‘Techniques and Exercises’ and, in the Preface, this is exactly how the author describes the strategy in developing this manual – ‘to devise exercises’. ‘What kind of statistical analysis is appropriate for this data?’ was asked in connection with a tobacco callus growthcurve exercise. One of the few experiments in this book is on salt selection using carrot calluses. It tested a dose–response of 0.0, 0.2, 1.6 and 6.0 g NaCl per l medium. Further on, Brent McCown suggests the demonstration of a 2iP-response curve using 0.0, 0.1, 1.0, 10.0 and 40.0 µM 2iP for either rhododendron or azalea, and 0, 1, 4 and 10 µM benzyl adenine for roses and birch. These experiments should be designed with regression analyses in mind because growth response to many substances of interest is logarithmic. Regression analysis requires that equal increment doses be applied. Thus, a series might include 0.00, 1.00, 3.16, 10.00 and 31.62 µM of a growth regulator. The review by McCown on the tissue culture of woody shrubs and trees is disappointingly limited to certain species and, in particular, says nothing about the vast amount of work on temperate fruit trees and on Eucalyptus species. Disappointingly, students are still being taught to make culture media using mass units rather than mole units of measurement – and then we find that 0.9 µM kinetin and 11.4 µM indole acetic acid is to be used with tobacco callus. I was amazed to read instructions about melting agar over a hot plate. A long paragraph is devoted to this archaic procedure followed by a short paragraph
acknowledging, almost reluctantly, that agar ‘can also be melted in the autoclave’. Why not just autoclave it – it is far easier and does not involve standing around stirring the pot over a hot plate. Better still, cold dispense culture medium and avoid double heating. Mention is made of using Gelrite as an alternative to agar but no warning that it can only be used with media containing a minimum quantity of minerals – try adding Gelrite to water and you will get an insoluble product. Four of the five culture media described use FeSO4.7H2O and Na2EDTA.2H2O but there is no mention of how to mix these two compounds – later described as the NaFeEDTA stock. Why not use EDTA ferric monosodium salt instead? In spite of these shortcomings, I think that the book is well written and, bearing in mind that it is intended as an adjunct to lecture discussions of the various topics, is a useful contribution to teaching in plant tissue culture.
Book Review
Tissue-culture guide Plant Tissue Culture: Techniques and Experiments (2nd edn) by Roberta H. Smith Academic Press, 2000 US$49.95 pbk ISBN 0 12 650342 7
This manual delivers a tested set of exercises for teachers in colleges and universities for students that have taken courses in chemistry, plant anatomy and plant physiology. For teachers, it presents a structured programme using plant material that is available all year round. Furthermore, material, initiated early in a training course, becomes source cultures for later exercises. The book starts with an excellent chapter on the history of plant cell culture, contributed by Trevor Thorpe, and is followed by four chapters of a general background nature covering the organization of a tissue culture laboratory, the preparation of culture media, explant preparation and aseptic techniques. The final nine chapters cover most of the major objectives of plant tissue culture users. I was impressed with the gradual introduction of complex techniques and ideas. For example: introducing the effect of polarity on culture performance; the selection of salt-tolerant strains and growth curves (Chapter 6); the invitation to squash immature anthers in acetoorcein to determine the stage of development of the microspore mother cells and microspores; the use of toluidine blue on root-tip macerates to count chromosomes; and using epidermal leaf strips to count stomates (Chapter 9). The chapters on ‘Meristem culture for virus-free plants’, ‘Protoplast isolation and fusion’, and ‘Agrobacterium-mediated transformation of plants’, will be valuable starting points for any researcher using special-purpose tissue culture techniques for the first time. When I agreed to review this book, I looked forward eagerly to its arrival. I was
Ronald A. de Fossard XARMA Pty Ltd, PO Box 37, Eagle Heights, Queensland 4271, Australia. e-mail: [email protected]
Book reviews in Trends in Plant Science Recently published books and CD-ROMs featured in Trends in Plant Science are assessed by experts in the field, providing an open peerreview service to potential purchasers. If there is a book or CD-ROM that you think should be reviewed then please contact the Assistant Editor ([email protected]).
http://plants.trends.com 1360-1385/01/$ – see front matter © 2001 Elsevier Science Ltd. All rights reserved. PII: S1360-1385(00)01856-2
TRENDS in Plant Science Vol.6 No.2 February 2001
85
interested in how experimentation in this field was being taught by others and, in particular, how they were teaching experimental design and statistical analyses of data – plant tissue culturists have, for decades, been notoriously cavalier in their treatment of these matters. Imagine my disappointment on finding no emphasis on the selection of appropriate experimental designs, no discussion on statistical analyses and, few experiments. Discussing the selection of culture media for a plant, Roberta Smith states that ‘the development of a suitable medium is based on trial and error’ – why not on experimentation? The subtitle to this book should read ‘Techniques and Exercises’ and, in the Preface, this is exactly how the author describes the strategy in developing this manual – ‘to devise exercises’. ‘What kind of statistical analysis is appropriate for this data?’ was asked in connection with a tobacco callus growthcurve exercise. One of the few experiments in this book is on salt selection using carrot calluses. It tested a dose–response of 0.0, 0.2, 1.6 and 6.0 g NaCl per l medium. Further on, Brent McCown suggests the demonstration of a 2iP-response curve using 0.0, 0.1, 1.0, 10.0 and 40.0 µM 2iP for either rhododendron or azalea, and 0, 1, 4 and 10 µM benzyl adenine for roses and birch. These experiments should be designed with regression analyses in mind because growth response to many substances of interest is logarithmic. Regression analysis requires that equal increment doses be applied. Thus, a series might include 0.00, 1.00, 3.16, 10.00 and 31.62 µM of a growth regulator. The review by McCown on the tissue culture of woody shrubs and trees is disappointingly limited to certain species and, in particular, says nothing about the vast amount of work on temperate fruit trees and on Eucalyptus species. Disappointingly, students are still being taught to make culture media using mass units rather than mole units of measurement – and then we find that 0.9 µM kinetin and 11.4 µM indole acetic acid is to be used with tobacco callus. I was amazed to read instructions about melting agar over a hot plate. A long paragraph is devoted to this archaic procedure followed by a short paragraph
acknowledging, almost reluctantly, that agar ‘can also be melted in the autoclave’. Why not just autoclave it – it is far easier and does not involve standing around stirring the pot over a hot plate. Better still, cold dispense culture medium and avoid double heating. Mention is made of using Gelrite as an alternative to agar but no warning that it can only be used with media containing a minimum quantity of minerals – try adding Gelrite to water and you will get an insoluble product. Four of the five culture media described use FeSO4.7H2O and Na2EDTA.2H2O but there is no mention of how to mix these two compounds – later described as the NaFeEDTA stock. Why not use EDTA ferric monosodium salt instead? In spite of these shortcomings, I think that the book is well written and, bearing in mind that it is intended as an adjunct to lecture discussions of the various topics, is a useful contribution to teaching in plant tissue culture.
Book Review
Tissue-culture guide Plant Tissue Culture: Techniques and Experiments (2nd edn) by Roberta H. Smith Academic Press, 2000 US$49.95 pbk ISBN 0 12 650342 7
This manual delivers a tested set of exercises for teachers in colleges and universities for students that have taken courses in chemistry, plant anatomy and plant physiology. For teachers, it presents a structured programme using plant material that is available all year round. Furthermore, material, initiated early in a training course, becomes source cultures for later exercises. The book starts with an excellent chapter on the history of plant cell culture, contributed by Trevor Thorpe, and is followed by four chapters of a general background nature covering the organization of a tissue culture laboratory, the preparation of culture media, explant preparation and aseptic techniques. The final nine chapters cover most of the major objectives of plant tissue culture users. I was impressed with the gradual introduction of complex techniques and ideas. For example: introducing the effect of polarity on culture performance; the selection of salt-tolerant strains and growth curves (Chapter 6); the invitation to squash immature anthers in acetoorcein to determine the stage of development of the microspore mother cells and microspores; the use of toluidine blue on root-tip macerates to count chromosomes; and using epidermal leaf strips to count stomates (Chapter 9). The chapters on ‘Meristem culture for virus-free plants’, ‘Protoplast isolation and fusion’, and ‘Agrobacterium-mediated transformation of plants’, will be valuable starting points for any researcher using special-purpose tissue culture techniques for the first time. When I agreed to review this book, I looked forward eagerly to its arrival. I was
Ronald A. de Fossard XARMA Pty Ltd, PO Box 37, Eagle Heights, Queensland 4271, Australia. e-mail: [email protected]
Book reviews in Trends in Plant Science Recently published books and CD-ROMs featured in Trends in Plant Science are assessed by experts in the field, providing an open peerreview service to potential purchasers. If there is a book or CD-ROM that you think should be reviewed then please contact the Assistant Editor ([email protected]).
http://plants.trends.com 1360-1385/01/$ – see front matter © 2001 Elsevier Science Ltd. All rights reserved. PII: S1360-1385(00)01856-2