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Biochemical methods in cell culture and virology
TIBS - November 1977
BOOKS Plant tissue culture comes of age Plant Cell, Tissue and Organ Culture edited by J. Reinert and Y. P. S. Bajaj, published by Springer- Verlag, Berlin, Heidelberg, New York, 1977. DM190.- (approx. $83.60) (xvi + 803pages)
This very costly book attempts to combine the fundamental aspects of plant tissue culture with those of a more applied nature. This is a difficult objective, and if it falls short of this objective it is because the applied aspects of plant tissue culture are still in their infancy and many of the problems are economic, and not biological. Nevertheless it is an excellent reference book, and its publication marks the ‘coming of age’ of plant cell, tissue and organ culture investigations. Its great attraction is its encyclopaedic coverage of so many aspects. The Editors have been able, to convince more than thirty active workers to contribute. The result is very much a patchwork quilt of plant tissue culture knowledge. The book does not contain sufficient practical detail to really be of use at the bench. In a way this is a pity because plant tissue culture methods need to be utilised by biochemists, and workers in other disciplines. Cloning of plants through tissue culture is particularly attractive to the horticulturist and to the plant breeder, in the case of plants that are propagated vegetatively. Haploids are also of considerable interest to the plant breeder, and all these considerations, and also the limitations of anther culture, are well contrasted with the production of haploids by embryo culture of inter-species hybrids showing directional chromosome elimination. Insufficient attention is, however, given to the use of haploid and diploid cell cultures in mutant isolations. Incompatibility barriers in sexual hybridisation can sometimes be overcome by the various in vitro culture approaches that are described in detail. This helps to put in perspective the use of isolated protoplasts for fusion work which can reshlt in the production of interspecies somatic hybrids, thereby providing an alternative method of hybridisation of great potential use to the plant breeder. These new techniques will be particularly
useful when hybridisation is desired, but is not possible sexually. Secondary metabolites produced by plants are of increasing interest to biochemists, and several :ontributors provide critical assessment of :he limitations of the use of relatively undifferentiated plant tissue cultures for work in this area. Differentiation and tissue organisation are not the only factors tvhich influence secondary product synthesis, but there are definite indications that :hey are paramount considerations. The book is very well produced and adeJuately illustrated, but an annoying, and :ostly feature, is grouping of the references into five sections. This leads to considerable repetition of reference citations. There is a lack of standardisation of terms - a feature very confusing for the specialist from another discipline. A glossary would have greatly helped in this respect. Nevertheless this is a very useful book. If its publication further ensures that plant cell, tissue and organ culture is no longer a mysterious discipline with little meaning or purpose, it will have fulfilled a very important role. E. C. COCKING
E. C. Cocking is Head of the Department q/ Botany, University of Nottingham, Nottingham, U.K.
Better never than late Biochemical Methods and Virology
in Cell Culture
by Robert J. Kuchler, published by Dowden, Hutchinson & Ross, Inc., Stroudsburg, PA/ distributed in Europe by John Wiley & Sons, London, 1977. E22.50 (approx. $38.-) (ix i- 33 I pages)
It is difficult to recommend a book of biochemical methods in which almost no articles later than 1972 are cited. This volume does contain some information which is still useful concerning the growth of animal cells and viruses, and the isolation and characterization of DNA, RNA, and protein. It is too much to expect a single author to be knowledgeable about all of the recent advances in these diverse and expanding fields, and as a consequence this book will be of little use to the serious student or research scientist. There is, for instance, no reference to restriction enzyme analysis of DNA, a technique which has revolutionized DNA sequence studies.
l’he method for mapping of oligoribonu:leotides is basically that of Sanger et al. !965) and has been superseded by many .ecent advances. There is no mention of he importance of denaturing RNA sam>les before and during gel or gradient malysis. Cylindrical gels are recomtriend:d for analysis of protein; no mention is nade of the far more useful slab gels, nor )f the higher resolution stacking gels. Appearing at this late date this book ,erves no real purpose; I wonder why it was written. HARVEY
F. LODISH
Y. F. Lodish is Professor
of Biology at Massachusetts institute of Technology, Cambridge, MA, U.S.A.
Isotachophoresis: a monastic view Journal of Chromatography Library, Volume 6. Isotachophoresis : Theory, Instrumentation and Applications Edited by Frans M. Everaerts, Jo L. Beckers and Theo P. E. M. Verheggen, published by Elsevier, Amsterdam, 1976, Djl.160.- (approx. $65.40) (xiv + 418pages)
Isotachophoresis is an electrophoretic trick leading to alignment of molecules in order of electrophoretic mobility and, under the usual conditions, to very high macromolecular concentrations. These can be exploited either analytically, giving rise to very thin, concentrated starting zones in gel electrophoresis, or preparatively. For this process, the term isotachophoresis was coined a few years ago by Everaerts in consensus with a number of European laboratories and with the Swedish company, LKB, presumably in order to emphasize what we will designate below as the particular theoretical genealogy and orthodoxy of the physical-chemical treatment, and also the concern with the capillary apparatus marketed soon thereafter by LKB under the name of Tachophor. In fact, however, the electrophoretic method is exactly the same as that termed ‘steady-state stacking’ by Ornstein or ‘multiphasic zone electrophoresis’ by Jovin. The trick, in each case, consists of orienting a more rapidly migrating ionic species across a phase boundary in front (in the direction of electrophoresis) of a more slowly migrating species. When an electric field is applied to this situation, a ‘regulation’ of concentrations, mobilities and potential gradients behind the ‘leading’ species ensues, which leads to an alignment of all ‘slower’ species in the order of their electrophoretic mobility behind the ‘leading’ constituent, and therefore to separation on the basis of molecular net charge.
BOOKS Plant tissue culture comes of age Plant Cell, Tissue and Organ Culture edited by J. Reinert and Y. P. S. Bajaj, published by Springer- Verlag, Berlin, Heidelberg, New York, 1977. DM190.- (approx. $83.60) (xvi + 803pages)
This very costly book attempts to combine the fundamental aspects of plant tissue culture with those of a more applied nature. This is a difficult objective, and if it falls short of this objective it is because the applied aspects of plant tissue culture are still in their infancy and many of the problems are economic, and not biological. Nevertheless it is an excellent reference book, and its publication marks the ‘coming of age’ of plant cell, tissue and organ culture investigations. Its great attraction is its encyclopaedic coverage of so many aspects. The Editors have been able, to convince more than thirty active workers to contribute. The result is very much a patchwork quilt of plant tissue culture knowledge. The book does not contain sufficient practical detail to really be of use at the bench. In a way this is a pity because plant tissue culture methods need to be utilised by biochemists, and workers in other disciplines. Cloning of plants through tissue culture is particularly attractive to the horticulturist and to the plant breeder, in the case of plants that are propagated vegetatively. Haploids are also of considerable interest to the plant breeder, and all these considerations, and also the limitations of anther culture, are well contrasted with the production of haploids by embryo culture of inter-species hybrids showing directional chromosome elimination. Insufficient attention is, however, given to the use of haploid and diploid cell cultures in mutant isolations. Incompatibility barriers in sexual hybridisation can sometimes be overcome by the various in vitro culture approaches that are described in detail. This helps to put in perspective the use of isolated protoplasts for fusion work which can reshlt in the production of interspecies somatic hybrids, thereby providing an alternative method of hybridisation of great potential use to the plant breeder. These new techniques will be particularly
useful when hybridisation is desired, but is not possible sexually. Secondary metabolites produced by plants are of increasing interest to biochemists, and several :ontributors provide critical assessment of :he limitations of the use of relatively undifferentiated plant tissue cultures for work in this area. Differentiation and tissue organisation are not the only factors tvhich influence secondary product synthesis, but there are definite indications that :hey are paramount considerations. The book is very well produced and adeJuately illustrated, but an annoying, and :ostly feature, is grouping of the references into five sections. This leads to considerable repetition of reference citations. There is a lack of standardisation of terms - a feature very confusing for the specialist from another discipline. A glossary would have greatly helped in this respect. Nevertheless this is a very useful book. If its publication further ensures that plant cell, tissue and organ culture is no longer a mysterious discipline with little meaning or purpose, it will have fulfilled a very important role. E. C. COCKING
E. C. Cocking is Head of the Department q/ Botany, University of Nottingham, Nottingham, U.K.
Better never than late Biochemical Methods and Virology
in Cell Culture
by Robert J. Kuchler, published by Dowden, Hutchinson & Ross, Inc., Stroudsburg, PA/ distributed in Europe by John Wiley & Sons, London, 1977. E22.50 (approx. $38.-) (ix i- 33 I pages)
It is difficult to recommend a book of biochemical methods in which almost no articles later than 1972 are cited. This volume does contain some information which is still useful concerning the growth of animal cells and viruses, and the isolation and characterization of DNA, RNA, and protein. It is too much to expect a single author to be knowledgeable about all of the recent advances in these diverse and expanding fields, and as a consequence this book will be of little use to the serious student or research scientist. There is, for instance, no reference to restriction enzyme analysis of DNA, a technique which has revolutionized DNA sequence studies.
l’he method for mapping of oligoribonu:leotides is basically that of Sanger et al. !965) and has been superseded by many .ecent advances. There is no mention of he importance of denaturing RNA sam>les before and during gel or gradient malysis. Cylindrical gels are recomtriend:d for analysis of protein; no mention is nade of the far more useful slab gels, nor )f the higher resolution stacking gels. Appearing at this late date this book ,erves no real purpose; I wonder why it was written. HARVEY
F. LODISH
Y. F. Lodish is Professor
of Biology at Massachusetts institute of Technology, Cambridge, MA, U.S.A.
Isotachophoresis: a monastic view Journal of Chromatography Library, Volume 6. Isotachophoresis : Theory, Instrumentation and Applications Edited by Frans M. Everaerts, Jo L. Beckers and Theo P. E. M. Verheggen, published by Elsevier, Amsterdam, 1976, Djl.160.- (approx. $65.40) (xiv + 418pages)
Isotachophoresis is an electrophoretic trick leading to alignment of molecules in order of electrophoretic mobility and, under the usual conditions, to very high macromolecular concentrations. These can be exploited either analytically, giving rise to very thin, concentrated starting zones in gel electrophoresis, or preparatively. For this process, the term isotachophoresis was coined a few years ago by Everaerts in consensus with a number of European laboratories and with the Swedish company, LKB, presumably in order to emphasize what we will designate below as the particular theoretical genealogy and orthodoxy of the physical-chemical treatment, and also the concern with the capillary apparatus marketed soon thereafter by LKB under the name of Tachophor. In fact, however, the electrophoretic method is exactly the same as that termed ‘steady-state stacking’ by Ornstein or ‘multiphasic zone electrophoresis’ by Jovin. The trick, in each case, consists of orienting a more rapidly migrating ionic species across a phase boundary in front (in the direction of electrophoresis) of a more slowly migrating species. When an electric field is applied to this situation, a ‘regulation’ of concentrations, mobilities and potential gradients behind the ‘leading’ species ensues, which leads to an alignment of all ‘slower’ species in the order of their electrophoretic mobility behind the ‘leading’ constituent, and therefore to separation on the basis of molecular net charge.