2. IMPORTANCE OF NITROGEN METABOLISM 2. 1. Range of naturally occurring nitrogenous components in forest trees 2. 2. Gene expression and mapping 2. 3. Metabolic changes in organized and unorganized systems 2. 4. Nitrogen and nutrition 2. 5. Aspects of intermediary nitrogen metabolism 3. NITROGEN METABOLISM IN GROWTH AND DEVELOPMENT 3. 1. Precultural factors 3. 2. Callus formation 3. 3. Cell suspensions 3. 3. 1. Conifers 3. 3. 2. Acer 3. 4. Morphogenesis 3. 4. 1. Nitrogen metabolism of natural embryos 3. 4. 2. Somatic embryogenesis 3. 4. 2. 1. Sweetgum (Liquidambar styraciflua) 3. 4. 2. 2. Douglar-fir and loblolly pine 3. 4. 3. Organogenesis 4. OUTLOOK 11. CARBOHYDRATE UTILIZATION AND METABOLISM - T. A. Thorpe 325 1. INTRODUCTION 2. NUTRITIONAL ASPECTS 3. CARBOHYDRATE UPTAKE 4. CARBOHYDRATE METABOLISM 4. 1. Sucrose degradation 4. 2. Metabolism of other carbon sources 4. 3. Hexose mobilization and metabolism 4. 3. 1. Cell cycle studies 4. 3. 2. Growth studies 4. 3. 3. Organized development 4. 4. Cell wall biogenesis 4. 4. 1. Primary cell walls 4. 4. 2. Cell wall turnover 4. 4. 3. Secondary cell walls 4. 5. Carbon skeleton utilization 5. OSMOTIC ROLE 6. CONCLUDING THOUGHTS 369 12. THE USE OF IN VITRO TECHNIQUES FOR GENETIC MODIFICATIO~FOREST TREES - E. G. Kirby 1. INTRODUCTION 2. IN VITRO SELECTION 2. 1. Natural variation 2. 2. Induction of variation 2. 3. Selection techniques 2. 4. Plant regeneration 2 . • 5. Applications x 3. SOMATIC HYBRIDIZATION 3. 1.

This symposium is the third in a series featuring the propaga tion of higher plants through tissue culture. The first of these symposia, entitled "A Bridge Between Research and Application," was held at the University in 1978 and was published by the Technical Information Center, Department of Energy. The second symposium, on "Emerging Technologies and Strategies," was held in 1980 and pub lished as a special issue of Environmental and Experimental Botany. One of the aims of these symposia was to examine the current state of-the-art in tissue culture technology and to relate this state of technology to practical, applied, and commercial interests. Thus, the third of this series on development and variation focused on embryogenesis in culture: how to recognize it, factors which affect embryogenesis, use of embryogenic systems, etc.; and variability from culture. A special session on woody species again emphasized somatic embryogenesis as a means of rapid propagation. This volume emphasizes tissue culture of forest trees. All of these areas, we feel, are breakthrough areas in which significant progress is expected in the next few years.

Since the first edition of our book "Tissue Culture in Fores try" in 1982 we have witnessed remarkable advances in cell and tissue culture technologies with woody perennials. In addition to forest biologists in government, industry, and universities, we now have molecular biologists, genetic engineers, and biochemists using cell and tissue cultures of woody species routinely. There fore, the time has come for an update of the earlier edition. In our present effort to cover new developments we have expanded to three volumes: 1. General principles and Biotechnology 2. Specific Principles and Methods: Growth and Development 3. Case Histories: Gymnosperms, Angiosperms and Palms The scientific barriers to progress in tree improvement are not so much lack of foreign gene expression in plants but our current inabili ty to regenerate plants in true-to-type fashion on a mas sive and economic scale. To achieve this in the form of an appro pr iate biotechnology, cell and tissue culture will increasing ly require a better understanding of basic principles in chemistry and physics that determine structural and functional relationships among molecules and macromolecules (proteins, RNA, DNA) within cells and tissues. These principles and their relationship with the culture medium and its physical environment, principles of clonal propagation, and genetic variation and ultrastructure are discussed in volume one.

Woody plants provide many challenges to the tissue culturist. Although there are many excellent tissue culture books and manuals available, these are generally strongly biased towards herbacious crops. Consequently, they often do not pay sufficient attention to the problems that specifically apply to in vitro culture of tree species. Culture of the latter often poses problems which are either absent or of lesser significance when culturing herbacious species. When trees in the field are used as explant source, the problems can be especially severe. For example, the physiological condition of the explants is difficult to control because of variation in weather and biotic factors. Furthermore, it is often difficult to obtain explants free of contaminants from field grown trees. Lack of genetic uniformity and maturation are additional problems one often has to deal with when culturing tree cells or tissues. These problems are emphasized in this text. In vitro culture of trees is not viewed in isolation. It is considered in conjunction with breeding, traditional cloning and other common tree improvement techniques. The text discusses theoretical as well as practical aspects of the in vitro culture of trees.

2. 2. Plant materials 2. 3. Pregrowth conditions 2. 4. Cryoprotectant treatment 2. 5. Freezing 2. 5. 1. Slow freezing 2. 5. 2. Rapid freezing 2. 5. 3. Droplet freezing 2. 6. Storage 2. 7. Thawing 2. 8. Viability testing 2. 9. Post-thaw regrowth 3. EXAMPLES OF CRYOPRESERVATION OF WOODY PLANT MATERIAL 4. POTENTIAL APPLICATION OF CRYOPRESERVATION IN TREE IMPROVEMENT 17. NURSERY HANDLING OF PROPAGULES - J. A. Driver, and 320 G. R. L. Suttle 1. INTRODUCTION 2. COMMERCIAL NURSERY NEEDS VS. LABORATORY PRACTICE 3. SEASONALITY OF GROWTH AND PRODUCTION CYCLES 4. MICROPROPAGATION OPTIONS 4. 1. Trends in commercial micropropagation 4. 1. 1. Contract micropropagation 5. FACTORS AFFECTING SURVIVAL AND GROWTH 5. 1. Hardening of propagules in vitro 5. 2. Greenhouse considerationS------ 5. 3. Field planting 5. 4. New approaches: Direct field rooting 5. 4. 1. Pretreatment in vitro 5. 4. 2. Root induction 5. 4. 3. Field placement 18. MYCORRHIZAE - R. K. Dixon, and D. H. Marx 336 1. INTRODUCTION 2. ROLE OF MYCORRHIZAE IN TREE GROWTH AND DEVELOPMENT 3. PRODUCTION AND APPLICATION OF ECTOMYCORRHIZAL FUNGUS INOCULUM 3. 1. Bareroot stock 3. 2. Container-grown stock 4. FIELD TRIALS WITH ECTOMYCORRHIZAL PLANTING STOCK 5. PRODUCTION AND APPLICATION OF ENDOMYCORRHIZAL INOCULUM 6. FIELD TRIALS WITH ENDOMYCORRHIZAL 7. RESEARCH OPPORTUNITIES 8. SUMMARY 351 19. TISSUE CULTURE APPLICATIUN TO FOREST PATHOLOGY AND PEST CONTROL - A. M. Diner, and D. F. Karnosky 1. INTRODUCTION 2. HOST AND PATHOGEN: CULTURE AND CHALLENGE 2. 1.

This volume covers recent advances in the vegetative propagation of woody plants by tissue culture. A wide range of topics relevant to micropropagation of woody plants are discussed by renowned international scientists. These include cellular contro of morphogenesis, light regimes in tissue culture, maturation and rejuvenation, synthetic seed, genetics of micropropagated plants, haploid embryogenesis, protoplast culture, and acclimatization of ex vitro woody plants. In addition to micropropagation of selected woody plants, both gymnosperms and angiosperms, this volume also includes in vitro genetic selection, strategic planning for application of biotechnology for genetics and breeding, and clonal options for woody plant improvement. A balanced view of both perspectives and limitations of woody plant micropropagation is presented.

Plant Cell and Tissue Culture gives an exhaustive account of plant cell culture and genetic transformation, including detailed chapters on all major field and plantation crops. Part A presents a comprehensive coverage of all necessary laboratory techniques for the initiation, nutrition, maintenance and storage of plant cell and tissue cultures, including discussions on these topics, as well as on morphogenesis and regeneration, meristem and shoot tip culture, plant protoplasts, mutant cell lines, variation in tissue cultures, isogenic lines, fertilization control, cryopreservation, transformation, and the production of secondary metabolites. Part B then proceeds into detail on the specific in vitro culture of specific crops, including cereals, legumes, vegetables, potatoes, other roots and tubers, oilseeds, temperate fruits, tropical fruits, plantation crops, forest trees and ornamentals. Plant Cell and Tissue Culture is, and is likely to remain, the laboratory manual of choice, as well as a source of inspiration and a guide to all workers in the field.

This book presents latest work in the field of plant biotechnology regarding high-efficiency micropropagation for commercial exploitation at low labor and equipment costs. The book consists of 18 chapters on establishing advanced culture systems, techniques as well as latest modification protocols on a variety of crops. It also discusses new methods such as nylon film culture system, light-emitting diode and wireless light-emitting diode system, stem elongation, wounding manipulation and shoot tip removal, in vitro hydroponic and microponic culture system, thin cell layer culture system etc. Plant cell tissue has been developed more than fifty years ago. Since then applications of in vitro plant propagation expanded rapidly all around the world and played as an important role in agricultural and horticultural systems. This book will be of interest to teachers, researchers, scientists, capacity builders and policymakers. Also the book serves as additional reading material for undergraduate and graduate students of agriculture, forestry, ecology, soil science, and environmental sciences.

This Volume contains the papers presented by twenty-eight invited speakers at the symposium entitled, "Genetic Manipulation of Woody Plants," held at Michigan State University, East Lansing, Michigan, from June 21-25, 1987. Also included are abstracts of contributed poster papers presented during the meeting. That the molecular biology of woody plants is a rapidly expanding field is attested to by the large attendance and high level of enthusiasm generated at the conference. Leading scientists from throughout the world discussed challenging problems and presented new insights into the devel opment of in vitro culture systems, techniques for DNA analysis and manipulation, gene vector systems, and experimental systems that will lead to a clearer understanding of gene expression and regulation for woody plant species. The presence at the conference of both invited speakers and other scientists who work with nonwoody plant species also added depth to the discussions and applicability of the information presented at the conference. The editors want to commend the speakers for their well-organized and informative talks, and feel particularly indebted to the late Dr. Alexander Hollaender and others on the planning committee who assist ed in the selection of the invited speakers. The committee consisted of David Burger (University of California, Davis), Don J. Durzan (University of California, Davis) , Bruce Haissig (U. S. Department of Agriculture Forest Service), Stanley Krugman (U. S. Department of Agriculture Forest Service) , Ralph Mott (North Carolina State University), Otto Schwarz (Univer.sity of Tennessee, Knoxville), and Roger Timmis (Weyerhaeuser Company).