Section A 1 Somatic Embryogenesis in White Spruce: Studies of Embryo Development and Cell Biology L Kong, et al 2 Proliferative Somatic Embryogenesis in Woody Species K Raemakers, et al 3 Somatic Embryo Germination and Desiccation Tolerance in Conifers EI Hay, PJ Charest 4 Performance of Conifer Stock Produced Through Somatic Embryogenesis SC Grossnickle 5 Apoptosis During Early Somatic Embryogenesis in Picea spp L Havel, DJ Durzan 6 Water Relation Parameters in Conifer Embryos: Methods and Results N Dumont-Beboux, et al 7 Image Analysis for Sorting Somatic Embryos Y Ibaraki 8 Somatic Embryogenesis in Woody Legumes RN Trigiano, et al 9 Cold Storage and Cryopreservation of Camellia Embryogenic Cultures AM Vieitez, A Ballester 10 Cryopreservation of Embryogenic Cultures of Conifers and Its Application to Clonal Forestry DR Cyr 11 Commercialization of Plant Somatic Embryogenesis BCS Sutton, DR Polonenko Section B 12 Somatic Embryogenesis in Myrtaceous Plants JM Canhoto, et al 13 Somatic Embryogenesis Induction in Bay Laurel (Laurus nobilis L) JM Canhoto, et al 14 Somatic Embryogenesis in Simarouba glauca Linn GR Rout, P Das 15 Somatic Embryogenesis in Magnolia spp SA Merkle 16 Somatic Embryogenesis and Evaluation of Variability in Somatic Seedlings of Quercus serata by RAPD Markers K Ishii, et al 17 Somatic Embryogenesis from Immature Fruit of Juglans cinerea PM Pijut Section C 18 Somatic Embrygonesis in Pinus patula Scheide et Deppe NB Jones, J van Staden 19 Somatic Embryogenesis in African Cycads (Encephalartos) AK Jager, J van Staden 20 Somatic Embryogenesis in Picea wilsonii Y Yang, Z Guo 21 Somatic Embryogenesis in Jack Pine (Pinus banksiana Lamb) YS Park, et al 22 Somatic Embryogenesis in Hybrid Firs J Jasik, et al 23 Somatic Embryogenesis in Taxus SR Wann, et al

Somatic embryogenesis in woody plants

Synthetic seed technology requires the inexpensive production of large numbers of high-quality somatic embryos. Proliferating embryogenic cultures from conifers consist of immature embryos, which undergo synchronous maturation in the presence of abscisic acid and elevated osmoticum. Improvements in conifer somatic embryo quality have been achieved by identifying the conditions in vitro that resemble the conditions during in ovulo development of zygotic embryos. One normal aspect of zygotic embryo development for conifers is maturation drying, which allows seeds to be stored and promotes normal germination. Conditions of culture are described that yield mature conifer somatic embryos that possess normal storage proteins and fatty acids and which survive either partial drying, or full drying to moisture contents similar to those achieved by mature dehydrated zygotic embryos. Large numbers of quiescent somatic embryos can be produced throughout the year and stored for germination in the spring, which simplifies production and provides plants of uniform size. This review focuses on recent advances in conifer somatic embryogenesis and synthetic seed technology, particularly in areas of embryo development, maturation drying, encapsulation and germination. Comparisons of conifer embryogeny are made with other gymnosperms and angiosperms.

Embryogeny of gymnosperms: advances in synthetic seed technology of conifers

There is currently considerable interest in developing techniques for producing somatic embryos from conifers. Somatic embryos could be used for micropropagation of trees with desirable characteristics such as improved dimension increment, better quality wood and disease resistance. The availability of somatic embryos would also provide excellent experimental material for basic studies of conifer embryo development. Finally, embryogenic callus and suspension cultures should facilitate the development of a protoplast regeneration system to be used in future genetic manipulation studies. The use of embryogenic material has previously proved successful for developing protoplast systems with other recalcitrant species (e.g. Vasil and Vasil 1986).

Somatic embryogenesis in conifers

Somatic embryogenesis of coniferous species was first reported more than 20 years ago. Since then, there has been an explosion of research aimed at developing and optimizing protocols for efficient regeneration of plantlets. Although routinely used both as a means of propagation, as well as a valuable model system for investigating the structural, physiological, and molecular events occurring during embryo development, in vitro embryogenesis is still problematic for some coniferous species. Major problems include: low number of embryos generated; and low frequency of mature embryos able to convert into viable plantlets. Until recent years, despite the fact that embryogenesis is comprised of a sequence of defined steps which include proliferation of embryogenic tissue, embryo maturation, and germination, attempts at improving the whole procedure have been made almost exclusively during the maturation stage. This strategy was based on the assumption that successful regeneration is related to treatments provided during the development of the embryos. Major optimizations of the maturation medium have involved judicious selections of type and concentration of growth regulators, namely abscisic acid, and adjustments of the osmoticum of the culture medium. Extensive work has been conducted in defining the effects of plasmolysing and non-plasmolysing osmoticum agents during maturation, as well as in improving desiccation techniques required for the completion of the maturation program. In the last 2 years, however, work on spruce has clearly demonstrated that the early events in embryogenesis are crucial for the successful completion of the overall embryogenic program. The use of cell tracking techniques, implemented by physiological and molecular studies, has revealed that manipulations of the culture conditions early in the process can increase both number and quality of embryos produced in culture. Additional manipulations of the germination medium can also enhance germination and conversion frequency of somatic embryos matured in a sub-optimal environment. These new findings, together with the unraveling of molecular mechanisms involved in the control/regulation of embryo development hold considerable promise for clonal propagation in conifers.

Recent advances in conifer somatic embryogenesis: improving somatic embryo quality

1. Introduction. 2. Laboratory Organization and Equipment. 3. Media Preparation. 4. Collection, Sterilization, Excision and Culture. 5. Clonal Propagation. 6. Special Cultures. 7. Secondary Metabolite Production. 8. Pathology. 9. Nitrogen Fixation. 10. Storage of Cultures. 11. Genetic Modification. 12. Epilogue. 13. References. 14. Indices.

In Vitro Culture of Trees

Somatic embryo maturation, germination, and soil establishment frequencies were compared for two conifer species, white and black spruce (Picea glauca and Picea mariana). The comparison of the two species regenerated and established in soil under the same conditions showed black spruce to be the most responsive. Shorter exposure times to 32 μM abscisic acid were not as effective as maturation on a medium containing 16 μM abscisic acid for 28 days. This gave similar maturation frequencies for the two species (6–8%), and germination frequencies of 64% for white spruce and over 73% for black spruce. Over 1800 black and white spruce plantlets were recovered, and more than 400 were transferred from in vitro to nonsterile conditions. Sixty percent (160) of the black spruce plantlets survived transfer and continued to grow vigorously. By comparison only 18% (29) of the white spruce plantlets survived, and half of these rapidly produced dormant buds and underwent no further shoot growth. White spruce plants that ...

Somatic embryo maturation, germination, and soil establishment of plants of black and white spruce (Picea mariana and Picea glauca)

Protoplasts were isolated either from the cotyledons of 8- to 10-day-old seedlings, or from cell suspensions, initiated from seedlings of the same open-pollinated seedlot of jack pine (Pinus banksiana Lamb.). Protoplast yields were 2.0 – 7.0 × 105/g fresh wt. for cotyledon-derived, and 0.3–5 × 105/g fresh wt. for suspension-derived protoplasts. Cotyledon protoplasts (6.8%) divided after 5 to 8 days incubation in liquid Laine medium C with 450 mM glucose as osmoticum. Small clusters of 6 to 10 cells, which were observed after approximately 2 weeks, formed microcalli upon further dilution of the culture medium. By 6 to 8 weeks, these microcalli could be transferred to solid medium using a nurse culture of jack pine cells. Protoplasts isolated from seven cell suspension lines differed in their responses to culture conditions. First divisions were only observed with two cell lines cultured in either Laine medium C or one-half Litvay with 370 mM glucose, but this did not lead to callus development. Key words: ...

Comparative studies of protoplast development in jack pine (Pinus banksiana)

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Somatic embryogenesis in conifers

Somatic embryo maturation, germination, and soil establishment of plants of black and white spruce (Picea mariana and Picea glauca)

Comparative studies of protoplast development in jack pine (Pinus banksiana)