Sant S. Bhojwani

Current trends in the embryology of angiosperms.

Introduction. 1. Male Gametogenesis Development and Structure of Sperm D. Southworth, S.C. Russell. 2. Sperm and Generative Cell Isolation and Manipulation D. Southworth. 3. Pollen Germination and Pollen Tube Growth Tip Growth Mechanism in Sexual Plant Reproduction A. Moscatelli, M. Cresti. 4. Female Gametogenesis Ontogenesis of the Embryo Sac and Female Gametes S.D. Russell. 5. Embryo Sac Isolation and Manipulation D.D. Cass, J.D. Laurie. 6. In Vivo Fertilization T.B. Batygina, V.E. Vasilyeva. 7. In Vitro Fertilization E. Kranz. 8. Sexual Incompatibility F. Cruz-Garcia, B.A. MacClure. 9. Zygotic Embryogenesis Structural Aspects R. Czapik, R. Izmailow. 10. Zygotic Embryogenesis Hormonal Control of Embryo Development B. Fischer-Iglesias, G. Neuhaus. 11. Zygotic Embryogenesis: Developmental Genetics The Formation of an Embryo from a Fertilized Egg K. Schrick, T. Laux. 12. Somatic Embryogenesis T.A. Thorpe, C. Stasolla. 13. Synthetic Seeds of Asparagus officinalis L. K. Mamiya, et al. 14. Endosperm Development P.W. Becraft, et al. 15. Seed Maturation, Germination, and Dormancy A.B. Downie. 16. Gametophytic Apomixis A Successful Mutation of the Female Gametogenesis Y. Savidan. 17. Parthenocarpy State of the Art A. Spena, G.L. Rotino. 18. Androgenesis in Brassica A Model System to Study the Initiation of Plant Embryogenesis J.B.M. Custers, et al. 19. Androgenesis in Cereals S.K. Datta. 20. In Vitro Gynogenesis S.S. Bhojwani, T.D. Thomas. 21. Inheritance of Cytoplasmic Traits -- Embryological Perspectives T. Kuroiwa, et al. Subject Index.

Plant cell reactors—A perspective

Abstract The potential of plant cell culture for the production of commercially valuable secondary metabolites is reviewed. The techniques employed for large-scale plant cell suspension culture are largely those developed for microbial culture, so the basic characteristics of the two types of culture are compared. Technological problems associated with plant cell culture—mixing, oxygen transfer, and cell aggregation and adhesion—are then considered in detail. Various bioreactors currently used for plant cell cultivation are analyzed according to the dynamics of growth and product formation in microbial systems exhibiting no cellular differentiation. Criteria for choosing a bioreactor type for optimal growth and metabolite production are also discussed. Finally a brief economic outlook and future prospects for plant cell culture processes are presented.

Production of podophyllotoxin by plant cell cultures of Podophyllum hexandrum in bioreactor.

Submerged cultivation of Podophyllum hexandrum for the production of podophyllotoxin was carried out in a 3l stirred tank bioreactor fitted with a low-shear Setric impeller. The specific requirements of the medium, such as carbon source (sugar) and light, were established for the growth of and podophyllotoxin production by P. hexandrum in suspension cultures. Substitution of sucrose by glucose resulted in higher growth and podophyllotoxin production. The biosynthesis of podophyllotoxin was favored when plant cells were cultivated in the dark. An agitation speed of 100 rpm was sufficient to mix the culture broth in the bioreactor without causing any significant cell damage. Biomass and podophyllotoxin accumulation in 3 l bioreactor under batch growth conditions were 6.5 g/l and 4.26 mg/l, respectively, in 22 d. This resulted in an overall podophyllotoxin productivity of 0.19 mg/(l.d), which represented an increase of 27% in comparison to its productivity in a shake flask. Podophyllotoxin production was found to be a combined growth-associated and non-growth associated process.

In vitro propagation and low temperature storage ofSaussurea lappa C.B. Clarke - An endangered, medicinal plant.

A procedure forin vitro multiplication ofSaussurea lappa (Asteraceae) is described. On Murashige and Skoogs medium (MS) containing benzylaminopurine and gibberellin 3.5-fold shoot multiplication occurred every three weeks. Shoots rooted on MS containing 0.5 μM naphthaleneacetic acid with 90% efficiency. The shoot cultures stored at 5°C in the dark for 12 months without an intervening subculture survived with 100% viability. The shoots cold stored for 6 months or more showed higher rates of multiplication under culture room conditions than the untreated shoots.

In vitro propagation of garlic by shoot proliferation

Abstract Shoot buds (5–8 mm long), excised from dormant cloves of the New Zealand commercial garlic (Allium sativum L.) and a virus-free French cultivar ‘Rose-de-Kakylis’, proliferated both axillary and adventitious shoots on B-5 basal medium supplemented with 0.5 mg l−1 isopentenyladenine (2-ip) and 0.1 mg l−1 naphthaleneacetic acid (NAA). An 8-fold increase in shoot number occurred every 6 weeks. Shoots were readily rooted in B-5 + 0.01 mg l−1 2-ip + 0.2 mg l−1 NAA and transferred to pots, where about 70% of the shoots formed established plants. The plants raised by this shoot-proliferation method retained the diploid condition of the parents.

Factors affecting high frequency differentiation of shoots and roots from cotyledon explants of Brassica juncea (L.) czern

Abstract Studies were undertaken to optimize conditions for high frequency shoot regeneration from excised cotyledons of Brassica juncea cv. RIK-81-1. Maximum differentiation of adventitious shoot buds occurred when the explants derived from 5-day-old seedlings were cultured on Murashige and Skoog medium (MS) containing 5 μM N6-benzyladenine (BA). On MS alone only roots were formed. Shoot or root formation was restricted to 1–2 mm of tissue at the cut end of the petiole. Organogenesis occured only if the proximal cut end of the petiole was in contact with the medium. The lamina did not exhibit organogenesis. Cotyledons cultured for up to 3 days on root induction medium (MS), still retained their full potential to form shoots upon transfer to MS + BA. With longer incubation on MS medium the shoot-forming capacity of the explants declined, and after 8 days it was completely lost. When applied through the agar medium, BA (5 μM) was required for at least 7 days for shoot bud induction. The possible usefulness of B. juncea cotyledons as an experimental system for detailed studies of morphogenesis and genetic transformation is discussed.

Somatic embryogenesis and regeneration of triploid plants in endosperm cultures of Acacia nilotica.

Immature endosperm of Acacia nilotica formed a nodular callus on MS medium supplemented with 2,4-D, BAP and CH. In the third passage on this medium, in the dark, the callus differentiated somatic embryos. The embryos germinated on MS only after 15 d pre-treatment on modified MS medium in which major salts were replaced by those of major salts of B5 medium and supplemented with glutamine, CH and CW. Triploid nature of the somatic embryos was confirmed by Feulgen cytophotometry.

In vitro clonal multiplication of 4-year-old plants of the bamboo,Dendrocalamus longispathus kurz

SummaryA complete protocol for micropropagation of 4-yr-old plants of the bambooDendrocalamus longispathus is described. Culture initiation was strongly influenced by the nature of the explant and the season. In vitro multiplication was achieved through forced axillary branching. Single node segments from the young lateral branches produced multiple shoots on agar-solidified Murashige and Skoog (MS) medium supplemented with 12µM benzylaminopurine (BAP) and 3µM kinetin. The shoots have been multiplied for 15 passages in liquid and thereafter for over 5 passages on semisolid MS+15µM BAP+1µM indolebutyric acid (IBA)+10% coconut water at a rate of 3.2- and 2.8-fold, every 4 wk, respectively. The nature of the propagule was a critical factor for shoot multiplication and rooting. Seventy-three percent of the shoots rooted on a modified MS medium (major salts reduced to half strength) containing 1µM indoleacetic acid, 1µM IBA, and 68µM coumarin. Through a simple in vitro hardening step, more than 85% of the tissue culture-raised plants were successfully transferred to soil.

In vitro corm formation and field evaluation of corm-derived plants of Gladiolus

Abstract Shoot cultures of Gladiolus cultivar ‘Friendship’, initiated from axillary buds, exhibited best elongation on liquid Murashige and Skoogs medium (MS) containing 0.5 mg 1 −1 BAP, in light. On liquid MS medium containing 6% sucrose, in light, 96% shoots formed corms measuring 19.6 mm across and with a fresh weight of 2767 mg. Except for their small size, the in vitro formed corms were comparable with those formed in vivo. Cytokinins (6-benzylaminopurine, kinetin), abscisic acid, CCC, gibberellic acid or activated charcoal either inhibited or had no effect on corm formation. The in vitro formed corms of cultivars ‘Friendship’, ‘Gold Finchs’ and ‘Her Majesty’, treated at 5 °C for 8 weeks showed 100% germination in the field. The micropropagated plants were morphologically and cytologically comparable to the control plants raised from in vivo formed corms.

Morphogenesis in plant tissue cultures

Introduction. Part I: Basic Studies. 1. Differentiation of Vascular Elements in Tissue Culture. 2. Plant Regeneration from Cultured Protoplasts. 3. Morphogenesis in Haploid Cell Cultures. 4. Light and Electron Microscopic Studies of Somatic Embryogenesis in Spruce. 5. Physiological and Morphological Aspects of Somatic Embryogenesis. 6. Developmental and Structural Aspects of Root Organogenesis. 7. Shoot Morphogenesis: Structure, Physiology, Biochemistry and Molecular Biology. 8. Floral and Vegetative Differentiation In Vitro and In Vivo. 9. Developmental and Structural Patterns of In Vitro Plants. 10. Morphogenesis in Cell and Tissue Cultures. 11. Regulation of Morphogenesis by Bacterial Auxin and Cytokinin Biosynthesis Transgenes. Part II: Applications. 12. In Vitro Induced Haploids in Plant Genetics and Breeding. 13. Somatic Hybridization for Plant Improvement. 14. Germination of Synthetic Seeds. 15. Morphogenesis in Micropropagation. 16. Cell Differentiation and Secondary Metabolite Production. Index.