Physiology of the Development of Somatic Embryos in Cell Cultures of Alfalfa and Celery

Stuart, David A.; Nelsen, Janet; McCall, Carol M.; Strickland, Steven G.; Walker, K.

doi:10.1016/b978-0-12-775310-2.50009-0

Cited by 16 publications

(6 citation statements)

References 22 publications

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“…It has been claimed that development of a system for alfalfa somatic embryogenesis is Limited by a number of factors such as initial genotype and exptant (Brown and Atanassov 1985;Kao and Michayluk 1981), type and duration of treatment with a morphogenesis inducing factor (Stuart et at. 1985) and composition of the culture medium (Dos Santos et at.…”

Section: Resultsmentioning

confidence: 99%

A new approach to direct somatic embryogenesis in Medicago

1991

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A highly efficient system for direct somatic embryogenesis is described. Leaf sections originating from young trifoliate leaves of Medicago falcata line 47/1-5 and Medicago sativa line No2/9R, directly produced embryos after cultivation in liquid B5IV induction medium. In comparison with indirect somatic embryogenesis the system omits the callus stage and thus allows shortening of the process of somatic embryogenesis in alfalfa by 35-40 days. It permits the avoidance of secondary changes occurring during the process of dedifferentiation. A modified B5/3H medium containing Polyethylene Glycol 6000 promoted embryo development from globular up to torpedo stage. It was clearly shown that 2.5% Polyethylene Glycol stimulated this process for both H. falcata 47/1-5 and M. sativa No 2/9R. Maturation of torpedo stage embryos was carried out on solidified or liquid abscisic acidcontaining medium. A 30μM abscisic acid concentration was optimal in allowing one embryo to yield one plant. Somatic embryo conversion to plants and plant regeneration was performed on Murashige and Skoog medium. Regenerated plants showed a normal morphology.

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Section: Resultsmentioning

confidence: 99%

A new approach to direct somatic embryogenesis in Medicago

1991

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“…The pro-embryogenic mass within the culture containing auxins is now generally believed to synthesize all the genes necessary to complete the globular stage of embryogenesis (Zimmerman, 1993). However, if embryos are exposed to too much auxin during development, they fail to accumulate storage protein and germinate at a lower frequency (Stuart et al, 1985). Removal of auxin from the culture, therefore, is considered to inactivate several genes or synthesize new gene products for completion of embryo.development (Zimmerman, 1993).…”

mentioning

confidence: 99%

Plant regeneration from somatic embryos in black pepper

Joseph

Philip

1996

Plant Cell Tiss Organ Cult

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Embryogenic callus derived from zygotic embryos of black pepper (Piper nigrum Linn.) were induced to form somatic embryos on solid and liquid Schenk and Hildebrandt basal medium. Callus proliferation, somatic embryogenesis and germination of embryos were achieved in about 8 months in static cultures while it took only 8 weeks in liquid suspension cultures. The highest number of embryos and plantlets was produced from cells grown as suspension cultures raised in half-strength medium without growth regulators and sucrose level reduced from 3% to 1.5%. Regenerated plants were established in soil.Abbreviations: 2,4-D -2,4-dichlorophenoxy acetic acid; SH -Schenk and Hildebrandt Black pepper is one of the most widely used spices in culinary preparations, food processing, perfumery and as a condiment. A major constraint in cultivation of the crop is its low productivity due to poor genetic stock and incidence of pests and diseases. All the 64 cultivats of Piper nigrum identified in India produce seeds.However, seed propagation in the species is cumbersome, uncertain and yields only a few heterogeneous progenies due to their short viability and high sterility in post-fertilization stages (Kanta, 1962). Black pepper is, therefore, conventionally propagated through cuttings with 2-6 nodes for nursery and field plantations. Plantlets have also been obtained from micropropagated shoot tips of mature vines (Philip et al., 1992) and seedlings (Mathews and Rao, 1984). However, to our knowledge, black pepper plants have not been raised from somatic embryos. Callus-mediated somatic embryos with dependable growth characteristics and plant regeneration are beneficial for raising improved varieties through in vitro selection from high-yielding callus lines and possibly also for genetic transformation. The present study reports an effective protocol for whole plant regeneration from somatic embryos of black pepper in static and suspension cultures.Ripe seeds of Piper nigrum Linn. C.V. Karimunda and Panniyur-1 were collected from plants grown in the Botanical gardens at the University of Calicut. The outer fleshy mesocarp was washed off under slow running water before being surface sterilized with 0.1% mercuric chloride for 5 minutes followed by three rinses with sterile distilled water for 5 min each. The embryo within the seed was dissected out aseptically with a sterile scalpel. The dissected embryos were incubated singly on filter paper bridges in 60 ml culture tubes containing 15 ml liquid SH (Schenk and Hildebrandt, 1972) medium, 20% without and 80% with 2,4-D (0.5-5 mg 1-1 ). The pH of the medium was adjusted to 5.7 prior to autaclaving. The cultures were maintained in dark at 24 -4-2°C with subculture carried out bimonthly to full-strength or half-strength SH medium without growth regulators. Suspension cultures were initiated by transferring 5 mg callus into 100 ml Erlenmeyer flasks containing 25 ml full or half-strength medium. The flasks were wrapped in with black paper and cultures were maintained at 110 rpm on a g...

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“…However, seed proteins have also been observed at low levels in in vitro cultures of alfalfa (Stuart et al, 1985) and zygotic and somatic Brassicu embryos in culture (Crouch, 1982). In those studies the levels of seed proteins accumulated in vitro were one-tenth of the level observed in vivo.…”

Section: Citrin Accumulation In Somatic Embryos In Vitromentioning

confidence: 90%

“…In those studies the levels of seed proteins accumulated in vitro were one-tenth of the level observed in vivo. Levels of seed protein accumulation in cultured embryos of Brassica and alfalfa (ex-albuminous seeds) were observed to be influenced by 2,4-D concentration (Stuart et al, 1985), ABA, and osmoticum (Crouch and Sussex, 1981;Crouch, 1982). In Citrus, a change in the carbon source from Suc to Gal and sorbitol is necessary to induce and maintain efficient embryogenesis from nucellar callus in vitro (Hidaka and Omura, 1989).…”

Section: Citrin Accumulation In Somatic Embryos In Vitromentioning

confidence: 99%

Polyembryony in Citrus (Accumulation of Seed Storage Proteins in Seeds and in Embryos Cultured in Vitro)

1996

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Cifrus exhibits polyembryonic seed development, an apomictic process i n which many maternally derived embryos arise from the nucellus surrounding the developing zygotic embryo. Cifrus seed storage proteins were used as markers t o compare embryogenesis in developing seeds and somatic embryogenesis in vitro. The saltsoluble, globulin protein fraction (designated citrin) was purified from Citrus sinensis cv Valencia seeds. Citrins separated into two subunits averaging 22 and 33 k D under denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A cDNA clone was isolated representing a citrin gene expressed in seeds when the majority of embryos were at the early globular stage of embryo development. l h e predicted protein sequence was most related to the globulin seed storage proteins of pumpkin and cotton. Accumulation of 33-kD polypeptides was first detected in polyembryonic Valencia seeds when the majority of embryos were at the globular stage of development. Somatic Citrus embryos cultured i n vivo were observed to initiate 33-kD polypeptide accumulation later in embryo development but accumulated these peptides at only 10 t o 20% of the leve1 observed in polyembryonic seeds. Therefore, factors within the seed environment must influence the higher quantitative levels of citrin accumulation in nucellar embryos developing in vivo, even though nucellar embryos, like somatic embryos, are not derived from fertilization events.

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Physiology of the Development of Somatic Embryos in Cell Cultures of Alfalfa and Celery

Cited by 16 publications

References 22 publications

A new approach to direct somatic embryogenesis in Medicago

A new approach to direct somatic embryogenesis in Medicago

Plant regeneration from somatic embryos in black pepper

Polyembryony in Citrus (Accumulation of Seed Storage Proteins in Seeds and in Embryos Cultured in Vitro)

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