Regeneration, Cultivation and Differentiation of Plant Protoplasts Immobilized in Ca-alginate Beads

Draget, Kurt I.; Myhre, Sigrid; Skjåk-Br˦k, Gudmund; Østgaard, Kjetill

doi:10.1016/s0176-1617(88)80252-9

Cited by 54 publications

(18 citation statements)

References 14 publications

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“…The commonly used method of sterilization of heat and high pressure treatment (autoclaving) can modify such properties of the gel as mechanical strength, abrasion resistance, diffusion within the gel, and development of entrapped biomass. It seems that only few authors working with alginate-embedded protoplasts took this fact into consideration, applying filter-sterilization (Draget et al 1988;Hall et al 1997;Dovzhenko et al 1998;Dovzhenko et al 2003). Very often, the method of alginate solution sterilization is not given in the papers.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, it is difficult to find numerical data confirming the beneficial effect of filter-sterilized versus autoclave-sterilized alginate on protoplast development. Nevertheless, several authors suggested the positive role of filter-sterilized alginate on plating efficiency, e.g., for Brassica napus (Draget et al 1988), B. vulgaris (Hall et al 1997), or N. tabacum (Dovzhenko et al 1998. Although filter sterilization of alginate solution is quite a difficult and long process (preceded by overnight stirring on a gyratory shaker to dissolve), the values noted for plating efficiency in the present study confirmed its stimulatory effect on mitotic activity of protoplast-derived cells.…”

Section: Discussionmentioning

confidence: 99%

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Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

Maćkowska

Jarosz

Grzebelus

2014

Plant Cell Tiss Organ Cult

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Protoplasts of four Daucus carota subspecies and three wild Daucus species were isolated from 2-weekold shoot cultures during overnight incubation in an enzyme mixture composed of 1 % (w/v) cellulase Onozuka R-10 and 0.1 % (w/v) pectolyase Y-23. Before the culture, they were embedded in autoclave-or filter-sterilized alginate solution. Modified thin alginate layer (TAL) and extra thin alginate film (ETAF) techniques were applied for protoplast immobilization. A rich mineral-organic medium based on the formulation of Kao and Michayluk supplemented with 0.1 mg l -1 2,4-dichlorophenoxyacetic acid, 0.2 mg l -1 zeatin, and optionally 100 nM phytosulfokine (PSK), a peptidyl plant growth factor, was used for protoplast culture. Plating efficiency was genotype-dependent and in 40-day-old cultures, it varied from 10 % for Daucus pusillus to 73 % for D. carota subsp. sativus. A considerably higher ability in colony formation was observed in the modified TAL culture system using filter-sterilized alginate and in the presence of PSK in the protoplast culture medium. Plant regeneration through somatic embryogenesis stimulated by PSK application occurred for five out of the seven Daucus accessions used in the present study. We believe our data may facilitate the use of wild Daucus in somatic hybridization with cultivated carrot.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

Maćkowska

Jarosz

Grzebelus

2014

Plant Cell Tiss Organ Cult

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“…In the egg-box model of alginate postulated by Grant et al [8], it is primarily the guluronic acid chains of alginate, which ionically bind each other via the calcium ions. Other factors which can affect the properties of the gel include the concentration of the alginate solution, the duration of gelation, the molarity of the gelation solution (usually CaCL) [15], and method of sterilization [4]. (2001 J 193-499 In identifying factors which contribute to the mechanical stability of alginate, we firstly investigated the effect of guluronic acid content and the sterilization method on the compressive properties of alginate cylinders.…”

mentioning

confidence: 99%

Development of mechanically stable alginate/chondrocyte constructs: effects of guluronic acid content and matrix synthesis

Wong¹,

Siegrist²,

Wang³

et al. 2001

Journal Orthopaedic Research

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M E hlullc7r Institute AbstractThe purpose of this study was to investigate factors which enhanced the compressive properties of alginatekhondrocyte constructs. Firstly, we studied the effect of biochemical composition (high, mid and low guluronic acid content) and sterilization method on alginate properties. Secondly, we studied the biosynthetic characteristics of chondrocytes in three different alginate compositions and performed mechanical tests to determine whether the synthesis of cartilage matrix components could significantly enhance the compressive properties. 2% alginate solutions containing an initial cell density of 4 Y lo6 cells/ml were cast into cylinders and cultured for seven weeks. Compression tests, biochemistry, immunohistochemistry and electron microscopy were performed at fixed intervals during the seven-week culture period. The dynamic modulus, peak strain, and peak stress were maximum for alginate with the highest guluronic acid content. The presence of cells and their respective matrix components enhanced the equilibrium modulus of the constructs for all types of alginate, though this effect was small. Alginate containing the middle amount of guluronic acid resulted in constructs which were both mechanically stable and which promoted synthesis of cartilage matrix proteins. In experiments and applications in which the mechanical integrity of the alginate is important, the composition and purity of the alginate and its method of sterilization should be selected with care.

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“…Alginate gels have an open pore structure, allowing free diffusion of small molecules (Tricoli et al, 1986;Draget et al, 1988;Smidsrød and Skjåk-Braek, 1990). Cells entrapped in alginate are easily manipulated with less risk of physical damage (Draget et al, 1988;Niedz, 1993).…”

Section: Introductionmentioning

confidence: 99%

Embedding in Filter-Sterilized Alginate Enhances Brassica Oleracea L. Protoplast Culture

Kiełkowska¹,

Adamus²

2015

Acta Biologica Cracoviensia S. Botanica

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The influence of sodium alginate sterilization on the viability and mitotic activity of embedded protoplasts was studied in protoplasts of Brassica oleracea subsp. alba and rubra isolated from hypocotyl tissue and leaves of seedlings or plants grown in vitro. Both leaf and hypocotyl-derived protoplasts were more viable and divided more frequently when embedded in filtrated alginate. Division frequency was highest in cv. Reball F1 and the mitotic activity of its protoplasts was three times higher when embedded in filtrated alginate (36.1 ± 6.8%) than when cultured in autoclaved alginate (10.9 ± 5.0%). Protoplast-derived calli colonies were transferred to solid regeneration media and plants of all tested accessions were obtained.K Ke ey y w wo or rd ds s: : Brassica, calcium alginate layers, mitotic division, protoplast viability, regeneration.ACTA BIOLOGICA CRACOVIENSIA Series Botanica 56

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Regeneration, Cultivation and Differentiation of Plant Protoplasts Immobilized in Ca-alginate Beads

Cited by 54 publications

References 14 publications

Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

Development of mechanically stable alginate/chondrocyte constructs: effects of guluronic acid content and matrix synthesis

Embedding in Filter-Sterilized Alginate Enhances Brassica Oleracea L. Protoplast Culture

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