Sugar beet guard cell protoplasts demonstrate a remarkable capacity for cell division enabling applications in stomatal physiology and molecular breeding

Hall, Robert D.; Riksen-Bruinsma, T.; Weyens, Guy; Lefebvre, Marc; Dunwell, Jim M.; Tunen, Arjen van; Krens, Frans A.

doi:10.1093/jxb/48.2.255

Cited by 19 publications

(16 citation statements)

References 35 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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“…For this species two alternative protoplast sources were devised. Hall et al (1996Hall et al ( , 1997 reported that sugar beet stomatal guard cells exhibited totipotency and provided protoplasts for which the mean plating efficiency of the best responsive accession reached almost 22%. Similar plating efficiencies were obtained by Dovzhenko and Koop (2003) with protoplasts isolated from friable regenerable callus.…”

Section: Discussionmentioning

confidence: 99%

“…Major improvements were introduced with the use of (1) n-propyl gallate (nPG)-a lipoxygenase inhibitor included both in the isolation and culture media (Krens et al 1990), (2) protoplast embedding-especially in calcium alginate (Schlangstedt et al 1992;Hall et al 1993), (3) feeder systems exploiting nurse protoplasts or cells from suspension culture (Hall et al 1993) and (4) stomatal guard cells or hypocotyl-derived friable callus as protoplast sources (Hall et al 1997;Dovzhenko and Koop 2003). However, the latter two improvements involve rather complex manipulations and therefore, they are not well-suited for the routine laboratory practice.…”

Section: Introductionmentioning

confidence: 99%

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

et al. 2012

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The aim of this work was to improve plating efficiency of sugar beet mesophyll protoplast cultures. Preliminary experiments showed that cultures of good quality, viable protoplasts were obtained in rich media based on the Kao and Michayluk formulation and with the calcium alginate as an embedding matrix. Nevertheless, in these cultures cell divisions were either not observed or very seldom confirming earlier reported recalcitrance of sugar beet protoplasts. The recalcitrant status of these cultures was reversed upon application of exogenous phytosulfokine (PSK)-a peptidyl plant growth factor. The highest effectiveness of PSK was observed at 100 nM concentration. Plating efficiencies obtained in the presence of PSK reached approximately 20% of the total cultured cells. The stimulatory effect of phytosulfokine was observed for all tested breeding stocks of sugar beet. Our data indicate that PSK is a powerful agent able to overcome recalcitrance of plant protoplast cultures.

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“…We also determined that ethylene can induce division in terminally differentiating guard cells. Division of formed guard cells has recently been observed using other treatment regimes including rapid temperature shifts (Hall et al, 1997;H. Kazama, unpublished data).…”

Section: Ethylene May Suspend Cell Fate Determinationmentioning

confidence: 89%

Transient Exposure to Ethylene Stimulates Cell Division and Alters the Fate and Polarity of Hypocotyl Epidermal Cells

et al. 2004

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After transient exposure to the gaseous hormone ethylene, dark-grown cucumber (Cucumis sativus) hypocotyls developed unusual features. Upon ethylene's removal, the developing epidermis showed significant increases in cell division rates, producing an abundance of guard cells and trichomes. These responses to ethylene depended on the stage of development at the time of ethylene exposure. In the upper region of the hypocotyl, where cells were least differentiated at the onset of ethylene treatment, complex, multicellular protuberances formed. Further down the hypocotyl, where stomata and trichomes were beginning to develop at the onset of ethylene exposure, an increase in the number of stomata and trichomes was observed. Stomatal complexes developing after the ethylene treatment had a significant increase in the number of stomatal subsidiary cells and the number of cells per trichome increased. Analysis of division patterns in stomatal complexes indicated that exposure to ethylene either suspended or altered cell fate. Ethylene also altered cell division polarity, resulting in aberrant stomatal complexes and branched trichomes. To our knowledge, the results of this study demonstrate for the first time that transient treatment with physiological concentrations of ethylene can alter cell fate and increase the propensity of cells to divide.Ethylene regulates a variety of physiological and biochemical processes in plants, such as fruit ripening, senescence, abscission, sex determination, root initiation, and cell elongation (Abeles et al., 1992). Ethylene also regulates the size and stature of plants in response to various environmental cues (Abeles, 1973;Kieber, 1997). Its effects on dark-grown seedling development, described initially as the triple response, include increased radial growth of the root and stem, reduced root and stem elongation, and abnormal horizontal stem growth (Neljubow, 1901;Knight et al., 1910). The term triple response, as it is now commonly used, includes exaggeration in the curvature of the apical hook instead of abnormal horizontal growth (Bleecker et al., 1988;Guzman and Ecker, 1990 and references therein). The triple response of Arabidopsis has been exploited for characterizing the ethylene signal transduction system by identifying numerous mutants that either phenocopy the triple response in the absence of exogenous ethylene or fail to respond properly to ethylene (Guzman and Ecker, 1990).The dramatic stimulation of radial swelling of stems and roots by ethylene has led to physiological studies and cellular analyses of this phenomenon (Burg and Burg, 1966;Lang et al., 1982;Bleecker et al., 1988;Abeles et al., 1992;Baskin and Williamson, 1992;Kieber, 1997). Depending on the timing of ethylene's application, the physiological condition, seedling age, and the plant species concerned, either promotive (Ku et al., 1970;Lehman et al., 1996;Smalle et al., 1997) or inhibitory (Kieber et al., 1993) effects on organ elongation have been reported. For young developing dicot seedlings, ethylene has ...

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Sugar beet guard cell protoplasts demonstrate a remarkable capacity for cell division enabling applications in stomatal physiology and molecular breeding

Cited by 19 publications

References 35 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

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

Transient Exposure to Ethylene Stimulates Cell Division and Alters the Fate and Polarity of Hypocotyl Epidermal Cells

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