Plant protoplasts: status and biotechnological perspectives

Davey, M. R.; Anthony, Paul; Power, J. B.; Lowe, K. C.

doi:10.1016/j.biotechadv.2004.09.008

Cited by 331 publications

(226 citation statements)

References 224 publications

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“…Although the reason for such negative correlation remains unclear; it may be directly related to protoplast recalcitrance of the examined species. Such a reaction may be a result of the unbalanced osmotic status leading to osmotic stress, and for that reason the reduction of medium osmolarity is usually essential for proper regeneration processes (Davey et al 2005). However, our previous studies on yellow lupin protoplasts revealed that gradual osmolarity reduction had no effect on the number of viable protoplasts and the frequency of mitoses (unpublished).…”

Section: Discussionmentioning

confidence: 97%

Studies on cell wall regeneration in protoplast culture of legumes - the effect of organic medium additives on cell wall components

Wiszniewska¹,

Piwowarczyk²

2014

Czech J. Genet. Plant Breed.

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Wiszniewska A., Piwowarczyk B. (2014): Studies on cell wall regeneration in protoplast culture of legumesthe effect of organic medium additives on cell wall components. Czech J. Genet. Plant Breed., 50: 84-91.The cell wall regeneration in mesophyll protoplasts of yellow lupin and grass pea was studied. The occurrence of cell wall components: cellulose, callose and arabinogalactan proteins was analysed during 15 days of culture. Protoplasts were cultured in different media to test the effect of culture environment on the cell wall regeneration. Medium supplementation with 2 mg/l chitosan resulted in prolonged viability, more balanced cellulose resynthesis, increased callose formation and induction of mitotic divisions in protoplast-derived cells of both examined legumes. In chitosan-enriched medium arabinogalactan proteins were detected in cell plates of divided cells. Medium rich in additional organic compounds, i.e. free amino acids, organic acids and monosaccharides, was inferior to media of simpler composition. In both species the relatively quick cellulose resynthesis negatively affected the viability of protoplast-derived cells. In grass pea cellulose appeared during 24 h of culture. In yellow lupin the process started significantly later and after 10 days the frequency of walled cells did not exceed 50%. Callose was detected in cultures of both species and its pattern suggested that the synthesis was unlikely to be a result of protoplast wounding. Arabinogalactan proteins were localized in cell walls of different types of cells: dividing, elongating, but predominantly in degenerating ones. Occurrence and organization of the cell wall components studied were discussed in relation to recalcitrance of grass pea and yellow lupin protoplasts.

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Section: Discussionmentioning

confidence: 97%

Studies on cell wall regeneration in protoplast culture of legumes - the effect of organic medium additives on cell wall components

Wiszniewska¹,

Piwowarczyk²

2014

Czech J. Genet. Plant Breed.

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“…Plant cell protoplasts are known to maintain their differentiated state without dedifferentiation in isotonic solutions and are simple experimental systems, much like mammalian cell lines, for studying specific cellular activities (Davey et al, 2005). SDX protoplasts are expected to retain specific cellular and transcriptomic potentials for wood formation.…”

Section: Discussionmentioning

confidence: 99%

“…Plant protoplasts can be cell-or tissue-specific populations of single cells used to study a broad spectrum of processes from physiology to gene function/regulation (Abel and Theologis, 1994;Chiu et al, 1996;Davey et al, 2005;Thorpe, 2007;Yoo et al, 2007). Freshly isolated protoplasts retain cell and transcriptome identity, differentiated state (without dedifferentiation), and original biochemical and regulatory activity (Cocking, 1972;Sheen, 2001;Birnbaum et al, 2003;Yoo et al, 2007;Faraco et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

SND1 Transcription Factor-Directed Quantitative Functional Hierarchical Genetic Regulatory Network in Wood Formation in Populus trichocarpa

et al. 2013

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Wood is an essential renewable raw material for industrial products and energy. However, knowledge of the genetic regulation of wood formation is limited. We developed a genome-wide high-throughput system for the discovery and validation of specific transcription factor (TF)-directed hierarchical gene regulatory networks (hGRNs) in wood formation. This system depends on a new robust procedure for isolation and transfection of Populus trichocarpa stem differentiating xylem protoplasts. We overexpressed Secondary Wall-Associated NAC Domain 1s (Ptr-SND1-B1), a TF gene affecting wood formation, in these protoplasts and identified differentially expressed genes by RNA sequencing. Direct Ptr-SND1-B1-DNA interactions were then inferred by integration of timecourse RNA sequencing data and top-down Graphical Gaussian Modeling-based algorithms. These Ptr-SND1-B1-DNA interactions were verified to function in differentiating xylem by anti-PtrSND1-B1 antibody-based chromatin immunoprecipitation (97% accuracy) and in stable transgenic P. trichocarpa (90% accuracy). In this way, we established a Ptr-SND1-B1-directed quantitative hGRN involving 76 direct targets, including eight TF and 61 enzyme-coding genes previously unidentified as targets. The network can be extended to the third layer from the second-layer TFs by computation or by overexpression of a second-layer TF to identify a new group of direct targets (third layer). This approach would allow the sequential establishment, one two-layered hGRN at a time, of all layers involved in a more comprehensive hGRN. Our approach may be particularly useful to study hGRNs in complex processes in plant species resistant to stable genetic transformation and where mutants are unavailable.

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“…They have also been used for organellar transplantation-in most cases uptake of chloroplasts and nuclei was reported (reviewed in Razdan 2003). The necessary prerequisite for these applications is an ability of plant protoplasts to reconstitute their cell wall and undergo mitotic divisions which results in callus formation and eventually shoot regeneration (Davey et al 2005a). The problem is that for many species this developmental program is hard to achieve.…”

Section: Discussionmentioning

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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Plant protoplasts: status and biotechnological perspectives

Cited by 331 publications

References 224 publications

Studies on cell wall regeneration in protoplast culture of legumes - the effect of organic medium additives on cell wall components

Studies on cell wall regeneration in protoplast culture of legumes - the effect of organic medium additives on cell wall components

SND1 Transcription Factor-Directed Quantitative Functional Hierarchical Genetic Regulatory Network in Wood Formation in Populus trichocarpa

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

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