Protoplasts: a useful research system for plant cell biology, especially dedifferentiation

Jiang, Fangwei; Zhu, Jian Kang; Liu, Hailiang

doi:10.1007/s00709-013-0513-z

Cited by 35 publications

(17 citation statements)

References 41 publications

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“…Thirdly, a very large number of protoplasts can be isolated from one preparation, allowing testing of many variables simultaneously in one assay. Finally, protoplast transformation is now well established (Yoo et al, 2007), with a higher efficiency than any other plant transformation system (Jiang et al, 2013), and is therefore very well suited for high throughput platforms. The use of protoplast-based systems also allows access to other types of high throughput approaches, such as flow cytometry and cell sorting, which open a wide new range of screening possibilities.…”

Section: Introductionmentioning

confidence: 99%

A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role of ABI3 in Lipid Metabolism Regulation

et al. 2020

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Traditional functional genetic studies in crops are time consuming, complicated and cannot be readily scaled up. The reason is that mutant or transformed crops need to be generated to study the effect of gene modifications on specific traits of interest. However, many crop species have a complex genome and a long generation time. As a result, it usually takes several months to over a year to obtain desired mutants or transgenic plants, which represents a significant bottleneck in the development of new crop varieties. To overcome this major issue, we are currently establishing a versatile plant genetic screening platform, amenable to high throughput screening in almost any crop species, with a unique workflow. This platform combines protoplast transformation and fluorescence activated cell sorting. Here we show that tobacco protoplasts can accumulate high levels of lipid if transiently transformed with genes involved in lipid biosynthesis and can be sorted based on lipid content. Hence, protoplasts can be used as a predictive tool for plant lipid engineering. Using this newly established strategy, we demonstrate the major role of ABI3 in plant lipid accumulation. We anticipate that this workflow can be applied to numerous highly valuable metabolic traits other than storage lipid accumulation. This new strategy represents a significant step toward screening complex genetic libraries, in a single experiment and in a matter of days, as opposed to years by conventional means.

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Section: Introductionmentioning

confidence: 99%

A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role of ABI3 in Lipid Metabolism Regulation

et al. 2020

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“…Plant protoplast technology can be widely used in various aspects of plant science to study such fundamental processes as differentiation, dedifferentiation, and pluripotency of cells (reviewed in Jiang et al 2013 ), and to introduce novel characteristics in commercial crops. Conventional breeding tools may incorporate different techniques exploring protoplast culture, including micropropagation, in vitro selection, genetic transformation, and, particularly, somatic hybridization (reviewed in Davey et al 2005a ).…”

Section: Introductionmentioning

confidence: 99%

Effect of β-lactam antibiotics on plant regeneration in carrot protoplast cultures

Grzebelus

Skop

2014

In Vitro Cell.Dev.Biol.-Plant

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Protoplasts of three carrot cultivars were isolated from in vitro-grown plantlets by overnight incubation in an enzyme mixture composed of 1% (w/v) cellulase Onozuka R-10 and 0.1% (w/v) pectolyase Y-23. After cell immobilization in modified thin alginate layers, three types of β-lactam antibiotics (cefotaxime, carbenicillin, or timentin) at five different concentrations (100, 200, 300, 400, or 500 mg L−1) were added to the culture medium. In 20-d-old cultures, a different number of cell colonies had formed and varied on average from 27 to 56% in carbenicillin- and cefotaxime-containing media, respectively. Supplementation of the culture media with antibiotics at concentrations higher than 100 mg L−1 resulted in a decrease in plating efficiency in comparison with the controls. However, from all antibiotic treatments, except carbenicillin at concentrations of 400–500 mg L−1, efficient plant regeneration occurred. For this reason, we believe that cefotaxime and timentin in the concentrations analyzed here may be used in complex in vitro procedures or valuable carrot cultures as a prophylactic agent for prevention against occasional contaminations.

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“…In plant cells, dedifferentiation can occur after enzymatic degradation of the cell wall, which leads to the formation of protoplasts. The most common sources of protoplasts are leaf mesophyll cells (Jiang et al 2013 ). After the degradation of their cell wall, these specialised mesophyll cells become unspecialised, totipotent protoplasts, which can regenerate the entire plant with an intermediate microcalli stage in the appropriate conditions (Takebe et al 1971 ; Guri et al 1987 ; Kim and Lee 1996 ; Chupeau et al 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Dedifferentiation of Arabidopsis thaliana cells is accompanied by a strong decrease in RNA polymerase II transcription activity and poly(A+) RNA and 25S rRNA eradication from the cytoplasm

et al. 2014

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The mechanisms of plant cell dedifferentiation and the acquisition of totipotency are poorly understood. One of the methods to induce the dedifferentiation process in plant cells is simple and requires the removal of the cell wall. After cell wall removal in protoplasts, large-scale chromatin decondensation is observed (Tessadori et al. in J Cell Sci 120:1200–1208, 2007). Here, we show that in Arabidopsis thaliana protoplasts, despite chromatin decondensation, RNA polymerase II transcriptional activity is reduced. The subsequent investigated stages displayed a clear decrease in the quantity of 25S ribosomal RNA (rRNA) first and then poly(A+) RNA, particularly in the cytoplasm. Therefore, the reduced transcription activity and the removal of these RNA transcripts from the cytoplasm is a crucial process in obtaining totipotency in plant cells. After the cytoplasm cleaning of transcripts derived from mesophyll cells, we observed the resynthesis of these RNAs. An increase in the amount of examined molecules to a level similar to that in differentiated mesophyll cells precedes the divisions of already undifferentiated cells. In this work, we show changes in RNA polymerase II transcription dynamics and the quantity of poly(A+) RNA and 25S rRNA during dedifferentiation and re-entry into the cell cycle.

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Protoplasts: a useful research system for plant cell biology, especially dedifferentiation

Cited by 35 publications

References 41 publications

A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role of ABI3 in Lipid Metabolism Regulation

A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role of ABI3 in Lipid Metabolism Regulation

Effect of β-lactam antibiotics on plant regeneration in carrot protoplast cultures

Dedifferentiation of Arabidopsis thaliana cells is accompanied by a strong decrease in RNA polymerase II transcription activity and poly(A+) RNA and 25S rRNA eradication from the cytoplasm

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