Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

doi:10.3390/proteomes4040034

Cited by 12 publications

(13 citation statements)

References 107 publications

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“…62 Pectinase, cellulose, and other enzymes related to glucose metabolism play a vital role in this process. 63 The amount of endoglucanase (DN13670_c0_g1_i1) involved in the degradation of cellulose was signicantly reduced in all four treatments, which indicated that stressful reactions that soen fruit are produced in cells. Pectinesterase (DN37718_c0_g2_i3), which slows the degradation of pectin in strawberries, was signicantly less abundant in the 2.144 mg m À3 treatment group.…”

Section: Proteins Contribute To Fruit Rmnessmentioning

confidence: 91%

Label-free quantitative proteomics to investigate the response of strawberry fruit after controlled ozone treatment

Chen

Zhang²,

Zhang

et al. 2019

RSC Adv.

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Section: Proteins Contribute To Fruit Rmnessmentioning

confidence: 91%

Label-free quantitative proteomics to investigate the response of strawberry fruit after controlled ozone treatment

Chen

Zhang²,

Zhang

et al. 2019

RSC Adv.

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“…Rapid isolation of large amounts of protoplasts will facilitate the isolation and analysis of specific endomembrane components. Finally, protoplasts have become a major means in higher plants to approach questions related to the architectural dynamics of cell wall construction and hybridization [20,96]. Such technology has been successfully used in the moss Physcomitrella patens, where techniques such as somatic hybridization and transformation have been achieved using protoplasts [97][98][99].…”

Section: Future Applications Of Protoplasts In Penium Studiesmentioning

confidence: 99%

Isolation and manipulation of protoplasts from the unicellular green alga Penium margaritaceum

et al. 2018

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Background: The unicellular charophycean green alga Penium margaritaceum has emerged as an appealing experimental organism in plant cell wall and cell biology research. Innovative practical approaches in the manipulation and maintenance of this unicellular model alga are needed in order to probe the complexities of its subcellular and molecular machinery. Protoplast isolation and manipulation expedites a new range of experimental possibilities for Penium-based studies. These include enhanced means of isolation of subcellular components and macromolecules, application of intracellular probes for high resolution microscopy of live cells, transformation studies and analysis of the fundamental mechanisms of plant cell expansion and wall polymer deposition. Results: We present a methodology for enzyme-based digestion of the Penium cell wall and the isolation of protoplasts. The subcellular events associated with this technology are presented using multiple microscopy-based techniques. We also provide protocols for applying an array of intracellular dyes that can be used as markers for specific organelles and membrane microdomains in live cells. Finally, we present a protocol for the purification of a nuclei-rich fraction from protoplasts, which can be used for the isolation of nuclear DNA. Conclusion: Protoplast isolation, culturing and manipulation provide valuable means for molecular and cellular studies of Penium. The protocol described here offers a rapid and effective mechanism for fast and effective production of protoplasts. Subsequently, the protoplasts may be used for microscopy-based studies of specific subcellular components and the isolation of organelles and nuclear DNA. These methods offer a new practical methodology for future studies of this model organism in cell and molecular biology.

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“…For example, staining with Pontamine Fast Scarlet 4B, a dye that fluoresces in the presence of cellulose, facilitates the imaging of cellulose dynamics, and has revealed that cellulose bundles rotate in a transverse to longitudinal direction during cell expansion [ 28 , 29 ]. Additionally, we recently developed an imaging technique that can be used to quantitatively evaluate the network of cellulose microfibrils [ 30 , 31 ]. By combining this quantitative imaging technique with a high-yield cell-wall regeneration procedure, we successfully quantified the total length, mean intensity, skewness of intensity distribution, and coefficient of variation of regenerating cellulose microfibrils in protoplasts derived from Arabidopsis leaf mesophyll cells [ 31 ].…”

Section: Introductionmentioning

confidence: 99%

A Genomic Perspective on the Evolutionary Diversity of the Plant Cell Wall

Yokoyama

2020

Plants

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The plant cell wall is a complex and dynamic structure composed of numerous different molecules that play multiple roles in all aspects of plant life. Currently, a new frontier in biotechnology is opening up, which is providing new insights into the structural and functional diversity of cell walls, and is thus serving to re-emphasize the significance of cell wall divergence in the evolutionary history of plant species. The ever-increasing availability of plant genome datasets will thus provide an invaluable basis for enhancing our knowledge regarding the diversity of cell walls among different plant species. In this review, as an example of a comparative genomics approach, I examine the diverse patterns of cell wall gene families among 100 species of green plants, and illustrate the evident benefits of using genome databases for studying cell wall divergence. Given that the growth and development of all types of plant cells are intimately associated with cell wall dynamics, gaining a further understanding of the functional diversity of cell walls in relation to diverse biological events will make significant contributions to a broad range of plant sciences.

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Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

Cited by 12 publications

References 107 publications

Label-free quantitative proteomics to investigate the response of strawberry fruit after controlled ozone treatment

Label-free quantitative proteomics to investigate the response of strawberry fruit after controlled ozone treatment

Isolation and manipulation of protoplasts from the unicellular green alga Penium margaritaceum

A Genomic Perspective on the Evolutionary Diversity of the Plant Cell Wall

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