Using Chemical Genomics to Study Cell Wall Formation and Cell Growth in Arabidopsis thaliana and Penium margaritaceum

Worden, Natasha; Esteve, V. Esteva; Domozych, David S.; Drakakaki, Georgia

doi:10.1007/978-1-4939-1902-4_2

Cited by 5 publications

(4 citation statements)

References 36 publications

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“…Given the loadbearing role of cellulose, we then examined the effect of cellulose compared to callose inhibition in our experimental conditions. Cellulose inhibition by isoxaben (IXB) treatment led to strong reduction of root growth ( Scheible et al, 2001 ; Worden et al, 2015 ) and a root swollen phenotype compared to ES7 ( Supplemental Figure S12 ). However, while cytokinesis defects in the form of cell plate stubs, were observed with ES7 treatment ( Figure 5, E and Supplemental Figure S12, E and J ), this effect was not detectable in IXB treatment ( Figure 5 and Supplemental Figure S12, G– J ).…”

Section: Resultsmentioning

confidence: 99%

A biophysical model for plant cell plate maturation based on the contribution of a spreading force

et al. 2021

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Plant cytokinesis, a fundamental process of plant life, involves de novo formation of a ‘cell plate’ partitioning the cytoplasm of dividing cells. Cell plate formation is directed by orchestrated delivery, fusion of cytokinetic vesicles, and membrane maturation to form a nascent cell wall by timely deposition of polysaccharides. During cell plate maturation, the fragile membrane network transitions to a fenestrated sheet and finally a young cell wall. Here, we approximated cell plate sub-structures with testable shapes and adopted the Helfrich free energy model for membranes, including a stabilizing and spreading force, to understand the transition from a vesicular network to a fenestrated sheet and mature cell plate. Regular cell plate development in the model was possible, with suitable bending modulus, for a two-dimensional late stage spreading force of 2-6 pN/nm, an osmotic pressure difference of 2-10 kPa, and spontaneous curvature between 0-0.04 nm−1. With these conditions, stable membrane conformation sizes and morphologies emerged in concordance with stages of cell plate development. To reach a mature cell plate, our model required the late-stage onset of a spreading/stabilizing force coupled with a concurrent loss of spontaneous curvature. Absence of a spreading/stabilizing force predicts failure of maturation. The proposed model provides a framework to interrogate different players in late cytokinesis and potentially other membrane networks that undergo such transitions. Callose, is a polysaccharide that accumulates transiently during cell plate maturation. Callose related observations were consistent with the proposed model’s concept, suggesting that it is one of the factors involved in establishing the spreading force.

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Section: Resultsmentioning

confidence: 99%

A biophysical model for plant cell plate maturation based on the contribution of a spreading force

et al. 2021

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“…Over the past decade, the desmid, Penium margaritaceum , has also become a valuable model organism ( Domozych et al, 2009 , 2014 ; Sørensen et al, 2014 ; Rydahl et al, 2015 ; Worden et al, 2015 ). Unlike Micrasterias, Penium has a simple cylindrical shape, possesses only a primary cell wall and deposits wall polymers at two specific loci of the cell surface during expansion ( Domozych et al, 2011 ).…”

Section: Charophytes As Model Organismsmentioning

confidence: 99%

“…After labeling of live cells, these can be returned to culture where subsequent cell expansion and wall deposition events can be monitored ( Domozych et al, 2009 ; Rydahl et al, 2015 ). Penium is also easily maintained in the laboratory and its fast growth rate under precisely controlled conditions makes it an excellent specimen for large-scale concurrent microarray screenings of many chemical agents (by growth in multi-well plates) and for assessment of their specific effects on expansion/wall development ( Worden et al, 2015 ). This significantly aids in revealing the role of specific subcellular components and processes in the expansion/differentiation.…”

Section: Charophytes As Model Organismsmentioning

confidence: 99%

Charophytes: Evolutionary Giants and Emerging Model Organisms

2016

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Charophytes are the group of green algae whose ancestral lineage gave rise to land plants in what resulted in a profoundly transformative event in the natural history of the planet. Extant charophytes exhibit many features that are similar to those found in land plants and their relatively simple phenotypes make them efficacious organisms for the study of many fundamental biological phenomena. Several taxa including Micrasterias, Penium, Chara, and Coleochaete are valuable model organisms for the study of cell biology, development, physiology and ecology of plants. New and rapidly expanding molecular studies are increasing the use of charophytes that in turn, will dramatically enhance our understanding of the evolution of plants and the adaptations that allowed for survival on land. The Frontiers in Plant Science series on "Charophytes" provides an assortment of new research reports and reviews on charophytes and their emerging significance as model plants.

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“…Penium margaritaceum (Zygnematophyceae; Streptophyta) is a unicellular green alga that belongs to the charophycean green algae (CGA), or basal Streptophyta [1][2][3], i.e., the assemblage of extant green algae most closely related and ancestral to land plants [4,5]. This species has become increasingly valuable as a tool for studies of the primary cell wall of plants [6][7][8][9]. Penium has a simple geometric form consisting of an elongate cylinder with rounded edges, and produces only a primary cell wall that contains several of the polymers that are typically found in the cell walls of land plants, namely cellulose, pectins (homogalacturonan (HG) and rhamnogalacturonan I (RGI)) and proteoglycans [10,11].…”

Section: Introductionmentioning

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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Using Chemical Genomics to Study Cell Wall Formation and Cell Growth in Arabidopsis thaliana and Penium margaritaceum

Cited by 5 publications

References 36 publications

A biophysical model for plant cell plate maturation based on the contribution of a spreading force

A biophysical model for plant cell plate maturation based on the contribution of a spreading force

Charophytes: Evolutionary Giants and Emerging Model Organisms

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

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