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Proteomic Analysis of Plasmodesmata From Populus Cell Suspension Cultures in Relation With Callose Biosynthesis.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience.ORCID iD: 0000-0002-5541-7853
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience.ORCID iD: 0000-0001-9832-027X
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience.ORCID iD: 0000-0003-1877-4154
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2018 (English)In: Frontiers in Plant Science, ISSN 1664-462X, E-ISSN 1664-462X, Vol. 9, article id 1681Article in journal (Refereed) Published
Abstract [en]

Plasmodesmata are channels that link adjacent cells in plant tissues through which molecular exchanges take place. They are involved in multiple processes vital to plant cells, such as responses to hormonal signaling or environmental challenges including osmotic stress, wounding and pathogen attack. Despite the importance of plasmodesmata, their proteome is not well-defined. Here, we have isolated fractions enriched in plasmodesmata from cell suspension cultures of Populus trichocarpa and identified 201 proteins that are enriched in these fractions, thereby providing further insight on the multiple functions of plasmodesmata. Proteomics analysis revealed an enrichment of proteins specifically involved in responses to stress, transport, metabolism and signal transduction. Consistent with the role of callose deposition and turnover in the closure and aperture of the plasmodesmata and our proteomic analysis, we demonstrate the enrichment of callose synthase activity in the plasmodesmata represented by several gene products. A new form of calcium-independent callose synthase activity was detected, in addition to the typical calcium-dependent enzyme activity, suggesting a role of calcium in the regulation of plasmodesmata through two forms of callose synthase activities. Our report provides the first proteomic investigation of the plasmodesmata from a tree species and the direct biochemical evidence for the occurrence of several forms of active callose synthases in these structures. Data are available via ProteomeXchange with identifier PXD010692.

Place, publisher, year, edition, pages
2018. Vol. 9, article id 1681
Keywords [en]
Populus, callose, callose synthase, mass spectrometry, plasmodesmata, spectral counting
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-240964DOI: 10.3389/fpls.2018.01681ISI: 000450425100001PubMedID: 30510561Scopus ID: 2-s2.0-85058796619OAI: oai:DiVA.org:kth-240964DiVA, id: diva2:1275523
Note

QC 20190107

Available from: 2019-01-07 Created: 2019-01-07 Last updated: 2019-10-14Bibliographically approved
In thesis
1.
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2. Understanding and manipulating primary cell walls in plant cell suspension cultures
Open this publication in new window or tab >>Understanding and manipulating primary cell walls in plant cell suspension cultures
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The cell wall is required for many aspects of plant function and development. It is also an accessible and renewable resource utilized both in unrefined forms and as raw material for further development. Increased knowledge regarding cell wall structure and components will contribute to better utilization of plants and the resources they provide. In this thesis aspects of the primary cell wall of Populus trichocarpa and Nicotiana tabacum are explored.

In Publication I a method for isolation and biochemical characterization of plant glycosyltransferases using a spectrophotometric or a radiometric assay was optimized. The radiometric assay was applied in Publication II where the proteome of the plasmodesmata isolated from P. trichocarpa was analyzed. Proteins identified belonged to functional classes such as “transport”, “signalling” and “stress responses”. Plasmodesmata-enriched fractions had high levels of callose synthase activity under ion depleted conditions as well as with calcium present.

The second part of the thesis comprises the alteration of the cell wall of N. tabacum cells and A. thaliana plants through in vivo expression of a carbohydrate binding module (CBM) (Publication III). In tobacco this resulted in cell walls with loose ultrastructure containing an increased proportion of 1,4-β-glucans. The cell walls were more susceptible to saccharification, possibly due to changes in the structure of cellulose or xyloglucan. Arabidopsis plants showed increased saccharification after mild pretreatment, suggesting that heterologous expression of CBMs is a promising method for cell wall engineering. In Publication IV cellulose microfibrils (CMFs) and nanocrystals (CNCs) were extracted from the transgenic cells. CNC preparation resulted in higher yields and longer CNCs. Nanopapers prepared from the CMFs of the CBM line demonstrated enhanced strength and toughness. Thus, changes to the ordered regions of cellulose were suggested to take place due to CBM expression.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2019. p. 82
Series
TRITA-CBH-FOU ; 2
Keywords
Callose synthase, carbohydrate-binding module, cell wall engineering, cellulose microfibril, cellulose nanocrystal, glycosyltransferase, mass spectrometry, plasmodesmata, Populus, primary cell wall, radiometric assay, spectrophotometric assay
National Category
Natural Sciences
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-251039 (URN)978-91-7873-074-2 (ISBN)
Public defence
2019-06-10, FA32, Roslagstullsbacken 21, AlbaNova, Stockholm, 10:00 (English)
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Note

QC 2019-05-08

Available from: 2019-05-08 Created: 2019-05-08 Last updated: 2019-05-08Bibliographically approved

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