67891011129 of 17
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Understanding and manipulating primary cell walls in plant cell suspension cultures
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience.ORCID iD: 0000-0002-5541-7853
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 ; 2019:2
Keywords [en]
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
Biochemistry and Molecular Biology
Research subject
Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-240980ISBN: 978-91-7873-074-2 (print)OAI: oai:DiVA.org:kth-240980DiVA, id: diva2:1275594
Public defence
2019-02-01, FA32, Roslagstullsbacken 21, AlbaNova, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 20190107

Available from: 2019-01-07 Created: 2019-01-07 Last updated: 2019-01-07Bibliographically approved
List of papers
1. Radiometric and spectrophotometric in vitro assays of glycosyltransferases involved in plant cell wall carbohydrate biosynthesis
Open this publication in new window or tab >>Radiometric and spectrophotometric in vitro assays of glycosyltransferases involved in plant cell wall carbohydrate biosynthesis
2012 (English)In: Nature Protocols, ISSN 1754-2189, E-ISSN 1750-2799, Vol. 7, no 9, p. 1634-1650Article in journal (Refereed) Published
Abstract [en]

Most of the glycosyltransferases (GTs) that catalyze the formation of plant cell wall carbohydrates remain to be biochemically characterized. This can be achieved only if specific assays are available for these enzymes. Here we present a protocol for in vitro assays of processive and nonprocessive membrane-bound GTs. The assays are either based on the use of radioactive nucleotide sugars (NDP sugars; e.g., UDP-[U-C-14] glucose) and the quantification of the radiolabeled monosaccharides incorporated into soluble or insoluble carbohydrates, or on the coupling of the GT reaction with that of pyruvate kinase (PK) and the oxidation of NADH by lactate dehydrogenase (LDH). The radiometric assays are more suitable for exploratory work on poorly characterized enzymes, whereas the spectrophotometric assays require the availability of highly enriched GTs. Both assays can be performed within 1 d, depending on the number of fractions to be assayed or reaction mixtures to be tested.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-103376 (URN)10.1038/nprot.2012.089 (DOI)000308526300006 ()
Note

QC 20121016

Available from: 2012-10-16 Created: 2012-10-11 Last updated: 2019-01-07Bibliographically approved
2. Proteomic Analysis of Plasmodesmata From Populus Cell Suspension Cultures in Relation With Callose Biosynthesis.
Open this publication in new window or tab >>Proteomic Analysis of Plasmodesmata From Populus Cell Suspension Cultures in Relation With Callose Biosynthesis.
Show others...
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.

Keywords
Populus, callose, callose synthase, mass spectrometry, plasmodesmata, spectral counting
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-240964 (URN)10.3389/fpls.2018.01681 (DOI)000450425100001 ()30510561 (PubMedID)2-s2.0-85058796619 (Scopus ID)
Note

QC 20190107

Available from: 2019-01-07 Created: 2019-01-07 Last updated: 2019-01-07Bibliographically approved
3. The effect of carbohydrate-binding modules (CBMs) on plant cell wall properties: an in vivo approach
Open this publication in new window or tab >>The effect of carbohydrate-binding modules (CBMs) on plant cell wall properties: an in vivo approach
Show others...
(English)Manuscript (preprint) (Other academic)
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-240965 (URN)
Note

QC 20190107

Available from: 2019-01-07 Created: 2019-01-07 Last updated: 2019-01-07Bibliographically approved
4. Stronger cellulose microfibrils network structure through the expression of cellulose-binding modules in plant primary cell walls
Open this publication in new window or tab >>Stronger cellulose microfibrils network structure through the expression of cellulose-binding modules in plant primary cell walls
(English)Manuscript (preprint) (Other academic)
National Category
Other Chemistry Topics
Identifiers
urn:nbn:se:kth:diva-240966 (URN)
Note

QC 20190107

Available from: 2019-01-07 Created: 2019-01-07 Last updated: 2019-01-07Bibliographically approved

Open Access in DiVA

fulltext(1963 kB)15 downloads
File information
File name FULLTEXT01.pdfFile size 1963 kBChecksum SHA-512
13fdd64336d9a7ce73ca3383fa167253d25d1ea29071697662d9a937befb8e71d1293a094a740c74a9206d31f73c6f0fb3939c129e3272ee46a517c2a767d2b6
Type fulltextMimetype application/pdf

Search in DiVA

By author/editor
Leijon, Felicia
By organisation
Glycoscience
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 15 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 105 hits
67891011129 of 17
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf