| 1. |
- Chowdhury, Jamil, et al.
(författare)
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Laccaria bicolor pectin methylesterases are involved in ectomycorrhiza development with Populus tremula × Populus tremuloides
- 2022
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Ingår i: New Phytologist. - : John Wiley & Sons. - 0028-646X .- 1469-8137. ; 236:2, s. 639-655
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Tidskriftsartikel (refereegranskat)abstract
- The development of ectomycorrhizal (ECM) symbioses between soil fungi and tree roots requires modification of root cell walls. The pectin-mediated adhesion between adjacent root cells loosens to accommodate fungal hyphae in the Hartig net, facilitating nutrient exchange between partners. We investigated the role of fungal pectin modifying enzymes in Laccaria bicolor for ECM formation with Populus tremula × Populus tremuloides.We combine transcriptomics of cell-wall-related enzymes in both partners during ECM formation, immunolocalisation of pectin (Homogalacturonan, HG) epitopes in different methylesterification states, pectin methylesterase (PME) activity assays and functional analyses of transgenic L. bicolor to uncover pectin modification mechanisms and the requirement of fungal pectin methylesterases (LbPMEs) for ECM formation.Immunolocalisation identified remodelling of pectin towards de-esterified HG during ECM formation, which was accompanied by increased LbPME1 expression and PME activity. Overexpression or RNAi of the ECM-induced LbPME1 in transgenic L. bicolor lines led to reduced ECM formation. Hartig Nets formed with LbPME1 RNAi lines were shallower, whereas those formed with LbPME1 overexpressors were deeper.This suggests that LbPME1 plays a role in ECM formation potentially through HG de-esterification, which initiates loosening of adjacent root cells to facilitate Hartig net formation.
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| 2. |
- Derba-Maceluch, Marta, et al.
(författare)
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Suppression of xylan endotransglycosylase PtxtXyn10A affects cellulose microfibril angle in secondary wall in aspen wood
- 2015
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Ingår i: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 205:2, s. 666-681
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Tidskriftsartikel (refereegranskat)abstract
- Certain xylanases from family GH10 are highly expressed during secondary wall deposition, but their function is unknown. We carried out functional analyses of the secondary-wall specific PtxtXyn10A in hybrid aspen (Populus tremulaxtremuloides).PtxtXyn10A function was analysed by expression studies, overexpression in Arabidopsis protoplasts and by downregulation in aspen.PtxtXyn10A overexpression in Arabidopsis protoplasts resulted in increased xylan endotransglycosylation rather than hydrolysis. In aspen, the enzyme was found to be proteolytically processed to a 68kDa peptide and residing in cell walls. Its downregulation resulted in a corresponding decrease in xylan endotransglycosylase activity and no change in xylanase activity. This did not alter xylan molecular weight or its branching pattern but affected the cellulose-microfibril angle in wood fibres, increased primary growth (stem elongation, leaf formation and enlargement) and reduced the tendency to form tension wood. Transcriptomes of transgenic plants showed downregulation of tension wood related genes and changes in stress-responsive genes. The data indicate that PtxtXyn10A acts as a xylan endotransglycosylase and its main function is to release tensional stresses arising during secondary wall deposition. Furthermore, they suggest that regulation of stresses in secondary walls plays a vital role in plant development.
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| 3. |
- Ibatullin, Farid M., et al.
(författare)
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A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta
- 2009
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Ingår i: Plant Physiology. - : Oxford University Press (OUP). - 0032-0889 .- 1532-2548. ; 151:4, s. 1741-1750
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Tidskriftsartikel (refereegranskat)abstract
- There currently exists a diverse array of molecular probes for the in situ localization of polysaccharides, nucleic acids, and proteins in plant cells, including reporter enzyme strategies (e. g. protein-glucuronidase fusions). In contrast, however, there is a paucity of methods for the direct analysis of endogenous glycoside hydrolases and transglycosidases responsible for cell wall remodeling. To exemplify the potential of fluorogenic resorufin glycosides to address this issue, a resorufin beta-glycoside of a xylogluco-oligosaccharide (XXXG-beta-Res) was synthesized as a specific substrate for in planta analysis of XEH activity. The resorufin aglycone is particularly distinguished for high sensitivity in muro assays due to a low pK(a) (5.8) and large extinction coefficient (epsilon 62,000 M-1 cm(-1)), long-wavelength fluorescence (excitation 571 nm/emission 585 nm), and high quantum yield (0.74) of the corresponding anion. In vitro analyses demonstrated that XXXG-beta-Res is hydrolyzed by the archetypal plant XEH, nasturtium (Tropaeolum majus) NXG1, with classical Michaelis-Menten substrate saturation kinetics and a linear dependence on both enzyme concentration and incubation time. Further, XEH activity could be visualized in real time by observing the localized increase in fluorescence in germinating nasturtium seeds and Arabidopsis (Arabidopsis thaliana) inflorescent stems by confocal microscopy. Importantly, this new in situ XEH assay provides an essential complement to the in situ xyloglucan endotransglycosylase assay, thus allowing delineation of the disparate activities encoded by xyloglucan endotransglycosylase/hydrolase genes directly in plant tissues. The observation that XXXG-beta-Res is also hydrolyzed by diverse microbial XEHs indicates that this substrate, and resorufin glycosides in general, may find broad applicability for the analysis of wall restructuring by polysaccharide hydrolases during morphogenesis and plant-microbe interactions.
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| 4. |
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| 5. |
- Lindberg, A., et al.
(författare)
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Cold stress stimulates algae to produce value-added compounds
- 2022
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Ingår i: Bioresource Technology Reports. - : Elsevier. - 2589-014X. ; 19
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Tidskriftsartikel (refereegranskat)abstract
- Two cold-tolerant microalgae, Chlorella vulgaris and Scenedesmus sp., were grown at 22 and 5 °C. At the lower temperature, the microalgae showed substantial biochemical and morphological changes. The soluble sugar profile in response to low-temperature cultivation was very different in the two strains. C. vulgaris increased both the sucrose and raffinose family oligosaccharides (RFOs) content at 5 °C while Scenedesmus sp. drastically reduced the sucrose content. Both strains increased the total fatty acid methyl ester (FAME) content when grown at 5 °C. However, the FAME profiles were very different: C. vulgaris mainly increased C18:1 and less so C18:3, while Scenedesmus sp. decreased C18:1 but greatly increased C18:3. The morphology of C. vulgaris changed slightly at the lower temperature, while Scenedesmus sp. showed substantial changes in the size and shape. Low temperature triggered the synthesis of unsaturated fatty acids that are essential for human nutrition.
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| 6. |
- Niemi, Calle, et al.
(författare)
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Quantitative and qualitative saccharide analysis of North Atlantic brown seaweed by gas chromatography/mass spectrometry and infrared spectroscopy
- 2024
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Ingår i: International Journal of Biological Macromolecules. - : Elsevier. - 0141-8130 .- 1879-0003. ; 254
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Tidskriftsartikel (refereegranskat)abstract
- Brown seaweeds contain a variety of saccharides which have potential industrial uses. The most abundant polysaccharide in brown seaweed is typically alginate, consisting of mannuronic (M) and guluronic acid (G). The ratio of these residues fundamentally determines the physicochemical properties of alginate. In the present study, gas chromatography/mass spectrometry (GC/MS) was used to give a detailed breakdown of the monosaccharide species in North Atlantic brown seaweeds. The anthrone method was used for determination of crystalline cellulose. The experimental data was used to calibrate multivariate prediction models for estimation of total carbohydrates, crystalline cellulose, total alginate and alginate M/G ratio directly in dried, brown seaweed using three types of infrared spectroscopy, using relative error (RE) as a measure of predictive accuracy. Diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) performed well for the estimation of total alginate (RE = 0.12, R2 = 0.82), and attenuated total reflectance (ATR) showed good prediction of M/G ratio (RE = 0.14, R2 = 0.86). Both DRIFTS, ATR and near infrared (NIR) were unable to predict crystalline cellulose and only DRIFTS performed better in determining total carbohydrates. Multivariate spectral analysis is a promising method for easy and rapid characterization of alginate and M/G ratio in seaweed.
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| 7. |
- Nishikubo, Nobuyuki, et al.
(författare)
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Xyloglucan endo-Transglycosylase-Mediated Xyloglucan Rearrangements in Developing Wood of Hybrid Aspen
- 2011
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Ingår i: Plant Physiology. - : Oxford University Press (OUP). - 0032-0889 .- 1532-2548. ; 155:1, s. 399-413
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Tidskriftsartikel (refereegranskat)abstract
- Xyloglucan endo-transglycosylases (XETs) encoded by xyloglucan endo-transglycosylases/hydrolase (XTH) genes modify the xyloglucan-cellulose framework of plant cell walls, thereby regulating their expansion and strength. To evaluate the importance of XET in wood development, we studied xyloglucan dynamics and XTH gene expression in developing wood and modified XET activity in hybrid aspen (Populus tremula X tremuloides) by overexpressing PtxtXET16-34. We show that developmental modifications during xylem differentiation include changes from loosely to tightly bound forms of xyloglucan and increases in the abundance of fucosylated xyloglucan epitope recognized by the CCRC-M1 antibody. We found that at least 16 Populus XTH genes, all likely encoding XETs, are expressed in developing wood. Five genes were highly and ubiquitously expressed, whereas PtxtXET16-34 was expressed more weakly but specifically in developing wood. Transgenic up-regulation of XET activity induced changes in cell wall xyloglucan, but its effects were dependent on developmental stage. For instance, XET overexpression increased abundance of the CCRC-M1 epitope in cambial cells and xylem cells in early stages of differentiation but not in mature xylem. Correspondingly, an increase in tightly bound xyloglucan content was observed in primary-walled xylem but a decrease was seen in secondary-walled xylem. Thus, in young xylem cells, XET activity limits xyloglucan incorporation into the tightly bound wall network but removes it from cell walls in older cells. XET overexpression promoted vessel element growth but not fiber expansion. We suggest that the amount of nascent xyloglucan relative to XET is an important determinant of whether XET strengthens or loosens the cell wall.
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| 8. |
- Oliveira de Sousa, Dannylo, et al.
(författare)
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Effects of grass species on cell wall components and milk production of dairy cows
- 2023
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Ingår i: Report from the Department of Crop Production Ecology (VPE). - 1653-5375. ; , s. 105-108
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- There is a balance between dry matter (DM) yield and feed value when choosing types of grasses on a farm depending on the acreages of farmland and types of ruminants to be fed. Tall fescue (TF, Festuca arundinacea Schreb.) appears to be an alternative to traditional grasses, such as timothy (TI, Phleum pretense L.) and meadow fescue (MF, Festuca pratensis L.), in northern Europe. This is especially important in a changing climate due to its high DM yield, drought tolerance and adaptability to a wide range of climates (Richard et al., 2020). Our previous study (Sousa et al., 2021) compared the effects of TI and TF on cell wall composition and their relationship to dairy cow performance. Besides the greater concentration of fiber components observed in TI, concentrations of most of the hydroxylcinnamic acids (p-coumaric acid, ferulic acid, and its respective cis isomerisation) in the cell walls were greater in TF silages than in TI silages. Consequently, cows receiving TI silages showed greater energy corrected milk (ECM) yield than cows receiving TF silages, suggesting that the concentration of hydroxycinnamic acids was the main factor regulating milk production. To our knowledge, there is no study comparing TF and TI with MF on performance of dairy cows. Therefore, the objective of this study was to evaluate the effect of these grass species on cell wall components and milk production of dairy cows.
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| 9. |
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| 10. |
- Takahashi Schmidt, Junko, et al.
(författare)
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ELIMAKI Locus Is Required for Vertical Proprioceptive Response in Birch Trees
- 2020
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Ingår i: Current Biology. - : Elsevier BV. - 0960-9822 .- 1879-0445. ; 30, s. 589-599
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Tidskriftsartikel (refereegranskat)abstract
- Tree architecture has evolved to support a top-heavy above-ground biomass, but this integral feature poses a weight-induced challenge to trunk stability. Maintaining an upright stem is expected to require vertical proprioception through feedback between sensing stem weight and responding with radial growth. Despite its apparent importance, the principle by which plant stems respond to vertical loading forces remains largely unknown. Here, by manipulating the stem weight of downy birch (Betula pubescens) trees, we show that cambial development is modulated systemically along the stem. We carried out a genetic study on the underlying regulation by combining an accelerated birch flowering program with a recessive mutation at the ELIMAKI locus (EKI), which causes a mechanically defective response to weight stimulus resulting in stem collapse after just 3 months. We observed delayed wood morphogenesis in eki compared with WT, along with a more mechanically elastic cambial zone and radial compression of xylem cell size, indicating that rapid tissue differentiation is critical for cambial growth under mechanical stress. Furthermore, the touch-induced mechanosensory pathway was transcriptionally misregulated in eki, indicating that the ELIMAKI locus is required to integrate the weight-growth feedback regulation. By studying this birch mutant, we were able to dissect vertical proprioception from the gravitropic response associated with reaction wood formation. Our study provides evidence for both local and systemic responses to mechanical stimuli during secondary plant development.
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