scholarly journals Targeted Cell Wall Degradation at the Penetration Site of Cowpea Rust Basidiosporelings

1997 ◽  
Vol 10 (1) ◽  
pp. 87-94 ◽  
Author(s):  
Haixin Xu ◽  
Kurt Mendgen
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Rust Fungus ◽  
Wall Material ◽  
Wall Structure ◽  
Polygalacturonic Acid ◽  
Similar Reduction ◽  
Nonhost Plant ◽  
Wall Apposition

Basidiospore germlings of the cowpea rust fungus (Uromyces vignae) penetrate the epidermal cell wall of the nonhost plant Vicia faba. In order to characterize the wall structure of the penetration site, leaves were high pressure frozen, freeze substituted, and embedded in appropriate resins. With antibodies against epitopes present in pectin, polygalacturonic acid, xyloglucan, and callose, we studied the modification of these wall components during infection. The density of epitopes was determined at the penetration site and compared with noninfected areas of the epidermal wall. Along the fungal penetration hypha, a zone of the plant wall, 0.2 μm wide, exhibited a reduced density of pectin and xyloglucan epitopes. A similar reduction of epitope density was also found for xyloglucan after treatment of sections from noninoculated plants with cellulase and xylanase and for pectin after treatment with pectinase. The density of polygalacturonic acid epitopes remained unchanged in the outer layer of the epidermal wall, but increased over the inner layer. A high density of polygalacturonic acid epitopes was found over a collarlike wall apposition produced by the plant cell along the penetration hypha. These results indicate that the fungus degrades the plant cell wall at the penetration site and that the plant cell secretes new wall material into this area to form the wall apposition.

scholarly journals Insights into plant cell wall structure, architecture, and integrity using glycome profiling of native and AFEXTM-pre-treated biomass

2015 ◽  
Vol 66 (14) ◽  
pp. 4279-4294 ◽  
Author(s):  
Sivakumar Pattathil ◽  
Michael G. Hahn ◽  
Bruce E. Dale ◽  
Shishir P. S. Chundawat
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Cell Wall Structure ◽  
Wall Structure ◽  
Glycome Profiling

scholarly journals Fermentation and subsequent disposition of 14C-labelled plant cell wall material in the rat

1993 ◽  
Vol 69 (1) ◽  
pp. 189-197 ◽  
Author(s):  
D. F. Gray ◽  
M. A. Eastwood ◽  
W. G. Brydon ◽  
S. C. Fry
Keyword(s):  
Cell Wall ◽  
Gastrointestinal Tract ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Spinacia Oleracea ◽  
Wall Material ◽  
Fermentation Products ◽  
Bacterial Fermentation ◽  
Cell Wall Preparation

A 14C-Iabelled plant cell wall preparation (I4C-PCW) produced from spinach (Spinacia oleracea L.) cell culture exhibits uniform labelling of the major polysaccharide groups (%): pectins 53, hemicellulose 13, cellulose 21, starch 3. This 14C-PCW preparation has been used in rat studies as a marker for plant cell wall metabolism. Metabolism of the 14C-PCW occurred largely over the first 24 h. This was due to fermentation in the caecum. The pectic fraction of the plant cell walls was degraded completely in the rat gastrointestinal tract, but some [14C-]cellulose was still detected after 24 h in the colon. Of the 14C,22% was recovered in the host liver, adipose tissue and skin, 26% excreted as 14CO2 and up to 18%was excreted in the faeces. There was no urinary excretion of 14C. In vitro fermentation using a caecal inocuium showed reduced 14CO2 production, 12% compared with 26% in the intact rat. 14C-PCW is auseful marker to investigate the fate of plant cell wall materials in the gastrointestinal tract. These studies show both bacterial fermentation of the 14C-PCW and host metabolism of the 14C-labelled fermentation products.

scholarly journals Direct Target Network of the Neurospora crassa Plant Cell Wall Deconstruction Regulators CLR-1, CLR-2, and XLR-1

mBio ◽  
2015 ◽  
Vol 6 (5) ◽  
Author(s):  
James P. Craig ◽  
Samuel T. Coradetti ◽  
Trevor L. Starr ◽  
N. Louise Glass
Keyword(s):  
Gene Expression ◽  
Cell Wall ◽  
Transcription Factors ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Nutrient Sensing ◽  
Wall Material ◽  
Content Type ◽  
Genes Encoding

ABSTRACTFungal deconstruction of the plant cell requires a complex orchestration of a wide array of intracellular and extracellular enzymes. InNeurospora crassa, CLR-1, CLR-2, and XLR-1 have been identified as key transcription factors regulating plant cell wall degradation in response to soluble sugars. The XLR-1 regulon was defined using a constitutively active mutant allele, resulting in hemicellulase gene expression and secretion under noninducing conditions. To define genes directly regulated by CLR-1, CLR-2, and XLR-1, we performed chromatin immunoprecipitation and next-generation sequencing (ChIPseq) on epitope-tagged constructs of these three transcription factors. WhenN. crassais exposed to plant cell wall material, CLR-1, CLR-2, and XLR-1 individually bind to the promoters of the most strongly induced genes in their respective regulons. These include promoters of genes encoding cellulases for CLR-1 and CLR-2 (CLR-1/CLR-2) and promoters of genes encoding hemicellulases for XLR-1. CLR-1 bound to its regulon under noninducing conditions; however, this binding alone did not translate into gene expression and enzyme secretion. Motif analysis of the bound genes revealed conserved DNA binding motifs, with the CLR-2 motif matching that of its closest paralog inSaccharomyces cerevisiae, Gal4p. Coimmunoprecipitation studies showed that CLR-1 and CLR-2 act in a homocomplex but not as a CLR-1/CLR-2 heterocomplex.IMPORTANCEUnderstanding fungal regulation of complex plant cell wall deconstruction pathways in response to multiple environmental signals via interconnected transcriptional circuits provides insight into fungus/plant interactions and eukaryotic nutrient sensing. Coordinated optimization of these regulatory networks is likely required for optimal microbial enzyme production.

scholarly journals Bundling of cellulose microfibrils in native and polyethylene glycol-containing wood cell walls revealed by small-angle neutron scattering

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Paavo A. Penttilä ◽  
Michael Altgen ◽  
Muhammad Awais ◽  
Monika Österberg ◽  
Lauri Rautkari ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neutron Scattering ◽  
Plant Cell ◽  
Small Angle ◽  
Cell Walls ◽  
Small Angle Neutron Scattering ◽  
Plant Cell Wall ◽  
Raw Materials ◽  
Cellulose Microfibrils ◽  
Wall Structure

AbstractWood and other plant-based resources provide abundant, renewable raw materials for a variety of applications. Nevertheless, their utilization would greatly benefit from more efficient and accurate methods to characterize the detailed nanoscale architecture of plant cell walls. Non-invasive techniques such as neutron and X-ray scattering hold a promise for elucidating the hierarchical cell wall structure and any changes in its morphology, but their use is hindered by challenges in interpreting the experimental data. We used small-angle neutron scattering in combination with contrast variation by poly(ethylene glycol) (PEG) to identify the scattering contribution from cellulose microfibril bundles in native wood cell walls. Using this method, mean diameters for the microfibril bundles from 12 to 19 nm were determined, without the necessity of cutting, drying or freezing the cell wall. The packing distance of the individual microfibrils inside the bundles can be obtained from the same data. This finding opens up possibilities for further utilization of small-angle scattering in characterizing the plant cell wall nanostructure and its response to chemical, physical and biological modifications or even in situ treatments. Moreover, our results give new insights into the interaction between PEG and the wood nanostructure, which may be helpful for preservation of archaeological woods.

Microsurgical isolation of intact plant cell wall appositions for microanalyses

1979 ◽  
Vol 57 (12) ◽  
pp. 1349-1353 ◽  
Author(s):  
Hitoshi Kunoh ◽  
James R. Aist ◽  
Herbert W. Israel
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Analytical Techniques ◽  
Chemical Components ◽  
Specimen Preparation ◽  
Low Frequencies ◽  
Wall Appositions ◽  
Wall Apposition ◽  
Mother Cells

Little is known about the chemical components of the plant cell wall apposition as they relate to its structure and function. The small sizes (5–15 μm diameter) of the appositions, their low frequencies in the cells, and their intimate connections to the cell wall have almost precluded meaningful cytochemical and (or) biochemical analyses. With the development of analytical techniques using the electron microprobe it is now feasible to discover the elemental composition of minute cellular structures, such as the wall apposition, but problems in specimen preparation remain. As a necessary and critical first step in microprobe analysis we have found it best to microsurgically remove fresh wall appositions from their mother cells and then admit them directly into the scanning electron microscope (SEM) after air drying and gold coating. This paper describes the requisite technologies of specimen preparation, microtool fabrication, specimen selection and isolation which are involved. The technique described could find ready application in the microanalyses of many other subcellular structures.

scholarly journals Adaptor Scaffoldins: An Original Strategy for Extended Designer Cellulosomes, Inspired from Nature

mBio ◽  
2016 ◽  
Vol 7 (2) ◽  
Author(s):  
Johanna Stern ◽  
Sarah Moraïs ◽  
Raphael Lamed ◽  
Edward A. Bayer
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Soluble Sugars ◽  
Single Type ◽  
Wall Material ◽  
Type I ◽  
Type Ii ◽  
Recombinant Enzymes ◽  
Designer Cellulosome

ABSTRACTDesigner cellulosomes consist of chimeric cohesin-bearing scaffoldins for the controlled incorporation of recombinant dockerin-containing enzymes. The largest designer cellulosome reported to date is a chimeric scaffoldin that contains 6 cohesins. This scaffoldin represented a technical limit of sorts, since adding another cohesin proved problematic, owing to resultant low expression levels, instability (cleavage) of the scaffoldin polypeptide, and limited numbers of available cohesin-dockerin specificities—the hallmark of designer cellulosomes. Nevertheless, increasing the number of enzymes integrated into designer cellulosomes is critical, in order to further enhance degradation of plant cell wall material. Adaptor scaffoldins comprise an intermediate type of scaffoldin that can both incorporate various enzymes and attach to an additional scaffoldin. Using this strategy, we constructed an efficient form of adaptor scaffoldin that possesses three type I cohesins for enzyme integration, a single type II dockerin for interaction with an additional scaffoldin, and a carbohydrate-binding module for targeting to the cellulosic substrate. In parallel, we designed a hexavalent scaffoldin capable of connecting to the adaptor scaffoldin by the incorporation of an appropriate type II cohesin. The resultant extended designer cellulosome comprised 8 recombinant enzymes—4 xylanases and 4 cellulases—thereby representing a potent enzymatic cocktail for solubilization of natural lignocellulosic substrates. The contribution of the adaptor scaffoldin clearly demonstrated that proximity between the two scaffoldins and their composite set of enzymes is crucial for optimized degradation. After 72 h of incubation, the performance of the extended designer cellulosome was determined to be approximately 70% compared to that of native cellulosomes.IMPORTANCEPlant cell wall residues represent a major source of renewable biomass for the production of biofuels such as ethanol via breakdown to soluble sugars. The natural microbial degradation process, however, is inefficient for achieving cost-effective processes in the conversion of plant-derived biomass to biofuels, either from dedicated crops or human-generated cellulosic wastes. The accumulation of the latter is considered a major environmental pollutant. The development of designer cellulosome nanodevices for enhanced plant cell wall degradation thus has major impacts in the fields of environmental pollution, bioenergy production, and biotechnology in general. The findings reported in this article comprise a true breakthrough in our capacity to produce extended designer cellulosomes via synthetic biology means, thus enabling the assembly of higher-order complexes that can supersede the number of enzymes included in a single multienzyme complex.

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