scholarly journals Improvement in Saccharification Yield of Mixed Rumen Enzymes by Identification of Recalcitrant Cell Wall Constituents Using Enzyme Fingerprinting

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Ajay Badhan ◽  
Yu-Xi Wang ◽  
Robert Gruninger ◽  
Donald Patton ◽  
Justin Powlowski ◽  
...  
Keyword(s):  
Cell Wall ◽  
Experimental Design ◽  
Plant Cell Wall ◽  
Structural Information ◽  
Barley Straw ◽  
Statistical Experimental Design ◽  
Enzyme Pretreatment ◽  
Barley Silage ◽  
Ruminal Digestion

Identification of recalcitrant factors that limit digestion of forages and the development of enzymatic approaches that improve hydrolysis could play a key role in improving the efficiency of meat and milk production in ruminants. Enzyme fingerprinting of barley silage fed to heifers and total tract indigestible fibre residue (TIFR) collected from feces was used to identify cell wall components resistant to total tract digestion. Enzyme fingerprinting results identified acetyl xylan esterases as key to the enhanced ruminal digestion. FTIR analysis also suggested cross-link cell wall polymers as principal components of indigested fiber residues in feces. Based on structural information from enzymatic fingerprinting and FTIR, enzyme pretreatment to enhance glucose yield from barley straw and alfalfa hay upon exposure to mixed rumen-enzymes was developed. Prehydrolysis effects of recombinant fungal fibrolytic hydrolases were analyzed using microassay in combination with statistical experimental design. Recombinant hemicellulases and auxiliary enzymes initiated degradation of plant structural polysaccharides upon application and improved thein vitrosaccharification of alfalfa and barley straw by mixed rumen enzymes. The validation results showed that microassay in combination with statistical experimental design can be successfully used to predict effective enzyme pretreatments that can enhance plant cell wall digestion by mixed rumen enzymes.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

EFFECT OF STRUCTURAL AND CHEMICAL FACTORS OF FORAGES ON POTENTIALLY DIGESTIBLE FIBER, INTAKE, AND TRUE DIGESTIBILITY BY RUMINANTS

1988 ◽  
Vol 68 (3) ◽  
pp. 787-799 ◽  
Author(s):  
V. GIRARD ◽  
G. DUPUIS
Keyword(s):  
Cell Wall ◽  
Dry Matter ◽  
Plant Cell Wall ◽  
Neutral Detergent Fiber ◽  
Optimum Number ◽  
Apparent Digestibility ◽  
Cool Season ◽  
Dry Matter Digestibility ◽  
True Digestibility

In view of the large variation found in plant cell wall digestibilities with ruminants, an attempt was made to group 124 feeds into different lignification classes (clusters) on the basis of chemical characteristics. Each feed cluster was described using a structural coefficient [Formula: see text] that related the potentially digestible fiber (PDF, %) to the ratio between lignin and cell wall volume. The optimum number of clusters was determined iteratively by performing a regression of the apparent digestibility of dry matter at maintenance level (DDM1, %) against the PDF and cell soluble (SOL, %) contents of feeds. The [Formula: see text] coefficients varied from 0.05 (grains, N = 13) to 1.85 (corn silage, N = 3) and increased with the maturity of the grasses from 0.88 (legumes, vegetative cool season grasses, N = 26) to 1.33 (mature, cool season grasses, N = 19). Predicted PDF were closely correlated (r > 0.9, P < 0.01) to in vitro cell wall disappearances (IVCWD). Apparently digestible cell wall in four grasses and four legumes increased linearly with 96-h IVCWD and standard error (SE) was similar to the SE of predicted apparent digestible SOL from SOL concentrations. Assuming that similarity between SE could be also observed in larger samples, PDF and SOL were used in summative equations to predict apparent dry matter digestibility. DDM1 discounted for intake (DDM1 – 4, %) was regressed against SOL and PDF concentrations of 87 feeds:[Formula: see text]with ds and df, the true digestibilities of SOL and PDF. Estimates of ds and df were 0.98 and 0.95 for a zero-production (maintenance) level of intake, and 0.91 and 0.79 for an intake level four times maintenance. Since the true digestibility of the PDF component was only 4% – 13% lower than that of the cell soluble component, the concentration of PDF in cell wall was the major determinant in the variation in apparent digestibility of forages. Key words: lignin, neutral detergent fiber, true digestibility, cluster analysis, feeds

scholarly journals The Role of Brachypodium distachyon Wall-Associated Kinases (WAKs) in Cell Expansion and Stress Responses

Cells ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 2478
Author(s):  
Xingwen Wu ◽  
Antony Bacic ◽  
Kim L. Johnson ◽  
John Humphries
Keyword(s):  
Cell Wall ◽  
Stress Response ◽  
Plant Cell ◽  
Stress Responses ◽  
Cell Expansion ◽  
Plant Cell Wall ◽  
Brachypodium Distachyon ◽  
Critical Role ◽  
Cell Integrity

The plant cell wall plays a critical role in signaling responses to environmental and developmental cues, acting as both the sensing interface and regulator of plant cell integrity. Wall-associated kinases (WAKs) are plant receptor-like kinases located at the wall—plasma membrane—cytoplasmic interface and implicated in cell wall integrity sensing. WAKs in Arabidopsis thaliana have been shown to bind pectins in different forms under various conditions, such as oligogalacturonides (OG)s in stress response, and native pectin during cell expansion. The mechanism(s) WAKs use for sensing in grasses, which contain relatively low amounts of pectin, remains unclear. WAK genes from the model monocot plant, Brachypodium distachyon were identified. Expression profiling during early seedling development and in response to sodium salicylate and salt treatment was undertaken to identify WAKs involved in cell expansion and response to external stimuli. The BdWAK2 gene displayed increased expression during cell expansion and stress response, in addition to playing a potential role in the hypersensitive response. In vitro binding assays with various forms of commercial polysaccharides (pectins, xylans, and mixed-linkage glucans) and wall-extracted fractions (pectic/hemicellulosic/cellulosic) from both Arabidopsis and Brachypodium leaf tissues provided new insights into the binding properties of BdWAK2 and other candidate BdWAKs in grasses. The BdWAKs displayed a specificity for the acidic pectins with similar binding characteristics to the AtWAKs.

Interactions between Treponema bryantii and cellulolytic bacteria in the in vitro degradation of straw cellulose

1987 ◽  
Vol 33 (3) ◽  
pp. 244-248 ◽  
Author(s):  
H. Kudo ◽  
K.-J. Cheng ◽  
J. W. Costerton
Keyword(s):  
Plant Cell Wall ◽  
Bacterial Species ◽  
Barley Straw ◽  
Cellulolytic Bacteria ◽  
Morphological Examination ◽  
Cellulose Digestion ◽  
Acid Generation ◽  
Pure Cultures ◽  
Treponema Bryantii

To assess the contribution of individual bacterial species to the overall process of cellulose digestion in the rumen, cellulolytic bacteria (Bacteroides succinogenes and Ruminococcus albus) were tested as pure cultures and as cocultures with noncellulolytic Treponema bryantii. In studies of in vitro barley straw digestion, Treponema cocultures surpassed pure cultures of the cellulolytic organisms in dry matter disappearance, volatile fatty acid generation, and in the production of succinic acid, lactic acid, and ethanol. Morphological examination, by electron microscopy, showed that cells of T. bryantii associate with the plant cell wall materials in straw, but that cellulose digestion occurs only when these organisms are present with cellulolytic species such as B. succinogenes. These results show that cellulolytic bacteria interact with noncellulolytic Treponema to promote the digestion of cellulosic materials.

scholarly journals Direct evidence for a new mode of plant defense against insects via a novel polygalacturonase-inhibiting protein expression strategy

2020 ◽  
Vol 295 (33) ◽  
pp. 11833-11844
Author(s):  
Wiebke Haeger ◽  
Jana Henning ◽  
David G. Heckel ◽  
Yannick Pauchet ◽  
Roy Kirsch
Keyword(s):  
Cell Wall ◽  
Plant Defense ◽  
Plant Cell Wall ◽  
Direct Evidence ◽  
Larval Growth ◽  
Gene Families ◽  
Preliminary Evidence ◽  
Cell Wall Polysaccharide ◽  
In Planta

Plant cell wall–associated polygalacturonase-inhibiting proteins (PGIPs) are widely distributed in the plant kingdom. They play a crucial role in plant defense against phytopathogens by inhibiting microbial polygalacturonases (PGs). PGs hydrolyze the cell wall polysaccharide pectin and are among the first enzymes to be secreted during plant infection. Recent studies demonstrated that herbivorous insects express their own PG multi-gene families, raising the question whether PGIPs also inhibit insect PGs and protect plants from herbivores. Preliminary evidence suggested that PGIPs may negatively influence larval growth of the leaf beetle Phaedon cochleariae (Coleoptera: Chrysomelidae) and identified BrPGIP3 from Chinese cabbage (Brassica rapa ssp. pekinensis) as a candidate. PGIPs are predominantly studied in planta because their heterologous expression in microbial systems is problematic and instability and aggregation of recombinant PGIPs has complicated in vitro inhibition assays. To minimize aggregate formation, we heterologously expressed BrPGIP3 fused to a glycosylphosphatidylinositol (GPI) membrane anchor, immobilizing it on the extracellular surface of insect cells. We demonstrated that BrPGIP3_GPI inhibited several P. cochleariae PGs in vitro, providing the first direct evidence of an interaction between a plant PGIP and an animal PG. Thus, plant PGIPs not only confer resistance against phytopathogens, but may also aid in defense against herbivorous beetles.

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 Ruminal and total plant cell-wall digestibility estimated by a combinedin situmethod utilizing mathematical models

1997 ◽  
Vol 78 (4) ◽  
pp. 583-598 ◽  
Author(s):  
Pekka Huhtanen ◽  
Aila Vanhatalo
Keyword(s):  
Cell Wall ◽  
Incubation Period ◽  
Plant Cell Wall ◽  
Intestinal Cell ◽  
Cell Wall Digestibility ◽  
Stages Of Growth ◽  
Cell Wall Digestion ◽  
Intestinal Digestion ◽  
Ruminal Digestion

Three ruminally and duodenally cannulated non-lactating Finnish Ayrshire cows were used to investigate ruminal and intestinal digestion of cell-wall carbohydrates by a combinedin situmethod. Five grasses cut at 10 d intervals were incubated in the rumen for 0, 6, 12, 24, 48, 72 and 96 h, and the undegraded residues were exposed to intestinal digestion. With advancing maturity of grass both the rate and extent of cell-wall digestion decreased. At early stages of growth the decreases were faster for the rate of digestion and at late stages of growth for the extent of digestion. Applying a passage rate of 0.02/h in one compartmental rumen model resulted in digestibility values markedly lower than typically observedin vivo.However, applying a rumen model incorporating a selective retention of particles and time-dependent release of particles from the non-escapable pool resulted in much higher digestibility values. Recovery of lignin after 96 h ruminal incubation with a subsequent mobile-bag incubation was very low (from 244 to 460 mg/g). Intestinal disappearance of neutral-detergent fibre (NDF) and hemicellulose decreased with advancing maturity of grass and with increasing length of preceding ruminal incubation period, i.e. with decreasing potential digestibility of the material. Disappearance of hemicellulose was much greater than that of cellulose for intact grasses but the difference diminished with increasing length of preceding rumen incubation period. On average, 195 mg/g of potentially digestible NDF disappeared from the mobile bags in the intestines. The post-ruminal digestion as a proportion of the total NDF digestibility varied between 0.034 and 0.058. Despite methodological problems both in ruminalin situand intestinal mobile bag techniques, these methods can be used to investigate ruminal and intestinal cell-wall digestion and to partition cell-wall digestibility between ruminal and post-ruminal digestion providing that appropriate rumen models are used.

scholarly journals Immunocytochemical Localization of HrpA and HrpZ Supports a Role for the Hrp Pilus in the Transfer of Effector Proteins from Pseudomonas syringae pv. tomato Across the Host Plant Cell Wall

2001 ◽  
Vol 14 (3) ◽  
pp. 394-404 ◽  
Author(s):  
Ian R. Brown ◽  
John W. Mansfield ◽  
Suvi Taira ◽  
Elina Roine ◽  
Martin Romantschuk
Keyword(s):  
Cell Wall ◽  
Pseudomonas Syringae ◽  
Plant Cell ◽  
Type Iii Secretion ◽  
Plant Cell Wall ◽  
Immunogold Labeling ◽  
Effector Proteins ◽  
Basal Bodies ◽  
Type Iii

The Hrp pilus, composed of HrpA subunits, is an essential component of the type III secretion system in Pseudomonas syringae. We used electron microscopy (EM) and immunocytochemistry to examine production of the pilus in vitro from P. syringae pv. tomato strain DC3000 grown under hrp-inducing conditions on EM grids. Pili, when labeled with antibodies to HrpA, developed rapidly in a nonpolar manner shortly after the detection of the hrpA transcript and extended up to 5 μm into surrounding media. Structures at the base of the pilus were clearly differentiated from the basal bodies of flagella. The HrpZ protein, also secreted via the type III system, was found by immunogold labeling to be associated with the pilus in vitro. Accumulation and secretion of HrpA and HrpZ were also examined quantitatively after the inoculation of wild-type DC3000 and hrpA and hrpZ mutants into leaves of Arabidopsis thaliana. The functional pilus crossed the plant cell wall to generate tracks of immunogold labeling for HrpA and HrpZ. Mutants that produced HrpA but did not assemble pili were nonpathogenic, did not secrete HrpA protein, and were compromised for the accumulation of HrpZ. A model is proposed in which the rapidly elongating Hrp pilus acts as a moving conveyor, facilitating transfer of effector proteins from bacteria to the plant cytoplasm across the formidable barrier of the plant cell wall.

scholarly journals The infection cushion: a fungal “weapon” of plant-biomass destruction

2020 ◽  
Author(s):  
Mathias Choquer ◽  
Christine Rascle ◽  
Isabelle R Gonçalves ◽  
Amélie de Vallée ◽  
Cécile Ribot ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell Wall ◽  
Plant Biomass ◽  
Ornamental Plants ◽  
Sugar Transport ◽  
Differential Staining ◽  
List Type ◽  
In Planta ◽  
Cell Wall Degrading Enzymes

SummaryGrey mold disease affects fruits, vegetables and ornamental plants around the world, causing considerable losses every year. Its causing agent, the necrotrophic fungus Botrytis cinerea, produces infection cushions (IC) that are compound appressorial structures dedicated to the penetration of the plant tissues.A microarray analysis was performed to identify genes up-regulated in mature IC. The expression data were supported by RT-qPCR analysis performed in vitro and in planta, proteomic analysis of the IC secretome and mutagenesis of two candidate genes.1,231 up-regulated genes and 79 up-accumulated proteins were identified. They highlight a secretion of ROS, secondary metabolites including phytotoxins, and proteins involved in virulence: proteases, plant cell wall degrading enzymes and necrosis inducers. The role in pathogenesis was confirmed for two up-regulated fasciclin genes. DHN-melanin pathway and chitin deacetylases genes are up-regulated and the conversion of chitin into chitosan was confirmed by differential staining of the IC cell wall. In addition, up-regulation of sugar transport and sugar catabolism encoding genes was found.These results support a role for the B. cinerea IC in plant penetration and suggest other unexpected roles for this fungal organ, in camouflage, necrotrophy or nutrition of the pathogen.

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