Enhancement of growth by expression of poplar cellulase in <i>Arabidopsis thaliana</i>

Park, Yong Woo; Tominaga, Rumi; Sugiyama, Junji; Furuta, Yuzo; Tanimoto, Eiichi; Samejima, Masahiro; Sakai, Fukumi; Hayashi, Takahisa

doi:10.1046/j.1365-313x.2003.01696.x

Cited by 95 publications

(60 citation statements)

References 30 publications

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“…Prior heat treatment of the sections prevented liberation of the aglycone due to enzyme denaturation. In the case of GG-b-Res substrate, the accumulation of resorufinyl anion was much more quickly observed, consistent with high expression levels of KORRIGAN1 (Takahashi et al, 2009) and other cellulases from the family GH9 (Park et al, 2003) in the stem tissues, as well as the combined action of endogenous b-glucosidases (Takahashi et al, 2009). The analysis shows that in Arabidopsis stem, the xyloglucanase and cellulase/ b-glucosidase activities are most readily observed in secondary cell walls of xylem cells and interfascicular fibers (Fig.…”

Section: Xeh and Cellulase Activity In Arabidopsis Vegetative Tissuesupporting

confidence: 64%

A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta

Ibatullin

Banasiak²,

Baumann³

et al. 2009

Plant Physiol.

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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 b-glycoside of a xylogluco-oligosaccharide (XXXG-b-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 (« 62,000 M 21 cm 21 ), 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-b-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-b-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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Section: Xeh and Cellulase Activity In Arabidopsis Vegetative Tissuesupporting

confidence: 64%

A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta

Ibatullin

Banasiak²,

Baumann³

et al. 2009

Plant Physiol.

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“…Plant genomes encode glycosyl hydrolases from 34 other enzyme families, two of which (families 9 and 16) may have endo-b-1,4-glucanase activity (Lopez-Casado et al, 2008), but it is unclear whether any of them exert the biomechanical effects of Cel12A. The plant growth effects observed with ectopic expression of plant cellulases may be mediated by a similar mechanism or by some other route (Park et al, 2003;Shani et al, 2004;Hartati et al, 2008). Auxin treatments result in slow increases in wall plastic and elastic compliances (Cleland, 1984), so the cell wall changes caused by Cel12A might be similar to those that occur during long-term auxin-induced growth.…”

Section: Implications For Plant Cell Wall Looseningmentioning

confidence: 99%

A Revised Architecture of Primary Cell Walls Based on Biomechanical Changes Induced by Substrate-Specific Endoglucanases

2012

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Xyloglucan is widely believed to function as a tether between cellulose microfibrils in the primary cell wall, limiting cell enlargement by restricting the ability of microfibrils to separate laterally. To test the biomechanical predictions of this “tethered network” model, we assessed the ability of cucumber (Cucumis sativus) hypocotyl walls to undergo creep (long-term, irreversible extension) in response to three family-12 endo-β-1,4-glucanases that can specifically hydrolyze xyloglucan, cellulose, or both. Xyloglucan-specific endoglucanase (XEG from Aspergillus aculeatus) failed to induce cell wall creep, whereas an endoglucanase that hydrolyzes both xyloglucan and cellulose (Cel12A from Hypocrea jecorina) induced a high creep rate. A cellulose-specific endoglucanase (CEG from Aspergillus niger) did not cause cell wall creep, either by itself or in combination with XEG. Tests with additional enzymes, including a family-5 endoglucanase, confirmed the conclusion that to cause creep, endoglucanases must cut both xyloglucan and cellulose. Similar results were obtained with measurements of elastic and plastic compliance. Both XEG and Cel12A hydrolyzed xyloglucan in intact walls, but Cel12A could hydrolyze a minor xyloglucan compartment recalcitrant to XEG digestion. Xyloglucan involvement in these enzyme responses was confirmed by experiments with Arabidopsis (Arabidopsis thaliana) hypocotyls, where Cel12A induced creep in wild-type but not in xyloglucan-deficient (xxt1/xxt2) walls. Our results are incompatible with the common depiction of xyloglucan as a load-bearing tether spanning the 20- to 40-nm spacing between cellulose microfibrils, but they do implicate a minor xyloglucan component in wall mechanics. The structurally important xyloglucan may be located in limited regions of tight contact between microfibrils.

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“…Other cellulases in GH9, lacking the membrane-spanning domain, had ESTs that were broadly distributed among poplar tissues (Table III). These cellulases may act on amorphous cellulose in the cell wall and, by doing so, increase cell wall plasticity (Ohmiya et al, 2000(Ohmiya et al, , 2003Park et al, 2003).…”

Section: Cellulosementioning

confidence: 99%

Poplar Carbohydrate-Active Enzymes. Gene Identification and Expression Analyses

et al. 2006

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Over 1,600 genes encoding carbohydrate-active enzymes (CAZymes) in the Populus trichocarpa (Torr. & Gray) genome were identified based on sequence homology, annotated, and grouped into families of glycosyltransferases, glycoside hydrolases, carbohydrate esterases, polysaccharide lyases, and expansins. Poplar (Populus spp.) had approximately 1.6 times more CAZyme genes than Arabidopsis (Arabidopsis thaliana). Whereas most families were proportionally increased, xylan and pectin-related families were underrepresented and the GT1 family of secondary metabolite-glycosylating enzymes was overrepresented in poplar. CAZyme gene expression in poplar was analyzed using a collection of 100,000 expressed sequence tags from 17 different tissues and compared to microarray data for poplar and Arabidopsis. Expression of genes involved in pectin and hemicellulose metabolism was detected in all tissues, indicating a constant maintenance of transcripts encoding enzymes remodeling the cell wall matrix. The most abundant transcripts encoded sucrose synthases that were specifically expressed in wood-forming tissues along with cellulose synthase and homologs of KORRIGAN and ELP1. Woody tissues were the richest source of various other CAZyme transcripts, demonstrating the importance of this group of enzymes for xylogenesis. In contrast, there was little expression of genes related to starch metabolism during wood formation, consistent with the preferential flux of carbon to cell wall biosynthesis. Seasonally dormant meristems of poplar showed a high prevalence of transcripts related to starch metabolism and surprisingly retained transcripts of some cell wall synthesis enzymes. The data showed profound changes in CAZyme transcriptomes in different poplar tissues and pointed to some key differences in CAZyme genes and their regulation between herbaceous and woody plants.

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Enhancement of growth by expression of poplar cellulase in Arabidopsis thaliana

Cited by 95 publications

References 30 publications

A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta

A Real-Time Fluorogenic Assay for the Visualization of Glycoside Hydrolase Activity in Planta

A Revised Architecture of Primary Cell Walls Based on Biomechanical Changes Induced by Substrate-Specific Endoglucanases

Poplar Carbohydrate-Active Enzymes. Gene Identification and Expression Analyses

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