Arabidopsis thaliana β-glucosidase BGLU15 attacks flavonol 3-O-β-glucoside-7-O-α-rhamnosides

Roepke, Jonathon; Bozzo, Gale G.

doi:10.1016/j.phytochem.2014.10.028

Cited by 44 publications

(36 citation statements)

References 66 publications

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“…S4). The pH optimum is characteristic of other plant ␤-glucosidases (22). The effect of various additives, which included several metal ions and chaotropic agents on in vitro enzyme activity of recombinant CsBGlu12, was also evaluated.…”

Section: Resultsmentioning

confidence: 99%

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Functional Characterization of CsBGlu12, a β-Glucosidase from Crocus sativus, Provides Insights into Its Role in Abiotic Stress through Accumulation of Antioxidant Flavonols

Baba

Vishwakarma

Ashraf

2017

Journal of Biological Chemistry

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Edited by Joseph JezGlycosylation and deglycosylation are impressive mechanisms that allow plants to regulate the biological activity of an array of secondary metabolites. Although glycosylation improves solubility and renders the metabolites suitable for transport and sequestration, deglycosylation activates them to carry out biological functions. Herein, we report the functional characterization of CsBGlu12, a ␤-glucosidase from Crocus sativus. CsBGlu12 has a characteristic glucoside hydrolase 1 family (␣/␤) 8 triose-phosphate isomerase (TIM) barrel structure with a highly conserved active site. In vitro enzyme activity revealed that CsBGlu12 catalyzes the hydrolysis of flavonol ␤-glucosides and cello-oligosaccharides. Site-directed mutagenesis of any of the two conserved catalytic glutamic acid residues (Glu 200 and Glu 414 ) of the active site completely abolishes the ␤-glucosidase activity. Transcript analysis revealed that Csbglu12 is highly induced in response to UV-B, dehydration, NaCl, methyl jasmonate, and abscisic acid treatments indicating its possible role in plant stress response. Transient overexpression of CsBGlu12 leads to the accumulation of antioxidant flavonols in Nicotiana benthamiana and confers tolerance to abiotic stresses. Antioxidant assays indicated that accumulation of flavonols alleviated the accretion of reactive oxygen species during abiotic stress conditions. ␤-Glucosidases are known to play a role in abiotic stresses, particularly dehydration through abscisic acid; however, their role through accumulation of reactive oxygen species (ROS) scavenging flavonols has not been established. Furthermore, only one ␤-glucosidase 12 homolog has been characterized so far. Therefore, this work presents an important report on characterization of CsBGlu12 and its role in abiotic stress through ROS scavenging.Plants are sessile organisms that cannot escape their predators and the harsh environmental conditions. However, during the course of evolution, plants have learned to defend themselves and adapt to different types of biotic and abiotic stresses by synthesizing a diverse assortment of secondary metabolites. Many of these secondary metabolites are stored in inactive glycosylated forms. The glycosylation chemically stabilizes and enhances the solubility of the metabolites and renders them fit for storage in the vacuole and more so to protect the plant from the noxious effects of its own defense system (1). These glyconjugates are activated by the hydrolysis of the ␤-glucosidic bond by a class of enzymes called ␤-glucosidases. ␤-Glucosidases (EC 3.2.1.2.1) belong to family 1 of glycoside hydrolases and perform diverse and vital functions in plants, which include, but are not limited to, the activation of lignin precursors (2), release of glucose from oligosaccharides (3), release of phytohormones from inactive glycosides (4), and activation of several defense compounds (5-9). This process of glycosylation and deglycosylation offers an efficient mechanism that regulates the homeostasis of secon...

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Section: Resultsmentioning

confidence: 99%

“…analysis was carried as described previously (22,60) with slight modifications using Nexera UHPLC (130 megapascals) equipped with MS-8030 (Shimadzu). Enable RP-C18 column (250 ϫ 4.6 mm, 5 m) was used for the analysis.…”

Section: Lc-esi-ms Analysis-lc-msmentioning

confidence: 99%

Functional Characterization of CsBGlu12, a β-Glucosidase from Crocus sativus, Provides Insights into Its Role in Abiotic Stress through Accumulation of Antioxidant Flavonols

Baba

Vishwakarma

Ashraf

2017

Journal of Biological Chemistry

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“…For example, MtBGLU6 and MtBGLU7, were clustered with AtBGLU45 and AtBGLU46 in cluster IV, which hydrolyzed lignin precursors in Arabidopsis [13], suggesting that they may be involved in lignin metabolism in Medicago. AtBGLU6 and AtBGLU10 in cluster V catalyzed the synthesis of flavonoid glycosides in Arabidopsis as glucosyltransferase [34,35], indicating the six MtBGLUs in the same clusters might have similar transglucosidase function. Notably, cluster I contain nearly half (25/51) of MtBGLU family members, but only six members of Arabidopsis.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive identification and characterization of abiotic stress and hormone responsive glycosyl hydrolase family 1 genes in Medicago truncatula

Yang

Jiang

et al. 2020

Preprint

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Background: β-glucosidases (BGLUs) hydrolyze the β-D-glycosidic bond with retention of anomeric configuration. BGLUs were associated with many aspects of plant physiological processes, in particular biotic and abiotic stresses through the activation of phytohormones and defense compounds. However, no comprehensive and systematic investigation on the stress- or hormone-responsive BGLU proteins had ever been reported in plant.Results: In this study, total 51 BGLU genes of the glycoside hydrolase family 1 were identified in one of the model legume plant Medicago truncatula genome, and they were classified into five distinct clusters. Sequence alignments revealed several conserved and characteristic motifs among these MtBGLU proteins. Analyses of their putative signal peptides and N-glycosylation site suggested that the majority of MtBGLU members have dual targeting to the vacuole/chloroplast. Many regulatory elements possibly related with phytohormones and/or abiotic stresses were identified in MtBGLU genes. Moreover, Microarray and qPCR analyses showed that these MtBGLU genes exhibited distinct expression patterns in various tissues, and in response to different abiotic stress and hormonal treatments. Notably, MtBGLU21, MtBGLU22, MtBGLU28, and MtBGLU30 in cluster I were dramatically activated by NaCl, PEG, IAA, ABA, SA and GA3 treatments.Conclusion: Collectively, our genome-wide characterization, evolutionary analysis, and expression pattern analysis of MtBGLU genes suggested that BGLU proteins play crucial roles in response to various abiotic stresses and hormonal cues in M. truncatula. This systematic analysis provided valuable information for the functional characterization and utilization of these MtBGLU genes in improving stress tolerance in M. truncatula and/or other plant species.

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“…A total of 47 BGLUs were identi ed in Arabidopsis, of which AtBGLU1-6 was used for the accumulation of avonoids in Arabidopsis [1,2], AtBGLU9 and AtBGLU10 catalyzed the nal step of anthocyanin formation in Arabidopsis [3]. AtBGLU15 displayed the highest catalytic e ciency for Q3G7R and K3G7R yielding their respective 7-O-rhamnosides as products [4]. AtBGLU18 and AtBGLU33 regulate ABA responses by increasing ABA levels through the hydrolysis of glucose-conjugated ABA (ABA-GE) [5,6].…”

Section: Introductionmentioning

confidence: 99%

“…PbBGLUs Group II were clustered together with Poplar PtrBGLU9 and PtrBGLU11, however the functions of these two genes are unclear. Arabidopsis AtBGLU12-17 was related to the use of Arabidopsis avonoids, AtBGLU15 can catalyze the hydrolysis of Q3G7R, K3G7R and avonol 3-O-glucosides [4], these genes and PbBGLU19 (group III) were clustered together, and it is speculated that PbBGLU19 may be involved in the hydrolysis of avonoid glycosides in pear. PbBGLUs IV-A was clustered together with AtBGLU (40,41,43,44) and some poplar BGLU genes, but the functions of these genes have not yet been explored.…”

Section: Phylogenetic Conserved Motif and Exon-intron Structures Analmentioning

confidence: 99%

Genome-Wide Comparative Analysis of the β-glucosidase Family in Five Rosaceae Species and their Potential Role on Lignification of Stone Cells in Chinese White Pear

Wang

Yang

Han

et al. 2020

Preprint

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Background: The β-glucosidase BGLU in the glycoside hydrolase family 1 (GH1) is involved in the sugar metabolism of the plant and plays an important role in maintaining the normal physiological function of the plant. Recent studies had shown that β-glucosidase was involved in plant lignification. The lignification in pear fruit is closely related to the formation of pear stone cells, but the BGLU genes family has not been identified in pears.Result: A total of 343 BGLU genes were identified from five species of Rosaceae (Pyrus bretschneideri, Prunus mume, Malus domestica, Prunus avium, Fragaria vesca). According to phylogenetic analysis, 50 PbBGLUs were divided into 8 groups. 298 syntenic pairs were found in intra- and inter-species collinear analysis of five Rosaceae species, found that pears and apples had more syntenic pairs than pear and the other three Rosaceae species. The Ka/Ks analysis of duplication PbBGLU genes in pear indicated that the main mode of expansion of the PbBGLUs was segmental replication and was mainly affected by purification. qRT-PCR showed that the three gene expression patterns of PbBGLU1, PbBGLU15 and PbBGLU16 were basically consistent with the change trend of pear fruit lignin and stone cell content, and may be involved in lignification and stone cell development of pear fruit. Subcellular localization showed that these three candidate genes were all located on the cell wall.Conclusion: In this study, a genome-wide analysis of BGLU genes in five Rosaceae species was carried out, and three candidate genes related to lignification and stone cell development of pear fruits were identified, which laid the foundation for a deeper understanding of the function of BGLU genes in pear fruits and potential in changing pear fruit quality.

show abstract

Arabidopsis thaliana β-glucosidase BGLU15 attacks flavonol 3-O-β-glucoside-7-O-α-rhamnosides

Cited by 44 publications

References 66 publications

Functional Characterization of CsBGlu12, a β-Glucosidase from Crocus sativus, Provides Insights into Its Role in Abiotic Stress through Accumulation of Antioxidant Flavonols

Functional Characterization of CsBGlu12, a β-Glucosidase from Crocus sativus, Provides Insights into Its Role in Abiotic Stress through Accumulation of Antioxidant Flavonols

Comprehensive identification and characterization of abiotic stress and hormone responsive glycosyl hydrolase family 1 genes in Medicago truncatula

Genome-Wide Comparative Analysis of the β-glucosidase Family in Five Rosaceae Species and their Potential Role on Lignification of Stone Cells in Chinese White Pear

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