Identification of four <i>Arabidopsis</i> genes encoding hydroxycinnamate glucosyltransferases

Milkowski, Carsten; Baumert, A.; Strack, Dieter

doi:10.1016/s0014-5793(00)02270-5

Cited by 30 publications

(27 citation statements)

References 10 publications

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“…Minute amounts of p-hydroxyphenylacetaldoxime detoxification products did accumulate and their presence may explain the reduced content of sinapoyl glucose (Table 1). The reduction of sinapoylmalate and sinopoylglucose and the three kaempferol glucosides in the 2x line was not accompanied by a reduction in steady-state transcript levels of UGT84s, UGT73C6, and UGT78D1, the corresponding sinapate and kaempferol glucosyltransferases (32,33).…”

Section: Discussionmentioning

confidence: 80%

Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome

Kristensen

Morant

Olsen

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

200

149

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Focused and nontargeted approaches were used to assess the impact associated with introduction of new high-flux pathways in Arabidopsis thaliana by genetic engineering. Transgenic A. thaliana plants expressing the entire biosynthetic pathway for the tyrosine-derived cyanogenic glucoside dhurrin as accomplished by insertion of CYP79A1, CYP71E1, and UGT85B1 from Sorghum bicolor were shown to accumulate 4% dry-weight dhurrin with marginal inadvertent effects on plant morphology, free amino acid pools, transcriptome, and metabolome. In a similar manner, plants expressing only CYP79A1 accumulated 3% dry weight of the tyrosine-derived glucosinolate, p-hydroxybenzylglucosinolate with no morphological pleitropic effects. In contrast, insertion of CYP79A1 plus CYP71E1 resulted in stunted plants, transcriptome alterations, accumulation of numerous glucosides derived from detoxification of intermediates in the dhurrin pathway, and in loss of the brassicaceae-specific UV protectants sinapoyl glucose and sinapoyl malate and kaempferol glucosides. The accumulation of glucosides in the plants expressing CYP79A1 and CYP71E1 was not accompanied by induction of glycosyltransferases, demonstrating that plants are constantly prepared to detoxify xenobiotics. The pleiotrophic effects observed in plants expressing sorghum CYP79A1 and CYP71E1 were complemented by retransformation with S. bicolor UGT85B. These results demonstrate that insertion of high-flux pathways directing synthesis and intracellular storage of high amounts of a cyanogenic glucoside or a glucosinolate is achievable in transgenic A. thaliana plants with marginal inadvertent effects on the transcriptome and metabolome.DNA microarrays ͉ metabolic engineering ͉ metabolite profiling ͉ channeling ͉ substantial equivalence M etabolic engineering offers the possibility to design plants that produce desired compounds (1-3). Knowledge of the impact of insertion of metabolic pathways on preexisting metabolic pathways is typically scarce, and is assessed by targeted approaches, but not by more global and unbiased approaches. We have previously reported transfer of the entire pathway for synthesis of the tyrosine-derived cyanogenic glucoside dhurrin from Sorghum bicolor to Arabidopsis thaliana by coexpression of three S. bicolor cDNAs: two multifunctional cytochromes P450, CYP79A1 and CYP71E1, and a family 1 glucosyltransferase, UGT85B1 (Fig. 1). The transgenic A. thaliana plants accumulated up 4% of plant dry matter of dhurrin (4). Transgenic A. thaliana plants in which only CYP79A1 was introduced accumulated up to 3% dry matter of the tyrosine derived p-hydroxybenzylglucosinolate, a glucosinolate that does not occur naturally in A. thaliana (5). The accumulation of tyrosine derived p-hydroxybenzylglucosinolate in A. thaliana is the result of metabolic crosstalk, where the p-hydroxyphenylacetaldoxime produced by CYP79A1 is efficiently used by the post-oximemetabolizing enzymes CYP83B1 or CYP83A1 in the endogenous glucosinolate biosynthetic pathway (5, 6) (Fig. 1).In this study,...

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Section: Discussionmentioning

confidence: 80%

Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome

Kristensen

Morant

Olsen

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

200

149

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“…SGT is a member of a family of UDP-glucosyltransferases (UGTs) in Arabidopsis, which contains 107 members divided into 12 major groups on the basis of sequence similarity . Many Arabidopsis UGTs exhibit activity towards secondary metabolites , including the phenylpropanoids (Milkowski et al, 2000;Lim et al, 2001;Lim et al, 2005;Lanot et al, 2006;Sinlapadech et al, 2007;Lanot et al, 2008;Meißner et al, 2008), synthesizing both glucosides and glucose esters. Initially, a single member of this UGT superfamily was identified as the likely Arabidopsis SGT: UGT84A2 (At3g21560; Lim et al, 2001).…”

Section: Sinapate:udp-glucose Glucosyltransferase (Sgt)mentioning

confidence: 99%

“…Kinetic analyses showed that UGT84A2 exhibits a much greater affinity for sinapic acid than any other UGT in Arabidopsis. Nonetheless, UGT84A2 is one of four UGTs (UGT84A1, UG-T84A2, UGT84A3 and UGT84A4) with affinities for the hydroxycinnamic acids (Milkowski et al, 2000;Lim et al, 2001), and there appears to be some functional overlap between the four enzymes (Milkowski et al, 2000;Lim et al, 2001;Sinlapadech et al, 2007;Meißner et al, 2008). The close proximity of of UGT84A1, UG-T84A3, and UGT84A4 in the Arabidopsis genome has prevented the generation of multiple mutants that would help to elucidate the biochemical roles of each individual protein and reveal any potential genetic redundancy (Meißner et al, 2008).…”

Section: Sinapate:udp-glucose Glucosyltransferase (Sgt)mentioning

confidence: 99%

The Phenylpropanoid Pathway in Arabidopsis

Fraser

Chapple

2011

The Arabidopsis Book

624

489

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The phenylpropanoid pathway serves as a rich source of metabolites in plants, being required for the biosynthesis of lignin, and serving as a starting point for the production of many other important compounds, such as the flavonoids, coumarins, and lignans. In spite of the fact that the phenylpropanoids and their derivatives are sometimes classified as secondary metabolites, their relevance to plant survival has been made clear via the study of Arabidopsis and other plant species. As a model system, Arabidopsis has helped to elucidate many details of the phenylpropanoid pathway, its enzymes and intermediates, and the interconnectedness of the pathway with plant metabolism as a whole. These advances in our understanding have been made possible in large part by the relative ease with which mutations can be generated, identified, and studied in Arabidopsis. Herein, we provide an overview of the research progress that has been made in recent years, emphasizing both the genes (and gene families) associated with the phenylpropanoid pathway in Arabidopsis, and the end products that have contributed to the identification of many mutants deficient in the phenylpropanoid metabolism: the sinapate esters.

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“…AB033758; 68.3% identity; Kita et al, 2000), to several esterforming Arabidopsis (Arabidopsis thaliana) GTs (GenBank accession nos. AB019232, Z97339; 49.6%-55.0% identity; Milkowski et al, 2000a;Sato et al, 2000), and to a rapeseed (Brassica napus) ester-forming GT (GenBank accession no. AF287143; 50.3% identity; Milkowski et al, 2000b;Figs.…”

Section: Identification Of Putative Glycosyltransferases From Strawbementioning

confidence: 99%

Cinnamate Metabolism in Ripening Fruit. Characterization of a UDP-Glucose:Cinnamate Glucosyltransferase from Strawberry

et al. 2006

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Strawberry (Fragaria 3 ananassa) fruit accumulate (hydroxy)cinnamoyl glucose (Glc) esters, which may serve as the biogenetic precursors of diverse secondary metabolites, such as the flavor constituents methyl cinnamate and ethyl cinnamate. Here, we report on the isolation of a cDNA encoding a UDP-Glc:cinnamate glucosyltransferase (Fragaria 3 ananassa glucosyltransferase 2 [FaGT2]) from ripe strawberry cv Elsanta that catalyzes the formation of 1-O-acyl-Glc esters of cinnamic acid, benzoic acid, and their derivatives in vitro. Quantitative real-time PCR analysis indicated that FaGT2 transcripts accumulate to high levels during strawberry fruit ripening and to lower levels in flowers. The levels in fruits positively correlated with the in planta concentration of cinnamoyl, p-coumaroyl, and caffeoyl Glc. In the leaf, high amounts of Glc esters were detected, but FaGT2 mRNA was not observed. The expression of FaGT2 is negatively regulated by auxin, induced by oxidative stress, and by hydroxycinnamic acids. Although FaGT2 glucosylates a number of aromatic acids in vitro, quantitative analysis in transgenic lines containing an antisense construct of FaGT2 under the control of the constitutive 35S cauliflower mosaic virus promoter demonstrated that the enzyme is only involved in the formation of cinnamoyl Glc and p-coumaroyl Glc during ripening.

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Identification of four Arabidopsis genes encoding hydroxycinnamate glucosyltransferases

Cited by 30 publications

References 10 publications

Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome

Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome

The Phenylpropanoid Pathway in Arabidopsis

Cinnamate Metabolism in Ripening Fruit. Characterization of a UDP-Glucose:Cinnamate Glucosyltransferase from Strawberry

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