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1.
J Genet Genomics ; 51(3): 279-291, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37451336

ABSTRACT

Protein post-translational modifications (PTMs), such as ubiquitination, phosphorylation, and small ubiquitin-like modifier (SUMO)ylation, are crucial for regulating protein stability, activity, subcellular localization, and binding with cofactors. Such modifications remarkably increase the variety and complexity of proteomes, which are essential for regulating numerous cellular and physiological processes. The regulation of auxin signaling is finely tuned in time and space to guide various plant growth and development. Accumulating evidence indicates that PTMs play critical roles in auxin signaling regulations. Thus, a thorough and systematic review of the functions of PTMs in auxin signal transduction will improve our profound comprehension of the regulation mechanism of auxin signaling and auxin-mediated various processes. This review discusses the progress of protein ubiquitination, phosphorylation, histone acetylation and methylation, SUMOylation, and S-nitrosylation in the regulation of auxin signaling.


Subject(s)
Protein Processing, Post-Translational , Sumoylation , Indoleacetic Acids/metabolism , Signal Transduction , Ubiquitination
2.
Plant Cell Rep ; 42(1): 17-28, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36224499

ABSTRACT

KEY MESSAGE: Rice glycosyltransferase gene UGT2 was identified to play a crucial role in salt tolerance. The transcription factor OsbZIP23 was demonstrated to regulate the UGT2 expression under stress conditions. UDP-glycosyltransferases (UGTs) play key roles in modulating plant responses to environmental challenges. In this study, we characterized a novel glycosyltransferase, UGT2, which plays an important role in salt stress responses in rice (Oryza sativa L). We found that seedlings overexpressing UGT2 exhibited better growth than wild type in shoot and root under hydroponic culture with salt stress treatments, while ugt2ko mutant lines suffered much more growth inhibition. When the soil-grown UGT2 transgenic plants were subjected to salt stress, we also found that ugt2ko mutant lines were severely withered and most of them died, while the overexpression lines grew well and had higher survival rate. Compared with wild-type plants, UGT2 overexpression greatly increased the expression levels of the reactive oxygen species scavenging genes and stress-responsive genes. Furthermore, the upstream regulatory mechanism of the UGT2 gene was identified and we found that a bZIP transcription factor, OsbZIP23, can bind to the UGT2 promoter and enhance the UGT2 transcription levels. This work reveals that OsbZIP23-UGT2 module may play a major role in regulating the salt stress tolerance in rice.


Subject(s)
Oryza , Transcription Factors , Transcription Factors/genetics , Transcription Factors/metabolism , Salt Tolerance/genetics , Oryza/metabolism , Stress, Physiological/genetics , Salt Stress/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Gene Expression Regulation, Plant/genetics , Plant Proteins/genetics , Plant Proteins/metabolism
3.
Front Plant Sci ; 12: 738261, 2021.
Article in English | MEDLINE | ID: mdl-34630489

ABSTRACT

Maize is one of the major crops in the world; however, diseases caused by various pathogens seriously affect its yield and quality. The maize Rp1-D21 mutant (mt) caused by the intragenic recombination between two nucleotide-binding, leucine-rich repeat (NLR) proteins, exhibits autoactive hypersensitive response (HR). In this study, we integrated transcriptomic and metabolomic analyses to identify differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) in Rp1-D21 mt compared to the wild type (WT). Genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) were enriched among the DEGs. The salicylic acid (SA) pathway and the phenylpropanoid biosynthesis pathway were induced at both the transcriptional and metabolic levels. The DAMs identified included lipids, flavones, and phenolic acids, including 2,5-DHBA O-hexoside, the production of which is catalyzed by uridinediphosphate (UDP)-dependent glycosyltransferase (UGT). Four maize UGTs (ZmUGTs) homologous genes were among the DEGs. Functional analysis by transient co-expression in Nicotiana benthamiana showed that ZmUGT9250 and ZmUGT5174, but not ZmUGT9256 and ZmUGT8707, partially suppressed the HR triggered by Rp1-D21 or its N-terminal coiled-coil signaling domain (CCD21). None of the four ZmUGTs interacted physically with CCD21 in yeast two-hybrid or co-immunoprecipitation assays. We discuss the possibility that ZmUGTs might be involved in defense response by regulating SA homeostasis.

5.
Plant J ; 107(1): 149-165, 2021 07.
Article in English | MEDLINE | ID: mdl-33866633

ABSTRACT

Recent studies have shown that global metabolic reprogramming is a common event in plant innate immunity; however, the relevant molecular mechanisms remain largely unknown. Here, we identified a pathogen-induced glycosyltransferase, UGT73C7, that plays a critical role in Arabidopsis disease resistance through mediating redirection of the phenylpropanoid pathway. Loss of UGT73C7 function resulted in significantly decreased resistance to Pseudomonas syringae pv. tomato DC3000, whereas constitutive overexpression of UGT73C7 led to an enhanced defense response. UGT73C7-activated immunity was demonstrated to be dependent on the upregulated expression of SNC1, a Toll/interleukin 1 receptor-type NLR gene. Furthermore, in vitro and in vivo assays indicated that UGT73C7 could glycosylate p-coumaric acid and ferulic acid, the upstream metabolites in the phenylpropanoid pathway. Mutations that lead to the loss of UGT73C7 enzyme activities resulted in the failure to induce SNC1 expression. Moreover, glycosylation activity of UGT73C7 resulted in the redirection of phenylpropanoid metabolic flux to biosynthesis of hydroxycinnamic acids and coumarins. The disruption of the phenylpropanoid pathway suppressed UGT73C7-promoted SNC1 expression and the immune response. This study not only identified UGT73C7 as an important regulator that adjusts phenylpropanoid metabolism upon pathogen challenge, but also provided a link between phenylpropanoid metabolism and an NLR gene.


Subject(s)
Arabidopsis Proteins/immunology , Arabidopsis/physiology , Glycosyltransferases/metabolism , Plant Immunity/physiology , Arabidopsis/drug effects , Arabidopsis Proteins/genetics , Coumaric Acids/metabolism , Disease Resistance/immunology , Gene Expression Regulation, Plant , Glycosylation , Glycosyltransferases/genetics , Glycosyltransferases/immunology , Host-Pathogen Interactions/physiology , Isonicotinic Acids/pharmacology , Plant Diseases/immunology , Plants, Genetically Modified , Pseudomonas syringae/pathogenicity
7.
Sci Rep ; 11(1): 1154, 2021 01 13.
Article in English | MEDLINE | ID: mdl-33441983

ABSTRACT

Leaf angle is an important agronomic trait affecting photosynthesis efficiency and crop yield. Although the mechanisms involved in the leaf angle control are intensively studied in monocots, factors contribute to the leaf angle in dicots are largely unknown. In this article, we explored the physiological roles of an Arabidopsis glucosyltransferase, UGT74D1, which have been proved to be indole-3-acetic acid (IAA) glucosyltransferase in vitro. We found that UGT74D1 possessed the enzymatic activity toward IAA glucosylation in vivo and its expression was induced by auxins. The ectopically expressed UGT74D1 obviously reduced the leaf angle with an altered IAA level, auxin distribution and cell size in leaf tissues. The expression of several key genes involved in the leaf shaping and leaf positioning, including PHYTOCHROME KINASE SUBSTRATE (PKS) genes and TEOSINTE BRANCHED1, CYCLOIDEA, and PCF (TCP) genes, were dramatically changed by ectopic expression of UGT74D1. In addition, clear transcription changes of YUCCA genes and other auxin related genes can be observed in overexpression lines. Taken together, our data indicate that glucosyltransferase UGT74D1 could affect leaf positioning through modulating auxin homeostasis and regulating transcription of PKS and TCP genes, suggesting a potential new role of UGT74D1 in regulation of leaf angle in dicot Arabidopsis.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Ectopic Gene Expression , Gene Expression Regulation, Plant , Glucosyltransferases/genetics , Indoleacetic Acids/metabolism , Arabidopsis/physiology , Arabidopsis Proteins/metabolism , Glucosyltransferases/metabolism , Homeostasis , Plant Leaves/genetics , Plant Leaves/physiology
8.
Plant Physiol Biochem ; 159: 226-233, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33387851

ABSTRACT

Plants can reduce or eliminate the damage caused by herbicides and gain herbicide resistance, which is an important theoretical basis for the development of herbicide-resistant crops at this stage. Thus, discovering novel herbicide-resistant genes to produce diverse herbicide-resistant crop species is of great value. The glycosyltransferases that commonly exist in plant kingdom modify the receptor molecules to change their physical characteristics and biological activities, and thus possess an important potential to be used in the herbicide-resistance breeding. Here, we identified a novel herbicide-induced UDP-glycosyltransferase 91C1 (UGT91C1) from Arabidopsis thaliana and demonstrated its glucosylating activity toward sulcotrione, a kind of triketone herbicides widely used in the world. Overexpression of UGT91C1 gene enhanced the Arabidopsis tolerance to sulcotrione. While, ugt91c1 mutant displayed serious damage and reduced chlorophyll contents in the presence of sulcotrione, suggesting an important role of UGT91C1 in herbicide detoxification through glycosylation. Moreover, it was also noted that UGT91C1 can affect tyrosine metabolism by reducing the sulcotrione toxicity. Together, our identification of glycosyltransferase UGT91C1, as a potential gene conferring herbicide detoxification through glucosylation, may open up a new possibility for herbicide resistant breeding of crop plants and environmental phytoremediation.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis , Glycosyltransferases/metabolism , Herbicide Resistance , Inactivation, Metabolic , Arabidopsis/drug effects , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Glycosyltransferases/genetics , Herbicide Resistance/genetics , Herbicides/metabolism , Herbicides/toxicity , Inactivation, Metabolic/genetics
9.
Front Plant Sci ; 12: 790195, 2021.
Article in English | MEDLINE | ID: mdl-35003178

ABSTRACT

Drought is one of the most important environmental constraints affecting plant growth and development and ultimately leads to yield loss. Uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) are believed to play key roles in coping with environmental stresses. In rice, it is estimated that there are more than 200 UGT genes. However, most of them have not been identified as their physiological significance. In this study, we reported the characterization of a putative glycosyltransferase gene UGT85E1 in rice. UGT85E1 gene is significantly upregulated by drought stress and abscisic acid (ABA) treatment. The overexpression of UGT85E1 led to an enhanced tolerance in transgenic rice plants to drought stress, while the ugt85e1 mutants of rice showed a more sensitive phenotype to drought stress. Further studies indicated that UGT85E1 overexpression induced ABA accumulation, stomatal closure, enhanced reactive oxygen species (ROS) scavenging capacity, increased proline and sugar contents, and upregulated expression of stress-related genes under drought stress conditions. Moreover, when UGT85E1 was ectopically overexpressed in Arabidopsis, the transgenic plants showed increased tolerance to drought as well as in rice. Our findings suggest that UGT85E1 plays an important role in mediating plant response to drought and oxidative stresses. This work may provide a promising candidate gene for cultivating drought-tolerant crops both in dicots and monocots.

10.
Front Plant Sci ; 11: 560696, 2020.
Article in English | MEDLINE | ID: mdl-33224159

ABSTRACT

Drought and salt stresses are common environmental threats that negatively affect rice development and yield. Here we report that the overexpression of AtUGT76C2, a cytokinin glycosyltransferase, in rice modulates cytokinin homeostasis and confers the plants an eminent property in drought and salt tolerance. The transgenic plants exhibit sensitivity to salt and drought stress as well as abscisic acid during the germination stage and the postgermination stage while showing enhanced tolerance to drought and salinity at the young seedling stage and the mature stage. The overexpression of UGT76C2 decreases the endogenous cytokinin level and enhances root growth, which greatly contributes to stress adaptation. In addition, the transgenic plants also show enhanced ROS scavenging activity, reduced ion leakage under salt stress, smaller stomatal opening, and more proline and soluble sugar accumulation, which demonstrate that UGT76C2 acts as an important player in abiotic stress response in rice. To explore the molecular mechanism of UGT76C2 in response to stress adaptation, the expressions of eight stress-responsive genes including OsSOS1, OsPIP2.1, OsDREB2A, OsCOIN, OsABF2, OsRAB16, OsP5CR, and OsP5CS1 were detected, which showed notable upregulation in UGT76C2 overexpression plants under salt and drought stresses. Our results reveal that the ectopic expression of AtUGT76C2 confers the transgenic rice many traits in improving drought and salt stress tolerance in both developmental and physiological levels. It is believed that AtUGT76C2 could be a promising candidate gene for cultivating saline- and drought-tolerant rice.

11.
Int J Mol Sci ; 21(19)2020 Sep 30.
Article in English | MEDLINE | ID: mdl-33008047

ABSTRACT

UDP-glycosyltransferases (UGTs) play key roles in modulating plant development and responses to environmental challenges. Previous research reported that the Arabidopsis UDP-glucosyltransferase 74E2 (AtUGT74E2), which transfers glucose to indole-3-butyric acid (IBA), is involved in regulating plant architecture and stress responses. Here, we show novel and distinct roles of UGT74E2 in rice. We found that overexpression of AtUGT74E2 in rice could enhance seed germination. This effect was also observed in the presence of IBA and abscisic acid (ABA), as well as salt and drought stresses. Further investigation indicated that the overexpression lines had lower levels of free IBA and ABA compared to wild-type plants. Auxin signaling pathway gene expression such as for OsARF and OsGH3 genes, as well as ABA signaling pathway genes OsABI3 and OsABI5, was substantially downregulated in germinating seeds of UGT74E2 overexpression lines. Consistently, due to reduced IBA and ABA levels, the established seedlings were less tolerant to drought and salt stresses. The regulation of rice seed germination and stress tolerance could be attributed to IBA and ABA level alterations, as well as modulation of the auxin/ABA signaling pathways by UGT74E2. The distinct roles of UGT74E2 in rice implied that complex and different molecular regulation networks exist between Arabidopsis and rice.


Subject(s)
Abscisic Acid/metabolism , Arabidopsis/genetics , Germination/genetics , Glycosyltransferases/genetics , Oryza/genetics , Arabidopsis/growth & development , Droughts , Gene Expression Regulation, Plant , Indoleacetic Acids/metabolism , Oryza/growth & development , Plant Growth Regulators/genetics , Plant Growth Regulators/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/growth & development , Seedlings/genetics , Seedlings/growth & development , Seeds/genetics , Seeds/growth & development , Stress, Physiological/genetics
12.
Plant Signal Behav ; 15(8): 1777377, 2020 08 02.
Article in English | MEDLINE | ID: mdl-32491966

ABSTRACT

The signaling network formed by external environmental signals and endogenous hormone signals is an important basis for the adaptive growth of plants. We recently identified a UDP-glucosyltransferase gene, UGT76F1, which controls the glucosylation of auxin precursor IPyA and mediates light-temperature signaling to regulate auxin-dependent hypocotyl elongation in Arabidopsis. However, it is unclear whether UGT76F1 is involved in the adaptive growth of other tissues and whether it is related to the signaling of other hormones besides auxin. Here we investigated the petiole elongation of UGT76F1 overexpression lines and knockout mutant lines, and also studied the effects of UGT76F1 on BR signaling. Experimental results indicated that UGT76F1 is involved in the PIF4-mediated petiole growth under high temperature and that UGT76F1 is also related to the BR signaling in controlling hypocotyl growth. These results suggest that UGT76F1 may have a wider significance in the plant adaptations to surrounding environments.


Subject(s)
Arabidopsis/metabolism , Glycosyltransferases/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Glycosyltransferases/genetics , Hypocotyl/genetics , Hypocotyl/metabolism , Signal Transduction/genetics , Signal Transduction/physiology
13.
Proc Natl Acad Sci U S A ; 117(12): 6910-6917, 2020 03 24.
Article in English | MEDLINE | ID: mdl-32152121

ABSTRACT

Auxin is a class of plant hormone that plays a crucial role in the life cycle of plants, particularly in the growth response of plants to ever-changing environments. Since the auxin responses are concentration-dependent and higher auxin concentrations might often be inhibitory, the optimal endogenous auxin level must be closely controlled. However, the underlying mechanism governing auxin homeostasis remains largely unknown. In this study, a UDP-glycosyltransferase (UGT76F1) was identified from Arabidopsis thaliana, which participates in the regulation of auxin homeostasis by glucosylation of indole-3-pyruvic acid (IPyA), a major precursor of the auxin indole-3-acetic acid (IAA) biosynthesis, in the formation of IPyA glucose conjugates (IPyA-Glc). In addition, UGT76F1 was found to mediate hypocotyl growth by modulating active auxin levels in a light- and temperature-dependent manner. Moreover, the transcription of UGT76F1 was demonstrated to be directly and negatively regulated by PIF4, which is a key integrator of both light and temperature signaling pathways. This study sheds a light on the trade-off between IAA biosynthesis and IPyA-Glc formation in controlling auxin levels and reveals a regulatory mechanism for plant growth adaptation to environmental changes through glucosylation of IPyA.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/growth & development , Gene Expression Regulation, Plant , Glucose/metabolism , Hypocotyl/growth & development , Indoleacetic Acids/pharmacology , Indoles/metabolism , Arabidopsis/drug effects , Arabidopsis/metabolism , Arabidopsis/radiation effects , Arabidopsis Proteins/genetics , Free Radical Scavengers/chemistry , Free Radical Scavengers/metabolism , Glucosyltransferases/metabolism , Glycosylation , Hypocotyl/drug effects , Hypocotyl/metabolism , Hypocotyl/radiation effects , Indoles/chemistry , Light , Plant Growth Regulators/pharmacology , Seedlings , Temperature
14.
Plant Mol Biol ; 102(4-5): 389-401, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31894456

ABSTRACT

KEY MESSAGE: This study revealed that the Arabidopsis UGT75B1 plays an important role in modulating ABA activity by glycosylation when confronting stress environments. The cellular ABA content and activity can be tightly controlled in several ways, one of which is glycosylation by family 1 UDP-glycosyltransferases (UGTs). Previous analysis has shown UGT75B1 activity towards ABA in vitro. However, the biological role of UGT75B1 remains to be elucidated. Here, we characterized the function of UGT75B1 in abiotic stress responses via ABA glycosylation. GUS assay and qRT-PCR indicated that UGT75B1 is significantly upregulated by adverse conditions, such as osmotic stress, salinity and ABA. Overexpression of UGT75B1 in Arabidopsis leads to higher seed germination rates and seedling greening rates upon exposure to salt and osmotic stresses. In contrast, the big UGT75B1 overexpression plants are more sensitive under salt and osmotic stresses. Additionally, the UGT75B1 overexpression plants showed larger stomatal aperture and more water loss under drought condition, which can be explained by lower ABA levels examined in UGT75B1 OE plants in response to water deficit conditions. Consistently, UGT75B1 ectopic expression leads to downregulation of many ABA-responsive genes under stress conditions, including ABI3, ABI5 newly germinated seedlings and RD29A, KIN1, AIL1 in big plants. In summary, our results revealed that the Arabidopsis UGT75B1 plays an important role in coping with abiotic stresses via glycosylation of ABA.


Subject(s)
Abscisic Acid/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Gene Expression Regulation, Plant , Glucosyltransferases/physiology , Glycosyltransferases/metabolism , Stress, Physiological , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Catalysis , Droughts , Genes, Plant , Germination , Glucosyltransferases/genetics , Glycosylation , Glycosyltransferases/genetics , Osmotic Pressure , Plants, Genetically Modified/genetics , Salinity , Seedlings/genetics , Seedlings/physiology , Sodium Chloride , Transcription Factors/genetics , Transcription Factors/physiology
15.
Rice (N Y) ; 12(1): 92, 2019 Dec 18.
Article in English | MEDLINE | ID: mdl-31853664

ABSTRACT

BACKGROUND: In cereal crop rice, auxin is known as an important class of plant hormone that regulates a plethora of plant growth and development. Glycosylation of auxin is known to be one of the important mechanisms mediating auxin homeostasis. However, the relevant auxin glucosyltransferase (GT) in rice still remains largely unknown. RESULTS: In this study, using known auxin glucosyltransferases from other species as queries, twelve putative auxin UDP-glycosyltransferase (UGT) genes were cloned from rice and the one showing highest sequence similarity, named as OsIAGT1, was expressed as recombinant protein. In vitro enzymatic analysis showed that recombinant OsIAGT1 was capable of catalyzing glucosylation of IAA, IBA and other auxin analogs, and that OsIAGT1 is quite tolerant to a broad range of reaction conditions with peak activity at 30 °Ð¡ and pH 8.0. OsIAGT1 showed favorite activity towards native auxins over artificially synthesized ones. Further study indicated that expression of OsIAGT1 can be upregulated by auxin in rice, and with OsIAGT1 overexpressing lines we confirmed that OsIAGT1 is indeed able to glucosylate IAA in vivo. Consistently, ectopic expression of OsIAGT1 leads to declined endogenous IAA content, as well as upregulated auxin synthesis genes and reduced expression of auxin-responsive genes, which likely leads to the reduced plant stature and root length in OsIAGT1 overexpression lines. CONCLUSION: Our result indicated that OsIAGT1 plays an important role in mediating auxin homeostasis by catalyzing auxin glucosylation, and by which OsIAGT1 regulates growth and development in rice.

16.
Plant Physiol ; 180(4): 2167-2181, 2019 08.
Article in English | MEDLINE | ID: mdl-30962291

ABSTRACT

Plant systemic acquired resistance (SAR) provides an efficient broad-spectrum immune response to pathogens. SAR involves mobile signal molecules that are generated by infected tissues and transported to systemic tissues. Methyl salicylate (MeSA), a molecule that can be converted to salicylic acid (SA), is an essential signal for establishing SAR, particularly under a short period of exposure to light after pathogen infection. Thus, the control of MeSA homeostasis is important for an optimal SAR response. Here, we characterized a uridine diphosphate-glycosyltransferase, UGT71C3, in Arabidopsis (Arabidopsis thaliana), which was induced mainly in leaf tissue by pathogens including Pst DC3000/avrRpt2 (Pseudomonas syringae pv tomato strain DC3000 expressing avrRpt2). Biochemical analysis indicated that UGT71C3 exhibited strong enzymatic activity toward MeSA to form MeSA glucosides in vitro and in vivo. After primary pathogen infection by Pst DC3000/avrRpt2, ugt71c3 knockout mutants exhibited more powerful systemic resistance to secondary pathogen infection than that of wild-type plants, whereas systemic resistance in UGT71C3 overexpression lines was compromised. In agreement, after primary infection of local leaves, ugt71c3 knockout mutants accumulated significantly more systemic MeSA and SA than that in wild-type plants. whereas UGT71C3 overexpression lines accumulated less. Our results suggest that MeSA glucosylation by UGT71C3 facilitates negative regulation of the SAR response by modulating homeostasis of MeSA and SA. This study unveils further SAR regulation mechanisms and highlights the role of glucosylation of MeSA and potentially other systemic signals in negatively modulating plant systemic defense.


Subject(s)
Arabidopsis/metabolism , Salicylates/metabolism , Salicylic Acid/isolation & purification , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant , Plant Diseases/microbiology , Plant Leaves/metabolism , Plant Leaves/microbiology , Pseudomonas syringae/pathogenicity , Signal Transduction
17.
Rice (N Y) ; 12(1): 19, 2019 Mar 29.
Article in English | MEDLINE | ID: mdl-30923923

ABSTRACT

BACKGROUND: Cytokinins are one of the five major hormones families in plants and are important for their normal growth and environmental adaptability. In plants, cytokinins are mostly present as glycosides in plants, and their glycosylation modifications are catalyzed by family 1 glycosyltransferases. Current research on cytokinin glycosylation has focused on the biochemical identification of enzymes and the analysis of metabolites in Arabidopsis. There are few studies that examine how cytokinin glycosylation affects its synthesis and accumulation in plants. It is particularly important to understand these processes in food crops such as rice (Oryza sativa); however, to date, cytokinin glycosyltransferase genes in rice have not been reported. RESULTS: In this study, we identified eight rice genes that were functionally homologous to an Arabidopsis cytokinin glycosyltransferase gene. These genes were cloned and expressed in a prokaryotic system to obtain their purified proteins. Through enzymatic analysis and liquid chromatography-mass spectrometry, a single rice glycosyltransferase, Os6, was identified that glycosylated cytokinin in vitro. Os6 was overexpressed in Arabidopsis, and the extraction of cytokinin glycosides showed that Os6 is functionally active in planta. CONCLUSIONS: The identification and characterization of the first cytokinin glycosyltransferase from rice is important for future studies on the cytokinin metabolic pathway in rice. An improved understanding of rice cytokinin glycosyltransferases may facilitate genetic improvements in rice quality.

18.
Ann Bot ; 122(7): 1203-1217, 2018 12 31.
Article in English | MEDLINE | ID: mdl-29982479

ABSTRACT

Background and Aims: Nowadays, the plant family 1 glycosyltransferases (UGTs) are attracting more and more attention since members of this family can improve the properties of secondary metabolites and have significantly enriched the chemical species in plants. Over the past decade, most studies on UGTs have been conducted in Arabidopsis thaliana and they were proved to play diverse roles during the plant life cycle. The Zea mays (maize) GT1 family comprises a large number of UDP-glycosyltransferase (UGT) members. However, their enzyme activities and the biological functions are rarely revealed. In this study, a maize flavonol glycosyltransferase, UFGT2, is identified and its biological role is characterized in detail. Methods: The UFGT2 enzyme activity, the flavonol and glycoside levels in planta were examined by high- performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). The functions of UFGT2 in modifying flavonols, mediating flavonol accumulation and improving stress tolerance were analysed using two ufgt2 mutants and transgenic arabidopsis plants. Key Results: By in vitro enzyme assay, the maize UFGT2 was found to show strong activity towards two flavonols: kaemferol and quercetin. Two ufgt2 knockout mutants, Mu689 and Mu943, exhibited obvious sensitivity to salt and drought stresses. The endogenous quercetin and kaempferol glycosides, as well as the total flavonol levels were found to be substantially decreased in the two ufgt2 mutants, with declined H2O2-scavenging capacity. In contrast, ectopic expression of UFGT2 in arabidopsis led to increased flavonol contents and enhanced oxidative tolerance. Moreover, expression of typical stress-related genes in arabidopsis and maize were affected in UFGT2 overexpression plants or knockout mutants in response to abiotic stresses. UFGT2 was also transferred into the arabidopsis ugt78d2 mutant and it was found to recover the deficient flavonol glycoside pattern in the ugt78d2 mutant, which confirmed its catalysing activity in planta. Conclusion: It is demonstrated in our study that a maize glycosyltransferase, UFGT2, involved in modifying flavonols, contributes to improving plant tolerance to abiotic stresses.


Subject(s)
Acclimatization/genetics , Droughts , Flavonols/metabolism , Glycosyltransferases/genetics , Plant Proteins/genetics , Salt Stress/physiology , Zea mays/physiology , Glycosyltransferases/metabolism , Plant Proteins/metabolism , Secondary Metabolism , Zea mays/genetics
19.
Plant Physiol ; 176(4): 3103-3119, 2018 04.
Article in English | MEDLINE | ID: mdl-29483147

ABSTRACT

Salicylic acid (SA) plays a crucial role in plant innate immunity. The deployment of SA-associated immune responses is primarily affected by SA concentration, which is determined by a balance between SA biosynthesis and catabolism. However, the mechanisms regulating SA homeostasis are poorly understood. In this study, we characterized a unique UDP-glycosyltransferase, UGT76D1, which plays an important role in SA homeostasis and associated immune responses in Arabidopsis (Arabidopsis thaliana). Expression of UGT76D1 was induced by treatment with both the pathogen Pseudomonas syringae pv. tomato (Pst) DC3000 and SA. Overexpression of UGT76D1 resulted in high SA accumulation, significant up-regulation of pathogen-related genes, and a hypersensitive response (HR)-like lesion mimic phenotype. This HR-like phenotype was not observed following UGT76D1 overexpression in SA-deficient NahG transgenic or sid2 plants, suggesting that the phenotype is SA dependent. Biochemical assays showed that UGT76D1 glycosylated 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA), the major catabolic forms of SA, to their Glc and Xyl conjugates in vitro and in vivo. Moreover, in a mutant background blocked in the formation of 2,3-DHBA and 2,5-DHBA, UGT76D1 overexpression did not cause a HR-like lesion mimic phenotype. Following infection with Pst DC3000, UGT76D1 knockout mutants displayed a delayed immune response, with reduced levels of DHBA glycosides and SA, and down-regulated SA synthase expression. By contrast, UGT76D1 overexpression lines showed an enhanced immune response and increased SA biosynthesis before and after pathogen infection. Thus, we propose that UGT76D1 plays an important role in SA homeostasis and plant immune responses by facilitating glycosylation of dihydroxybenzoic acids.


Subject(s)
Arabidopsis/metabolism , Gentisates/metabolism , Hydroxybenzoates/metabolism , Salicylic Acid/metabolism , Arabidopsis/genetics , Arabidopsis/microbiology , Disease Resistance/genetics , Gene Expression Regulation, Plant/drug effects , Glycosylation , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Homeostasis , Plant Diseases/genetics , Plant Diseases/microbiology , Plants, Genetically Modified , Pseudomonas syringae/physiology , Salicylic Acid/pharmacology
20.
Plant Biol (Stuttg) ; 20(1): 10-19, 2018 Jan.
Article in English | MEDLINE | ID: mdl-28902451

ABSTRACT

Although plant glycosyltransferases are thought to play important roles in growth and interaction with the environment, little is known about their physiological roles for most members of the plant glycosyltransferase family. We cloned and characterised an Arabidopsis glycosyltransferase gene, UGT76E11. Its in vivo physiological effects on flavonoid accumulation and plant tolerance to abiotic stresses were investigated. The UGT76E11 gene was up-regulated in transcription expression under stress conditions of salinity, drought and H2 O2 treatment. Transgenic plants ectopically overexpressing UGT76E11 showed substantially enhanced tolerance to salinity and drought at germination and during post-germination growth. Enzyme activity of UGT76E11 to glucosylate quercetin and other flavonoids was confirmed. Ectopic expression of UGT76E11 resulted in significantly increased flavonoid content in transgenic plants compared to wild type, suggesting a contribution of UGT76E11 to modulation of flavonoid metabolism. Consistent with this result, several biosynthesis genes in the flavonoid pathway were clearly up-regulated in transgenic plants. Furthermore, overexpression of UGT76E11 also enhanced the scavenging capacity for ROS and increased expression levels of a number of stress-related genes. Based on these results, we suggest that the glycosyltransferase UGT76E11 plays an important role in modulating flavonoid metabolism and enhancing plant adaptation to environmental stresses. Our findings might allow use of glycosyltransferase UGT76E11 in crop improvement, towards both enhanced stress tolerance and increased flavonoid accumulation.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Flavonoids/metabolism , Stress, Physiological , Adaptation, Physiological , Arabidopsis Proteins/physiology , Dehydration , Ectopic Gene Expression , Gene Expression Regulation, Plant , Plants, Genetically Modified/metabolism , Salt Tolerance/physiology
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