生长素及苯丙烷代谢相关糖基转移酶基因克隆与酶活性鉴定
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摘要
糖基化是一种普遍的生理现象,是植物细胞维持代谢平衡的主要机制之一糖基转移酶是专门负责糖基化的酶,它将活性糖从核苷糖,通常是从尿嘧啶核苷二磷酸-葡萄糖,转移到植物小分子化合物上,从而改变植物小分子的生物活性、水溶性、稳定性、亚细胞定位以及与受体的结合特性。
1.水稻生长素糖基转移酶基因克隆及鉴定
生长素是植物五大类激素之一,控制着植物根的生长发育、顶端优势、向重性、向光性、叶形态建成等过程,在植物体内发挥重要作用。而植物体内大部分生长素是以结合态形式存在的,生长素的糖基化是生长素结合态的重要形式。以往的研究发现过量表达生长素糖基转移酶基因后表现出向地性减弱,影响植株分枝,叶的形态发育。虽然在模式植物拟南芥中有了一定研究,但在水稻这一重要粮食作物中一直未见报道,研究水稻中生长素糖基转移酶在生长素修饰和代谢的中的关系,具有很重要的理论和现实意义。本论文首次从水稻中鉴定出了可以糖基化生长素的糖基转移酶基因,为进一步研究水稻生长素糖基化修饰奠定了良好的基础。论文取得了以下方面的进展：
(1)预测、克隆并原核系统表达了11个生长素糖基转移酶候选基因并得到纯化蛋白。据拟南芥和玉米中生长素糖基转移酶基因,预测和选定水稻中生长素糖基转移酶的候选基因11个,发现其中一个基因与玉米中糖基转移酶基因iaglu相似度最大。将这11个候选基因连接到pGEX系统的原核表达载体上,在大肠杆菌XL1-Blue中表达并得到了纯化的酶蛋白。
(2)发现糖基转移酶OsIAGTl对多个生长素及其前体有催化活性。用得到的11个候选基因的融合蛋白分别与生长素及其前体物质反应,反应产物经过HPLC鉴定,发现OsIAGTl对IAA,IBA,IPA有较强的催化活性,分别催化这些生长素形成相应的糖酯类物质。
(3)研究了糖基转移酶基因OsIAGTl所编码蛋白的酶学性质。在相同条件下,分析了6种底物的相对转化率大小,发现OsIAGTl对IAA、IBA、IPA的催化活性要高于其他ICA、2,4-D、NAA。以底物IAA为例,研究了温度对酶活性的影响,发现30℃时活性最高；研究了pH对酶活性的影响,发现pH=8.0时活性最高；最后对内源生理活性生长素IAA、IBA进行了酶学动力学分析,结果显示它们的Km值差异比较小,推测它们在植物体内会竞争性的结合糖基转移酶OsIAGT1。
本论文主要的研究成果是从水稻中首次鉴定出了生长素的糖基转移酶基因OsIAGT1。该发现为进一步研究OsIAGT1基因在水稻体内的功能以及对生长素的调控作用奠定了基础。
2.水稻和杨树中苯丙烷代谢相关糖基转移酶基因克隆鉴定
植物体中,芥子酸的糖基化形式芥子酰葡萄糖酯是芥子酸转变成其他芥子酰酯类的中间物质。通过芥子酰苹果酸转移酶将芥子酰葡萄糖酯转化成芥子酰苹果酸,通过芥子酰胆碱转移酶将芥子酰葡萄糖酯转变成芥子酰胆碱。其中芥子酰葡萄糖酯和芥子酰苹果酸具有抗紫外辐射的作用。拟南芥中芥子酰糖基转移酶基因突变后表现出对大豆锈病的抗性。同时芥子酸与木质素合成前体松柏醇,芥子醇具有相同的前期合成途径,芥子酸的修饰有可能影响木质素的合成。
基于此种考虑,本实验在水稻和杨树中克隆芥子酸糖基转移酶基因,为研究芥子酸等苯丙烷化合物的糖基化修饰在水稻和杨树中的作用奠定基础。论文取得了以下方面的进展：
(1)水稻苯丙烷代谢相关糖基转移酶基因克隆及鉴定。用进化树分析水稻候选基因与拟南芥中苯丙烷代谢相关糖基转移酶基因的关系,预测、克隆了多个水稻的候选基因,并在原核系统中表达和纯化了它们的蛋白。
(2)发现糖基转移酶OsSGT1对多个苯丙烷代谢途径物质有催化活性。用得到的3个候选基因的融合蛋白分别与苯丙烷代谢相关物质反应,反应产物经过HPLC鉴定,发现对芥子酸、阿魏酸以及其他苯丙烷类物质都有较强的催化活性,分别催化形成相应的糖酯类物质。
(3)同时对杨树中苯丙烷代谢相关糖基转移酶基因进行预测和克隆,经过原核表达获得融合蛋白。发现杨树糖基转移酶PtSGT1,PtSGT3对多个苯丙烷代谢途径物质有糖基化催化活性。
3.另外还发现拟南芥糖基转移酶UGT76C2酶活性与突变碱基的关系,对突变基因进行了初步的生化与生理学分析。
Glycosylation is a wide-spread physiological phenomenon, and is thought to be one of the most important mechanisms in maintaining plant cell homeostasis. Glycosyltransferases (GTs) are the enzymes responsible for glycosylation. They can typically transfer single or multiple activated sugars from nucleotide sugar donors, especially uridine5'-diphosopho glucose (UDP-glucose), to a wide range of small molecular acceptors, thus change their bioactivity, solubility, stability, subcellular localization and binding property to the acceptors. This study is focused on the gene cloning, prokaryotic expression and enzyme activity identification of glucosyltransferase genes of auxins and phenylpropanoids from rice and poplar. The main contents and results of this study are listed as follows.
1. The cloning and identification of auxin glucosyltransferase from rice
Auxin is one of the most important phytohormones in plant, which controls the development of roots, apical sominance, gravitroism, phototropism, leaf morphology. Auxin combines with other small molecules, such as glucose and methyl. The glycosylation of auxin is an mportant form of bound auxin. Previous sdudies have revealed that the overexpression of auxin glycosyltransferase genes in plants exhibits phenomenon of weak gravitropism, influencing plant branches,leaves morphogenesis. The auxin glucosytransferase has been sdudied in the model plant Arabidopsis, but the gene of auxin glucosyltransferase in Oryza sativa has not yet been isolated now. In this study, an auxin glycosyltransferase of rice was cloned and characterized for the first time.
(1) Eleven candidate genes of auxin glycosyltransferases were predicted, cloned, and expressed using prokaryotic system and their recombinant proteins were purified. A phylogenetic tree was constructed using eleven putative auxin glycosyltransferases of rice and all published Arabidopsis and Zea may family1GTs. It was found that rice llgenes were located on a unique branch with Arabidopsis and zea may genes which glycosylate auxins to their glucosides in Arabidopsis and Zea may, suggesting that the rice11genes might be involved in the glycosylation modification of auxin. These eleven genes were constructed to pGEX prokaryotic expression vectors, expressed in E.coli XL1-Blue, and then recombinant proteins were purified.
(2) Glycosyltransferase OsIAGT1was found to have high enzyme activity responsible for the glycosylation of auxins. The enzyme activities of eleven recombinant proteins were analyzed by incubating them separately with six auxin. Following HPLC and LC-MS analyses for the reaction products, it was found that OsIAGT1glycosylated6sbustance to produce their corresponding glucose ester. These6substrates include IAA, IBA, ICA, IP A, NAA,2,4-D.
(3) The enzymatic properties of glycosyltranferase OsIAGT1were studied. Relative conversion rates of these6substrates were calculated under the same condition, and it was found that OsIAGTl had a higher enzymatic activity toward IAA, IBA and IP A than toward ICA, NAA, and2,4-D. Taking IAA as an example, the influence factors of enzymatic activity including temperature, pH were tested. The results showed that30degrees centigrade was the best temperature for enzyme activity. The pH optimum was8.0. Finally, the kinetic parameters of this enzyme catalyzing IAA, IBA were also calculated. Results showed that the Km values of these two auxins were almost identical, indicating that they may competitively bind glycosyltransferase OsIAGTl in planta.
In summary, this study identified, for the first time, a rice glycosyltransferase OsIAGT1responsible for the glycosylation of auxins. This discovery lays a foundation for the further study on the function of OsIAGTl in vivo and its role in regulating auxin metabolism.
2. The cloning and identification of glucosyltransferase genes of phenylpropanoids in Oryza sativa and Poplus tometentosa.
Phenylpropanoids can be converted into a broad spectrum of O-ester conjugates. The abundant sinapate esters in Brassica napus and Arabidopsis thaliana reflect a well-known metabolic network, including UDPglucose:sinapate glucosyltransferase (SGT), sinapoylglucose:choline sinapoyltransferase (SCT), sinapoylglucose:L-malate sinapoyltransferase (SMT) and sinapoylcholine (sinapine) esterase (SCE).1-O-Sinapoylglucose, produced by SGT during seed development, is converted to sinapine by SCT and hydrolyzed by SCE in germinating seeds. The released sinapate feeds via sinapoylglucose into the biosynthesis of sinapoylmalate in the seedlings catalyzed by SMT. Sinapoylmalate is involved in protecting the leaves against the deleterious enflects of UV-B radiation. Sinapine might function as storage vehicle for ready supply of choline for phosphatidylcholine biosynthesis in young seedlings. And the sinapate has same pathway with the synthsis of lignins.The A. thaliana brtl (bright trichome1) mutant impaired in UGT84A2enhances resistance of Soybean Rust. In this study, the glucosyltransferase genes of phenylpropanoids in Oryza sativa and Poplus tometentosa were cloned and identified for the first time.
(1) Three rice candidate genes of phenylpropanoid glycosyltransferases were cloned and expressed using prokaryotic system and recombinant proteins were purified. A phylogenetic tree was constructed using three putative rice glycosyltransferases of phenylpropanoids, and all published Arabidopsis family1GTs about phenylpropanoids.
It was found that rice three rice genes were located on a unique branch with Arabidopsis genes which glycosylate phenylpropanoids to their glucose ester suggesting that the three genes might be involved in the glycosylation modification of phenylpropanoids. These three genes were constructed to pGEX prokaryotic expression vectors, expressed in E.coli XL1-Blue, and then recombinant proteins were purified.
(2) Glycosyltransferase OsSGT was found to have high enzyme activity responsible for the glycosylation of phenylpropanoids. The enzyme activities of three recombinant proteins were analyzed by incubating them separately with sinapate. Following HPLC and LC-MS analyses for the reaction products, it was found that OsSGT glycosylated sinapate acid to produce its corresponding glucose ester.
(3) The cloning and identification glucosyltransferase genes of phenylpropanoids in Populus tometentosa. The candidate genes of phenylpropanoid glycosyltransferases were cloned from poplar and expressed using prokaryotic system and recombinant proteins were purified. Glycosyltransferase PtSGT1and PtSGT3were found to have high enzyme activity responsible for the glycosylation of phenylpropanoids.
3. In addition, the relationship of Arabidopsis UGT76C2glycosyltransferase activity and the single nucleotide substitution was studied in this thesis. The effects of mutated gene on the enzyme activity and in vivo physiological role were also investigated.
1.水稻生长素糖基转移酶基因克隆及鉴定
生长素是植物五大类激素之一,控制着植物根的生长发育、顶端优势、向重性、向光性、叶形态建成等过程,在植物体内发挥重要作用。而植物体内大部分生长素是以结合态形式存在的,生长素的糖基化是生长素结合态的重要形式。以往的研究发现过量表达生长素糖基转移酶基因后表现出向地性减弱,影响植株分枝,叶的形态发育。虽然在模式植物拟南芥中有了一定研究,但在水稻这一重要粮食作物中一直未见报道,研究水稻中生长素糖基转移酶在生长素修饰和代谢的中的关系,具有很重要的理论和现实意义。本论文首次从水稻中鉴定出了可以糖基化生长素的糖基转移酶基因,为进一步研究水稻生长素糖基化修饰奠定了良好的基础。论文取得了以下方面的进展：
(1)预测、克隆并原核系统表达了11个生长素糖基转移酶候选基因并得到纯化蛋白。据拟南芥和玉米中生长素糖基转移酶基因,预测和选定水稻中生长素糖基转移酶的候选基因11个,发现其中一个基因与玉米中糖基转移酶基因iaglu相似度最大。将这11个候选基因连接到pGEX系统的原核表达载体上,在大肠杆菌XL1-Blue中表达并得到了纯化的酶蛋白。
(2)发现糖基转移酶OsIAGTl对多个生长素及其前体有催化活性。用得到的11个候选基因的融合蛋白分别与生长素及其前体物质反应,反应产物经过HPLC鉴定,发现OsIAGTl对IAA,IBA,IPA有较强的催化活性,分别催化这些生长素形成相应的糖酯类物质。
(3)研究了糖基转移酶基因OsIAGTl所编码蛋白的酶学性质。在相同条件下,分析了6种底物的相对转化率大小,发现OsIAGTl对IAA、IBA、IPA的催化活性要高于其他ICA、2,4-D、NAA。以底物IAA为例,研究了温度对酶活性的影响,发现30℃时活性最高；研究了pH对酶活性的影响,发现pH=8.0时活性最高；最后对内源生理活性生长素IAA、IBA进行了酶学动力学分析,结果显示它们的Km值差异比较小,推测它们在植物体内会竞争性的结合糖基转移酶OsIAGT1。
本论文主要的研究成果是从水稻中首次鉴定出了生长素的糖基转移酶基因OsIAGT1。该发现为进一步研究OsIAGT1基因在水稻体内的功能以及对生长素的调控作用奠定了基础。
2.水稻和杨树中苯丙烷代谢相关糖基转移酶基因克隆鉴定
植物体中,芥子酸的糖基化形式芥子酰葡萄糖酯是芥子酸转变成其他芥子酰酯类的中间物质。通过芥子酰苹果酸转移酶将芥子酰葡萄糖酯转化成芥子酰苹果酸,通过芥子酰胆碱转移酶将芥子酰葡萄糖酯转变成芥子酰胆碱。其中芥子酰葡萄糖酯和芥子酰苹果酸具有抗紫外辐射的作用。拟南芥中芥子酰糖基转移酶基因突变后表现出对大豆锈病的抗性。同时芥子酸与木质素合成前体松柏醇,芥子醇具有相同的前期合成途径,芥子酸的修饰有可能影响木质素的合成。
基于此种考虑,本实验在水稻和杨树中克隆芥子酸糖基转移酶基因,为研究芥子酸等苯丙烷化合物的糖基化修饰在水稻和杨树中的作用奠定基础。论文取得了以下方面的进展：
(1)水稻苯丙烷代谢相关糖基转移酶基因克隆及鉴定。用进化树分析水稻候选基因与拟南芥中苯丙烷代谢相关糖基转移酶基因的关系,预测、克隆了多个水稻的候选基因,并在原核系统中表达和纯化了它们的蛋白。
(2)发现糖基转移酶OsSGT1对多个苯丙烷代谢途径物质有催化活性。用得到的3个候选基因的融合蛋白分别与苯丙烷代谢相关物质反应,反应产物经过HPLC鉴定,发现对芥子酸、阿魏酸以及其他苯丙烷类物质都有较强的催化活性,分别催化形成相应的糖酯类物质。
(3)同时对杨树中苯丙烷代谢相关糖基转移酶基因进行预测和克隆,经过原核表达获得融合蛋白。发现杨树糖基转移酶PtSGT1,PtSGT3对多个苯丙烷代谢途径物质有糖基化催化活性。
3.另外还发现拟南芥糖基转移酶UGT76C2酶活性与突变碱基的关系,对突变基因进行了初步的生化与生理学分析。
Glycosylation is a wide-spread physiological phenomenon, and is thought to be one of the most important mechanisms in maintaining plant cell homeostasis. Glycosyltransferases (GTs) are the enzymes responsible for glycosylation. They can typically transfer single or multiple activated sugars from nucleotide sugar donors, especially uridine5'-diphosopho glucose (UDP-glucose), to a wide range of small molecular acceptors, thus change their bioactivity, solubility, stability, subcellular localization and binding property to the acceptors. This study is focused on the gene cloning, prokaryotic expression and enzyme activity identification of glucosyltransferase genes of auxins and phenylpropanoids from rice and poplar. The main contents and results of this study are listed as follows.
1. The cloning and identification of auxin glucosyltransferase from rice
Auxin is one of the most important phytohormones in plant, which controls the development of roots, apical sominance, gravitroism, phototropism, leaf morphology. Auxin combines with other small molecules, such as glucose and methyl. The glycosylation of auxin is an mportant form of bound auxin. Previous sdudies have revealed that the overexpression of auxin glycosyltransferase genes in plants exhibits phenomenon of weak gravitropism, influencing plant branches,leaves morphogenesis. The auxin glucosytransferase has been sdudied in the model plant Arabidopsis, but the gene of auxin glucosyltransferase in Oryza sativa has not yet been isolated now. In this study, an auxin glycosyltransferase of rice was cloned and characterized for the first time.
(1) Eleven candidate genes of auxin glycosyltransferases were predicted, cloned, and expressed using prokaryotic system and their recombinant proteins were purified. A phylogenetic tree was constructed using eleven putative auxin glycosyltransferases of rice and all published Arabidopsis and Zea may family1GTs. It was found that rice llgenes were located on a unique branch with Arabidopsis and zea may genes which glycosylate auxins to their glucosides in Arabidopsis and Zea may, suggesting that the rice11genes might be involved in the glycosylation modification of auxin. These eleven genes were constructed to pGEX prokaryotic expression vectors, expressed in E.coli XL1-Blue, and then recombinant proteins were purified.
(2) Glycosyltransferase OsIAGT1was found to have high enzyme activity responsible for the glycosylation of auxins. The enzyme activities of eleven recombinant proteins were analyzed by incubating them separately with six auxin. Following HPLC and LC-MS analyses for the reaction products, it was found that OsIAGT1glycosylated6sbustance to produce their corresponding glucose ester. These6substrates include IAA, IBA, ICA, IP A, NAA,2,4-D.
(3) The enzymatic properties of glycosyltranferase OsIAGT1were studied. Relative conversion rates of these6substrates were calculated under the same condition, and it was found that OsIAGTl had a higher enzymatic activity toward IAA, IBA and IP A than toward ICA, NAA, and2,4-D. Taking IAA as an example, the influence factors of enzymatic activity including temperature, pH were tested. The results showed that30degrees centigrade was the best temperature for enzyme activity. The pH optimum was8.0. Finally, the kinetic parameters of this enzyme catalyzing IAA, IBA were also calculated. Results showed that the Km values of these two auxins were almost identical, indicating that they may competitively bind glycosyltransferase OsIAGTl in planta.
In summary, this study identified, for the first time, a rice glycosyltransferase OsIAGT1responsible for the glycosylation of auxins. This discovery lays a foundation for the further study on the function of OsIAGTl in vivo and its role in regulating auxin metabolism.
2. The cloning and identification of glucosyltransferase genes of phenylpropanoids in Oryza sativa and Poplus tometentosa.
Phenylpropanoids can be converted into a broad spectrum of O-ester conjugates. The abundant sinapate esters in Brassica napus and Arabidopsis thaliana reflect a well-known metabolic network, including UDPglucose:sinapate glucosyltransferase (SGT), sinapoylglucose:choline sinapoyltransferase (SCT), sinapoylglucose:L-malate sinapoyltransferase (SMT) and sinapoylcholine (sinapine) esterase (SCE).1-O-Sinapoylglucose, produced by SGT during seed development, is converted to sinapine by SCT and hydrolyzed by SCE in germinating seeds. The released sinapate feeds via sinapoylglucose into the biosynthesis of sinapoylmalate in the seedlings catalyzed by SMT. Sinapoylmalate is involved in protecting the leaves against the deleterious enflects of UV-B radiation. Sinapine might function as storage vehicle for ready supply of choline for phosphatidylcholine biosynthesis in young seedlings. And the sinapate has same pathway with the synthsis of lignins.The A. thaliana brtl (bright trichome1) mutant impaired in UGT84A2enhances resistance of Soybean Rust. In this study, the glucosyltransferase genes of phenylpropanoids in Oryza sativa and Poplus tometentosa were cloned and identified for the first time.
(1) Three rice candidate genes of phenylpropanoid glycosyltransferases were cloned and expressed using prokaryotic system and recombinant proteins were purified. A phylogenetic tree was constructed using three putative rice glycosyltransferases of phenylpropanoids, and all published Arabidopsis family1GTs about phenylpropanoids.
It was found that rice three rice genes were located on a unique branch with Arabidopsis genes which glycosylate phenylpropanoids to their glucose ester suggesting that the three genes might be involved in the glycosylation modification of phenylpropanoids. These three genes were constructed to pGEX prokaryotic expression vectors, expressed in E.coli XL1-Blue, and then recombinant proteins were purified.
(2) Glycosyltransferase OsSGT was found to have high enzyme activity responsible for the glycosylation of phenylpropanoids. The enzyme activities of three recombinant proteins were analyzed by incubating them separately with sinapate. Following HPLC and LC-MS analyses for the reaction products, it was found that OsSGT glycosylated sinapate acid to produce its corresponding glucose ester.
(3) The cloning and identification glucosyltransferase genes of phenylpropanoids in Populus tometentosa. The candidate genes of phenylpropanoid glycosyltransferases were cloned from poplar and expressed using prokaryotic system and recombinant proteins were purified. Glycosyltransferase PtSGT1and PtSGT3were found to have high enzyme activity responsible for the glycosylation of phenylpropanoids.
3. In addition, the relationship of Arabidopsis UGT76C2glycosyltransferase activity and the single nucleotide substitution was studied in this thesis. The effects of mutated gene on the enzyme activity and in vivo physiological role were also investigated.
引文
Biener, J., A. Wittstock, L. A. Zepeda-Ruiz, M. M. Biener, V. Zielasek, D. Kramer, R. N. Viswanath, J. Weissmuller, M. Baumer and A. V. Hamza (2009). "Surface-chemistry-driven actuation in nanoporous gold." Nat Mater 8(1):47-51.
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Creelman, R. A. and J. E. Mullet (1997). "Biosynthesis And Action Of Jasmonates In Plants." Annu Rev Plant Physiol Plant Mol Biol 48:355-381.
Fraissinet-Tachet, L., R. Baltz, J. Chong, S. Kauffmann, B. Fritig and P. Saindrenan (1998). "Two tobacco genes induced by infection, elicitor and salicylic acid encode glucosyltransferases acting on phenylpropanoids and benzoic acid derivatives, including salicylic acid." FEBS Letters 437(3):319-323.
Franke, R., J. M. Humphreys, M. R. Hemm, J. W. Denault, M. O. Ruegger, J. C. Cusumano and C. Chapple (2002). "The ArabidopsisREF8 gene encodes the 3-hydroxylase of phenylpropanoid metabolism." The Plant Journal 30(1):33-45.
Frydman, A., O. Weisshaus, M. Bar-Peled, D. V. Huhman, L. W. Sumner, F. R. Marin, E. Lewinsohn, R. Fluhr, J. Gressel and Y. Eyal (2004). "Citrus fruit bitter flavors:isolation and functional characterization of the gene Cml,2RhaT encoding a 1,2 rhamnosyltransferase, a key enzyme in the biosynthesis of the bitter flavonoids of citrus." Plant J 40(1):88-100.
Fujioka, S. and T. Yokota (2003). "Biosynthesis and metabolism of brassinosteroids." Annu Rev Plant Biol 54:137-164.
Hou, B., E.-K. Lim, G. S. Higgins and D. J. Bowles (2004). "N-glucosylation of cytokinins by glycosyltransferases of Arabidopsis thaliana." Journal of Biological Chemistry 279(46):47822-47832.
Jackson, R. G., M. Kowalczyk, Y. Li, G. Higgins, J. Ross, G. Sandberg and D. J. Bowles (2002). "Over-expression of an Arabidopsis gene encoding a glucosyltransferase of indole-3-acetic acid:phenotypic characterisation of transgenic lines." The Plant Journal 32(4):573-583.
Jin,S.,Ma,X.,Han,P.,Wang,B.,Sun,Y.,Zhang,G.,Li,Y.,Hou,B.(2013)."UGT74D1 Is a Novel Auxin Glucosytransferase frome Arabidopsis thaliana."Plos one8(4):e61705.
Jones, P., B. Messner, J.-I. Nakajima, A. R. Schaffner and K. Saito (2003). "UGT73C6 and UGT78D1, glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana." Journal of Biological Chemistry 278(45): 43910-43918.
Kajita, S., S. Hishiyama, Y. Tomimura, Y. Katayama and S. Omori (1997). "Structural Characterization of Modified Lignin in Transgenic Tobacco Plants in Which the Activity of 4-Coumarate:Coenzyme A Ligase Is Depressed." Plant Physiol 114(3):871-879.
Kajita, S., Y. Katayama and S. Omori (1996). "Alterations in the biosynthesis of lignin in transgenic plants with chimeric genes for 4-coumarate:coenzyme A ligase." Plant Cell Physiol 37(7):957-965.
Kudo, T., N. Makita, M. Kojima, H. Tokunaga and H. Sakakibara (2012). "Cytokinin activity of cis-zeatin and phenotypic alterations induced by overexpression of putative cis-Zeatin-O-glucosyltransferase in rice." Plant Physiol 160(1):319-331.
Langenbach, C., R. Campe, U. Schaffrath, K. Goellner and U. Conrath (2013). "UDP-glucosyltransferase UGT84A2/BRT1 is required for Arabidopsis nonhost resistance to the Asian soybean rust pathogen Phakopsora pachyrhizi." New Phytol 198(2):536-545.
Lee, D. and C. J. Douglas (1996). "Two divergent members of a tobacco 4-coumarate:coenzyme A ligase (4CL) gene family. cDNA structure, gene inheritance and expression, and properties of recombinant proteins." Plant Physiol 112(1): 193-205.
Li, A. X. and J. C. Steffens (2000). "An acyltransferase catalyzing the formation of diacylglucose is a serine carboxypeptidase-like protein." Proc Natl Acad Sci U S A 97(12):6902-6907.
Li, X., J. Bergelson and C. Chapple (2010). "The ARABIDOPSIS accession Pna-10 is a naturally occurring sngl deletion mutant." Mol Plant 3(1):91-100.
Liang, M., E. Davis, D. Gardner, X. Cai and Y. Wu (2006). "Involvement of AtLAC15 in lignin synthesis in seeds and in root elongation of Arabidopsis." Planta 224(5):1185-1196.
Lim, E. K. and D. J. Bowles (2004). "A class of plant glycosyltransferases involved in cellular homeostasis." EMBO J 23(15):2915-2922.
Lim, E. K., Y. Li, A. Parr, R. Jackson, D. A. Ashford and D. J. Bowles (2001). "Identification of glucosyltransferase genes involved in sinapate metabolism and lignin synthesis in Arabidopsis." J Biol Chem 276(6):4344-4349.
Mittasch, J., D. Strack and C. Milkowski (2007). "Secondary product glycosyltransferases in seeds of Brassica napus." Planta 225(2):515-522.
Mok, D. W. and M. C. Mok (2001). "Cytokinin Metabolism And Action." Annu Rev Plant Physiol Plant Mol Biol 52:89-118.
Moraga, A. R., P. F. Nohales, J. A. Perez and L. Gomez-Gomez (2004). "Glucosylation of the saffron apocarotenoid crocetin by a glucosyltransferase isolated from Crocus sativus stigmas." Planta 219(6):955-966.
Nair, R. B., Q. Xia, C. J. Kartha, E. Kurylo, R. N. Hirji, R. Datla and G. Selvaraj (2002). "Arabidopsis CYP98A3 mediating aromatic 3-hydroxylation. Developmental regulation of the gene, and expression in yeast." Plant Physiol 130(1):210-220.
Osmani, S. A., E. H. Hansen, C. Malien-Aubert, C. E. Olsen, S. Bak and B. L. Moller (2009). "Effect of glucuronosylation on anthocyanin color stability." J Agric Food Chem 57(8):3149-3155.
Paquette, S., B. L. M(?)ller and S. Bak (2003). "On the origin of family 1 plant glycosyltransferases." Phytochemistry 62(3):399-413.
Peer, W. A., A. Bandyopadhyay, J. J. Blakeslee, S. N. Makam, R. J. Chen, P. H. Masson and A. S. Murphy (2004). "Variation in expression and protein localization of the PIN family of auxin efflux facilitator proteins in flavonoid mutants with altered auxin transport in Arabidopsis thaliana." Plant Cell 16(7):1898-1911.
Poppenberger, B., F. Berthiller, D. Lucyshyn, T. Sieberer, R. Schuhmacher, R. Krska, K. Kuchler, J. Glossl, C. Luschnig and G. Adam (2003). "Detoxification of the Fusarium mycotoxin deoxynivalenol by a UDP-glucosyltransferase from Arabidopsis thaliana." J Biol Chem 278(48):47905-47914.
Quiel, J. A. and J. Bender (2003). "Glucose conjugation of anthranilate by the Arabidopsis UGT74F2 glucosyltransferase is required for tryptophan mutant blue fluorescence." J Biol Chem 278(8):6275-6281.
Sasaki, S., T. Nishida, Y. Tsutsumi and R. Kondo (2004). "Lignin dehydrogenative polymerization mechanism:a poplar cell wall peroxidase directly oxidizes polymer lignin and produces in vitro dehydrogenative polymer rich in beta-O-4 linkage." FEBS Lett 562(1-3):197-201.
Schoch, G., S. Goepfert, M. Morant, A. Hehn, D. Meyer, P. Ullmann and D. Werck-Reichhart (2001). "CYP98A3 from Arabidopsis thaliana is a 3'-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway." J Biol Chem 276(39):36566-36574.
Schuler, M. A. and D. Werck-Reichhart (2003). "Functional genomics of P450s." Annual review of plant biology 54(1):629-667.
Sewalt, V., W. Ni, J. W. Blount, H. G. Jung, S. A. Masoud, P. A. Howles, C. Lamb and R. A. Dixon (1997). "Reduced Lignin Content and Altered Lignin Composition in Transgenic Tobacco Down-Regulated in Expression of L-Phenylalanine Ammonia-Lyase or Cinnamate 4-Hydroxylase." Plant Physiol 115(1):41-50.
Shao, H., X. He, L. Achnine, J. W. Blount, R. A. Dixon and X. Wang (2005). "Crystal structures of a multifunctional triterpene/flavonoid glycosyltransferase from Medicago truncatula." Plant Cell 17(11):3141-3154.
Stepanova, A. N., J. Robertson-Hoyt, J. Yun, L. M. Benavente, D.-Y. Xie, K. Dolezal, A. Schlereth, G. Jurgens and J. M. Alonso (2008). "TAA1-Mediated Auxin Biosynthesis Is Essential for Hormone Crosstalk and Plant Development." Cell 133(1):177-191.
Stepanova, A. N., J. Yun, A. V. Likhacheva and J. M. Alonso (2007). "Multilevel interactions between ethylene and auxin in Arabidopsis roots." Plant Cell 19(7): 2169-2185.
Tao, Y., J. L. Ferrer, K. Ljung, F. Pojer, F. Hong, J. A. Long, L. Li, J. E. Moreno, M. E. Bowman, L. J. Ivans, Y. Cheng, J. Lim, Y. Zhao, C. L. Ballare, G. Sandberg, J. P. Noel and J. Chory (2008). "Rapid synthesis of auxin via a new tryptophan-dependent pathway is required for shade avoidance in plants." Cell 133(1): 164-176.
wang, J., Ma,X.,Kojima,M.,Sakakibara,H.,Hou,B (2011). "N-glucosytransferase UGT76C2 is involved in cytokinin homeostasis and cytokinin response in Arabidopsis thaliana."Plant cell physiol 52(12):2200-2213.
Weis, M., E.-K. Lim, N. C. Bruce and D. J, Bowles (2008). "Engineering and kinetic characterisation of two glucosyltransferases from Arabidopsis thaliana." Biochimie 90(5):830-834.
Woodward, A. W. and B. Bartel (2005). "Auxin:regulation, action, and interaction." Ann Bot 95(5):707-735.
Xu, Z. J., M. Nakajima, Y. Suzuki and I. Yamaguchi (2002). "Cloning and characterization of the abscisic acid-specific glucosyltransferase gene from adzuki bean seedlings." Plant Physiol 129(3):1285-1295.
Yonekura-Sakakibara, K. and K. Hanada (2011). "An evolutionary view of functional diversity in family 1 glycosyltransferases." Plant J 66(1):182-193.
Zhao, Y. (2010). "Auxin biosynthesis and its role in plant development." Annu Rev Plant Biol 61:49-64.
Bowles, D., J. Isayenkova, E.-K. Lim and B. Poppenberger (2005). "Glycosyltransferases:managers of small molecules." Current opinion in plant biology 8(3):254-263.
Buer, C. S. and G. K. Muday (2004). "The transparent testa4 mutation prevents flavonoid synthesis and alters auxin transport and the response of Arabidopsis roots to gravity and light." Plant Cell 16(5):1191-1205.
Chong, J., R. Baltz, C. Schmitt, R. Beffa, B. Fritig and P. Saindrenan (2002). "Downregulation of a pathogen-responsive tobacco UDP-Glc:phenylpropanoid glucosyltransferase reduces scopoletin glucoside accumulation, enhances oxidative stress, and weakens virus resistance." The Plant Cell Online 14(5):1093-1107.
Clauβ, K., A. Baumert, M. Nimtz, C. Milkowski and D. Strack (2008). "Role of a GDSL lipase-like protein as sinapine esterase in Brassicaceae." The Plant Journal 53(5):802-813.
Creelman, R. A. and J. E. Mullet (1997). "Biosynthesis And Action Of Jasmonates In Plants." Annu Rev Plant Physiol Plant Mol Biol 48:355-381.
Fraissinet-Tachet, L., R. Baltz, J. Chong, S. Kauffmann, B. Fritig and P. Saindrenan (1998). "Two tobacco genes induced by infection, elicitor and salicylic acid encode glucosyltransferases acting on phenylpropanoids and benzoic acid derivatives, including salicylic acid." FEBS Letters 437(3):319-323.
Franke, R., J. M. Humphreys, M. R. Hemm, J. W. Denault, M. O. Ruegger, J. C. Cusumano and C. Chapple (2002). "The ArabidopsisREF8 gene encodes the 3-hydroxylase of phenylpropanoid metabolism." The Plant Journal 30(1):33-45.
Frydman, A., O. Weisshaus, M. Bar-Peled, D. V. Huhman, L. W. Sumner, F. R. Marin, E. Lewinsohn, R. Fluhr, J. Gressel and Y. Eyal (2004). "Citrus fruit bitter flavors:isolation and functional characterization of the gene Cml,2RhaT encoding a 1,2 rhamnosyltransferase, a key enzyme in the biosynthesis of the bitter flavonoids of citrus." Plant J 40(1):88-100.
Fujioka, S. and T. Yokota (2003). "Biosynthesis and metabolism of brassinosteroids." Annu Rev Plant Biol 54:137-164.
Hou, B., E.-K. Lim, G. S. Higgins and D. J. Bowles (2004). "N-glucosylation of cytokinins by glycosyltransferases of Arabidopsis thaliana." Journal of Biological Chemistry 279(46):47822-47832.
Jackson, R. G., M. Kowalczyk, Y. Li, G. Higgins, J. Ross, G. Sandberg and D. J. Bowles (2002). "Over-expression of an Arabidopsis gene encoding a glucosyltransferase of indole-3-acetic acid:phenotypic characterisation of transgenic lines." The Plant Journal 32(4):573-583.
Jin,S.,Ma,X.,Han,P.,Wang,B.,Sun,Y.,Zhang,G.,Li,Y.,Hou,B.(2013)."UGT74D1 Is a Novel Auxin Glucosytransferase frome Arabidopsis thaliana."Plos one8(4):e61705.
Jones, P., B. Messner, J.-I. Nakajima, A. R. Schaffner and K. Saito (2003). "UGT73C6 and UGT78D1, glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana." Journal of Biological Chemistry 278(45): 43910-43918.
Kajita, S., S. Hishiyama, Y. Tomimura, Y. Katayama and S. Omori (1997). "Structural Characterization of Modified Lignin in Transgenic Tobacco Plants in Which the Activity of 4-Coumarate:Coenzyme A Ligase Is Depressed." Plant Physiol 114(3):871-879.
Kajita, S., Y. Katayama and S. Omori (1996). "Alterations in the biosynthesis of lignin in transgenic plants with chimeric genes for 4-coumarate:coenzyme A ligase." Plant Cell Physiol 37(7):957-965.
Kudo, T., N. Makita, M. Kojima, H. Tokunaga and H. Sakakibara (2012). "Cytokinin activity of cis-zeatin and phenotypic alterations induced by overexpression of putative cis-Zeatin-O-glucosyltransferase in rice." Plant Physiol 160(1):319-331.
Langenbach, C., R. Campe, U. Schaffrath, K. Goellner and U. Conrath (2013). "UDP-glucosyltransferase UGT84A2/BRT1 is required for Arabidopsis nonhost resistance to the Asian soybean rust pathogen Phakopsora pachyrhizi." New Phytol 198(2):536-545.
Lee, D. and C. J. Douglas (1996). "Two divergent members of a tobacco 4-coumarate:coenzyme A ligase (4CL) gene family. cDNA structure, gene inheritance and expression, and properties of recombinant proteins." Plant Physiol 112(1): 193-205.
Li, A. X. and J. C. Steffens (2000). "An acyltransferase catalyzing the formation of diacylglucose is a serine carboxypeptidase-like protein." Proc Natl Acad Sci U S A 97(12):6902-6907.
Li, X., J. Bergelson and C. Chapple (2010). "The ARABIDOPSIS accession Pna-10 is a naturally occurring sngl deletion mutant." Mol Plant 3(1):91-100.
Liang, M., E. Davis, D. Gardner, X. Cai and Y. Wu (2006). "Involvement of AtLAC15 in lignin synthesis in seeds and in root elongation of Arabidopsis." Planta 224(5):1185-1196.
Lim, E. K. and D. J. Bowles (2004). "A class of plant glycosyltransferases involved in cellular homeostasis." EMBO J 23(15):2915-2922.
Lim, E. K., Y. Li, A. Parr, R. Jackson, D. A. Ashford and D. J. Bowles (2001). "Identification of glucosyltransferase genes involved in sinapate metabolism and lignin synthesis in Arabidopsis." J Biol Chem 276(6):4344-4349.
Mittasch, J., D. Strack and C. Milkowski (2007). "Secondary product glycosyltransferases in seeds of Brassica napus." Planta 225(2):515-522.
Mok, D. W. and M. C. Mok (2001). "Cytokinin Metabolism And Action." Annu Rev Plant Physiol Plant Mol Biol 52:89-118.
Moraga, A. R., P. F. Nohales, J. A. Perez and L. Gomez-Gomez (2004). "Glucosylation of the saffron apocarotenoid crocetin by a glucosyltransferase isolated from Crocus sativus stigmas." Planta 219(6):955-966.
Nair, R. B., Q. Xia, C. J. Kartha, E. Kurylo, R. N. Hirji, R. Datla and G. Selvaraj (2002). "Arabidopsis CYP98A3 mediating aromatic 3-hydroxylation. Developmental regulation of the gene, and expression in yeast." Plant Physiol 130(1):210-220.
Osmani, S. A., E. H. Hansen, C. Malien-Aubert, C. E. Olsen, S. Bak and B. L. Moller (2009). "Effect of glucuronosylation on anthocyanin color stability." J Agric Food Chem 57(8):3149-3155.
Paquette, S., B. L. M(?)ller and S. Bak (2003). "On the origin of family 1 plant glycosyltransferases." Phytochemistry 62(3):399-413.
Peer, W. A., A. Bandyopadhyay, J. J. Blakeslee, S. N. Makam, R. J. Chen, P. H. Masson and A. S. Murphy (2004). "Variation in expression and protein localization of the PIN family of auxin efflux facilitator proteins in flavonoid mutants with altered auxin transport in Arabidopsis thaliana." Plant Cell 16(7):1898-1911.
Poppenberger, B., F. Berthiller, D. Lucyshyn, T. Sieberer, R. Schuhmacher, R. Krska, K. Kuchler, J. Glossl, C. Luschnig and G. Adam (2003). "Detoxification of the Fusarium mycotoxin deoxynivalenol by a UDP-glucosyltransferase from Arabidopsis thaliana." J Biol Chem 278(48):47905-47914.
Quiel, J. A. and J. Bender (2003). "Glucose conjugation of anthranilate by the Arabidopsis UGT74F2 glucosyltransferase is required for tryptophan mutant blue fluorescence." J Biol Chem 278(8):6275-6281.
Sasaki, S., T. Nishida, Y. Tsutsumi and R. Kondo (2004). "Lignin dehydrogenative polymerization mechanism:a poplar cell wall peroxidase directly oxidizes polymer lignin and produces in vitro dehydrogenative polymer rich in beta-O-4 linkage." FEBS Lett 562(1-3):197-201.
Schoch, G., S. Goepfert, M. Morant, A. Hehn, D. Meyer, P. Ullmann and D. Werck-Reichhart (2001). "CYP98A3 from Arabidopsis thaliana is a 3'-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway." J Biol Chem 276(39):36566-36574.
Schuler, M. A. and D. Werck-Reichhart (2003). "Functional genomics of P450s." Annual review of plant biology 54(1):629-667.
Sewalt, V., W. Ni, J. W. Blount, H. G. Jung, S. A. Masoud, P. A. Howles, C. Lamb and R. A. Dixon (1997). "Reduced Lignin Content and Altered Lignin Composition in Transgenic Tobacco Down-Regulated in Expression of L-Phenylalanine Ammonia-Lyase or Cinnamate 4-Hydroxylase." Plant Physiol 115(1):41-50.
Shao, H., X. He, L. Achnine, J. W. Blount, R. A. Dixon and X. Wang (2005). "Crystal structures of a multifunctional triterpene/flavonoid glycosyltransferase from Medicago truncatula." Plant Cell 17(11):3141-3154.
Stepanova, A. N., J. Robertson-Hoyt, J. Yun, L. M. Benavente, D.-Y. Xie, K. Dolezal, A. Schlereth, G. Jurgens and J. M. Alonso (2008). "TAA1-Mediated Auxin Biosynthesis Is Essential for Hormone Crosstalk and Plant Development." Cell 133(1):177-191.
Stepanova, A. N., J. Yun, A. V. Likhacheva and J. M. Alonso (2007). "Multilevel interactions between ethylene and auxin in Arabidopsis roots." Plant Cell 19(7): 2169-2185.
Tao, Y., J. L. Ferrer, K. Ljung, F. Pojer, F. Hong, J. A. Long, L. Li, J. E. Moreno, M. E. Bowman, L. J. Ivans, Y. Cheng, J. Lim, Y. Zhao, C. L. Ballare, G. Sandberg, J. P. Noel and J. Chory (2008). "Rapid synthesis of auxin via a new tryptophan-dependent pathway is required for shade avoidance in plants." Cell 133(1): 164-176.
wang, J., Ma,X.,Kojima,M.,Sakakibara,H.,Hou,B (2011). "N-glucosytransferase UGT76C2 is involved in cytokinin homeostasis and cytokinin response in Arabidopsis thaliana."Plant cell physiol 52(12):2200-2213.
Weis, M., E.-K. Lim, N. C. Bruce and D. J, Bowles (2008). "Engineering and kinetic characterisation of two glucosyltransferases from Arabidopsis thaliana." Biochimie 90(5):830-834.
Woodward, A. W. and B. Bartel (2005). "Auxin:regulation, action, and interaction." Ann Bot 95(5):707-735.
Xu, Z. J., M. Nakajima, Y. Suzuki and I. Yamaguchi (2002). "Cloning and characterization of the abscisic acid-specific glucosyltransferase gene from adzuki bean seedlings." Plant Physiol 129(3):1285-1295.
Yonekura-Sakakibara, K. and K. Hanada (2011). "An evolutionary view of functional diversity in family 1 glycosyltransferases." Plant J 66(1):182-193.
Zhao, Y. (2010). "Auxin biosynthesis and its role in plant development." Annu Rev Plant Biol 61:49-64.