Journal of Shandong University (Health Sciences) ›› 2019, Vol. 57 ›› Issue (4): 27-33.doi: 10.6040/j.issn.1671-7554.0.2019.027

Previous Articles    

Cloning and functional characterization of two O-methyltransferases from Adiantum capillus-veneris L.

NI Rong, ZHANG Xiaoshuang, CHENG Aixia   

  1. Key Laboratory of Chemical Biology of Natural Products, Ministry of Education;
    School of Pharmaceutical Sciences, Shandong University, Jinan 250012, Shandong, China
  • Published:2022-09-27

PDF (PC)

13

Abstract: Objective To isolate and characterize the O-methyltransferase genes involved in the biosynthesis of flavonoids and phenypropanoids from Adiantum capillus-veneris L. Methods The full-length cDNA sequences of two O-methyltransferase genes were cloned by the RT-PCR method using the total RNA isolated from the Adiantum capillus-veneris L. as template, and designated as AcOMT1 and AcOMT2, respectively. The amplified ORFs were ligated into the pET-32a vector, and then were transformed into E.coli BL21 for expression. The recombinant proteins were purified using the Ni-NTA Sefinose His-bind column and used for enzyme assays. The protein homology models of AcOMT1 and AcOMT2 were generated by Swiss-model on-line server, and the key amino acids which were postulated to affect protein activity were predicted by PyMOL software. The analyses of phylogenetic tree and multiple sequence alignment were performed using MEGA v5.1 and DNAMAN 7.0.2 software, respectively. Results Two genes encoding putative O-methyltransferase were identified from the transcriptome sequencing database of Adiantum capillus-veneris L. and cloned from the plants. The sequence alignment and phylogenetic analysis indicated that AcOMTs belong to the family of type Ⅰ OMTs. Meanwhile, the enzymatic assays showed that AcOMT1 and AcOMT2 had the similar catalytic characteristics and broad substrate selectivety. Quercetin was the favorite substrate. Conclusion Two OMT genes were isolated and characterized from Adiantum capillus-veneris L., which would provide theoretical significance to elucidate the methylation modification of flavonoids in ferns.

Key words: Adiantum capillus-veneris L., Flavonoids, O-methyltransferase, Gene cloning, Functional characterization

CLC Number: 

  • Q789
[1] 严欢欢,肖娟,梁永书,等. 基于SCI-E的我国蕨类植物研究进展分析[J]. 安徽农业科学, 2016, 44(9): 280-283. YAN Huanhuan, XIAO Juan, LIANG Yongshu, et al. Research progress of ferns in China basel on SCI-E database[J]. Journal of Anhui Agricultural Sciences, 2016, 44(9): 280-283.
[2] Chen J, Zhao DG. Research advances on the enzymes and their coding gene involved in plant terpene biosynthesis[J]. Molecular Plant Breeding, 2004, 2(6):757-764.
[3] 郭治友. 药用植物乌蕨配子体发育的观察[J].基因组学与应用生物学, 2009, 28(5): 961-964. GUO Zhiyou. Observation on the gametophyte development of fairy fern(Stenoloma chusanum)[J]. Genomics & Applied Biology, 2009, 28(5): 961-964.
[4] Wellman CH, Osterloff PL, Mohiuddin U. Fragments of the earliest land plants[J]. Nature, 2003, 425(6955): 282-285.
[5] Christensen AB, Gregersen PL, Olsen CE, et al. A flavonoid 7-O-methyltransferase is expressed in barley leaves in response to pathogen attack[J]. Plant Mol Biol, 1998, 36(2): 219-227.
[6] Zhu BT, Ezell EL, Liehr JG. Catechol-O-methyltransferase-catalyzed rapid O-methylation of mutagenic flavonoids. Metabolic inactivation as a possible reason for their lack of carcinogenicity in vivo[J]. J Biol Chem, 1994, 269(1): 292-299.
[7] Joshi CP, Chiang VL. Conserved sequence motifs in plant S-adenosyl-L-methionine-dependent methyltransferases[J]. Plant Mol Biol, 1998, 37(4): 663-674.
[8] Ferrer JL, Zubieta C, Dixon RA, et al. Crystal structures of alfalfa caffeoyl coenzyme A 3-O-methyltransferase[J]. Plant Physiol, 2005, 137(3): 1009-1017.
[9] Raes J, Rohde A, Christensen JH, et al. Genome-wide characterization of the lignification toolbox in Arabidopsis[J]. Plant Physiol, 2003, 133(3): 1051-1071.
[10] Lee YJ, Kim BG, Chong Y, et al. Cation dependent O-methyltransferases from rice[J]. Planta, 2008, 227(3): 641-647.
[11] Wagner A, Tobimatsu Y, Phillips L, et al. CCoAOMT suppression modifies lignin composition in Pinus radiata[J]. Plant J, 2011, 67(1): 119-129.
[12] Wanner LA, Li G, Ware D, et al. The phenylalanine ammonia-lyase gene family in Arabidopsis thaliana[J]. Plant Mol Biol, 1995, 27(2): 327-338.
[13] Gambino G, Perrone I, Gribaudo I. A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants[J]. Phytochem Anal, 2008, 19(6): 520-525.
[14] Xu RX, Zhao Y, Gao S, et al. Functional characterization of a plastidal cation-dependent O-methyltransferase from the liverwort Plagiochasma appendiculatum[J]. Phytochemistry, 2015, 118: 33-41. doi:10.1016/j.phytochem.2015.08.002.
[15] Xu RX, Gao S, Zhao Y, et al. Functional characterization of a Mg2+-dependent O-methyltransferase with coumarin as preferred substrate from the liverwort Plagiochasma appendiculatum[J]. Plant Physiol Biochem, 2016, 106: 269-277. doi:10.1016/j.plaphy.2016.05.018.
[16] Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular Evolutionary Genetics Analysis(MEGA)software version 4.0[J]. Mol Biol Evol, 2007, 24(8): 1596-1599.
[17] Bordoli L, Kiefer F, Arnold K, et al. Protein structure homology modeling using SWISS-MODEL workspace[J]. Nat Protoc, 2009, 4(1): 1-13.
[18] 李丹丹,刘慧,程爱霞. 一个蛇苔氧甲基转移酶基因的克隆与功能鉴定[J]. 山东大学学报(医学版), 2016, 54(12): 20-26. LI Dandan, LIU Hui, CHENG Aixia. Cloning and functional characterization of an O-methyltransferase gene from Conocephalum conicum[J]. Journal of Shandong University(Health Sciences), 2016, 54(12): 20-26.
[19] 张玉莹. 苔类植物两个bHLH转录因子和一个甲氧基转移酶的功能研究[D]. 济南:山东大学, 2017.
[20] Raghavan V. Developmental biology of fern gametophytes[M]. Oxford City: Cambridge University Press, 1989.
[21] 蒋云,秦华,顾丽. 观赏蕨类在园林中的应用[J]. 江西农业学报, 2008, 20(5): 44-45, 48. JIANG Yun, QIN Hua, GU Li. Application of ornamental pteridophyte in landscape[J]. Acta Agriculturae Jiangxi, 2008, 20(5): 44-45, 48.
[22] 吴彩霞,张勇,顾雪竹,等.铁线蕨化学成分和药理作用研究进展[J].中国医药导报, 2014, 11(2): 39-40, 46. WU Caixia, ZHANG Yong, GU Xuezhu, et al. Phytochemical and phamacological research progress in Adiantum capillus-veneris L.[J]. China Medical Herald, 2014, 11(2): 39-40, 46.
[23] 李珊,姚默,刘向辉,等.铁线蕨属药学研究概况[J].宁夏农林科技, 2012, 53(7): 71-73. LI Shan, YAO Mo, LIU Xianghui, et al. Overview of Pharmacological research on Adiantum L.[J]. Ningxia Journal of Agriculture & Forestry Science & Technology, 2012, 53(7): 71-73.
[24] Nonato FR, Nogueira M, Barros A, et al. Antinociceptive and antiinflammatory activities of Adiantum latifolium Lam.: evidence for a role of IL-1β inhibition[J]. J Ethnopharmacol, 2011, 136(3): 518-524.
[25] Guo D, Chen F, Inoue K, et al. Downregulation of caffeic acid 3-O-methyltransferase and caffeoyl CoA 3-O-methyltransferase in transgenic alfalfa. impacts on lignin structure and implications for the biosynthesis of G and S lignin[J]. Plant Cell, 2001, 13(1): 73-88.
[26] Ibrahim RK. A forty-year journey in plant research: original contributions to flavonoid biochemistry[J]. Canadian Journal of Botany, 2005, 83(5): 433-450.
[1] SHANG Yuying, MA Yingjuan, WU Xiaofan, HOU Xunyao, LUO Dingzhen, CHEN Jian, HONG Yan, SHEN Chao, LIU Xueping. Synapse-protective effect of flavonoids extracted from the leaves of Diospyros kaki in APP/PS1 transgenic mice [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(2): 33-37.
[2] LI Dandan, LIU Hui, CHENG Aixia. Cloning and functional characterization of an O-methyltransferase gene from Conocephalum conicum [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(12): 20-26.
[3] LIU Yiqing,WANG Lai-cheng,JIAO Yu-lian,ZHANG Jie,MA Chunyan,CUI Bin,GAO Xinpu,LIU Zheng-in,HANG Xue,ZHAO Yue-ran,. 〖KH+9.2mmD]Cloning of human interleukin26 and constructing ofits eukaryotic expression vector [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2006, 44(6): 541-544.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd