Cloning and Expression Analysis of HMGR Gene from Chamaemelum nobile

doi:10.7668/hbnxb.2016.06.011

Abstract

Abstract: 3-hydroxy-3-methyl-glutaryl-CoA reductase (3-hydroxy-3-methyl glutaryl coenzyme A reductase,HMGR) is one of the key rate-limiting enzymes in mevalonic (MVA) pathway of terpenoid biosynthesis.To analysis the function of HMGR gene in terpenoid biosynthesis in Chamaemelum nobile,a HMGR gene (designated as CnHMGR,GenBank accession number KU589282) was cloned from C.nobile using RT-PCR method.The full-length cDNA of CnHMGR gene was 1 856 bp and contained an open reading frame (ORF) of 1 746 bp,which encodes a 582 amino-acid protein.The theoretical molecular weight and PI of the CnHMGR were 62.5 kDa and 6.80,respectively.Multi-alignment comparison analysis showed the protein sequence of CnHMGR had high similarity with those of HMGR proteins from other plants.Furthermore,CnHMGR has two HMG-CoA-binding motifs:TTEGCLVA,EMPVGYVQIP,and two NADP(H)-binding motifs:TVGGGT,DAMGMMM,suggesting CnHMGR is one of HMGR family mumbers in C.nobile.The results of tissue expression analysis showed that CnHMGR was constituently expressed indifferent tissues of C.nobile,with highest expression level in flowers and lowest expression level in stems.The present study cloned,characterized and analyzed the tissue expression pattern of CnHMGR in C.nobile,providing basic data for further studying the function of CnHMGR in sesquiterpene biosynthesis of C.nobile.

Key words: Chamaemelum nobile, CnHMGR, Gene cloning, Tissue expression

CLC Number:

Q78
S682.03

MENG Xiangxiang, YAN Jiaping, LIU Xiaomeng, LIAO Yongling, CHANG Jie, XU Feng. Cloning and Expression Analysis of HMGR Gene from Chamaemelum nobile[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(6): 68-75. doi: 10.7668/hbnxb.2016.06.011.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2016.06.011

http://www.hbnxb.net/EN/Y2016/V31/I6/68

References

[1] Lemberkovics E, Kery A, Marczal G, et al. Phytochemical evaluation of essential oils,medicinal plants and their preparations[J]. Acta Pharmaceutica Hungarica, 1998, 68(3):141-149.
[2] Carnat A, Carnat A P, Fraisse D, et al. The aromatic and polyphenolic composition of Roman chamomile tea[J]. Fitoterapia, 2004, 75(1):32-38.
[3] Srivastava J K, Shankar E, Gupta S. Chamomile:A herbal medicine of the past with a bright future (Review)[J]. Molecular Medicine Reports, 2010, 3(6):895-901.
[4] 张长波, 孙红霞, 巩中军, 等. 植物萜类化合物的天然合成途径及其相关合酶[J]. 植物生理学通讯, 2007, 43:779-786.
[5] 何云飞, 高伟, 刘塔斯, 等. 二萜合酶的研究进展[J]. 药学学报, 2011, 46(9):1019.
[6] Doblas V G, Amorim-Silva V, Posé D, et al. The SUD1 gene encodes a putative E3 ubiquitin ligase and is a positive regulator of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in Arabidopsis[J]. Plant Cell, 2013, 25(2):728-743.
[7] Chappell J, Wolf F, Proulx J, et al. Is the reaction catalyzed by 3-hydroxy-3-methylglutaryl coenzyme A reductase a rate-limiting step for isoprenoid biosynthesis in plants[J]. Plant Physiology, 1995, 109(4):1337-1343.
[8] Chappell J, Nable R. Induction of sesquiterpenoid biosynthesis in tobacco cell suspension cultures by fungal elicitor[J]. Plant Physiology, 1987, 85(2):469-473.
[9] Ha S H, Kim J B, Hwang Y S, et al. Molecular characterization of three 3-hydroxy-3-methylglutaryl-CoA reductase genes including pathogen-induced Hmg2 from pepper(Capsicum annuum)[J]. Biochimica et Biophysica acta, 2003, 1625(3):253-260.
[10] Shen G, Pang Y, Wu W, et al. Cloning and characterization of a root-specific expressing gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from Ginkgo biloba[J]. Molecular Biology Reports, 2006, 33(2):117-127.
[11] Cao X Y, Li C G, Miao Q, et al. Molecular cloning and expression analysis of a leaf-specific expressing 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase gene from Michelia chapensis Dandy[J]. Journal of Medicinal Plant Research, 2011, 5(16):3868-3875.
[12] Dai Z, Cui G, Zhou S F, et al. Cloning and characterization of a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Salvia miltiorrhiza involved in diterpenoid tanshinone accumulation[J]. Journal of Plant Physiology, 2011, 168(2):148-157.
[13] Xia R, Lu W J, Wang Z H, et al. Cloning and characterisation of two genes for 3-hydroxy-3-methylglutaryl coenzyme A reductase and their possible roles during fruit development in Dimocarpus longan[J]. The Journal of Horticultural Science and Biotechnology, 2011, 86(1):25-30.
[14] Van D N, Bachmann A L, Schmidt T, et al. Molecular cloning of mevalonate pathway genes from Taraxacum brevicorniculatum and functional characterisation of the key enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase[J]. Molecular Biology Reports, 2012, 39(4):4337-4349.
[15] Wang Q J, Zheng L P, Zhao P F, et al. Cloning and characterization of an elicitor-responsive gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase involved in 20-hydroxyecdysone production in cell cultures of Cyanotis arachnoidea[J]. Plant Physiology and Biochemistry, 2014, 84:1-9.
[16] Kalita R, Patar L, Shasany A K, et al. Molecular cloning, characterization and expression analysis of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Centella asiatica L.[J]. Molecular Biology Reports, 2015, 42(9):1431-1439.
[17] Livak K J, Schmittgen T D. Analysis of relative gene expression data using Real-time quantitative PCR and the 2^-ΔΔCt method[J]. Methods, 2001, 25(4):402-408.
[18] Wang Y, Darnay B G, Rodwell V W. Identification of the principal catalytically important acidic residue of 3-hydroxy-3-methylglutaryl coenzyme A reductase[J]. The Journal of Biological Chemistry, 1990, 265(35):21634-21641.
[19] 朴英花, 朴惠顺. 倍半萜类化合物生物活性研究进展[J]. 职业与健康, 2012, 28(18):2291-2293.
[20] Chappell J. Biochemistry and molecular biology of the isoprenoid biosynthetic pathway in plants[J]. Annual Review of Plant Biology, 1995, 46(1):521-547.
[21] Darabi M, Masoudi-Nejad A, Nemat-Zadeh G. Bioinformatics study of the 3-hydroxy-3-methylglotaryl-coenzyme A reductase (HMGR) gene in Gramineae[J]. Molecular Biology Reports,2012,42(9):1243-1243.
[22] Yang Z, Park H, Lacy G H, et al. Differential activation of potato 3-hydroxy-3-methylglutaryl coenzyme A reductase genes by wounding and pathogen challenge[J]. The Plant Cell, 1991, 3(4):397-405.
[23] Choi D, Ward B L, Bostock R M. Differential induction and suppression of potato 3-hydroxy-3-methylglutaryl coenzyme A reductase genes in response to Phytophthora infestans and to its elicitor arachidonic acid[J]. The Plant Cell, 1992, 4(10):1333-1344.
[24] Aquil S, Husaini A M, Abdin M Z, et al. Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants[J]. Planta Medica, 2009, 75(13):1453-1458.
[25] Alam P, Abdin M Z. Over-expression of HMG-CoA reductase and amorpha-4,11-diene synthase genes in Artemisia annua L. and its influence on artemisinin content[J]. Plant Cell Reports, 2011, 30(10):1919-1928.
[26] Ruiz-Albert J, Cerdá-Olmedo E, Corrochano L M. Genes for mevalonate biosynthesis in Phycomyces[J]. Molecular Genetics and Genomics, 2002, 266(5):768-777.
[27] Akhtar N, Gupta P, Sangwan N S, et al. Cloning and functional characterization of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Withania somnifera:an important medicinal plant[J]. Protoplasma, 2013, 250(2):613-622.
[28] 罗红梅, 宋经元, 李雪莹, 等. 人参皂苷合成生物学关键元件HMGR基因克隆与表达分析[J].药学学报, 2013, 48(2):219-227.
[29] 徐艳红, 杨欣, 张争, 等. 白木香3-羟基-3-甲基戊二酰辅酶A还原酶基因-AsHMGR2的克隆及表达分析[J]. 药学学报, 2013, 48(6):953.
[30] Agatonovic-Kustrin S, Ortakand D B, Morton D W. Migraine headaches:Feverfew or Chamomile leaves[J]. Modern Chemistry Applications, 2015, 3(169):2.

Please choose a citation manager

Content to export