Abscisic Acid Regulates the 3-Hydroxy-3-methylglutaryl CoA Reductase Gene Promoter and Ginsenoside Production in Panax quinquefolium Hairy Root Cultures

被引:28
作者
Kochan, Ewa [1 ]
Balcerczak, Ewa [2 ]
Szymczyk, Piotr [1 ]
Sienkiewicz, Monika [3 ]
Zielinska-Blizniewska, Hanna [3 ]
Szymanska, Grazyna [1 ]
机构
[1] Med Univ Lodz, Dept Pharmaceut Biotechnol, Muszynskiego l, PL-90151 Lodz, Poland
[2] Med Univ Lodz, Interfaculty Cathedral Lab & Mol Diagnost, Dept Pharmaceut Biochem & Mol Diagnost, Lab Mol Diagnost & Pharmacogen, Muszynskiego 1, PL-90151 Lodz, Poland
[3] Med Univ Lodz, Dept Allergol & Resp Rehabil, Chair Otolaryngol 2, Zeligowskiego 7-9, PL-90725 Lodz, Poland
关键词
3-hydroxy-3-methylglutaryl CoA reductase gene promoter; ginsenosides; hairy roots; Panax quinquefolium; FUNCTIONAL-ANALYSIS; EXPRESSION ANALYSIS; PLANT-GROWTH; REAL-TIME; STRESS; ABA; IDENTIFICATION; ARABIDOPSIS; ELEMENTS; METHYLATION;
D O I
10.3390/ijms20061310
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Panax quinquefolium hairy root cultures synthesize triterpenoid saponins named ginsenosides, that have multidirectional pharmacological activity. The first rate-limiting enzyme in the process of their biosynthesis is 3-hydroxy-3-methylglutaryl CoA reductase (HMGR). In this study, a 741 bp fragment of the P. quinquefolium HMGR gene (PqHMGR), consisting of a proximal promoter, 5 ' UTR (5 ' untranslated region) and 5 ' CDS (coding DNA sequence) was isolated. In silico analysis of an isolated fragment indicated a lack of tandem repeats, miRNA binding sites, and CpG/CpNpG elements. However, the proximal promoter contained potential cis-elements involved in the response to light, salicylic, and abscisic acid (ABA) that was represented by the motif ABRE (TACGTG). The functional significance of ABA on P. quinquefolium HMGR gene expression was evaluated, carrying out quantitative RT-PCR experiments at different ABA concentrations (0.1, 0.25, 0.5, and 1 mg center dot L-1). Additionally, the effect of abscisic acid and its time exposure on biomass and ginsenoside level in Panax quinquefolium hairy root was examined. The saponin content was determined using HPLC. The 28 day elicitation period with 1 mg center dot L-1 ABA was the most efficient for Rg2 and Re (17.38 and 1.83 times increase, respectively) accumulation; however, the protopanaxadiol derivative content decreased in these conditions.
引用
收藏
页数:16
相关论文
共 70 条
[1]   Molecular cloning and expression profile of an abiotic stress and hormone responsive MYB transcription factor gene from Panax ginseng [J].
Afrin, Sadia ;
Zhu, Jie ;
Cao, Hongzhe ;
Huang, Jingjia ;
Xiu, Hao ;
Luo, Tiao ;
Luo, Zhiyong .
ACTA BIOCHIMICA ET BIOPHYSICA SINICA, 2015, 47 (04) :267-277
[2]   Gene-associated CpG islands in plants as revealed by analyses of genomic sequences [J].
Ashikawa, I .
PLANT JOURNAL, 2001, 26 (06) :617-625
[3]  
BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513
[4]   STaRRRT: a table of short tandem repeats in regulatory regions of the human genome [J].
Bolton, Katherine A. ;
Ross, Jason P. ;
Grice, Desma M. ;
Bowden, Nikola A. ;
Holliday, Elizabeth G. ;
Avery-Kiejda, Kelly A. ;
Scott, Rodney J. .
BMC GENOMICS, 2013, 14
[5]   Transcriptome Analysis of Methyl Jasmonate-Elicited Panax ginseng Adventitious Roots to Discover Putative Ginsenoside Biosynthesis and Transport Genes [J].
Cao, Hongzhe ;
Nuruzzaman, Mohammed ;
Xiu, Hao ;
Huang, Jingjia ;
Wu, Kunlu ;
Chen, Xianghui ;
Li, Jijia ;
Wang, Li ;
Jeong, Ji-Hak ;
Park, Sun-Jin ;
Yang, Fang ;
Luo, Junli ;
Luo, Zhiyong .
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2015, 16 (02) :3035-3057
[6]   Locus-specific control of asymmetric and CpNpG methylation by the DRM and CMT3 methyltransferase genes [J].
Cao, XF ;
Jacobsen, SE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 :16491-16498
[7]  
Cao XY, 2011, J MED PLANTS RES, V5, P3868
[8]   PlantPAN: Plant promoter analysis navigator, for identifying combinatorial cis-regulatory elements with distance constraint in plant gene groups [J].
Chang, Wen-Chi ;
Lee, Tzong-Yi ;
Huang, Hsien-Da ;
Huang, His-Yuan ;
Pan, Rong-Long .
BMC GENOMICS, 2008, 9 (1)
[9]   Seed genome hypomethylated regions are enriched in transcription factor genes [J].
Chen, Min ;
Lin, Jer-Young ;
Hur, Jungim ;
Pelletier, Julie M. ;
Baden, Russell ;
Pellegrini, Matteo ;
Harada, John J. ;
Goldberg, Robert B. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2018, 115 (35) :EB315-EB322
[10]   ABFs, a family of ABA-responsive element binding factors [J].
Choi, HI ;
Hong, JH ;
Ha, JO ;
Kang, JY ;
Kim, SY .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (03) :1723-1730