Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2019, Vol. 39 Issue (11): 22-30    DOI: 10.13523/j.cb.20191103
    
Flavonoids Metabolism and Antimicrobial Activity of Arabidopsis Heterologous Expressing CiRS Gene
YANG Fei-yun1,2,YANG Tian-rui1,LIU Kun1,CUI Shuang1,WANG Rui-gang1,LI Guo-jing1,**()
1 College of Life Sciences, Inner Mongolia Agricultural University, Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology, Hohhot 010011, China
2 College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China
Download: HTML   PDF(1361KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  

Resveratrol synthetase (RS), an important enzyme of chalcone synthase gene super family, catalyzes resveratrol production in plants. Resveratrol is a non-flavonoid polyphenol metabolite, which is a phytoalexin produced by plants under both biotic and abiotic stress. It has been proved that resveratrol has a variety of physiological activities.The CDS sequence annotated as CHS gene was obtained from the database of transcriptome, and the full length gene sequence was cloned using Caragana intermedia cDNA as template. Sequence, phylogenetic analysis and characteristic study of transgenic Arabidopsis indicated that the gene was a RS gene, then it was named CiRS. The expression of CiRS was induced by drought, NaCl and ultraviolet (UV) stresses as revealed by quantitative real-time PCR (qRT-PCR). Heterologous expression of CiRS gene inhibited the expression of endogenous AtCHS gene in transgenic Arabidopsis, meanwhile the antimicrobial activity of transgenic lines was stronger than that of wild type. All these results confirmed that the CiRS of C. intermedia played roles in transgenic Arabidopsis.

Key words： Resveratrol synthase      Caragana intermedia      Antimicrobial activity      Transgenic Arabidopsis     
Received: 29 April 2019      Published: 17 December 2019
ZTFLH:  Q786  
Corresponding Authors: Guo-jing LI     E-mail: liguojing@imau.edu.cn
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
Fei-yun YANG
Tian-rui YANG
Kun LIU
Shuang CUI
Rui-gang WANG
Guo-jing LI
Cite this article:

YANG Fei-yun,YANG Tian-rui,LIU Kun,CUI Shuang,WANG Rui-gang,LI Guo-jing. Flavonoids Metabolism and Antimicrobial Activity of Arabidopsis Heterologous Expressing CiRS Gene. China Biotechnology, 2019, 39(11): 22-30.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20191103     OR     https://manu60.magtech.com.cn/biotech/Y2019/V39/I11/22

Fig.1 Gel electrophoresis of amplified of CiRS M1: DL5000 marker; M2: DL2000 marker (a) CiRS 3'-RACE (b) CiRS cDNA (c) CiRS gDNA
Fig.2 The full-length DNA and protein sequence of CiRS The lowercase letter indicates the intron
Fig.3 The phylogenetic tree constructed with CiRS and RS and CHS from other species
Fig.4 Expression analysis of CiRS gene under different stress conditions
Fig.5 Expression analysis of AtCHS in transgenic Arabidopsis
Fig.6 Phytophthora infestans infection of the Arabidopsis leaves (a) Phenotype of P. infestans infection (b) Statistics of P. infestans infection rate The numbers represent serial number of independent heterologous expression lines
[1]   杨飞芸, 武燕燕, 崔爽 , 等. 异源表达CiRS基因通过生成白藜芦醇增强拟南芥的抗氧化能力. 中国生物工程杂志, 2017,37(12):27-33.
[1]   Yang F Y, Wu Y Y, Cui S , et al. Heterologous expression of CiRS gene enhances the antioxidant capacity of Arabidopsis by increasing the content of resveratrol. China Biotechnology, 2017,37(12):27-33.
[2]   Jeandet P, Delaunois B, Conreux A , et al. Biosynjournal, metabolism, molecular engineering and biological functions of stilbene phytoalexins in plants. Biofactors, 2010,36(5):331-341.
doi: 10.1002/biof.108 pmid: 20726013
[3]   Gambini J, Inglés M, Olaso G , et al. Properties of resveratrol: In vitro and in vivo studies about metabolism, bioavailability, and biological effects in animal models and humans. Oxidative Medicine and Cellular Longevity, 2015,2015:1-13.DOI: 10.1155/2015/837042.
doi: 10.1155/2015/837042 pmid: 29168580
[4]   Basholli-Salihu M, Schuster R, Mulla D , et al. Bioconversion of piceid to resveratrol by selected probiotic cell extracts. Bioprocess Biosyst Eng, 2016,39(12):1879-1885.
doi: 10.1007/s00449-016-1662-1 pmid: 27497981
[5]   Flieger J, Tatarczak-Michalewska M, Blicharska E . Characterization of the cis/trans isomerization of resveratrol by high-performance liquid chromatography. Analytical Letters, 2016,50(2):10.
doi: 10.1002/rcm.3316 pmid: 18023075
[6]   Lu Y, Shao D Y, Shi J L , et al. Strategies for enhancing resveratrol production and the expression of pathway enzymes. Applied Microbiology & Biotechnology, 2016,100(17):7407-7421.
doi: 10.1111/gcb.14962 pmid: 31838767
[7]   Shomura Y, Torayama I, Suh D Y , et al. Crystal structure of stilbene synthase from Arachis hypogaea. Proteins: Structure, Function, and Bioinformatics, 2005,60(4):803-806.
[8]   Yu C K Y, Karin S, Jürgen S , et al. A stilbene synthase gene (SbSTS1) is involved in host and nonhost defense responses in sorghum. Plant Physiology, 2005,138(1):393-401.
doi: 10.1104/pp.105.059337 pmid: 15821144
[9]   Hain R, Bieseler B, Kindl H , et al. Expression of a stilbene synthase gene in Nicotiana tabacum results in synjournal of the phytoalexin resveratrol. Plant Molecular Biology, 1990,15(2):325-335.
doi: 10.1007/bf00036918 pmid: 2103451
[10]   Shi J L, He M Y, Cao J L , et al. The comparative analysis of the potential relationship between resveratrol and stilbene synthase gene family in the development stages of grapes (Vitis quinquangularis and Vitis vinifera). Plant Physiology & Biochemistry, 2014,74:24-32.
doi: 10.1016/j.plaphy.2019.12.002 pmid: 31837557
[11]   Delaunois B, Cordelier S, Conreux A , et al. Molecular engineering of resveratrol in plants. Plant Biotechnology Journal, 2009,7(1):2-12.
doi: 10.1111/j.1467-7652.2008.00377.x pmid: 19021877
[12]   Bai J, Kang T, Wu H D , et al. Relative contribution of photorespiration and antioxidative mechanisms in Caragana korshinskii under drought conditions across the Loess Plateau. Functional Plant Biology, 2017,44(11):1111-1123.
doi: 10.1071/FP17060
[13]   Yang F Y, Yang T R, Liu K , et al. Analysis of metabolite accumulation related to pod color variation of Caragana intermedia. Molecules, 2019,24(4):717.
doi: 10.1038/sj.emboj.7600559 pmid: 15692567
[14]   Luan G X, Wang H L, Lv H H , et al. Separation and purification of five flavone glucosides and one lignan from Caragana korshinskii Kom. by the combination of HSCCC and semi-preparative RPLC. Chromatographia, 2016,79(13):823-831.
doi: 10.1007/s10337-016-3090-4
[15]   Zeng Z, Ji Z Y, Hu N , et al. A sensitive pre-column derivatization method for the analysis of free fatty acids by RP-HPLC with fluorescence detector and its application to Caragana species. Journal of Chromatography B, 2017,1064 : 151-159.
doi: 10.1016/j.jchromb.2017.07.037 pmid: 28927568
[16]   韩晓敏 . 中间锦鸡儿3个非生物胁迫相关转录因子的克隆与功能分析. 呼和浩特:内蒙古农业大学, 2015: 24-26.
[16]   Han X M . The cloning and analysis of three stress-related transcription factors from Caragana intermedia. Hohhot: Inner Mongolia Agricultural University, 2015: 24-26.
[17]   杨飞芸, 刘坤, 崔爽 , 等. 转CiCHS基因拟南芥的黄酮代谢及抗氧化能力分析. 西北植物学报, 2018,38(3):393-400.
[17]   Yang F Y, Liu K, Cui S , et al. Metabolism of flavonoids and the antioxidant capacity of transgenic Arabidopsis expressing CiCHS gene. Acta Botanica Boreali-Occidentalia Sinica, 2018,38(3):393-400.
[18]   O’Dell M, Metzlaff M, Flavell R B . Post-transcriptional gene silencing of chalcone synthase in transgenic petunias, cytosine methylation and epigenetic variation. The Plant Journal, 1999,18(1):33-42.
doi: 10.1099/00221287-18-1-33 pmid: 13525625
[19]   Shanker A K, Shanker C . Abiotic and biotic stress in plants: Recent advances and future perspectives. London: IntechOpen, 2016: 185-227.
doi: 10.1089/ars.2019.7819 pmid: 31701753
[20]   Chong J L, Poutaraud A, Hugueney P . Metabolism and roles of stilbenes in plants. Plant Science, 2009,177(3):143-155.
doi: 10.1186/s12870-019-1935-3 pmid: 31387524
[21]   Salehi B, Mishra A P, Nigam M , et al. Resveratrol: A double-edged sword in health benefits. Biomedicines , 2018,6(3):91.
doi: 10.3390/biomedicines6030091 pmid: 30205595
[22]   Cho Y B, Jones S I, Vodkin L O . Mutations in Argonaute5 illuminate epistatic interactions of the K1 and I loci leading to saddle seed color patterns in Glycine max. The Plant Cell, 2017,29(4):708-725.
doi: 10.1105/tpc.17.00162 pmid: 28351993
[23]   Espa?a L ,Heredia-Guerrero J A, Reina-Pinto J J, et al. Transient silencing of chalcone synthase during fruit ripening modifies tomato epidermal cells and cuticle properties. Plant Physiology, 2014,166(3):1371-1386.
doi: 10.1104/pp.114.246405
[24]   Suzuki N, Rivero R M, Shulaev V , et al. Abiotic and biotic stress combinations. New Phytologist, 2014,203(1):32-43.
doi: 10.1111/nph.12797
[25]   Tsuda K, Somssich I E . Transcriptional networks in plant immunity. New Phytologist, 2015,206(3):932-947.
doi: 10.1111/nph.13286 pmid: 25623163
[26]   Dao T T H, Linthorst H J M, Verpoorte R . Chalcone synthase and its functions in plant resistance. Phytochemistry Reviews, 2011,10(3):397-412.
doi: 10.1007/s11101-011-9211-7
[27]   Tang K, Zhan J C, Yang H R , et al. Changes of resveratrol and antioxidant enzymes during UV-induced plant defense response in peanut seedlings. Journal of Plant Physiology, 2010,167(2):95-102.
doi: 10.1016/j.jplph.2009.07.011 pmid: 19716623
[1] Wen-juan CHAI,Qi YANG,Guo-jing LI,Rui-gang WANG. CiMYB15 from Caragana Intermedia Positively Regulates Flavonoids Metabolism of Arabidopsis[J]. China Biotechnology, 2018, 38(10): 8-19.
[2] Fei-yun YANG,Yan-yan WU,Shuang CUI,Xiu-juan ZHANG,Rui-gang WANG,Guo-jing LI. Heterologous Expression of CiRS Gene Eenhances the Antioxidant Capacity of Arabidopsis by Increasing the Content of Resveratrol[J]. China Biotechnology, 2017, 37(12): 27-33.
[3] XIE Huan, YU Hui-min, SHEN Zhong-yao. Structure, Properties and Synthesis Reinforcement of Fengycins: Lipopeptide Biosurfactant[J]. China Biotechnology, 2015, 35(7): 102-110.
[4] SUN Li-jun, WANG Ya-ling, LIU Huan-min, XU De-feng, ZHANG Yong-pin, NIE Fang-hong. Identification of Antimicrobial Lipopetides Component Produced by Isolate from Douchi and Its Antimicrobial Properties[J]. China Biotechnology, 2013, 33(7): 50-56.
[5] . Bacillus natto TK-1产脂肽的纯化、抑菌活性及其表面活性剂特性的初步探讨[J]. China Biotechnology, 2008, 28(1): 44-48.
[6] . Overexpression of the fusion gene encoding bovine antimicrobial peptides Bac7-Bac5 in Escherichia coli, and purification and antimicrobial activity of the fusion protein[J]. China Biotechnology, 2007, 27(3): 65-70.
主管：中国科学院 　主办：中国科学院文献情报中心 　中国生物技术发展中心 　中国生物工程学会