盐胁迫诱导盐穗木Rev1、Rev3基因的表达分析

doi:10.7525/j.issn.1673-5102.2017.02.008

摘要/Abstract

摘要： 根据盐穗木盐胁迫下响应的转录组测序结果，参考盐穗木HcRev1、HcRev3基因的ESTs序列设计荧光定量PCR特异性引物，建立检测盐穗木Revs基因相对表达量的荧光定量PCR方法，分析Rev1和Rev3基因在盐穗木不同浓度盐胁迫处理不同时间的转录水平。结果表明，HcRev1、HcRev3基因具有相似的表达模式，在100 mmol·L^-1 NaCl低盐胁迫下表达稳定，在300、500、700 mmol·L^-1 NaCl胁迫下，随胁迫浓度增高、胁迫时间延长，表达量升高。其中HcRev1在700 mmol·L^-1 NaCl胁迫14 d后达到峰值，是对照组的4.63倍。HcRev3基因在300 mmol·L^-1 NaCl胁迫14 d时，表达量迅速升高，是对照组的15.55倍，表达差异极显著。研究结果说明HcRev1、HcRev3基因都受盐胁迫诱导表达，提示HcRev1、HcRev3基因虽然表达量存在差异，但在盐胁迫过程中参与了DNA损伤修复。研究有助于阐明Rev1、Rev3基因在DNA损伤修复和植物耐盐性间的调控功能作用。

关键词: 盐穗木, HcRev1基因, HcRev3基因, 盐胁迫, 表达分析

Abstract: According to the transcriptome of Halostachys caspica under salt stress, the real-time PCR primers designed about Rev1 and Rev3 were referred to the ESTs of transcriptome sequencing. We established a method for detecting of mRNA transcription of Revs by Real-time PCR, and the transcription level of Revs in different concentration of NaCl present. Two genes have similar expression pattern, the expression levels remain stable under 100 mmol·L^-1 NaCl treatment. As the concentration of NaCl was increased and the stress time was prolonged, the genes relative expression levels were also gradually increased. HcRev1 gene in 700 mmol·L^-1 NaCl stress for 14 d was rapidly up-regulated and reached the peak level, which was 4.63-fold of the control. HcRev3 gene in 300 mmol·L^-1 NaCl stress for 14 d was 15.55-fold of the control with significant differences. Both HcRev1 and HcRev3 genes were induced by salt stress, although there were different expression levels of HcRev1 and HcRev3 genes, which were involved in the DNA damage repair during salt stress.Our study could be used to clarify the regulatory function of HcRev1 and HcRev3 genes between DNA damage-repair and salt resistance in plants.

Key words: Halostachys caspica, HcRev1 gene, HcRev3 gene, salt stress, expression analysis

中图分类号:

Q786

杜驰, 张冀, 张富春. 盐胁迫诱导盐穗木Rev1、Rev3基因的表达分析[J]. 植物研究, 2017, 37(2): 211-215.

DU Chi, ZHANG Ji, ZHANG Fu-Chun. Expression Analysis of Rev1 and Rev3 of Halostachys caspica under Salt Stress[J]. Bulletin of Botanical Research, 2017, 37(2): 211-215.

参考文献

1. Zhu J K. Plant salt tolerance[J]. Trends in Plant Science,2001,6(2):66-71.
2. Chinnusamy V,Zhu J K. Epigenetic regulation of stress responses in plants[J]. Current Opinion in Plant Biology,2009,12(2):133-139.
3. Bridges B A. Error-prone DNA repair and translesion synthesis:focus on the replication fork[J]. DNA Repair,2005,4(5):618-634.
4. Ohashi E,Ogit,Kusumotor,et al. Error-prone bypass of certain DNA lesions by the human DNA polymerase κ[J]. Genes &Development,2000,14(13):1589-1594.
5. Ho T V,Schärer O D. Translesion DNA synthesis polymerases in DNA interstrand crosslink repair[J]. Environmental and Molecular Mutagenesis,2010,51(6):552-566.
6. Sousa R. Structural and mechanistic relationships between nucleic acid polymerases[J]. Trends in Biochemical Sciences,1996,21(5):186-190.
7. Guo C X,Sonoda E,Tang T S,et al. REV1 protein interacts with PCNA:significance of the REV1 BRCT domain in vitro and in vivo[J]. Molecular Cell,2006,23(2):265-271.
8. Gibbs PE M,Mcgregor W G,Maher V M,et al. A human homolog of the Saccharomyces cerevisiae REV3 gene,which encodes the catalytic subunit of DNA polymerase ζ[J]. Proceedings of the National Academy of Sciences of the United States of America,1998,95(12):6876-6880.
9. Szüts D,Marcus A P,Himoto M,et al. REV1 restrains DNA polymerase ζ to ensure frame fidelity during translesion synthesis of UV photoproducts in vivo[J]. Nucleic Acids Research,2008,36(21):6767-6780.
10. Pozo F M,Odat,Sekimotot,et al. Molecular chaperone Hsp90 regulates REV1-mediated mutagenesis[J]. Molecular and Cellular Biology,2011,31(16):3396-3409.
11. Lawrence C W. Cellular roles of DNA polymerase ζ and Rev1 protein[J]. DNA Repair,2002,1(6):425-435.
12. 张立新,李生秀. 甜菜碱与植物抗旱/盐性研究进展[J]. 西北植物学报,2004,24(9):1765-1771.Zhang L X,Li SX,Research progress on relationships betaine and drought/salt resistance of plants[J]. Acta Botanica Boreali-Occidentalia Sinica,2004,24(9):1765-1771.
13. 常丹,张霞,张富春. 盐穗木HcPEAMT基因的克隆及表达分析[J]. 西北植物学报,2014,34(8):1522-1528.Chang D,Zhang X,Zhang F C. Cloning and expression analysis of HcPEAMT gene from Halostachys caspica[J]. Acta Botanica Boreali-Occidentalia Sinica,2014,34(8):1522-1528.
14. 赵航,贾富强,张富春,等. 盐胁迫下盐穗木差异表达基因的转录组信息分析[J]. 生物信息学,2014,12(2):90-98.Zhao H,Jia FQ,Zhang F C,et al. The transcriptome information analysis of differentially expressed genes of Halostachys caspica under salt stress[J]. China Journal of Bioinformatics,2014,12(2):90-98.
15. 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.
16. Schmittgen T D,Zakrajsek B A. Effect of experimental treatment on housekeeping gene expression:validation by real-time,quantitative RT-PCR[J]. Journal of Biochemical and Biophysical Methods,2000,46(1-2):69-81.
17. 张吉宇,王彦荣,南志标. 相对定量和绝对定量——以CsSAMDC基因表达分析为例[J]. 中国生物工程杂志,2009,29(8):86-91.Zhang J Y,Wang Y R,Nan Z B. Relative and absolute quantification-expression analysis of CsSAMDC gene as a case[J]. China Biotechnology,2009,29(8):86-91.
18. Wan H J,Zhao Z G,Qian C T,et al. Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber[J]. Analytical Biochemistry,2010,399(2):257-261.
19. Schefe J H,Lehmann K E,Buschmann I R,et al. Quantitative real-time RT-PCR data analysis:current concepts and the novel "gene expression's CT difference" formula[J]. Journal of Molecular Medicine,2006,84(11):901-910.
20. 奚才华,王慧博,赵世光,等. SYBR Green实时定量PCR检测人原发脑胶质瘤中REV3和REV7基因的表达[J]. 中国神经精神疾病杂志,2008,34(8):491-493.Xi C H,Wang H B,Zhao S G,et al. Expression of REV3 and REV7 genes in human primary gliomasby Green SYBR real-time quantitative PCR[J]. China Journal of Nervous and Mental Diseases,2008,34(8):491-493.
21. Wang Z. Translesion synthesis by the UmuC family of DNA polymerases[J]. Mutation Research/fundamental & Molecular Mechanisms of Mutagenesis,2001,486(2):59-70.
22. Johnson R E,Washington M T,Haracska L,et al. Eukaryotic polymerases ι and ζ act sequentially to bypass DNA lesions[J]. Nature,2000,406(6799):1015-1019.
23. Ohashi E,Bebenek K,Matsuda T,et al.Fidelity and processivity of DNA synthesis by DNA polymerase κ,the product of the human DINB1 gene[J].Journal of Biological Chemistry,2000,275(50):39678-39684.
24. Xiao W,Lechler T,Chow B L,et al.Identification,chromosomal mapping and tissue-specific expression of hREV3 encoding a putative human DNA polymerase ζ[J].Carcinogenesis,1998,19(5):945-949.
25. López-Cabrera M,Santis A G,Fernández-Ruiz E,et al.Molecular cloning,expression,and chromosomal localization of the human earliest lymphocyte activation antigen AIM/CD69,a new member of the C-type animal lectin superfamily of signal-transmitting receptors[J].Journal of Experimental Medicine,1993,178(2):537-547.
26. Morelli C,Mungall A J,Negrini M,et al.Alternative splicing,genomic structure,and fine chromosome localization of REV3L[J].Cytogenetics and Cell Genetics,1998,83(1-2):18-20.
27. Murakumo Y,Ogura Y,Ishii H,et al.Interactions in the error-prone postreplication repair proteins hREV1,hREV3 and hREV7[J].Journal of Biological Chemistry,2001,276(38):35644-35651.
28. Lindahl T,Wood R D.Quality control by DNA repair[J].Science,1999,286(5446):1897-1905.
29. Nelson J R,Lawrence C W,Hinkle D C.Thymine-thymine dimer bypass by yeast DNA polymerase ζ[J].Science,1996,272(5268):1646-1649.
30. Nelson J R,Lawrence C W,Hinkle D C.Deoxycytidyl transferase activity of yeast REV1 protein[J].Nature,1996,382(6593):729-731.

[1]	隋德宗, 王保松. 盐胁迫下乌桕无性系叶片的比较蛋白组学研究[J]. 植物研究, 2023, 43(5): 679-689.
[2]	徐磊, 胥晓, 刘沁松. 外源水杨酸对盐胁迫下珙桐幼苗抗氧化系统和基因表达的影响[J]. 植物研究, 2023, 43(4): 572-581.
[3]	廖诗贤, 王宇婷, 董立本, 顾咏梅, 贾丰璘, 姜廷波, 周博如. 小黑杨转录因子PsnbZIP1应答盐胁迫功能分析[J]. 植物研究, 2023, 43(2): 288-299.
[4]	刘森尧, 贾丰璘, 国庆, 樊高锋, 周博如, 姜廷波. 小黑杨转录因子PsnbHLH162基因在盐和低温胁迫下应答分析[J]. 植物研究, 2023, 43(2): 300-310.
[5]	黄安瀛, 夏德安, 张洋, 那冬晨, 燕青, 魏志刚. PtrWRKY51基因的克隆及抗旱表达特性分析[J]. 植物研究, 2022, 42(6): 1005-1013.
[6]	陈华峰, 代龙军, 刘明洋, 郭冰冰, 杨洪, 王立丰. 橡胶树胶乳高表达热激蛋白HbHSP90.4基因抗逆功能分析[J]. 植物研究, 2022, 42(6): 1023-1032.
[7]	岳莉然, 刘颖婕, 刘晨旭, 周蕴薇. 响应盐胁迫调控的露地菊miR398a的克隆及功能研究[J]. 植物研究, 2022, 42(6): 986-996.
[8]	刘明洋, 肖化兴, 王立丰, 梁晓宇, 张宇, 王萌. 橡胶树热激蛋白HbHSP90.8-1基因的克隆与功能分析[J]. 植物研究, 2022, 42(5): 811-820.
[9]	李登高, 林睿, 穆青慧, 周娜, 张焱如, 白薇. 马铃薯StNPR4基因的克隆与功能分析[J]. 植物研究, 2022, 42(5): 821-829.
[10]	王宏鹏, 李一丹, 汪耀, 谭晓宇, 陈成彬, 张力鹏. 菊叶薯蓣DcPMK基因克隆及互作蛋白筛选[J]. 植物研究, 2022, 42(5): 855-865.
[11]	陈娇娆, 续旭, 胡章立, 杨爽. 植物感受盐胁迫及相关钙信号的研究进展[J]. 植物研究, 2022, 42(4): 713-720.
[12]	程赫, 田双慧, 张洋, 刘聪, 夏德安, 魏志刚. 毛果杨nsLTP基因家族全基因组水平鉴定及其表达特性分析[J]. 植物研究, 2022, 42(3): 412-423.
[13]	杨宇宁, 董昊, 董实伟, 王乃锐, 宋跃, 张含国, 李淑娟. 长白落叶松转录因子LobHLH34克隆及表达分析[J]. 植物研究, 2022, 42(1): 112-120.
[14]	马霜, 王博雅, 曹颖, 胡尚连, 高志民. 毛竹扩展蛋白基因的鉴定及其表达分析[J]. 植物研究, 2022, 42(1): 29-38.
[15]	田双慧, 程赫, 张洋, 刘聪, 夏德安, 魏志刚. 毛果杨类胡萝卜素裂解双加氧酶基因家族全基因组水平鉴定及其干旱与盐胁迫响应分析[J]. 植物研究, 2021, 41(6): 993-1005.