白桦CESA7基因启动子的克隆与表达分析

doi:10.7525/j.issn.1673-5102.2018.04.009

摘要/Abstract

摘要： 克隆得到了一个白桦纤维素合成酶基因（CESA7）GenBanK登录号（EU591531）启动子序列，通过序列分析发现该启动子含有多个不同功能的顺式作用元件，包括光响应元件、激素响应元件、叶片形态发育元件等，推测该启动子在白桦生长发育过程中具有关键作用。将BpCESA7启动子克隆至带有GUS报告基因的植物表达载体，命名为proBpCESA7-121-GUS，并利用农杆菌介导方法侵染白桦和拟南芥，然后通过GUS组织化学染色观察BpCESA7基因启动子的组织表达特性。结果在白桦的根、茎、叶和拟南芥的根，叶，萼片、雌蕊中检测到了GUS活性，说明BpCESA7基因启动子具有启动子活性，并且在白桦的根和叶中染色最深，表明BpCESA7基因在白桦根和叶中表达量较高，并且其存在组织表达特异性。

关键词: 白桦, CESA7, 启动子, 表达, GUS染色

Abstract: A cellulosic synthase gene Betula platyphylla(CESA7) GenBank ID(EU591531) promoter was cloned and analyzed. By sequence analysis, the promoter was found to contain multiple cis-acting elements with different functions, including response elements, hormone-responsive elements, leaf morphology development elements, etc. It was speculated that this promoter plays a key role in the growth and development of birch. We cloned the promoter of BpCESA7 into a plant expression vector with a GUS reporter gene, named proBpCESA7-121-GUS, and invaded B.platyphylla and Arabidopsis thaliana by Agrobacterium-mediated method, and then observed that BpCESA7 gene was activated by GUS histochemical staining. Sub-organizational expression characteristics. Results GUS activity was detected in the roots, stems and leaves of B.platyphylla and Arabidopsis roots, leaves, sepals, and stamens, indicating that the promoter of BpCESA7 gene had promoter activity and was stained the most in the roots and leaves of birch, indicating BpCESA7 genes are highly expressed in the birch roots and leaves, and their tissue-specific expression is present.

Key words: Betula platyphylla, CESA7, promoter, expression, GUS staining

中图分类号:

S792.153

朱德财, 胡晓晴, 赵桐, 田晶, 张勇, 刘雪梅. 白桦CESA7基因启动子的克隆与表达分析[J]. 植物研究, 2018, 38(4): 551-558.

ZHU De-Cai, HU Xiao-Qing, ZHAO Tong, TIAN Jing, ZHANG Yong, LIU Xue-Mei. Cloning and Expression Analysis of Promoter of CESA7 Gene in Betula platyphylla[J]. Bulletin of Botanical Research, 2018, 38(4): 551-558.

参考文献

1. Delmer D P. Cellulose biosynthesis:exciting times for a difficult field of study[J]. Annual Review of Plant Physiology and Plant Molecular Biology,1999,50:245-276.
2. Arioli T,Peng L C,Betzner A S,et al. Molecular analysis of cellulose biosynthesis in Arabidopsis[J]. Science,1998,279(5351):717-720.
3. Wang L,Guo K,Li Y,et al. Expression profiling and integrative analysis of the CESA/CSL superfamily in rice[J]. Bmc Plant Biology,2010,10(1):282-297.
4. Somerville C. Cellulose synthesis in higher plants[J]. Annual Review of Cell and Developmental Biology,2006,22:53-78.
5. Doblin M S,Kurek I,Jacob-Wilk D,et al. Cellulose biosynthesis in plants:from genes to rosettes[J]. Plant and Cell Physiology,2002,43(12):1407-1420.
6. Persson S,Paredez A,Carroll A,et al. Genetic evidence for three unique components in primary cell-wall cellulose synthase complexes in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America,2007,104(39):15566-15571.
7. Caño-Delgado A,Penfield S,Smith C,et al. Reduced cellulose synthesis invokes lignification and defense responses in Arabidopsis thaliana[J]. The Plant Journal,2003,34(3):351-362.
8. Scheible W R,Eshed R,Richmond T,et al. Modifications of cellulose synthase confer resistance to isoxaben and thiazolidinone herbicides in Arabidopsis Ixr1 mutants[J]. Proceedings of the National Academy of Sciences of the United States of America,2001,98(18):10079-10084.
9. Desprez T,Juraniec M,Crowell E F,et al. Organization of cellulose synthase complexes involved in primary cell wall synthesis in Arabidopsis thaliana[J]. Proceedings of the National Academy of Sciences of the United States of America,2007,104(39):15572-15577.
10. Bischoff V,Desprez T,Mouille G,et al. Phytochrome regulation of cellulose synthesis in Arabidopsis[J]. Current Biology,2011,21(21):1822-1827.
11. Taylor N G,Howells R M,Huttly A K,et al. Interactions among three distinct CesA proteins essential for cellulose synthesis[J]. Proceedings of the National Academy of Sciences of the United States of America,2003,100(3):1450-1455.
12. Turner S R,Somerville C R. Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall[J]. Plant Cell,1997,9(5):689-701.
13. 李田,孙景宽,刘京涛. 植物启动子研究进展[J]. 生物技术通报,2015,31(2):18-25.Li T,Sun J K,Liu J T. Research advances on plant promoter[J]. Biotechnology Bulletin,2015,31(2):18-25.
14. 文添龙,刘雪梅,冀亚萍,等. 高等植物胁迫诱导型启动子的研究进展[J]. 西北植物学报,2014,34(1):206-214.Wen T L,Liu X M,Ji Y P,et al. Research progress of stress-induced promoter in higher plant[J]. Acta Botanica Boreali-Occidentalia Sinica,2014,34(1):206-214.
15. 雒雅婧,李杰,张爽,等. 植物启动子研究进展[J]. 北方园艺,2015(22):186-189.Luo Y J,Li J,Zhang S,et al. Research progress on plant promoter[J]. Northern Horticulture,2015(22):186-189.
16. 李天芳,姜静,杨传平,等. 我国白桦育种研究概况[J]. 江苏林业科技,2008,35(2):47-49.Li T F,Jiang J,Yang C P,et al. Status of breeding research on Betula platyphylla in China[J]. Journal of Jiangsu Forestry Science & Technology,2008,35(2):47-49.
17. 郭勇,王玉成,王智博. 一种基于农杆菌介导的拟南芥瞬时转化技术优化[J]. 东北林业大学学报,2016,44(6):41-44,83.Guo Y,Wang Y C,Wang Z B. Optimizing transient genetic transformation method on Arabidopsis plants mediated by agrobacterium tumefaciens[J]. Journal of Northeast Forestry University,2016,44(6):41-44,83.
18. 李萌,哈努拉·塔斯肯,陈卓,等. 白桦4CL基因启动子克隆及表达分析[J]. 中国农学通报,2017,33(5):29-34.Li M,Hanula·Tusken,Chen Z,et al. Cloning and expression analysis of 4CL gene promoter of Betula platyphylla[J]. Chinese Agricultural Science Bulletin,2017,33(5):29-34.
19. 朱登峰,唐冬英,李明芳,等. 观察植物木质部的3种切片方法比较[J]. 湖南农业科学,2010(7):20-22.Zhu D F,Tang D Y,Li M F,et al. Comparison among three sectioning methods to observe plant xylem[J]. Hunan Agricultural Sciences,2010(7):20-22.
20. Liu X M,Wang Q Y,Chen P P,et al. Four novel cellulose synthase(CESA) Genes from Birch(Betula platyphylla Suk. ) involved in primary and secondary cell wall biosynthesis[J]. International Journal of Molecular Sciences,2012,13(10):12195-12212.
21. 江海霞,郭栋良,李玉环,等. 亚麻快速生长期细胞壁形成相关基因的表达分析[J]. 中国农业科学,2017,50(13):2442-2450.Jiang H X,Guo D L,Li Y H,et al. The expression of genes related to cell wall formation at fast growth stage in flax(Linum usitatissimum L. )[J]. Scientia Agricultura Sinica,2017,50(13):2442-2450.
22. Tanaka K,Murata K,Yamazaki M,et al. Three distinct rice cellulose synthase catalytic subunit genes required for cellulose synthesis in the secondary wall[J]. Plant Physiology,2003,133(1):73-83.
23. 王文芳,刘爱玲,李琦,等. 水稻CesA4基因启动子与GUS基因融合表达[J]. 现代生物医学进展,2009,9(8):1459-1461,1554.Wang W F,Liu A L,Li Q,et al. Localization of rice CesA4 promoter with GUS-fusion[J]. Progress in Modern Biomedicine,2009,9(8):1459-1461,1554.
24. Liang Y K,Xie X D,Lindsay S E,et al. Cell wall composition contributes to the control of transpiration efficiency in Arabidopsis thaliana[J]. Plant Journal for Cell & Molecular Biology,2010,64(4):679-686.

[1]	陈柄华, 张杰, 刘桂丰, 李思婷, 高元科, 李慧玉, 李天芳. 白桦半同胞家系纸浆材优良家系选择及选择方法评价[J]. 植物研究, 2023, 43(5): 690-699.
[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]	张昊楠, 陈珊珊, 徐建民, 罗萍, 王晓萍, 许志茹, 范春节. 巨桉EgrWAT1基因克隆和功能初步分析[J]. 植物研究, 2023, 43(4): 601-611.
[4]	王景哲, 牛朝奎, 梁馨元, 申晨静, 尹静. 水杨酸在白桦苗期抵御盐碱胁迫中的调控作用[J]. 植物研究, 2023, 43(3): 379-387.
[5]	宋海云, 张涛, 贺鹏, 郑树芳, 王立丰, 王文林. 澳洲坚果MibZIP1基因克隆及表达规律分析[J]. 植物研究, 2023, 43(1): 131-139.
[6]	杜金霞, 申婷婷, 王浩然, 林一萍, 李慧玉, 张连飞. 白桦BpSPL9基因抑制表达载体的构建及遗传转化研究[J]. 植物研究, 2023, 43(1): 30-35.
[7]	杨洪, 王立丰, 代龙军, 郭冰冰. 死皮对橡胶树树皮线粒体超微结构及活性氧代谢的影响[J]. 植物研究, 2023, 43(1): 69-75.
[8]	李梦烁, 刘莹泽, 鲁焕, 强胜. 基因转录介导同质园条件下拟南芥不同地理种群的光合能力分化[J]. 植物研究, 2023, 43(1): 90-99.
[9]	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.
[10]	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.
[11]	李登高, 林睿, 穆青慧, 周娜, 张焱如, 白薇. 马铃薯StCRKs基因家族的鉴定分析及响应逆境信号的表达[J]. 植物研究, 2022, 42(6): 1033-1043.
[12]	覃碧, 刘明洋, 王萌, 王立丰, 黄飞. 橡胶树DELLA基因HbRGL1的克隆与表达分析[J]. 植物研究, 2022, 42(6): 997-1004.
[13]	代龙军, 刘明洋, 阳江华, 周凯, 郭冰冰, 杨洪, 王立丰. 巴西橡胶树胶乳中DUF1262结构域蛋白结构与基因表达分析[J]. 植物研究, 2022, 42(5): 802-810.
[14]	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.
[15]	白向东, 顾宸瑞, 张潇月, 姜静. 欧洲白榆UlGLK基因克隆及组织表达特异性研究[J]. 植物研究, 2022, 42(5): 848-854.