84K杨PagC3H3基因表达模式分析

doi:10.7525/j.issn.1673-5102.2019.04.006

植物研究 ›› 2019, Vol. 39 ›› Issue (4): 521-528.doi: 10.7525/j.issn.1673-5102.2019.04.006

84K杨PagC3H3基因表达模式分析

樊二勤, 刘彩霞, 付鹏跃, 杨传平, 曲冠证

林木遗传育种国家重点实验室, 东北林业大学, 哈尔滨 150040

收稿日期:2019-01-03 出版日期:2019-07-05 发布日期:2019-07-03
通讯作者: 杨传平 E-mail:yangcp@nefu.edu.cn
作者简介:樊二勤(1992-),男,硕士研究生,主要从事林木遗传育种研究。
基金资助:
国家自然科学基金项目（31670675）；教育部新世纪优秀人才支持计划项目（NCET-12-0808）

Expression Pattern of PagC3H3 Gene in Populus alba×P.glandulosa

FAN Er-Qin, LIU Cai-Xia, FU Peng-Yue, YANG Chuan-Ping, QU Guan-Zheng

State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040

Received:2019-01-03 Online:2019-07-05 Published:2019-07-03
Supported by:
National Natural Science Foundation of China(31670675);Ministry of Education New Century Excellent Talent Support Program(NCET-12-0808)

摘要/Abstract

摘要： 木质素作为木材的主要组成成分，通常是由3种单体聚合而成，在其生物合成过程中，共有10个酶家族参与负责将苯丙胺酸转化为单体木质素，其中C3H是在对-香豆酰辅酶A（p-coumaroyl CoA）到咖啡酰辅酶A（caffeoyl CoA）的羟基化过程和G/S单体形成中的关键控制酶类，探究PagC3H3基因表达模式，对于进一步了解该基因功能具有重要意义。该研究通过定量PCR对PagC3H3基因的组织特异性表达进行分析；克隆得到了长度为2 035 bp的PagC3H3的启动子序列，预测含有多个顺式作用元件；同时，将获得的PagC3H3的启动子序列构建植物表达载体pBI121-PagC3H3pro：：GUS，进行拟南芥瞬时转化，结果显示PagC3H3基因在84K杨的根、中部茎节和基部茎节中的表达量较高；瞬时转化拟南芥，GUS染色表明：在下胚轴和根中GUS活性较强，由此推测PagC3H3基因在木质素合成过程中发挥作用。

关键词: 84K杨, PagC3H3, 启动子, 拟南芥, 瞬时转化, GUS检测

Abstract: As the main component of wood, lignin is usually polymerized from three monomers. During its biosynthesis, a total of ten enzyme families are involved in the conversion of phenylalanine to monomeric lignin, of which C3H is the key control enzymes in the hydroxylation process of p-coumaroyl CoA to caffeoyl CoA and in the formation of G/S monomer. Exploring the expression pattern of PagC3H3 gene is of great significance for further understanding of the function of this gene. In this study, the tissue-specific expression of PagC3H3 gene was analyzed by quantitative PCR; the promoter sequence of PagC3H3 with a length of 2 035 bp was cloned, and it was predicted to contain multiple cis-acting elements. At the same time, the promoter sequence of PagC3H3 was constructed to construct a plant expression vector pBI121-PagC3H3pro::GUS for transient transformation of Arabidopsis thaliana. The expression of PagC3H3 gene was higher in the roots, middle stem segments and basal stem segments of Populus alba×P. grandulos; transient transformation of A. thaliana showed that GUS activity was stronger in hypocotyls and roots, suggesting that PagC3H3 gene plays a role in lignin synthesis.

Key words: Populus alba×P. grandulos, PagC3H3 gene, promoter, Arabidopsis thaliana, transient transformation, GUS detection

中图分类号:

Q943

樊二勤, 刘彩霞, 付鹏跃, 杨传平, 曲冠证. 84K杨PagC3H3基因表达模式分析[J]. 植物研究, 2019, 39(4): 521-528.

FAN Er-Qin, LIU Cai-Xia, FU Peng-Yue, YANG Chuan-Ping, QU Guan-Zheng. Expression Pattern of PagC3H3 Gene in Populus alba×P.glandulosa[J]. Bulletin of Botanical Research, 2019, 39(4): 521-528.

参考文献

1. Sarkanen K V. Renewable resources for the production of fuels and chemicals[J]. Science,1976,191(4228):773-776.
2. Chiang V L. From rags to riches[J]. Nature Biotechnology,2002,20(6):557-558.
3. Lee B R,Kim K Y,Jung W J,et al. Peroxidases and lignification in relation to the intensity of water-deficit stress in white clover(Trifolium repens L.)[J]. Journal of Experimental Botany,2007,58(6):1271-1279.
4. Jubany-Marí T,Munné-Bosch S,López-Carbonell M,et al. Hydrogen peroxide is involved in the acclimation of the Mediterranean shrub,Cistus albidus L. to summer drought[J]. Journal of Experimental Botany,2009,60(1):107-120.
5. Xu Q,Yin X R,Zeng J K,et al. Activator-and repressor-type MYB transcription factors are involved in chilling injury induced flesh lignification in loquat via their interactions with the phenylpropanoid pathway[J]. Journal of Experimental Botany,2014,65(15):4349-4359.
6. Scurfield G. Reaction Wood:its structure and function:Lignification may generate the force active in restoring the trunks of leaning trees to the vertical[J]. Science,1973,179(4074):647-655.
7. Timell T E. Compression wood in gymnosperms. Volume 1:bibliography,historical background,determination,structure,chemistry,topochemistry,physical properties,origin,and formation of compression wood[J]. The Quarterly Review of Biology,1987,62(4):449.
8. Dixon R A. Natural products and plant disease resistance[J]. Nature,2001,411(6839):843-847.
9. 高原,陈信波,张志扬. 木质素生物合成途径及其基因调控的研究进展[J]. 生物技术通报,2007,(2):47-51.
Gao Y,Chen X B,Zhang Z Y. Advances in research on lignin biosynthesis and its molecular regulation[J]. Biotechnology Bulletin,2007,(2):47-51.
10. Hu W J,Harding S A,Lung J,et al. Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees[J]. Nature Biotechnology,1999,17(8):808-812.
11. 王雪霞,薛永常,赵文超. 木质素生物合成中C3H/HCT的研究进展[J]. 生命的化学,2008,28(5):650-653.
Wang X X,Xue Y C,Zhao W C. The research progress of C3H/HCT in lignin biosynthesis[J]. Chemistry of Life,2008,28(5):650-653.
12. Schoch G,Goepfert S,Morant M,et al. CYP98A3 from Arabidopsis thaliana is a 3'-hydroxylase of phenolic esters,a missing link in the phenylpropanoid pathway[J]. Journal of Biological Chemistry,2001,276(39):36566-36574.
13. Franke R,Humphreys J M,Hemm M R,et al. The Arabidopsis REF8 gene encodes the 3-hydroxylase of phenylpropanoid metabolism[J]. The Plant Journal,2002,30(1):13-45.
14. Anterola A M,Jeon J H,Davin L B,et al. Transcriptional control of monolignol biosynthesis in Pinus taeda:factors affecting monolignol ratios and carbon allocation in phenylpropanoid metabolism[J]. The Journal of Biological Chemistry,2002,277(21):18272-18280.
15. Brown G R,Gill G P,Kuntz R J,et al. Nucleotide diversity and linkage disequilibrium in loblolly pine[J]. Proceedings of the National Academy of Sciences of the United States of America,2004,101(42):15255-15260.
16. Gang D R,Beuerle T,Ullmann P,et al. Differential production of meta hydroxylated phenylpropanoids in sweet basil peltate glandular trichomes and leaves is controlled by the activities of specific acyltransferases and hydroxylases[J]. Plant Physiology,2002,130(3):1536-1544.
17. Fao(Food and Agriculture Organization of the United Nations). State of the World's Forests 2012[R]. Rome,Italy:FAO,2012.
18. Chen F,Dixon R A. Lignin modification improves fermentable sugar yields for biofuel production[J]. Nature Biotechnology,2007,25(7):759-761.
19. 丁浩,郑唐春,梁德洋,等. 一种简易高效提取多种植物纯净DNA的方法[J]. 植物研究,2015,35(3):457-461.
Ding H,Zheng T C,Liang D Y,et al. A simple and efficient method of DNA extraction from different plant samples[J]. Bulletin of Botanical Research,2015,35(3):457-461.
20. Ji X Y,Zheng L,Liu Y J,et al. A transient transformation system for the functional characterization of genes involved in stress response[J]. Plant Molecular Biology Reporter,2014,32(3):732-739.
21. Li L G,Cheng X F,Leshkevich J,et al. The last step of syringyl monolignol biosynthesis in angiosperms is regulated by a novel gene encoding sinapyl alcohol dehydrogenase[J]. The Plant Cell,2001,13(7):1567-1585.
22. Coleman H D,Park J Y,Nair R,et al. RNAi-mediated suppression of p-coumaroyl-CoA 3'-hydroxylase in hybrid poplar impacts lignin deposition and soluble secondary metabolism[J]. Proceedings of the National Academy of Sciences of the United States of America,2008,105(11):4501-4506.
23. 李杰,张福城,王文泉,等. 高等植物启动子的研究进展[J]. 生物技术通讯,2006,17(4):658-661.
Li J,Zhang F C,Wang W Q,et al. Advance in the study of higher plant promoter[J]. Letters in Biotechnology,2006,17(4):658-661.
24. 聂丽娜,夏兰琴,徐兆师,等. 植物基因启动子的克隆及其功能研究进展[J]. 植物遗传资源学报,2008,9(3):385-391.
Nie L N,Xia L Q,Xu Z S,et al. Progress on cloning and functional study of plant gene promoters[J]. Journal of Plant Genetic Resources,2008,9(3):385-391.
25. 李一琨,王金发. 高等植物启动子研究进展[J]. 植物学通报,1998,15(S1):1-6.
Li Y K,Wang J F. Advances of the studies on plant promoter[J]. Chinese Bulletin of Botany,1998,15(S1):1-6.

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84K杨PagC3H3基因表达模式分析

Expression Pattern of PagC3H3 Gene in Populus alba×P.glandulosa

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