Antisense CCoAOMT Gene Regulates Lignin Biosynthesis in Betula platyphylla

doi:10.7525/j.issn.1673-5102.2019.01.015

Abstract

Abstract: Betula platyphylla is an important forestation tree species in northern China, but its high lignin content severely restricts its development and utilization as a papermaking resource plant. In this paper, the full-length ORF sequence of CCoAOMT(Caffeoyl-coenzyme A-3-O-methyltransferase) gene was obtained using RACE technology. The antisense CCoAOMT expression vector was constructed and transformed into B.platyphylla by Agrobacterium infection method. The molecular detection indicated that the CCoAOMT antisense gene had been successfully integrated into B.platyphylla. By semi-quantitative PCR analysis of the transgenic plants, the CCoAOMT gene expression of the transgenic lines was decreased. Compared to the wild type, by the Wiesner staining, the lignin content of the transgenic plants was decreased. Chemical compositions of seven-year-old transgenic and wild-type birch were analyzed. The phenyl alcohol extracts and Klason lignin of transgenic birch were significantly reduced, and the pentosan content was increased. The BpCCoAOMT gene plays an important role in the synthesis of lignin, which lays a foundation for the cultivation of a new species of birch with low lignin content.

Key words: CCoAOMT gene, antisense RNA, birch, lignin, Wiesner stain, transgene

CLC Number:

Q786

YAO Lian-Mei, HU Xiao-Qing, ZHOU Fei, ZHENG Yao-Qiang, WANG Guo-Dong. Antisense CCoAOMT Gene Regulates Lignin Biosynthesis in Betula platyphylla[J]. Bulletin of Botanical Research, 2019, 39(1): 123-130.

References

1. 宋福南. 白桦木质素合成苯丙氨酸途径相关酶基因的表达和功能分析[D]. 哈尔滨:东北林业大学,2009. Song F N. Expression and functional analysis of correlative enzyme gene of lignin Phenylpropanoid Biosynthetic Pathway in Betula Platphylla[D]. Harbin:Northeast Forestry University,2009.
2. Ye Z H. Association of caffeoyl coenzyme A3-O-methyltransferase expression with lignifying tissues in several dicot plants[J]. Plant Physiology,1997,115(4):1341-1350.
3. 黄春琼,刘国道,郭安平. 反义CCoAOMT基因调控烟草木质素的生物合成[J]. 安徽农业科学,2008,36(19):8026-8027,8043. Huang C Q,Liu G D,Guo A P. Lignin biosynthesis regulated by antisense CCoAOMT gene in tobacco[J]. Journal of Anhui Agricultural Sciences,2008,36(19):8026-8027,8043.
4. 王玲,田敏,段红平,等. 反义CCoAOMT基因转化烟草调控木质素生物合成[J]. 江苏农业科学,2011,39(2):62-64. Wang L,Tian M,Duan H P,et al. Study on content and ratio of lignin in transformed tobacco plants changed by anti-sense CCoAOMT[J]. Jiangsu Agricultural Science,2011,39(2):62-64.
5. 倪志勇,吕萌,范玲. 棉花咖啡酰辅酶A-O-甲基转移酶基因的克隆及特征分析[J]. 西北植物学报,2010,30(6):1083-1091. Ni Z Y,Lü M,Fan L. Cloning and characterization of CCoAOMT gene from Gossypium hirsutum L.[J]. Acta Botanica Boreali-Occidentalia Sinica,2010,30(6):1083-1091.
6. Guo D J,Chen F,Inoue K,et al. Downregulation of caffeic acid 3-O-methyltransferase and caffeoyl CoA 3-O-methyltransferase in transgenic alfalfa. Impacts on lignin structure and implications for the biosynthesis of G and S lignin[J]. The Plant Cell,2001,13(1):73-88.
7. 刘红梅,胡尚连,卢学琴,等. 青杨CCoAOMT基因的克隆及其生物信息学分析[J]. 湖北农业科学,2014,53(11):2670-2674. Liu H M,Hu S L,Lu X Q,et al. Cloning and bioinformatics analysis of CCoAOMT gene of Populus cathayana Rehd[J]. Hubei Agricultural Science,2014,53(11):2670-2674.
8. 赵华燕,魏建华,路静,等. 利用反义CCoAOMT基因培育低木质素含量毛白杨的研究[J]. 自然科学进展,2004,14(9):1067-1071. Zhao H Y,Wei J H,Lu J,et al. Studies on the cultivation of Populus tomentosa with low lignin content by antisense CCoAOMT gene[J]. Progress in Natural Sciences,2004,14(9):1067-1071.
9. 魏建华,王彦珍,王宏芝,等. 抑制CCoAOMT表达的转基因杨树的制浆性能[J]. 科学通报,2008,53(21):2612-2616. Wei J H,Wang Y Z,Wang H Z,et al. Pulping performance of transgenic poplar with depressed Caffeoyl-CoA O-methyltransferase[J]. Chinese Science Bulletin,2008,53(22):3553-3558.
10. 张学科,刘文俊,朱明华,等. 木质素酚醛树脂的研究进展[J]. 植物研究,2012,32(6):760-764,769. Zhang X K,Liu W J,Zhu M H,et al. Research progress of phenol-formaldehyde resin[J]. Bulletin of Botanical Research, 2012,32(6):760-764,769.
11. 赵华燕. 利用反义RNA技术进行木质素生物合成调控的研究[D]. 北京:中国科学院研究生院,2004. Zhao H Y. Studies on the regulation of lignin biosynthesis by antisense RNA technique[D]. Beijing:Graduate School of Chinese Academy of Sciences:Institute of Botany,2004.
12. Shi R,Sun Y H,Li Q Z,et al. Towards a systems approach for lignin biosynthesis in Populus trichocarpa:transcript abundance and specificity of the monolignol biosynthetic genes[J]. Plant and Cell Physiology,2009,51(1):144-163.
13. 陈肃. 白桦4CL与CCoAOMT基因表达分析及蛋白预测[D]. 哈尔滨:东北林业大学,2009. Chen S. Expression and forecast of 4CL and CCoAOMT of Birch[D]. Harbin:Northeast Forestry University,2009.
14. 姜爽. CCoAOMT和4CLcDNA的克隆及转基因紫穗槐的研究[D]. 长春:吉林大学,2007. Jiang S. Cloning of two cDNA encoding CCoAOMT and 4CL and their transformation in Amorpha fruitcosa[D]. Changchun:Jilin University,2007.
15. 赵华燕,魏建华,宋艳茹. 木质素生物合成及其基因工程研究进展[J]. 植物生理与分子生物学学报,2004,30(4):361-370. Zhao H Y,Wei J H,Song Y R. Advances in research on lignin biosynthesis and its genetic engineering[J]. Journal of Plant Physiology and Molecular Biology,2004,30(4):361-370.
16. Wagner A,Tobimatsu Y,Phillips L,et al. CCoAOMT suppression modifies lignin composition in Pinus radiata[J]. The Plant Journal,2011,67(1):119-129.
17. 贾彩红. 低木质素含量的转基因毛白杨(Populus tomentosa)的获得与毛白杨4CL基因启动子的克隆[D]. 保定:河北农业大学,2004. Jia C H. Obtaining the transgenic poplars with low lignin content and cloning of 4CL promoter from Chinese white poplar(Populus tomentosa)[D]. Baoding:Hebei Agricultural University,2004.
18. 徐洋,蔺占兵,马庆虎. 木质素与人类生活[J]. 植物杂志,2002(4):40-41. Xu Y,Lin Z B,Ma Q H. Lignin and human life[J]. Plants,2002(4):40-41.
19. Umezawa T. The cinnamate/monolignol pathway[J]. Phytochemistry Reviews,2010,9(1):1-17.
20. 涂红艳,刘元风. 反义RNA的作用机理及其在植物基因工程领域的应用[J]. 生物磁学,2005(02):32-34. Tu H Y,Liu Y F. The mechanism of antisense RNA and its application in plant gene engineering[J]. Biomagnetism. 2005(02):32-34.
21. 马兴亮,刘耀光. 植物CRISPR/Cas9基因组编辑系统与突变分析[J]. 遗传,2016,38(2):118-125. Ma X L,Liu Y G. CRISPR/Cas9-based genome editing systems and the analysis of targeted genome mutations in plants[J]. Hereditas,2016,38(2):118-125.

[1]	Chaoran SHAN, Xiaohu CHEN, Yunfei DING, Wei ZHAO, Han LU, Shangzhu GAO, Fenghui QI, Yaguang ZHAN, Fansuo ZENG. Functional Analysis of FmCCoAOMT Gene in Fraxinus mandshurica During Lignin Synthesis and Abiotic Stress [J]. Bulletin of Botanical Research, 2023, 43(5): 768-778.
[2]	Ping LUO, Haonan ZHANG, Jianmin XU, Bing HU, Xiaoping WANG, Guangyou LI, Chunjie FAN. Establishment of Agrobacterium rhizogenes-mediated Genetic Transformation System of Eucalyptus urophylla × E. grandis [J]. Bulletin of Botanical Research, 2022, 42(3): 512-520.
[3]	Yibo Yin, Jixiang Li, Yingjie Guo, Ziting Lu, Ying Xiao, Hualing Liu, Yaguang Zhan, Fansuo Zeng. Variation of Lignin Content and Association Analysis of FmPAL Nucleotide Polymorphism in Progenies of Interspecific Hybrids of Fraxinus [J]. Bulletin of Botanical Research, 2022, 42(2): 191-199.
[4]	Dong-Lin LÜ, Teng LI, Yi-Wen GUO, Jing JIANG, Hai-Jiao HUANG. Determination of Seed Vigor and Genetic Analysis of Foreign Genes in Different Transgenic Birch Hybrids [J]. Bulletin of Botanical Research, 2021, 41(4): 564-572.
[5]	LI Ying, LI Lin-Xi, ZHANG Yu-Qi, WANG Yu, ZHAN Ya-Guang, YIN Jing. An Efficient Method for Separation and Purification of Betulin from Birch Bark [J]. Bulletin of Botanical Research, 2020, 40(3): 468-475.
[6]	REN Shuo-Qi, LIU Bing-Yang, LI Xue-Ying, XING Jian-Hui, LI Zheng-Lun, WANG Chu, GANG Hui-Xin, LIU Gui-Feng, JIANG Jing. Analysis of Leaf Color Variation and Height Growth Characteristics of Yellow-green Leaf Mutant in Birch [J]. Bulletin of Botanical Research, 2018, 38(6): 852-859.
[7]	JIANG Hai-Gang, WANG Shu-Gang, LIU Yang, SHEN Yan-He, LI Xiang-Ning, CAO Ying. Transcriptome-wide Identification of Lignin Gene Families of Bambusa emeiensis Shoots and Their Differential Expression [J]. Bulletin of Botanical Research, 2018, 38(3): 415-421.
[8]	SUN Feng-Kun, LI Si-Da, LI Ji-Xiang, CHEN Xiao-Hui, ZENG Fan-Suo. Effects of Exogenous Nitric Oxide on Exogenous Gene Expression and DNA Methylation in Transgenic Birch [J]. Bulletin of Botanical Research, 2017, 37(6): 870-875.
[9]	SHEN Ting-Ting, JIANG Jing, LIU Gui-Feng, XU Si-Jia, LI Hui-Yu, YUAN Hong-Mei. Bioinformatics and Expression Analysis of BpCesA Genes of Betula platyphylla [J]. Bulletin of Botanical Research, 2016, 36(6): 909-916.
[10]	WANG Yan-Hong1,2；JIA Qing-Bin1；ZHANG Lei1；ZHANG Han-Guo1. Allelic Variation in 4-coumarate:CoA ligase(4CL) Associated with Wood Properties of Larix olgensis* [J]. Bulletin of Botanical Research, 2016, 36(2): 242-251.
[11]	XU Lei1,2；MENG Yong-Bin1,2；ZHANG Zhi-Dong1,2；LU Qi1,2；LIU Ying1,2；MENG Qing-Huan1,2；NIE Si-Ming1,2；ZU Yuan-Gang1,2*. [J]. Bulletin of Botanical Research, 2016, 36(2): 316-320.
[12]	LI Wen-Gang；SUN Xiao-Li；GE Yun-Long；ZHAO Xiu-Hua；ZU Yuan-Gang*. Economic Dynamics Performance Evaluation of Lignin Diesel [J]. Bulletin of Botanical Research, 2016, 36(1): 146-150.
[13]	LU Qi1,2；MENG Yong-Bin1,2；XU Lei1,2；ZHANG Zi-Dong1,2；LIU Ying1,2；MENG Qing-Huan1,2；ZU Yuan-Gang1,2*. Hydroxymethylation of Lignin from Ethanol Pulping [J]. Bulletin of Botanical Research, 2016, 36(1): 151-155.
[14]	ZOU Li1；ZHANG Guo-Quan1；SAXI Yaertu2；YU Yang1；TANG Qing-Ming3. Plant Diversity of Virgin Broadleaved-Korean Pine Forest and Birch Secondary Forest in Liangshui [J]. Bulletin of Botanical Research, 2015, 35(6): 945-951.
[15]	LIU Yu1；XU Huan-Wen1；LI Zhi-Xin1；TENG Wen-Hua1；ZHANG Lian-Fei2；ZOU Jian-Jun3；SHANG Fu-Qiang4；LIU Gui-Feng1*. Growth Variation and Stability Analysis of Birch Crossbreed Families [J]. Bulletin of Botanical Research, 2015, 35(6): 937-944.