Expression Analysis of Lignin Biosynthesis Genes of Pennisetum purpureum Leaf and Stem

doi:10.7668/hbnxb.20190867

Abstract

Abstract: In order to systematically study the expression changes of lignin biosynthesis genes in the stems and leaves of elephant grass,the lignin content and lignin biosynthesis gene expressions were determined in the first,third and fifth stem internodes,as well as the top first,third,fifth,seventh and ninth leaf of N51 elephant grass. The contents of lignin in stem internodes and leaves increased from the head to the base of the mature elephant grass. Real-time quantitative PCR analysis showed that the expression of different lignin biosynthesis genes was tissue-specific. The expression of COMT, C3H and HCT4 genes in leaf tissue was significantly lower than that in stem tissues. For example, COMT had the highest expression level in the fifth internode,which was about 6 times that in leaves. The 4CL and F5H gene expressions were significantly higher than those in the stem tissues. For example, 4CL had the highest expression level in the ninth leaf,which was about 14 times the expression level of the first internode. Correlation analysis showed that the lignin content in stems was positively correlated with the expression of F5H, HCT4 and CCoAOMT genes,and negatively correlated with the expression of C3H gene,while the lignin content in leaves was positively correlated with the expression of 4CL gene,and negatively correlated with the expression of C4H, COMT, HCT4 and CCR genes. This result indicated that different lignin synthetases showed obvious expression differences in different tissues,and the main lignin synthesized was also different in different tissues.

Key words: Elephant grass, Lignin biosynthesis, Lignol, Real-time fluorescence quantitative PCR, Gene expression

CLC Number:

FU Chengcheng, HUANG Huaian, ZHOU Mengdie, ZHONG Tianxiu, ZHANG Xiangqian, CHEN Shu, XIE Xinming. Expression Analysis of Lignin Biosynthesis Genes of Pennisetum purpureum Leaf and Stem[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 87-95. doi: 10.7668/hbnxb.20190867.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20190867

http://www.hbnxb.net/EN/Y2020/V35/I4/87

References

[1] 冯鹤翔, 涂轶. 木质素生物合成的研究[J]. 青岛大学学报(自然科学版),2018,31(1):46-54.doi:10.3969/j.issn.1006-1037.2018.02.10. Feng H X, Tu Y.Research on lignin biosynthesis[J]. Journal of Qingdao University(Natural Science Edition),2018,31(1):46-54.
[2] 丁霄, 曹彩荣, 李朋波, 吴翠翠, 曹美莲, 杨六六. 植物木质素的合成与调控研究进展[J]. 山西农业科学,2016,44(9):1406-1411.doi:10.3969/j.issn.1002-2481.2016.09.43. Ding X, Cao C R, Li P B, Wu C C, Cao M L, Yang L L. Research progress on synthesis and regulation of plant lignin[J]. Journal of Shanxi Agricultural Sciences, 2016, 44(9):1406-1411.
[3] 陈磊. 石榴籽粒木质素合成途径关键基因的表达及功能分析[D]. 合肥:安徽农业大学, 2018. Chen L. Expession and functional analysis of pomegranate seed lignin biosynthesis key gene[D]. Hefei:Anhui Agricultural University, 2018.
[4] 李果. 油用牡丹‘凤丹’DFR基因克隆与功能验证及木质素合成基因发掘[D]. 杨凌:西北农林科技大学, 2017. Li G. Molecular cloning and functional analysis of dihydro flavonol 4-reductas (DFR) gene and screening of lignin genes in Paeonia ostii[D]. Yangling:Northwest A&F University, 2017.
[5] 张旭, 王小佳, 黎思辰, 董甜甜, 汪志辉. 柑橘果实粒化过程中木质素生物合成与调控研究进展[J]. 浙江农业学报, 2019, 31(12):2131-2140. doi:10.3969/j.issn.1004-1524.2019.12.22. Zhang X, Wang X J, Li S C, Dong T T, Wang Z H. Research progress of lignin biosynthesis and regulation during granulation of citrus[J]. Acta Agriculturae Zhejiangensis, 2019, 31(12):2131-2140.
[6] 丁忆然.甘蓝型油菜种皮木质素合成相关基因的差异表达分析[D]. 重庆:西南大学, 2019. Ding Y R. Differential expression analysis of lignin synthesis related genes in the seed coat of Brassica napus L.[D]. Chongqing:Southwest University, 2019.
[7] Ralph J, MacKay J J, Hatfield R D, O'Malley D M, Whetten R W, Sederoff R R. Abnormal lignin in a loblolly pine mutant[J]. Science, 1997, 277(5323):235-239. doi:10.1126/science.277.5323.235.
[8] Whetten R W, MacKay J J, Sederoff R R. Recent advances in understanding lignin biosynthesis[J]. Annual Review of Plant Physiology and Plant Molecular Biology, 1998, 49:585-609. doi:10.1146/annurev.arplant.49.1.585.
[9] 郭璇. 木质素模型化合物的合成及解聚研究[D].北京:北京化工大学, 2018. Guo X. Synthesis and depolymerization of lignin model compounds[D]. Beijing:Beijing University of Chemical Technology, 2018.
[10] Somerville C, Youngs H, Taylor C, Davis S C, Long S P. Feedstocks for lignocellulosic biofuels[J]. Science, 2010, 329(5993):790-792. doi:10.1126/science.1189268.
[11] 钟天秀, 李有涵, 李菲, 彭小群, 柯善文, 陈曙, 解新明. 华南象草 Pp4CL 基因的克隆及其转基因烟草木质素含量分析[J]. 西北植物学报, 2015, 35(12):2355-2364. doi:10.7606/j.issn.1000-4025.2015.12.2355. Zhong T X, Li Y H, Li F, Peng X Q, Ke S W, Chen S, Xie X M.Isolation of 4-coumarate COA ligase gene from Pennisetum purpureum cv. Huanan and lignin content analysis of transgenic tobacco plants[J].Acta Botanica Boreali-Occidentalia Sinica, 2015, 35(12):2355-2364.
[12] Liu X H, Shen Y X, Lou L Q, Ding C L, Cai Q S. Copper tolerance of the biomass crops elephant grass (Pennisetum purpureum Schumach), vetiver grass (Vetiveria zizanioides) and the upland reed(Phragmites australis)in soil culture[J]. Biotechnology Advances, 2009, 27(5):633-640. doi:10.1016/j.biotechadv.2009.04.017.
[13] 彭小群. 象草MYB4转录因子及CCR启动子的克隆与功能分析[D]. 广州:华南农业大学, 2016. Peng X Q. Cloning and functional analysis of PpMYB4 transcription factor and PpCCR gene promoter in Pennisetum purpureum[D]. Guangzhou:South China Agricultural University, 2016.
[14] 赵新坤, 李赫龙, 佘雕, 孙润仓. 木质素模型化合物合成研究的现状与展望[J]. 生物质化学工程, 2018,52(1):41-52. doi:10.3969/j.issn.1673-5854.2018.01.007. Zhao X K, Li H L, She D, Sun R C. Status and prospect of lignin model compounds synthesis[J]. Biomass Chemical Engineering, 2018, 52(1):41-52.
[15] 郭永翠, 秦江南, 张锐. 核桃内果皮木质素生物合成途径关键基因研究进展[J].现代园艺, 2019(7):5-8. doi:10.14051/j.cnki.xdyy.2019.07.002. Guo Y C, Qin J N, Zhang R. Research progress on key genes of lignin biosynthesis pathway in walnut endocarp[J]. Xiandai Horticulture, 2019(7):5-8.
[16] 陈永忠, 谭晓风, David Clapham. 木质素生物合成及其基因调控研究综述[J]. 江西农业大学学报(自然科学), 2003, 25(4):613-617. doi:10.13836/j.jjau.2003140. Chen Y Z, Tan X F, Clapham D.Lignin biosynthesis and genetic regulation[J].Acta Agriculturae Universitatis Jiangxiensis(Natural Sciences Edition), 2003,25(4):613-617.
[17] 彭小群, 张博雅, 刘洁雯, 王亚, 解新明. 象草PpCCR基因正、反义表达载体的构建及对烟草的转化[J]. 草业科学, 2015,9(10):1586-1593. doi:10.11829/j.issn.1001-0629.2014-0572. Peng X Q, Zhang B Y, Liu J W, Wang Y, Xie X M.Construction of sense and antisense expression vectors of PpCCR gene in Pennisetum purpureum and transformation to tobacco plants[J]. Pratacultural Science, 2015,9(10):1586-1593.
[18] 唐然, 张博雅, 彭小群, 张向前, 江院, 解新明. 华南象草PpCAD基因的亚细胞定位及其在转基因烟草中的异源表达[J]. 西北植物学报, 2015, 35(6):1069-1077. doi:10.7606/j.issn.1000-4025.2015.06.1069. Tang R, Zhang B Y, Peng X Q, Zhang X Q, Jiang Y, Xie X M. Subcellular localization of PpCAD from elephant grass (Pennisetum purpureum cv. Huanan) and its heterologous expression in transgenic tobacco[J]. Acta Botanica Boreali-Occidentalia Sinica, 2015, 35(6):1069-1077.
[19] 王建庆, 曹佃元, 张玉. 乙酰溴法测定棉籽壳中木质素的含量[J]. 纺织学报, 2013, 34(9):12-16. doi:10.13475/j.fzxb.2013.09.011. Wang J Q, Cao D Y, Zhang Y.Acetyl bromide test method for determining amount of lignin in cottonseed coat[J].Journal of Textile Research, 2013, 34(9):12-16.
[20] 陈博雯, 刘海龙, 肖玉菲, 覃子海, 张烨, 张晓宁. 尾叶桉COMT和CCoAOMT基因定向调控木质素单体合成的烟草转化研究[J]. 中国生物工程杂志, 2018, 38(3):24-32. doi:10.13523/j.cb.20180304. Chen B W, Liu H L, Xiao Y F, Qin Z H, Zhang Y, Zhang X N.Directional regulation of lignin monomer synthesis in tobacco by using COMT gene and CCo AOMT gene of Eucalyptus urophylla[J]. Chinese Biotechnology, 2018, 38(3):24-32.
[21] 张婷. 木质素合成酶基因 4CL和CCoAOMT 反义转化紫穗槐的实验研究[D]. 长春:吉林大学, 2008. Zhang T. Study on antisense transformation with lignin biosynthesis 4CL and CCoAOMT gene in Amorpha fruitcosa[D]. Changchun:Jilin University, 2008.
[22] 师竹娟. 甘蓝型油菜木质素单体合成基因 COMT 和 F5H 的克隆及表达调控[D]. 北京:中国农业科学院, 2006.doi:10.7666/d.Y880672. Shi Z J. Monolignol biosynthesis gene COMT and F5H cloning and expression regulation in Brassica napus L.[D]. Beijing:Chinese Academy of Agricultural Sciences, 2006.
[23] Meyer K, Shirley A M, Cusumano J C, Bell-Lelong D A, Chapple C. Lignin monomer composition is determined by the expression of a cytochrome P450-dependent monooxygenase in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(12):6619-6623. doi:10.1073/pnas.95.12.6619.
[24] Shuai L, Amiri M T, Questell-Santiago Y M, Héroguel F, Li Y D, Kim H, Meilan R, Chapple C, Ralph J, Luterbacher J S. Formaldehyde stabilization facilitates lignin monomer production during biomass depolymerization[J]. Science, 2016, 354(6310):329-333. doi:10.1126/science.aaf7810.
[25] Stewart J J, Akiyama T, Chapple C, Ralph J, Mansfield S D. The effects on lignin structure of overexpression of ferulate 5-Hydroxylase in hybrid poplar[J]. Plant Physiology, 2009, 150(2):621-635. doi:10.1104/pp.109.137059.
[26] Voelker S L, Lachenbruch B, Meinzer F C, Jourdes M, Ki C, Patten A M, Davin L B, Lewis N G, Tuskan G A, Gunter L, Decker S R, Selig M J, Sykes R, Himmel M E, Kitin P, Shevchenko O, Strauss S H. Antisense down-regulation of 4CL expression alters lignification, tree growth, and saccharification potential of field-grown poplar[J]. Plant Physiology, 2010, 154(2):874-886. doi:10.1104/pp.110.159269.
[27] Tu Y, Rochfort S, Liu Z Q, Ran Y D, Griffith M, Badenhorst P, Louie G V, Bowman M E, Smith K F, Noel J P, Mouradov A, Spangenberg G. Functional analyses of caffeic acid O-methyltransferase and Cinnamoyl-CoA-reductase genes from perennial ryegrass (Lolium perenne)[J]. The Plant Cell, 2010, 22(10):3357-3373. doi:10.1105/tpc.109.072827.
[28] Ong R G, Higbee A, Bottoms S, Dickinson Q, Xie D, Smith S A, Serate J, Pohlmann E, Jones A D, Coon J J, Sato T K, Sanford G R, Eilert D, Oates L G, Piotrowski J S, Bates D M, Cavalier D, Zhang Y P. Inhibition of microbial biofuel production in drought-stressed switchgrass hydrolysate[J]. Biotechnology for Biofuels, 2016, 9:237. doi:10.1186/s13068-016-0657-0.
[29] Vanholme R, Cesarino I, Rataj K, Xiao Y G,Sundin L,Goeminne G, Kim H,Cross J, Morreel K, Araujo P, Welsh L, Haustraete J,McClellan C, Vanholme B,Ralph J,Simpson G G,Halpin C,Boerjan W. Caffeoyl shikimate esterase (CSE) is an enzyme in the lignin biosynthetic pathway in Arabidopsis[J]. Science, 2013, 341(6150):1103-1106. doi:10.1126/science.1241602.
[30] Bonawitz N D, Kim J I, Tobimatsu Y, Ciesielski P N, Anderson N A, Ximenes E, Maeda J, Ralph J, Donohoe B S, Ladisch M, Chapple C. Disruption of mediator rescues the stunted growth of a lignin-deficient Arabidopsis mutant[J]. Nature, 2014, 509(7500):376-380. doi:10.1038/nature13084.

Please choose a citation manager

Content to export