五节芒CCoAOMT和4CL的克隆和表达分析

doi:10.3969/j.issn.1000-7091.2012.04.006

华北农学报 ›› 2012, Vol. 27 ›› Issue (4): 29-35. doi: 10.3969/j.issn.1000-7091.2012.04.006

五节芒CCoAOMT和4CL的克隆和表达分析

常瑞娜¹, 汪杏芬³, 陈鸿鹏²

1. 南京大学生命科学院, 江苏南京200091;
2. 中国科学院上海生命科学研究院植物生理生态研究所, 上海200032;
3. 中南林业科技大学, 经济林育种与栽培国家林业局重点实验室, 湖南长沙410004

出版日期:2012-08-28
作者简介:常瑞娜（1986-），女，山西临汾人，在读硕士，主要从事植物遗传改良研究。汪杏芬（1969-），女，浙江上虞人，副研究员，博士，主要从事植物分子生物学和能源植物遗传改良研究。
基金资助:
国家自然科学基金（30972329）

Cloning and Expression Analysis of CCoAOMT and 4CL Genes from Miscanthus floridulus

CHANG Rui-na¹, WANG Xing-fen³, CHEN Hong-peng²

1. Biology College of Nanjing University, Nanjing Ecology, Chinese Academy of Sciences, Shanghai 210091, China;
2. Shanghai Institute of Plant Physiology and 200032, China;
3. The Key Lab of Non-wood Forest Product of Forestry Ministry, Central South University of Forestry and Technology, Changsha 410004, China

Published:2012-08-28

摘要/Abstract

摘要： 木质素是细胞壁的重要组成部分，在植物的生长发育和抵御病原微生物的侵害方面发挥着重要的作用，但木质素结构稳定，成为生物质能源转化的障碍。以五节芒（Miscanthus floridulus）为材料克隆得到了木质素合成过程中的关键酶CCoAOMT基因全长CDS序列和4CL基因的CDS半长序列，并对其CDS和氨基酸序列与其他物种进行了比对分析。相应的qRT-PCR分析显示，2个基因在五节芒的幼苗期、抽穗期的茎节、成熟叶片、幼穗都有表达，且与木质素的合成有一定的相关性。间苯三酚溶液染色结果亦显示，2个基因的表达规律与茎秆的木质化进程一致。五节芒CCoAOMT和4CL基因的克隆和表达分析将为进一步研究其生物学功能和改良能源作物奠定基础

关键词: 五节芒, 木质素, 咖啡酰辅酶A-O-甲基转移酶（CCoAOMT）, 4-香豆酸辅酶A连接酶（4CL）

Abstract: Lignin,a major component of the cell wall,plays an important role in the development of plant and defense to pathogens, however,its stable structure makes lignin become a main barrier for transforming biomass into biofuels. In this study, the full and half length CDS of CCoAOMT（ Caffeoyl CoA 3-O-methyhransferase） and 4CL （4-Coumarate：coenzyme A ligase）,which are key enzymes in the phenylpropanoid metabolic pathways for monoli- gnol biosynthesis,were cloned respectively from Miscanthus floridulus, and named as MfCCoAOMT and Mf4CL. The expression patterns of these lignin genes were investigated in stalk/mature leaf/young panicle tissues at seedling/ early heading/late heading stages with qRT-PCR. Results indicated that MfCCoAOMT and Mf4CL.expressed in every different tissue at 3 developmental stages. The expression patterns of CCoAOMT and 4CL showed close correlation with the synthesis of lignin. The spasfon staining results also demonstrated that the expression of both genes consis- ted with the progress of stem lignification during plant development. The gone cloning and expression analysis of Mf- COA and Mf4CL provided reference for gene function study and modification of lignin content of miscanthus.

Key words: Miscanthus floridulus, Lignin, CCoAOMT, 4-Coumarate ： coenzyme A ligase （4CL）

中图分类号:

常瑞娜, 汪杏芬, 陈鸿鹏. 五节芒CCoAOMT和4CL的克隆和表达分析[J]. 华北农学报, 2012, 27(4): 29-35. doi: 10.3969/j.issn.1000-7091.2012.04.006.

CHANG Rui-na, WANG Xing-fen, CHEN Hong-peng. Cloning and Expression Analysis of CCoAOMT and 4CL Genes from Miscanthus floridulus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(4): 29-35. doi: 10.3969/j.issn.1000-7091.2012.04.006.

http://www.hbnxb.net/CN/Y2012/V27/I4/29

参考文献

[1] Clifton-Brown J C, Stample P F, Jones M B. Miscanthus biomass production for energy in Europe and its potential contribution to decreasing fossil fuel carbon emissions [J].Global Change Biol,2004,10:509 - 518.

[2] Bi C, Chen F, Jackson L,et al. Expression of lignin bio- synthesis genes in wheat during development and upon in- fection by fungal pathogens [J]. Plant Mol Biol Reports, 2011,29:149 - 161.

[3] 李尧臣,戚存扣.抗倒伏橄榄型油菜(Brassica napus L.)木质素含量及木质素合成关键酶基因的表达[J].江苏农业科学,2011,27:481-487.

[4] Vorwerk S, Somerville S, Somerville C. The role of plant cell wall polysaccharide composition in disease resistance [J]. Trends Plant Sci,2004,9:205 - 209.

[5] Marques G, Gutierrez A, Del Rio J. Chemical characteriza- tion of lignin and lipophilic fractions from leaf fibers of Curaua (Ananas erectifolius) [J]. J Agri Food Chem, 2007,55(4) :1327 - 1336.

[6] Guerra A,Filpponen I,Lucia L,et al. Comparative evalua- tion of three lignin isolation protocols for various wood spe- cies [J]. J Agri Food Chem,2006,54 ( 26 ) :9696 - 9705.

[7] Goncalves A, Schuchardt U, Bianchi M,et al. Piassava fi- bers (Attalea funifera) :NMR spectroscopy of their lignin [J]. J Brazil Chem Soci,2000,11 (5) :491 -494.

[8] Shi R,Sun YH,Li Q,et al. Towards a systems approach for lignin biosynthesis in Populus trichocarpa :transcript abundance and speciality of the monolignol biosynthetic genes [J]. Plant Cell Physiol,2010,51 ( 1 ) : 144 - 163.

[9] Weng J K, Akiyama T, Ralph J, et al. Independent re- cruitment of an O-Methyltransferase for syringyl lignin biosynthesis in Selaginella moellendorffii [J]. Plant Cell,2003,23:2708 - 2724.

[10] Li L,Cheng X,Lu S,et al. Clarification of cinnamoyl co-en- zyme A reductase catalysis in monolignol biosynthesis of Aspen [J]. Plant Cell Physiol,2005,46 (7) : 1073 - 1082.

[11] Wei J H,Zhao H Y,Zhang J Y,et al. Cloning of cDNA encoding CCoAOMT from Populus tomentosa and down- regulation of lignin content in transgenic plant expressing antisense gene [J]. Acta Bot Sin,2001,43 : 1179 - 1183.

[12] Li L, Zhou Y, Cheng X,et al. Combinatorial modification of multiple lignin traits in trees through multigene co- transformation[J]. Proc Natl Acad Sci USA, 2003,100 ( 8 ) :4939 - 4944.

[13] Wagner A,Tobimatsu Y,Phillips L,et al. CCoAOMT su- preesion modifies lignin composition in Pinus radiata [J]. Plant J.2011.67:119 - 129.
[14] Gui J,Shen J, Li L. Functional characterization of evolu- tionarily divergent 4-Coumarate: coenzyme A ligases in rice [J]. Plant Physiol, 2011,157 : 574 - 586.

[15] Stabile S S, Bodini A P, Jank L, et al. Expression of genes from the lignin synthasis pathway in guineagrass genotypes differing in cell-wall digestibility [J]. Grass Forage Sci,2011,67:43 - 54.

[16] Xu B, Escamilla-Trevino L L, Sathitsuksanoh N, et al. Silencing of 4-coumarate : Coenzyme A ligase in switch- grass leads to reduced lignin content and improved fer- mentable sugar yields for biofel production [J]. New Phytol,2011,192:611 - 625.

[17] Voelker S L, Lachenbruch B, Meinzer F C, et al. Anti- sense down-regulation of 4CL expression alters lignifica- tion,tree growth, and saeeharification potential of field- grown poplar[J]. Plant Physiol,2010,154 : 874 - 886.

[18] Stout A T. Eeophysiology and productivity of transgenie deereased-lignin Populus for use in short-rotation bioen- ergy cropping systems[D]. Master degree dissertation of North Carolina State University,2011.

[19] 《韧皮部特异性启动子研究概况》[J].中国麻业科学,2010(2).

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