Cloning and Expression Analysis of Partial Fragments from Lignin Biosynthesis F5H Genes in Switchgrass

doi:10.11733/j.issn.1007-0435.2012.05.021

›› 2012, Vol. 20 ›› Issue (5): 927-934.DOI: 10.11733/j.issn.1007-0435.2012.05.021

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Cloning and Expression Analysis of Partial Fragments from Lignin Biosynthesis F5H Genes in Switchgrass

LI Yuan, WANG Shan, FANG Cheng

Institute of Grassland Science, China Agricultural University, Beijing 100193, China

Received:2012-02-27 Revised:2012-04-12 Online:2012-10-15 Published:2012-11-01

柳枝稷木质素合成酶基因F5H核心片段的克隆与表达特性的分析

李园, 王珊, 方程

中国农业大学草地研究所, 北京 100193

通讯作者: 方程,E-mail:fangcheng@cau.edu.cn
作者简介:李园(1987-),女,河北邢台人,硕士研究生,研究方向为牧草分子技术,E-mail:liyuan2010kaoyan@163.com
基金资助:
边际土地能源草分子育种与新种质创制(2012AA101801);耐盐碱能源草筛选与新品种培育项目(2009BADA7B04);草业科学北京市重点实验室共建项目资助

Abstract

Abstract: Switchgrass (Panicum virgatum L.) is an important lignocellulosic crop. High lignin content seriously limits its utilization as an energy plant. The partial fragments of F5H genes involved in G/S lignin monomer conversion were amplified from switchgrass bark mRNA using RT-PCR and designated as PvF5H1 and PvF5H2 respectively (GenBank accession number: JQ430747 and JQ430748). DNAMAN sequence analysis of both genes indicated that the similarity of nucleic acid sequence reached 93% and the similarity of protein sequence reached 90%. NCBI Blast online was performed to analyze the biological information of both PvF5H1 and PvF5H2. Sequence analyzed through biological methods showed that encoded polypeptides were highly homologous to F5H proteins of Zea mays. Phylogenetic tree analysis also demonstrated that both genes were related to the F5H gene from Zea mays. Real-time quantitative PCR (qPCR) was performed to investigate the expression profiles of both PvF5H1 and PvF5H2 under wound stress. The results of qPCR analysis revealed that both PvF5H1 and PvF5H2 showed fast-response and up-regulated expression under wound stress. PvF5H1 and PvF5H2 had different expression level with similar changing tendency.

Key words: F5H gene, Panicum virgatum L., Lignin, Wound response, Expression analysis

摘要： 柳枝稷(Panicum virgatum L.)是一种重要的木质纤维作物,但其高木质素含量严重制约了其作为能源植物的开发和利用。以柳枝稷茎秆中的mRNA为模板,采用RT-PCR方法克隆得到控制G/S木质素单体相互转化的关键酶基因F5H家族的2个基因,分别命名为PvF5H1与PvF5H2,将这2个基因序列提交到GenBank,登录号分别为JQ430747与JQ430748。DNAMAN序列分析表明,这2个基因的核酸序列相似性为93%,氨基酸序列相似性为90%。利用NCBI在线比对功能对这2个基因的生物学信息进行分析,结果表明:该基因通过生物学方法分析的编码蛋白质与玉米(Zea mays)的同源性最高,系统进化树分析也表明,其与玉米的亲缘关系最近。利用实时荧光定量PCR技术研究这2个基因在机械伤害胁迫下的表达趋势,结果表明:在伤害胁迫下,PvF5H1与PvF5H2基因都快速响应,并且在一定时间内,这种响应呈现正相关,其变化趋势一致,但是表达量及其变化相差较大。推测它们在柳枝稷应答伤害胁迫的过程中起重要作用。

关键词: F5H基因, 柳枝稷, 木质素, 机械伤害胁迫, 表达分析

CLC Number:

Q943.2

LI Yuan, WANG Shan, FANG Cheng. Cloning and Expression Analysis of Partial Fragments from Lignin Biosynthesis F5H Genes in Switchgrass[J]. , 2012, 20(5): 927-934.

李园, 王珊, 方程. 柳枝稷木质素合成酶基因F5H核心片段的克隆与表达特性的分析[J]. , 2012, 20(5): 927-934.

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References

[1] Bransby D I, Ward C Y, Rose P A, et al. Biomass production from selected herbaceous species in the Southeastern USA[J]. Biomass,1989,20(1/2):187-197
[2] McLaughlin S B, Ugarte D G D L, Garten C T, et al. High-value renewable energy from prairie grasses[J]. Environmental Science and Technology,2002,36(10):2122-2129
[3] Schmer M R, Vogel K P, Mitchell R B, et al. Net energy of cellulosic ethanol from switchgrass[J]. Proceedings of the National Academy of Sciences of the United States of America,2008,105(2):464-469
[4] Sanderson M A, Reed R L, McLaughlin S B, et al. Switchgrass as a sustainable bioenergy crop[J].Bioresource Technology,1996,56(1):83-93
[5] 程序.能源牧草堪当未来生物能源之大任[J].草业学报,2008,17(3):1-5
[6] 刘吉利,程序,谢光辉,等. 能源作物柳枝稷研究进展[J].草业学报,2009,18(3):232-240
[7] Derikvand M, Sierra J B, Ruel K, et al. Redirection of the phenylpropanoid pathway to feruloyl malate in Arabidopsis mutants decient for cinnamoyl-CoA reductase 1[J]. Planta,2008,227(5):943-956
[8] 魏建华,宋艳茹.木质素生物合成途径及调控的研究进展[J].植物学报,2001,43(8):771-779
[9] Pauly M K. Physiology and metabolism'tear down this wall’[J]. Current Opinion in Plant Biology,2008,11(3):233-235
[10] Meyer K, Shirley A M, Cusumano J C, et al. Lignin monomer composition is determined by the expression of a cytochrome P450-dependent monooxygenase[J]. Proceedings of the National Academy of Sciences of the United States of America,1998,95(12):6619-6623
[11] Meyer K, Cusumano J C, Somerville C, et al.Ferulate-5-hydroxylase from Arabidopsis thaliana defines a new family of cytochrome P450-dependent monooxygenases[J].Proceedings of the National Academy of Sciences of the United States of America,1996,93(14):6869-6874
[12] Franke R, McMichael CM, Meyer K, et al. Modified lignin in tobacco and poplar plants over-expressing the Arabidopsis gene encoding ferulate5-hydroxylase [J]. The Plant Journal,2000,22(3):223-234
[13] Sibout R, Baucher M, Gatineau M, et al. Expression of a poplar cDNA encoding a ferulate-5-hydroxylase/coniferaldehyde 5-hydroxylase increases S lignin deposition in Arabidopsis thaliana[J]. Plant Physiology and Biochemistry,2002,40(12):1087-1096
[14] Smith C G, Rodgers M W, Zimmerlin A, et al. Tissue and subcellular immunolocalisation of enzymes of lignin synthesis in differentiating and wounded hypocotyls tissue of French bean (Phaseolus vulgaris L.) [J]. Planta,1994,192(2):155-164
[15] Hahlbrock K, Scheel D. Physiology and molecular biology of phenylpropanoid metabolism [J]. Annual Review of Plant Physiolgy and Plant Molecular Biology,1989,40(1):347-369
[16] Diallinas G, Kanellis A K. A phenylalanine ammonia-lyase gene from melon fruit: cDNA cloning, sequence and expression in response to development and wounding [J]. Plant Molecular Biology,1994,26(1):473-479
[17] Batard Y, Schalk M, Pierrel M A, et al. Regulation of the cinnamate 4-hydroxylase (CYP73A1) in Jerusalem artichoke tubers in response to wounding and chemical treatments [J]. Plant Physiology,1997,113(3):951-959
[18] Richard S, Lapointe G, Rutledge R G, et al. Induction of chalcone synthase expression in white spruce by wounding and jasmonate[J]. Plant and Cell Physiology,2000,41(8):982-987
[19] Kim Y J, Kim D K, Lee S H, et al. Wound-induced expression of ferulate 5-hydroxylase gene in Camptotheca acuminate[J].Biochimica et Biophysica Acta,2006,1760(2):182-190
[20] Kim Y J, Yang D H, Kim J S, et al. Cloning, characterization, and expression of two cDNA clones for a rice ferulate-5-hydroxylase gene, a cytochrome P450-dependent monooxygenase [J]. Journal of Plant Biology,2006,49(3):200-204
[21] Chapple C. Molecular genetic analysis of plant cytochrome P450-dependent monooxygenases [J]. Annual Review of Plant Physiolgy and Plant Molecular Biology,1998,49:311-343
[22] Frank M R, Deyneka J M, Schuler M A. Cloning of wound-induced cytochromeP450 monooxygenases expressed in pea [J]. Plant Physiology,1996,110(3):1035-1046
[23] Brown J W. A catalogue of splice junction and putative branch point sequences from plant introns [J]. Nucleic Acids Research,1986,14(24):9549-9559
[24] Dixon R A, Paiva N R. Stress-induced phenylpropanoid metabolism [J]. The Plant Cell,1995,7(7):1085-1097
[25] Ellard-Ivey M, Douglas C J. Role of jasmonates in the elicitor and wound-inducible expression of defense genes in parsley and transgenic tobacco [J]. Plant Physiology,1996,112(1):183-192
[26] 付月,李学龙,薛永常.木质素合成酶基因F5H的克隆及其鉴定[J].安徽农业科学,2006,34(8):1553-1555
[27] Chapple C C S, Vogt T, Ellis B E, et al. An Arabidopsis mutant defective in the general phenylpropanoid pathway [J].The Plant Cell,1992,4(11):1413-1424

Cloning and Expression Analysis of Partial Fragments from Lignin Biosynthesis F5H Genes in Switchgrass

柳枝稷木质素合成酶基因F5H核心片段的克隆与表达特性的分析

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