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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (03): 479-486.doi: 10.3724/SP.J.1006.2015.00479

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Chromolaena odorata Flavonoid 3’-hydroxylase Gene Cloning and Its Expression in Tobacco

HE Hai-Wang,PAN Hua-Qing,ZHANG Nao-Dan,HE Long-Fei*   

  1. Agricultural College of Guangxi University, Nanning 530004, China
  • Received:2014-08-12 Revised:2014-12-19 Online:2015-03-12 Published:2014-12-29
  • Contact: 何龙飞, E-mail: lfhe@gxu.edu.cn, Tel: 0771-3235212-801 E-mail:gxhhwn@gxaas.net


A flavonoid 3’-hydroxylase gene cDNA sequence was cloned by RACE and RT-PCR techniques from Chromolaena odorata. The obtained full-length cDNA was named as CoF3H with GenBank accession number HQ268505.1. It is 1628 bp in length, containing a 1524 bp open reading frame, encoding 507 amino acid residues. The amino acid sequence of CoF3H contains cytochrome P450 domain and cysteineheme binding region (F××G×R×C×G). Homology analysis by DNAMAN software showed that the deduced CoF3H protein was highly homologous to F3’H proteins from different species. CoF3H gene was transferred into tobacco by agrobacterium-mediated genetic transformation. There was higher content of flavonvoids in T2 transgenic tobacco than in the wild type. This result shows that CoF3H plays an important role in flavonoid biosynthesis, and provides a basis for researches on allelopathy and comprehensive utilization of C. odorata in the future.

Key words: Chromolaena odorata, Flavonoid 3’-hydroxylase, Clone, Expression

[1]VonSengerI, Barker N, Zachariades C. Preliminary phylogeography of Chromolaena odorata: finding the origin of a South African weed. In: Proceedings 2002 Fifth International Workshop on Biological Control and Management of Chromolaena odorata, Durban, South Africa, 2002: 90–99

[2]Witkowski E T F, Wilson M. Changes in density, biomass, seed production and soil seed banks of the non-native invasive plant, Chromolaena odorata, along a 15 year chronosequence. Plant Ecol, 2001, 152: 13–27

[3]Ye W H, Mu H P, Cao H L, Ge X J. Genetic structure of the invasive Chromolaena odorata in China. Weed Res, 2004, 44: 129–135

[4]Biswas K. Some foreign weeds and their distribution in India and Burma. Indian forest, 1934, 60: 861–865

[5]Hall J B, Kumar R, Enti A A. The obnoxious weed Eupatorium odoratum (Compositae) in Ghana. Ghana J Agric Sci, 1972, 5(1): 75–78

[6]刘金海, 黄必志, 罗富成. 飞机草的危害及防治措施简介. 草业科学, 2006, 23(10): 73–77

Liu J H, Huang B Z, Luo F C. The damage and control measures of Eupatorium odoratum. Pratac Sci, 2006, 23(10): 73–77 (in Chinese with English abstract)

[7]吴邦兴. 滇南飞机草群落的初步研究. 云南植物研究, 1982, 4: 177–184

Wu B X. The primary studies on Eupatorium odoratum community in Southern Yunnan. Acta Bot Yunnanica, 1982, 4: l77–184 (in Chinese with English abstract)

[8]Patane K. A, Setter S, Graham M. Effect of foliar herbicides on the germination and viability of Siam weed (Chromolaena odorata) seeds located on plants at the time of application. Plant Prot Quart, 2009, 24(4): 138–140

[9]侯杰, 佟玲, 崔国新, 许志茹, 李玉花. 植物类黄酮3’-羟化酶(F3’H)基因的研究进展. 植物生理学报, 2011, 47: 641–647

Hou J, Tong L, Cui G X, Xu Z R, Li Y H. Research advances of plant flavonoid 3’-hydroxylase (F3’H) gene. Plant Physiol J, 2011, 47: 641–647 (in Chinese with English abstract)

[10]Brugliera F, Barri-Rewell G, Holton T A, Mason J G. Isolation and characterization of a flavonoid 3’-hydroxylase cDNA clone corresponding to the Ht1 locus of petunia hybrid. Plant J, 1999, 19: 441–451

[11]Schoenbohm C, Martens S, Eder C, Forkmann G, Weisshaar B. Identification of the Arabidopsis thaliana flavonoid 3’-hydroxylase gene and functional expression of the encoded p450 enzyme. Biol Chem, 2005, 381: 749–753

[12]Seitz C, Eder C, Deiml B, Kellner S, Martens S, Forkmann G. Cloning, functional identification and sequence analysis of flavonoid 3’-hydroxylase and flavonoid 3’,5’-hydroxylase cDNAs reveals independent evolution of flavonoid 3’,5’-hydroxylase in the Asteraceae family. Plant Mol Biol, 2006, 61: 365–381

[13]Xu B B, Li J N, Zhang X K, Wang R, Xie L L, Chai Y R. Clone and molecular characterization of a functional flavonoid 3’-hydroxylase gene from Brassica napus. J Plant Physiol, 2007, 164: 350–363

[14]许志茹, 崔国新, 李春雷, 孙燕, 李玉花. 芜菁的类黄酮3’-羟化酶基因克隆和UV-A诱导表达特性. 植物生理学通讯, 2008, 44: 931–935

Xu Z R, Cui G X, Li C L, Sun Y, Li Y H. Cloning and UV-Ainduced expression trait of flavonoid 3’-hydroxylase gene in turnip (Brassica rapa L.). Plant Physiol Commun, 2008, 44: 931–935 (in Chinese with English abstract)

[15]陶吉寒, 招雪晴, 苑兆和, 尹燕雷, 冯立娟. 石榴花类黄酮3’-羟化酶基因的克隆及序列分析. 河南农业科学, 2013, 42(4): 121–124

Tao J H, Zhao X Q, Yuan Z H, Yin Y L, Feng L J. Cloning and sequence analysis of flavonoid 3’-hydroxylase gene in pomegranate flower. J Henan Agric Sci, 2013, 42(4): 121–124 (in Chinese with English abstract)

[16]Castellarin S D, Gaspero G D, Marconi R, Nonis A, Peterlunger E, Paillard S, Adam-Blondon A F, Testolin R. Colour variation in red grapevines (Vitis vinifera L.): genomic organization, expression of flavonoid 3’-hydroxylase, flavonoid 3’,5’-hydroxylase genes and related metabolite profiling of red cyaniding-/blue delphinidin-based anthocyanins in berry skin. BMC Genomic, 2006, 7: 12

[17]Toda K, Yand D, Yamanaka N, Watanabe S, Harada K, Takahashi R. A single-base deletion in soybean flavonoid 3’-hydroxylase gene if associated with gray pubescence color. Plant Mol Biol, 2002, 50: 187–196

[18]Jeong S T, Yamamoto N G, Hashizume K, Esaka M. Expression of the flavonoid 3’-hydroxylase and flavonoid 3’,5’-hydorxylase genes and flavonoid composition in grape (Vitis vinifera). Plant Sci, 2006, 170: 61–69

[19]Holton T A, Brugliera F, Lester D R, Tanaka Y, Hyland C D, Menting J G T, Lu C Y, Farcy E, Stevenson T W, Cornish E C. Cloning and expression of cytochrome P450 genes controlling flower colour. Nature, 1993, 366: 276–279

[20]张松焕, 郭惠明, 裴熙祥, 李春奇, 程红梅. 紫茎泽兰类黄酮3’-羟化酶在烟草中的表达. 中国农业科学, 2009, 44: 4182–4186

Zhang S H, Guo H M, Pei X X, Li C Q, Cheng H M. Expression of flavonoid 3’-hydrolyase from Eupatorium adenophorum in tobacco. Sci Agric Sin, 2009, 44: 4182–4186 (in Chinese with English abstract)

[21]何海旺, 许春燕, 李文, 何龙飞, 李创珍, 韦善清. 玻璃粉吸附核酸及其在植物核酸提取中应用的研究. 生物技术进展, 2012, 2: 359–365

He H W, Xu C Y, Li W, He L F, Li C Z, Wei S Q. The research of glass powder absorbing nucleic acid and its application of nucleic acid extraction from plant tissue. Curr Biotechnol, 2012, 2: 359–365 (in Chinese with English abstract)

[22]Holsters M, de Waele D, DepickerA, Messens E, van Montagu M, Schell J. Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet, 1978, 163: 181–187

[23]Horsch R B, Fry J E, Hoffmann N L, Eicholtz D, Rogers S G, Fraley R T. A simple and general method for transferring genes into plants. Science, 1985, 227: 1229–1231

[24]李增富, 吴荣锋. 葛根总黄酮的提取研究. 应用化工, 2008, 38: 1353–1358

Li Z F, Wu R F. Study on extracting total puerar in flavonoids. Appl Chem Ind, 2008, 38: 1353–1358 (in Chinese with English abstract)

[25]梁国兴, 赵缓, 王彩芳. 入侵物种飞机草(Eupatorium odoratum)化学成份及生物活性研究进展. 北京师范大学学报(自然科学版), 2011, 47: 611–617

Liang G X, Zhao Y, Wang C F. Chemical constituents and their bioactivity of the invasive species Eupatorium odoratum L. J Beijing Norm Univ (Nat Sci), 2011, 47: 611–617 (in Chinese with English abstract)

[26]陈向明, 马云飞. 山核桃外果皮黄酮提取液对小麦和绿豆幼苗的化感效应. 西北植物学报, 2010, 30 : 645–651

Chen X M, Ma Y F. Allelopathy of flavonoid extract from cary a cathayensisexocarp on wheat and mung bean seedlings. Acta Bot Boreal-occident Sin, 2010, 30: 645–651 (in Chinese with English abstract)

[27]郭亚姣, 郑书馨,李洁, 江莎. 黄顶菊叶中类黄酮对白菜种子萌发的影响. 内蒙古农业大学学报, 2011, 32(4): 105–109

Guo Y J, Zheng S X, Li J, Jing S. The effects of flavonoids extracted from flaveriabidentis leaves on seed germination of brassica pekinensi. J Inner Mongolia Agric Univ, 2011, 32(4): 105–109 (in Chinese with English abstract)

[28]雷磊. 飞机草中化感物质的提取分离与鉴定. 吉林农业大学硕士学位论文, 吉林长春, 2011. p 31

Lei L. Extraction separation and identification of allelochemicals in Chromolaena odoratum. MS Thesis of Jilin Agricultural University, Changchun, China, 2011. p 31 (in Chinese with English abstract)

[29]Phan T T, Wang L, See P, Grayer R J, Chan S Y, Lee S T. Phenolic compounds of Chromolaena odorata protect cultured skin cells from oxidative damage: implication for cutaneous wound healing. Biol Pharm Bull, 2001, 21: 1373–1379

[30]Suksamram A, Chotipong A, Suavansri T, Boongird S, Timsuksai P, Vimuttipong S, Chuaynugul A. Antimycobacterial activity and cytotoxicity of flavonoids from the flowers of Chromolaena odorata. Arch Pharm Res, 2004, 27: 507–511

[31]Owoyele B V, Oguntoye S O, Dare K, Ogunbiyi B A, Aruboula E A, Soladoye A O. Analegsic anti-inflammatory and antipyretic activities from flavonoid fractions of Chromolaena odorata. J Med Plants Res, 2008, 2: 219–225

[32]张松焕, 郭惠明, 裴熙祥, 李春奇, 程红梅. 紫茎泽兰类黄酮3’-羟化酶在烟草中的表达. 中国农业科学, 2009, 42: 4182–4186

Zhang S H, Guo H M, Pei X X, Li C Q, Cheng H M. Expression of Flavonoid 3’-hydrolyase from Eupatorium adenophorum in tobacco. Sci Agric Sin, 2009, 42: 4182–4186 (in Chinese with English abstract)

[33] Nakatsula T, Nishihara M, Mishiba K, Yamamura S. Heterologous expression of two gentian cytochrome P450 genes can modulate the intensity of ?ower pigmentation in transgenic tobacco plants. Mol Breed, 2006, 17: 91–99

[34]乔小燕, 马春雷, 陈亮. 植物类黄酮生物合成途径及重要基因的调控. 天然产物研究与开发, 2009, 21: 354–360

Qiao X Y, Ma C L, Chen L. Plant flavonoid biosynthesis pathway and regulation of its important genes. Nat Prod Res Dev, 2009, 21: 354–360 (in Chinese with English abstract)

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