观赏海棠McF3′H的克隆及不同品种间表达差异分析
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摘要
为了探究McF3′H基因与花色苷之间的关系,以苹果属观赏海棠‘王族’叶片总RNA为模板,通过RACE扩增,获得类黄酮3′-羟化酶(Flavonoid 3′-hydroxylase,F3′H)基因序列,其基因编码区共1 536bp,编码511个氨基酸,命名为McF3′H。McF3′H编码的氨基酸序列与其他物种的F3′H蛋白具有较高相似性。对3种不同叶色的观赏海棠品种‘火焰’、‘绚丽’和‘王族’叶片不同发育时期的McF3′H表达量、花青苷含量进行测定分析。结果表明McF3′H基因在3种不同叶色类型的观赏海棠品种叶片中均有表达,常色紫叶类‘王族’叶片McF3′H相对表达量明显高于新叶有色类‘绚丽’和绿色叶‘火焰’,McF3′H表达量与花色苷含量的变化规律较为一致。说明McF3′H在苹果属观赏海棠叶片花青苷代谢及色泽形成过程中具有重要作用。
In order to study the relationship between McF3′H gene and anthocyanins,the total RNA from the leaves of Malus crabapple 'Royalty' was used as the template and the full gene sequence of F3′H(Flavonoid 3′-hydroxylase) gene was cloned by rapid-amplification of cDNA end(RACE).The gene was named as McF3′H,containing an 1 536 bp open reading frame and encoding a protein of 511 amino acids.McF3′H protein was highly homologous to other species.In addition,McF3′H gene expression and anthocyanins content were detected in the leaves of 'Flame','Radiant' and 'Royalty' in different developmental stage.The results showed the expression level of McF3′H in'Royalty' is higher than in 'Flame' and 'Radiant'.The expression of McF3′H gene and anthocyanins content are almost consistent.Those results suggest that McF3′H may play an important role in crabapple anthocyanins metabolism and leaf color formation.
In order to study the relationship between McF3′H gene and anthocyanins,the total RNA from the leaves of Malus crabapple 'Royalty' was used as the template and the full gene sequence of F3′H(Flavonoid 3′-hydroxylase) gene was cloned by rapid-amplification of cDNA end(RACE).The gene was named as McF3′H,containing an 1 536 bp open reading frame and encoding a protein of 511 amino acids.McF3′H protein was highly homologous to other species.In addition,McF3′H gene expression and anthocyanins content were detected in the leaves of 'Flame','Radiant' and 'Royalty' in different developmental stage.The results showed the expression level of McF3′H in'Royalty' is higher than in 'Flame' and 'Radiant'.The expression of McF3′H gene and anthocyanins content are almost consistent.Those results suggest that McF3′H may play an important role in crabapple anthocyanins metabolism and leaf color formation.
引文
[1]Holton TA,Cornish EC.Genetics and biochemistry of anthocyanin bio-synthesis[J].Plant Cell,1995,7:1071-1083
[2]TanakaY.Flower colour and cytochromes P450[J].Phytochemistry Re-views,2006,5(2):283-291
[3]Tanaka Y,Sasaki N,Ohmiya A.Biosynthesis of plant pigments:antho-cyanins,betalains and carotenoids[J].The Plant Journal,2008,54(4):733-749
[4]Brugliera F,Barri-Rewell Q Holton T A,Mason J G.Isolation and charac-terization of a flavonoid3′-hydroxylase cDNA clone corresponding to the Htl locus of Petunia hybrida[J].Plant J,1999,19(4):441-451
[5]胡可,孟丽,韩科厅,等.瓜叶菊花青素合成关键结构基因的分离及表达分析[J].园艺学报,2009,36(7):1013-1022
[6]Ueyama Y,Suzuki K,Fukuchi M,et al.Molecular and biochemical char-acterization of torenia flavonoid3′-hydroxylase and flavone synthaseⅡand modification of flower color by modulating the expression of these genes[J].Plant Science,2002,63:253-263
[7]Fukuchi M M,Ohira K,Ueyama Y,et al.Flower color modification of Petunia hybrida commercial varieties by metabolic engineering[J].Plant Biotechnol,2004,21:377-386
[8]陆旺金,蒋跃明.荔枝果实两个膨大素基因的克隆与序列分析[J].中国农业科学,2003,36(12):1525-1529
[9]Wang LS,Shiraishi A,Hashimoto F,et al.Analysis of petal anthocyanins to investigate flower coloration of Zhongyuan(Chinese)and Daikon Island(Japanese)tree peony cultivars[J].J Plant Res,2001a,114:33-43
[10]Wang LS,Hashimoto F,Shiraishi A,et al.Phenetics in tree peony species from China by flower pigment cluster analysis[J].J Plant Res,2001b,114:213-221
[11]Zhang JJ,Wang LS,Shu QY,et al.Comparison of anthocyanins in non-blotches and blotches of the petals of Xibei tree peony[J].Sci Hortic,2007,114:104-111
[12]包满珠.植物花青素基因的克隆及应用[J].园艺学报,1997,24(3):279-284
[13]Han YP,Vimolmangkang S,Soria-Guerra RE,et al.Ectopic expres-sion of apple F3′H genes contributes to anthocyanin accumulation in the Arabidopsis tt7mutant grown under nitrogen stress[J].Plant Physiolo-gy,2010,153(2):806-820
[14]Chen WH,Hsu CY,Cheng HY,et al.Downregulation of putative UDP-glucose:flavonoid3-O-glucosyltransferase gene alters flower colo-ring in Phalaenopsis[J].Plant Cell Rep,2011,30(6):1007-1017
[15]ChagnéD,Lin-Wang K,Espley RV,et al.An ancient duplication of apple MYB transcription factors is responsible for novel red fruit-flesh phenotypes[J].Plant Physiol,2013,161(1):225-239
[2]TanakaY.Flower colour and cytochromes P450[J].Phytochemistry Re-views,2006,5(2):283-291
[3]Tanaka Y,Sasaki N,Ohmiya A.Biosynthesis of plant pigments:antho-cyanins,betalains and carotenoids[J].The Plant Journal,2008,54(4):733-749
[4]Brugliera F,Barri-Rewell Q Holton T A,Mason J G.Isolation and charac-terization of a flavonoid3′-hydroxylase cDNA clone corresponding to the Htl locus of Petunia hybrida[J].Plant J,1999,19(4):441-451
[5]胡可,孟丽,韩科厅,等.瓜叶菊花青素合成关键结构基因的分离及表达分析[J].园艺学报,2009,36(7):1013-1022
[6]Ueyama Y,Suzuki K,Fukuchi M,et al.Molecular and biochemical char-acterization of torenia flavonoid3′-hydroxylase and flavone synthaseⅡand modification of flower color by modulating the expression of these genes[J].Plant Science,2002,63:253-263
[7]Fukuchi M M,Ohira K,Ueyama Y,et al.Flower color modification of Petunia hybrida commercial varieties by metabolic engineering[J].Plant Biotechnol,2004,21:377-386
[8]陆旺金,蒋跃明.荔枝果实两个膨大素基因的克隆与序列分析[J].中国农业科学,2003,36(12):1525-1529
[9]Wang LS,Shiraishi A,Hashimoto F,et al.Analysis of petal anthocyanins to investigate flower coloration of Zhongyuan(Chinese)and Daikon Island(Japanese)tree peony cultivars[J].J Plant Res,2001a,114:33-43
[10]Wang LS,Hashimoto F,Shiraishi A,et al.Phenetics in tree peony species from China by flower pigment cluster analysis[J].J Plant Res,2001b,114:213-221
[11]Zhang JJ,Wang LS,Shu QY,et al.Comparison of anthocyanins in non-blotches and blotches of the petals of Xibei tree peony[J].Sci Hortic,2007,114:104-111
[12]包满珠.植物花青素基因的克隆及应用[J].园艺学报,1997,24(3):279-284
[13]Han YP,Vimolmangkang S,Soria-Guerra RE,et al.Ectopic expres-sion of apple F3′H genes contributes to anthocyanin accumulation in the Arabidopsis tt7mutant grown under nitrogen stress[J].Plant Physiolo-gy,2010,153(2):806-820
[14]Chen WH,Hsu CY,Cheng HY,et al.Downregulation of putative UDP-glucose:flavonoid3-O-glucosyltransferase gene alters flower colo-ring in Phalaenopsis[J].Plant Cell Rep,2011,30(6):1007-1017
[15]ChagnéD,Lin-Wang K,Espley RV,et al.An ancient duplication of apple MYB transcription factors is responsible for novel red fruit-flesh phenotypes[J].Plant Physiol,2013,161(1):225-239