Cinnamate-4-hydroxylase( C4H) is a key enzyme in phenylpropanoid pathway in plants. Its activity and abundance directly affect the biosynthesis of flavonoids and aromatic compounds. In this study,degenerate primers we...Cinnamate-4-hydroxylase( C4H) is a key enzyme in phenylpropanoid pathway in plants. Its activity and abundance directly affect the biosynthesis of flavonoids and aromatic compounds. In this study,degenerate primers were designed according to previously reported C4 H gene sequences to clone C4H cDNA sequence with 3'and 5'RACE-PCR from mango( Mangifera indica L). The full-length cD NA of M. indica C4H is 1 680 bp long. Its open reading frame( ORF)is 1 518 bp,encoding a protein of 505 amino acids with a predicted molecular weight of 58. 08 kDa. The isoelectric point of the predicted protein is 9. 52. Functional prediction showed that this gene is mainly located in mitochondria. In addition,the tertiary structure of the protein was built using SWISS-MODEL,and the results showed that the protein has three possible conformations. Phylogenetic analysis based on C4H protein sequences revealed that M. indica has a close genetic relationship with olive( Canarium album) and cocoa( Theobroma cacao). By analyzing the expression level of C4H gene in three colored mango cultivars,we found that that the expression level of C4 H gene in Guifei( with red peel) was the highest,and that in Guiqi( with green peel) was the lowest. The results provide a theoretical basis for studying the molecular mechanism of anthocyanin biosynthesis and C4H's impact on the color of mango fruit.展开更多
肉桂酸-4-羟基化酶是苯丙烷途径中继L-苯丙氨酸解氨酶(PAL)之后的第2个关键酶。为了从分子水平上了解C4H基因的功能特性,本文从植物进化的角度出发,根据Genebank中已有的豆科植物C4H氨基酸保守序列为基础,设计引物,通过RT-PCR和cDNA末...肉桂酸-4-羟基化酶是苯丙烷途径中继L-苯丙氨酸解氨酶(PAL)之后的第2个关键酶。为了从分子水平上了解C4H基因的功能特性,本文从植物进化的角度出发,根据Genebank中已有的豆科植物C4H氨基酸保守序列为基础,设计引物,通过RT-PCR和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)方法克隆膜荚黄芪C4H基因,对其序列进行分析,为进一步研究其功能奠定基础。展开更多
基金Supported by National Natural Science Foundation of China(31471850)the Fund for the Protection of Tropical Crops Genetic Resources(15RZZY-07)+1 种基金"948"Program of the Ministry of Agriculture of China(2011-G13)the Startup Fund for the Reform of Nonprofit Scientific Research Institutions(CATAS PZS-201225,CATAS-TCGRI 1630032013003)
文摘Cinnamate-4-hydroxylase( C4H) is a key enzyme in phenylpropanoid pathway in plants. Its activity and abundance directly affect the biosynthesis of flavonoids and aromatic compounds. In this study,degenerate primers were designed according to previously reported C4 H gene sequences to clone C4H cDNA sequence with 3'and 5'RACE-PCR from mango( Mangifera indica L). The full-length cD NA of M. indica C4H is 1 680 bp long. Its open reading frame( ORF)is 1 518 bp,encoding a protein of 505 amino acids with a predicted molecular weight of 58. 08 kDa. The isoelectric point of the predicted protein is 9. 52. Functional prediction showed that this gene is mainly located in mitochondria. In addition,the tertiary structure of the protein was built using SWISS-MODEL,and the results showed that the protein has three possible conformations. Phylogenetic analysis based on C4H protein sequences revealed that M. indica has a close genetic relationship with olive( Canarium album) and cocoa( Theobroma cacao). By analyzing the expression level of C4H gene in three colored mango cultivars,we found that that the expression level of C4 H gene in Guifei( with red peel) was the highest,and that in Guiqi( with green peel) was the lowest. The results provide a theoretical basis for studying the molecular mechanism of anthocyanin biosynthesis and C4H's impact on the color of mango fruit.
文摘肉桂酸-4-羟基化酶是苯丙烷途径中继L-苯丙氨酸解氨酶(PAL)之后的第2个关键酶。为了从分子水平上了解C4H基因的功能特性,本文从植物进化的角度出发,根据Genebank中已有的豆科植物C4H氨基酸保守序列为基础,设计引物,通过RT-PCR和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)方法克隆膜荚黄芪C4H基因,对其序列进行分析,为进一步研究其功能奠定基础。
