Construction of a Recombinant Plasmid for Petal-Specific Expression of HQT, a Key Enzyme in Chlorogenic Acid Biosynthesis
The flower buds of Lonicera japonica are widely used in Chinese medicine due to the anti-inflammatory properties. Chlorogenic acid (CGA) may be one of the most critical components in L. japonica. Previous studies suggest that HQT gene play a very important role in the synthesis of CGA. In order to overexpress HQT gene specifically in the petals, we successfully isolated the full-length cDNA of HQT gene from L. japonica. Then from Lilium orential ‘Sorbonne’, we cloned an 896-bp promoter (PLoCHS) which can drive chalcone synthase gene (CHS) specifically expression in flowers. After sequencing and bioinformatical analysis, two constructs based on pCAMBIA2300 were obtained, one was a constituently expression vector driving HQT by 35S promoter, and the other fused HQT and PLoCHS promoter. In the further research, we will transform the constructs into L. japonica callus. This study provides a means of improving the CGA content in L. japonica flowers, and will be helpful in understanding the expression pattern of HQT related with CGA and development of the plants.
KeywordsHQT PLoCHS CGA Construction Petal
Thanks to the support of China Three Gorges University for the preliminary study. This project was supported by the National Natural Science Foundation of China (31270389) and Natural Science Research of Hubei Education Department (D20121301).
- 1.Peng XX, Li WD, Wang WQ, Bai GB (2010) Cloning and characterization of a cDNA coding a hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase involved in chlorogenic acid biosynthesis in Lonicera japonica. Planta MedGoogle Scholar
- 3.State Pharmacopoeia Committee (2005) Pharmacopoeia of China, Part 1. Chemical Industry Press, Beijing, pp 152–153Google Scholar
- 4.Teng HL (2007) Synthetically study on medicinal materials of Lonicera japonica. J Chin Med Mat 30:744–748Google Scholar
- 6.Chang JL, Luo J, He GY (2009) Regulation of polyphenols accumulation by combined overexpression/silencing key enzymes of phyenylpropanoid pathway. Acta Biochim Biophys Sin 41(2):123–130Google Scholar
- 9.Lofty S, Fleuriet A, Macheix JJ (1992) Partial purification and characterization of hydroxycinnamoyl CoA: transferases from apple and date fruits. Biochemistry 13:767–772Google Scholar
- 11.Han YY, Ming F, Wang JW, Ye MM, Shen DL (2006) Molecular characterization and functional analysis of a novel chalcone synthase gene from Phalaenopsis Orchid in transgenic tobacco. Plant Mol Biol Rep 23:193a–193mGoogle Scholar
- 12.Kobayashi H, Oikawa Y, Koiwa H, Yamamura S (1998) Flower-specific gene expression directed by the promoter of a chalcone synthase gene from Gentiana triflora in Petunia hybrida. Plant Sci 131:173–180Google Scholar
- 13.Uimari A, Strommer J (1997) Myb26: a MYB-like protein of pea flowers with affinity for promoters of phenylpropanoid genes. Plant J 12:1273–1284Google Scholar
- 14.Van der Meer IM, Spelt CE, Mol JNM, Stuitje AR (1992) Promoter analysis of the chalcone synthase (CHSA) gene of Petunia hybrida: a 67 bp promoter region directs flower-specific expression. Plant Mol Biol 15:95–109Google Scholar
- 15.Huits HSM, Gerats AGM, Kreike MM, Mol JNM, Koes RE (1994) Genetic control of dihydroflavonol 4-reductase gene expression in Petunia hybrida. Plant J 6:295–310Google Scholar