Characterization and functional analysis of the genes encoding 1-deoxy-D-xylulose-5-phosphate reductoisomerase and 1-deoxy-D-xylulose-5-phosphate synthase, the two enzymes in the MEP pathway, from Amomum villosum Lour.
A DXR gene, AvDXR (GenBank accession no. FJ459894), and a DXS gene, AvDXS (GenBank accession no. FJ455512), were isolated from the leaves of Amomum villosum, one of the most well-known and authentic herbs in South China. The 1,749-bp full-length cDNA of AvDXR encoded a peptide of 472 amino acids, and the 2,347-bp full-length cDNA of AvDXS encoded a peptide of 715 amino acids. The deduced amino acid sequences of the AvDXR and AvDXS proteins share high homology with DXRs and DXSs from other plant species, and AvDXS belongs to class 1 plant DXS. The characterization based on bioinformatic analysis indicated that the AvDXR and AvDXS encoded functional proteins as DXR and DXS, respectively. The functional color assay in Escherichia coli with pAC-BETA implied that AvDXR and AvDXS encoded functional proteins that manipulated the biosynthesis of isoprenoid precursors. Both AvDXR and AvDXS were expressed extensively in the leaves, stems, roots, pericarps and seeds of A. villosum. AvDXS expression was similar in all tissues investigated, whereas higher levels of AvDXR were observed in the fruits, the main part for the accumulation of volatile oil in this plant. AvDXR was transformed into tobacco to confirm its function further. Overexpression of AvDXR in transgenic T1 generation tobacco increased DXR activity, photosynthetic pigment content and volatile isoprenoid components, and the increase of photosynthetic pigment content was consistent with the AvDXR transcription level. This study demonstrated that AvDXR plays important role in isoprenoid biosynthesis and it is useful for metabolic engineering.
Research Center of Chinese Herbal Resource Science and Engineering, Guangzhou University of Chinese Medicine, 232 Waihuandong Road, Guangzhou Higher Education Mega Center, Guangzhou 510006, People's Republic of China.
Jinfen
J
Yang
Megha Nath
MN
Adhikari
Hui
H
Liu
Hui
H
Xu
Guozhen
G
He
Ruoting
R
Zhan
Jieshu
J
Wei
Weiwen
W
Chen
D019747:Aldose-Ketose Isomerases / Q000235:genetics* / Q000378:metabolism*; D035801:Amomum / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004896:Erythritol / Q000031:analogs & derivatives / Q000378:metabolism; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D053858:Metabolic Networks and Pathways; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D010860:Pigments, Biological / Q000378:metabolism; D030821:Plants, Genetically Modified; D013403:Sugar Phosphates / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014166:Transferases / Q000235:genetics* / Q000378:metabolism*
C114232:2-C-methylerythritol 4-phosphate; D009097:Multienzyme Complexes; D010860:Pigments, Biological; D013403:Sugar Phosphates; D013729:Terpenes; D010088:Oxidoreductases; C114231:1-deoxy-D-xylulose 5-phosphate reductoisomerase; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase; D019747:Aldose-Ketose Isomerases; D004896:Erythritol