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Int. J. Mol. Sci. 2015, 16(10), 25516-25535; doi:10.3390/ijms161025516

Molecular Cloning and Characterization of DXS and DXR Genes in the Terpenoid Biosynthetic Pathway of Tripterygium wilfordii

1
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China
2
National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
*
Authors to whom correspondence should be addressed.
Academic Editor: Jianhua Zhu
Received: 7 September 2015 / Revised: 8 October 2015 / Accepted: 13 October 2015 / Published: 23 October 2015
(This article belongs to the Special Issue Molecular Research in Plant Secondary Metabolism 2015)
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Abstract

1-Deoxy-d-xylulose-5-phosphate synthase (DXS) and 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) genes are the key enzyme genes of terpenoid biosynthesis but still unknown in Tripterygium wilfordii Hook. f. Here, three full-length cDNA encoding DXS1, DXS2 and DXR were cloned from suspension cells of T. wilfordii with ORF sizes of 2154 bp (TwDXS1, GenBank accession no.KM879187), 2148 bp (TwDXS2, GenBank accession no.KM879186), 1410 bp (TwDXR, GenBank accession no.KM879185). And, the TwDXS1, TwDXS2 and TwDXR were characterized by color complementation in lycopene accumulating strains of Escherichia coli, which indicated that they encoded functional proteins and promoted lycopene pathway flux. TwDXS1 and TwDXS2 are constitutively expressed in the roots, stems and leaves and the expression level showed an order of roots > stems > leaves. After the suspension cells were induced by methyl jasmonate, the mRNA expression level of TwDXS1, TwDXS2, and TwDXR increased, and triptophenolide was rapidly accumulated to 149.52 µg·g−1, a 5.88-fold increase compared with the control. So the TwDXS1, TwDXS2, and TwDXR could be important genes involved in terpenoid biosynthesis in Tripterygium wilfordii Hook. f. View Full-Text
Keywords: Tripterygium wilfordii; TwDXS1; TwDXS2; TwDXR; color complementation Tripterygium wilfordii; TwDXS1; TwDXS2; TwDXR; color complementation
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Tong, Y.; Su, P.; Zhao, Y.; Zhang, M.; Wang, X.; Liu, Y.; Zhang, X.; Gao, W.; Huang, L. Molecular Cloning and Characterization of DXS and DXR Genes in the Terpenoid Biosynthetic Pathway of Tripterygium wilfordii. Int. J. Mol. Sci. 2015, 16, 25516-25535.

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