Skip to main content

Advertisement

SpringerLink
Log in

Molecular cloning, characterization and expression analysis of the gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Aquilaria sinensis (Lour.) Gilg

  • RESEARCH ARTICLE
  • Published:
Journal of Genetics Aims and scope Submit manuscript

Abstract

The major constituents of agarwood oils are sesquiterpenes that are obtained from isoprenoid precursors through the plastidial methylerythritol phosphate (MEP) pathway and the cytosolic mevalonate pathway. In this study, a novel full-length cDNA of 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR), which was the second key enzyme in the plastid MEP pathway of sesquiterpenes biosynthesis was isolated from the stem of Aquilaria sinensis (Lour.) Gilg by the methods of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technique for the first time, and named as AsDXR. The full-length cDNA of AsDXR was 1768 bp, containing a 1437 bp open reading frame (ORF) encoding a polypeptide of 478 amino acids with a molecular weight of 51.859 kD and the theoretical isoelectric point of 6.29. Comparative and bioinformatic analysis of the deduced AsDXR protein showed extensive homology with DXRs from other plant species, especially Theobroma cacao and Gossypium barbadense, and contained a conserved transit peptide for plastids, and extended pro-rich region and a highly conserved NADPH-binding motif owned by all plant DXRs. Southern blot analysis indicated that AsDXR belonged to a small gene family. Tissue expression pattern analysis revealed that AsDXR expressed strongly in root and stem, but weakly in leaf. Additionally, AsDXR expression was found to be activated by exogenous elicitor of MeJA (methyl jasmonate). The contents of three sesquiterpenes (α-guaiene, α-humulene and δ-guaiene) were significantly induced by MeJA. This study enables us to further elucidate the role of AsDXR in the biosynthesis of agarwood sesquiterpenes in A. sinensis at the molecular level.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10

Similar content being viewed by others

References

  • Brown A. C. and Parish T. 2008 DXR is essential in Mycobacterium tuberculosis and fosmidomycin resistance is due to a lack of uptake. BMC Microbial. 8, 78.

  • Carretero-Paulet L., Ahumada I., Cunillera N., Rodríguez-Concepción M., Ferrer A., Boronat A. et al. 2000 Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiol. 129, 1581–1591.

  • Chen H. Q., Wei J. H., Yang J. S., Zhang Z. and Yang Y. 2012 Chemical constituents of agarwood originating from the endemic genus Aquilaria plants. Chem. Biodivers. 9, 236–250.

  • Chen H. Q., Yang Y., Xue J., Wei J. H., Zhang Z. and Chen H. J. 2011 Comparison of compositions and antimicrobial activities of essential oils from chemically stimulated agarwood, wild agarwood and healthy Aquilaria sinensis (Lour.) Gilg trees. Molecules 16, 4884–4896.

  • China pharmacopoeia Committee 2010. The Pharmacopoeia of People’s Republic of China (I), pp. 172. China Medical Science Press, Beijing, China.

  • Dudareva N., Andersson S., Orlova I., Gatto N., Reichelt M., Rhodes D. et al. 2005 The nonmevalonate pathway supports both monoterpene and sesquiterpene formation in snapdragon flowers. Proc. Natl. Acad. Sci. USA 102, 933–938.

  • Engprasert S., Taura F. and Shoyama Y. 2005 Molecular cloning, expression and characterization of recombinant 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Coleus forskohlii Briq. Plant Sci. 169, 287–294.

  • Gao Z. H., Wei J. H., Yang Y., Zhang Z. and Zhao W. T. 2012 Selection and validation of reference genes for studying stress-related agarwood formation of Aquilaria sinensis. Plant Cell Rep. 31, 1759–1768.

  • Gong Y., Liao Z., Chen M., Zuo K., Guo L., Tan Q. et al. 2005 Molecular cloning and characterization of a 1-deoxy-D-xylulose 5-phosphate reductoisomerase gene from Ginkgo biloba. DNA Seq 16, 111–120.

  • Ito M., Okimoto K., Yagura T., Honda G., Kiuchi F. and Shimada Y. 2012 Induction of sesquiterpenoid production by Methyl Jasmonate in Aquilaria sinensis cell suspension culture. J. Essen. Oil Res. 17, 175–180.

  • Ito M. and Kumeta Y. 2010 Characterization of delta-guaiene synthases from cultured cells of Aquilaria, responsible for the formation of the sesquiterpenes in agarwood. Plant Physiol. 154, 1998–2007.

  • Kim S. M., Kuzuyama T., Chang Y. J., Song K. S. and Kim S. U. 2006 Identification of class 2 1-deoxy-D-xylulose 5-phosphate synthase and 1-deoxy-D-xylulose 5-phosphate reductoisomerase genes from Ginkgo biloba and their transcription in embryo culture with respect to ginkgolide biosynthesis. Planta Med. 72, 234–240.

  • Lu J., Wu W., Cao S., Zhao H., Zeng H., Lin L. et al. 2008 Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase gene from Ginkgo biloba. Mol. Biol. Rep. 35, 413–420.

  • Mahmoud S. S. and Croteau R. B. 2001 Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylulose phosphate reductoisomerase and menthofuran synthase. Proc. Natl. Acad. Sci. USA 98, 8915–8920.

  • Newman J. D. and Chappell J. 1999 Isoprenoid biosynthesis in plants: carbon partitioning within the cytoplasmic pathway. Crit. Rev. Biochem. Mol. Biol. 34, 11.

  • Okuder Y. and Ito M. 2009 Production of agarwood fragrant constituents in Aquilaria calli and cell suspension cultures. Plant Biotechnol 26, 307–315.

  • Regula N. 2011 The volatile and semi-volatile constituents of agarwood, the infected heartwood of Aquilaria species: a review. Flavour. Fragr. J. 26, 73–89.

  • Rodríguez-Concepción M. and Boronat A. 2002 Elucidation of the methylerythritol phosphate pathway for isoprenoid biosynthesis in bacteria and plastids: a metabolic milestone achieved through genomics. Plant Physiol. 130, 1079– 1089.

  • Rohmer M., Knani M., Simonin P., Sutter B. and Sahm H. 1993 Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphosphate. Biochem. J. 295, 517–524.

  • Sambrook J., Fritsh E. F. and Maniatis T. 1989 Molecular cloning: a laboratory manual, 2nd edition. Cold Spring Harbor Laboratory, New York, USA.

  • Schwender J., Müller C., Zeidler J. and Lichtenthaler H. K. 1999 Cloning and heterologous expression of a cDNA encoding 1-deoxy-D-xylulose-5-phosphate reductoisomerase of Arabidopsis thaliana. FEBS Lett. 455, 140–144.

  • Seetang-Nun Y., Sharkey T. D. and Suvachittanont W. 2008 Molecular cloning and characterization of two cDNAs encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Heave brasiliensis. J. Plant Physiol. 165, 991–1002.

  • Walter M. H., Fester T. and Strack D. 2000 Arbuscular mycorrhizal fungi induce the non-mevalonate methylerythritol phosphate pathway of isoprenoid biosynthesis correlated with accumulation of the ‘yellow pigment’ and other apocarotenoids. Plant J. 21, 571–578.

  • Xu Y., Liu J., Liang L., Yang X., Zhang Z., Gao Z. et al. 2014 Molecular cloning and characterization of three cDNAs encoding 1-deoxy-D-xylulose-5-phosphate synthase in Aquilaria sinensis (Lour.) Gilg. Plant Physiol. Biochem. 82, 133–141.

  • Xu Y., Zhang Z., Wang M., Wei J., Chen H., Gao Z. et al. 2013 Identification of genes related to agarwood formation: transcriptome analysis of healthy and wounded tissues Aquilaria sinensis. BMC Genomics 14, 227.

  • Yao H., Gong Y., Zuo K., Ling H., Qiu C., Zhang F. et al. 2008 Molecular cloning, expression profiling and functional analysis of a DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Camptotheca acuminate. J. Plant Physiol. 165, 203–213.

  • Yu F. and Utsumi R. 2009 Diversity, regulation and genetic manipulation of plant mono- and sesquiterpenoid biosynthesis. Cell Mol. Life Sci. 66, 3043–3052.

Download references

Acknowledgements

This work was funded by the National Natural Science Foundation of China (no. 81173481, no. 31100220, no. 31000136), 2013 Graduate Innovation Fund Project of Chinese Academy of Medical Sciences and Peking Union Medical College (2013-1007-07).

Author information

Authors and Affiliations

  1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 151, Malianwa North Road, Haidian District, Beijing, 100193, People’s Republic of China

    JUAN LIU, YANHONG XU & JIANHE WEI

  2. Hainan Branch Institute of Medicinal Plant (Hainan Provincial Key Laboratory of Resources Conservation and Development of Southern Medicine), Chinese Academy of Medical Sciences and Peking Union Medical College, Wanning, 571533, People’s Republic of China

    JIANHE WEI

  3. Shandong University of Traditional Chinese Medicine, Jinan, 250355, People’s Republic of China

    LIANG LIANG

Authors
  1. JUAN LIU
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. YANHONG XU
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. LIANG LIANG
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. JIANHE WEI
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to JIANHE WEI.

Additional information

[Liu J., Xu Y., Liang L. and Wei J. 2015 Molecular cloning, characterization and expression analysis of the gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Aquilaria sinensis (Lour.) Gilg. J. Genet. 94, xx–xx]

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

LIU, J., XU, Y., LIANG, L. et al. Molecular cloning, characterization and expression analysis of the gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Aquilaria sinensis (Lour.) Gilg. J Genet 94, 239–249 (2015). https://doi.org/10.1007/s12041-015-0521-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12041-015-0521-1

Keywords

Search

Navigation