Expression levels of hdr, fds and idi genes in aerial organs of Artemisia diffusa and their relationships with some terpene contents

Document Type : Research Paper

Authors

1 Former M.Sc. Student of Agriculture Biotechnology, Department of Agronomy, University College of Agriculture & Natural Resources of University of Tehran, Karaj, Iran

2 Professor of Agriculture Biotechnology, Department of Agronomy, University College of Agriculture & Natural Resources of University of Tehran, Karaj, Iran

3 Associate Professor, Department of Agronomy, University College of Agriculture & Natural Resources of University of Tehran, Karaj, Iran

Abstract

In view of the importance of terpene in human’s life, analyzing the relationship between them and the involved genes are of crucial necessity. The necessity is for exploring influential genes. Among different plants containing terpene, Artemisia is included. In this study, several genes Farnesyl diphosphate synthase (fds), Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (hdr) and Isopentenyl diphosphate isomerase (idi) in pathways 2-c-methyl-erythritol 4- phosphate (MEP) and Mevalonate (MVA) were analyzed in comparison to some monoterpene and sesquiterpene content in three tissues namely leaf, bud and flower of Artemisia diffusa. This study was conducted to compare RT-PCR data of genes with data from terpene GC-MS results. Results showed that it seemed for certain terpenes production in certain tissues, the above genes had limiting function (as idi in bud for 1,8- cineole and germacrene-D). These genes play important roles in terpene production and for some other terpenes, probably, other downstream genes such as terpene synthase had this limiting role (about beta-farnescene, for example, genes idi and fds had no limiting role in none of tissues mentioned above). Another point which casts doubt on fds, hdr and idi functions in terpene production is post-transcriptional regulation of these genes, the factor which was not evaluated in this study. 

Keywords


  1. Abad, M.J., Miguel Bedoya, L., Apaza, L. & Bermejo, P. (2012). The Artemisia L. genus: A review of bioactive essential oils. Molecules, 17, 2542-2566.
  2. Botella-Pavı´a, P., Besumbes, O., Phillips, M.A., Carretero-Paulet, L., Boronat, A. & Rodrı´guez-Concepcio´n, M. (2004). Regulation of carotenoid biosynthesis in plants: evidence for a key role of hydroxylmethylbutenyl diphosphate reductase in controlling the supply of plastidial isoprenoid precursors, The Plant Journal, 40, 188-199.
  3. Cheng, A.X., Lou, Y.G., Mao, Y.B., Lu, S., Wang, L.J. & Chen, X.Y. (2007). Plant terpenoids: Biosynthesis and ecological functions. Journal of Integrative Plant Biology, 49(2), 179-186.
  4. Huang, J. Z., Cheng, T.C., Wen, P.J., Hsieh, M.H., & Chen, F.C. (2009). Molecular characterization of the Oncidium orchid hdr gene encoding 1 hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, the last step of the methylerythritol phosphate pathway. Plant Cell Reports., 28, 1475-1486
  5. Iijima, Y., Davidovich-Rikanati, R., Fridman, E., Gang, D.R., Bar, E., Lewinsohn, E. & Pichersky, E. (2004). The biochemical and molecular basis for the divergent patterns in the biosynthesis of terpenes and phenylpropenes in the peltate glands of three cultivars of basil, Plant Physiology, 136, 3724-3736.
  6. Ma, Y., Yuan, L., Wu, B., Li, X., Chen, S. & Lu, S. (2012). Genome-wide identification and characterization of novel genes involved in terpenoid biosynthesis in Salvia miltiorrhiza, Journal of Experimental Botany. 63, 2809-2823.
  7. Munoz-Bertomeu, J., Arrillaga, I., Ros, R. & Segura, J. (2006). Up-regulation of 1-deoxy-dxylulose-5-phosphatesynthase enhances production of essential oils in transgenic Spike lavender. Plant Physiology 142, 890-900.
  8. Nagegowda, D.A. (2010). Plant volatile terpenoid metabolism: Biosynthetic genes, transcriptional regulation and sub-cellular compartmentalization, FEBS Letters, 584, 2965-2973.
  9. Olofsson, L., Engström, A., Lundgren, A. & Brodelius, P. E. (2011). Relative expression of genes of terpene metabolism in different tissues of Artemisia annua L, BMC Plant Biology, 11, 45.
  10. Pfaffl, M.W., Horgan, G.W. & Dempfle, L. (2002). Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research, 30(9), e36-e36.
  11. Page, J.E., Hause, G., Raschke, M., Gao, W., Schmidt, J., Zenk, M.H. & Kutchan, T.M. (2004). Functional analysis of the final steps of the 1-deoxy-d-xylulose 5-phosphate (DXP) Pathway to isoprenoids in plants using virus-induced gene silencing. Plant Physiology 134, 1401-1413.
  12. Phillips, M.A., Walter, M.H., Ralph, S.G., Dabrowska, P., Luck, K., Uro, E.M., Boland, W., Strack, D., Rodri´guez-Concepcio, M., Bohlmann, J. & Gershenzon, J. (2007).  Functional identification and differential expression of 1-deoxy D-xylulose 5-phosphate synthase in induced terpenoid resin formation of Norway spruce (Picea abies), Plant Molecular Biology, 65, 243-257.
  13. Wang, Q., Pi Y., Hou, R., Jiang, K., Huang, Z., Hsieh, M. & Sun, X. (2007). Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (CaHDR) from Camptotheca acuminata and its functional identification in Escherichia coliBMB Reports, 29, 112-118.
  14. Wen, W. & Yu, R. (2011). Artemisinin biosynthesis and its regulatory enzymes: Progress and perspective, Pharmacogn Review, 5(10), 189.
  15. Zare Mehrjerdi, M., Bihamta, M.R., Omidi, M., Naghavi, M.R., Soltanloo, H. & Ranjbar, M. (2013). Effects of exogenous methyl jasmonate and 2-isopentenyladenine on artemisinin production and gene expression in Artemisia annua, Turkish Journal of Botany 37, 499-505.
  16. Zeng, Q., Zhao, C., Yin, L., Yang, R., Zeng, X., Huang, Y., Feng, L. & Yang, X. (2008). Cloning of artemisinin biosynthetic cDNAs and novel ESTs and quantification of low temperature-induced gene over-expression. Science China C Life Science, 51(3), 232-4.