Abstract
This study aimed to determine the response of Satureja rechingeri to water deficit by quantifying the expression of three targeted genes and four traditional reference genes using quantitative real-time PCR analysis (RT-qPCR). Drought stress was imposed by withholding water 4 months after planting. Profiling of volatile and non-volatile compounds using gas chromatography/mass spectrometry (GC/MS) and high-performance thin layer chromatography (HPTLC) showed an increasing–decreasing trend of major phenolic and terpenoid compounds such as rosmarinic and caffeic acids, carvacrole, thymol and p-Cymene. Drought stress also lead to significant increases in oil yield, soluble sugars and proline as well as significant reductions in leaf water potential (LWP), relative water content (RWC), and pigments. Metabolite profiling revealed the strategies savory employed to generate different biochemical phenotypes. RT-qPCR analysis showed that up-regulation of the three genes [1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), 3-hydroxy-3-methylglutaryl-coenzyme. A reductase (HMGR) and rosmarinic acid synthase: 4-coumaroyl-CoA (RAS)] selected from the phenylpropanoid and terpenoid biosynthesis pathways were markedly enhanced at the transcript levels of the regulatory steps and directly increased the production of secondary metabolites, including phenolic and terpenoid compounds. Actin protein (ACT), elongation factor 1-α (EF1α), glyceraldehyde-3-phosphate dehydrogenase cytosolic (GAPC) and ubiquitin-conjugating enzyme (UBC) were used as traditional reference genes. UBC’s suitability as the reference genes were verified in S. rechingeri. The study’s results provide the foundation for gene expression analysis of savory and other species of Lamiaceae. Thus, the effective application of drought stress before harvesting can increase the quantity and quality of raw material.
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Authors gratefully acknowledge the support provided for this research project by the Tarbiat Modares University and by Iran National Science Foundation, Deputy of Science and Technology, Presidential Office.
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13205_2018_1491_MOESM1_ESM.pdf
The biosynthetic pathway of target compounds, selected enzymes in this study are depicted in red. (a) The isoprenoid pathway in plants (Mahmoud and Croteau <link rid="bib17">2001</link>). The indicated enzymes are: AACT, acetyl-CoA/acetyl-CoA C-acetyltransferase; HMGS, 3-hydroxy-3- methylglutaryl-CoA synthase; HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase; MK, mevalonate kinase; PMK, phosphomevalonate kinase; MDC, mevalonate-5-diphosphate decarboxylase; DXS, 1-deoxy-d-xylulose 5-phosphate synthase; DXR, 1-deoxyxylulose-5-phosphate reductoisomerase; MCT, 2-C- methylerythritol-4-phosphate (MEP) cytidyltransferase; CMK, 4-diphosphocytidyl-2Cmethyl-d-erythritol kinase; MECPS, 2-C-methylerythritol-2,4- cyclodiphosphate synthase (b) Biosynthetic. (b) Pathway for rosmarinic acid (a part of phenylpropanoid pathway) (Weitzel and Petersen <link rid="bib33">2011</link>). PAL = phenylalanine ammonia-lyase, C4H = cinnamic acid 4-hydroxylase, 4CL = 4-coumarate:CoAligase,TAT = tyrosine aminotransferase, HPPR = hydroxyphenylpyruvate reductase, RAS = rosmarinic acid synthase. (PDF 121 KB)
13205_2018_1491_MOESM2_ESM.pdf
Greenhouse pots. (a) Preliminary design layout of the greenhouse pot experiment (b) Experimental design using a completely randomized design (CRD) (PDF 59 KB)
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Shariat, A., Karimzadeh, G., Assareh, M.H. et al. Metabolite profiling and molecular responses in a drought-tolerant savory, Satureja rechingeri exposed to water deficit. 3 Biotech 8, 477 (2018). https://doi.org/10.1007/s13205-018-1491-9
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DOI: https://doi.org/10.1007/s13205-018-1491-9