The Expression of Limonene Biosynthesis Pathway Key Genes under Drought Stress in Dracocephalum kotschyi at Vegetative and Flowering Stages

Document Type : Research Paper

Authors

1 payame noor university, faculty of agriculture, iran, shahrekord, farokhshar

2 Professor, Department of Genetics and Plant Breeding, Faculty of Agriculture, Imam Khomeini International University, Qazvin, Iran.

3 Professor of Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran

4 Associate Professor, Department of Genetics and Plant Breeding, Faculty of Agriculture, Imam Khomeini International University, Qazvin, Iran

5 Assistant Professor, Department of Biology, School of Sciences, Razi University, Bagh-e-Abrisham Kermanshah, Iran

Abstract

Objective
 Dracocephalum kotschyi is an endemic medicinal plant in Iran belonging to the Lamiaceae family. One of the most important monoterpenes identified in this plant is limonene. Drought stress is one of the factors affecting the content of monoterpene in medicinal plants.  In this study, the effect of different levels of drought stress was investigated on the expression of some genes involved in limonene biosynthetic pathway in D. kotschyi at two growth stages.
 
Materials and Methods
 This experiment was conducted as a factorial experiment with two factor included drought stress and growth stages in a completely randomized design with three replications. Drought stress consisted of severe (25% FC), moderate (50% FC), low (75% FC) stress and non-stress or control (100% FC) and growth stages consisted of vegetative and flowering. Secondary metabolites were analyzed and the amount of limonene was measured by GC / MS using Head-space method and the relative expression of key genes involved in limonene biosynthesis was measured using Real Time PCR.
 
Results
 Analysis of secondary metabolites showed, one of the main monoterpenes was limonene whose amounts was 20.57 % and 13.41% during vegetative and flowering stages, respectively. Drought stress significantly increased HMGR and DXS gene expression during flowering stage, while during vegetative stage no significant change was observed on expression of both genes. In both growth stages, low drought stress increased HDR gene expression, although at flowering stage this increase was not significant but severe drought decreased HDR gene expression, although this decrease was not significant at vegetative stage. Drought stress decreased LS gene expression during flowering stage and increased it during vegetative stage.
 
Conclusion
 The results indicated that limonene content and expression of limonene biosynthesis key genes in D. kotschyi under drought stress are growth stage-specific.

Keywords

References

Anand A (2016) Major secondary metabolites and their biosynthesis in selected Ocimum sp. PhD thesis, Biochemical Sciences Division CSIR-National Chemical Laboratory. pp. 2-69.
Carretero-Paulet L, Cairo A, Botella-Pavía P et al. (2006) Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Mol Biol 62, 683-695.
Fan H, Wu Q, Wang X et al. (2016) Molecular cloning and expression of 1-deoxy-d-xylulose-5-phosphate synthase and 1-deoxy-d-xylulose-5-phosphate reductoisomerase in Dendrobium officinale. Plant Cell Tiss Org 125, 381-385.
Fattahi M, Bonfill M, Fattahi B et al. (2016) Secondary metabolites profiling of Dracocephalum kotschyi Boiss at three phenological stages using uni-and multivariate methods. J Appl Res Med Aroma 3, 177-185.
Fattahi M, Nazeri V, Torras-Claveria L et al. (2013) Identification and quantification of leaf surface flavonoids in wild-growing populations of Dracocephalum kotschyi by LC–DAD–ESI-MS. Food Chem 141, 139-146.
Ghasemi S, Kumleh HH, Kordrostami M (2018) Changes in the expression of some genes involved in the biosynthesis of secondary metabolites in Cuminum cyminum L. under UV stress. Protoplasma 256, 279-290.
Golshani S, Karamkhani F, Monsef-Esfehani HR, Abdollahi M (2004) Antinociceptive effects of the essential oil of Dracocephalum kotschyi in the mouse writhing test. J pharm pharm Sci 7, 76-79.
Hemmerlin A, Hoeffler JF, Meyer O et al. (2003) Cross-talk between the cytosolic mevalonate and the plastidial methylerythritol phosphate pathways in tobacco bright yellow-2 cells. J Biol Chem 278, 26666-26676.
Heydari P, Yavari M, Adibi P et al. (2019) Medicinal Properties and Active Constituents of Dracocephalum kotschyi and Its Significance in Iran: A Systematic Review. Evidence-Based Complementary and Alternative Medicine. https://doi.org/10.1155/2019/9465309
Jafari Darehdor AH, Mohammadabadi MR, Esmailizadeh AK, Riahi Madvar A (2016) Investigating expression of CIB4 gene in different tissues of Kermani Sheep using Real Time qPCR. J Rumin Res 4, 119-132 (In Persian).
Jalaei Z, Fattahi M, Aramideh S, (2015) Allelopathic and insecticidal activities of essential oil of Dracocephalum kotschyi Boiss. from Iran: A new chemotype with highest limonene-10-al and limonene. Industrial Crops and Products 73, 109-117.
Jongedijk E, Cankar K, Buchhaupt M et al. (2016) Biotechnological production of limonene in microorganisms. Appl Microbiol Biotechnol 100, 2927-2938.
Khorasaninejad, S, Mousavi A, Soltanloo H et al. (2011) The effect of drought stress on growth parameters, essential oil yield and constituent of Peppermint (Mentha piperita L.). J Med Plant Res 5, 5360-5365.
Kleinwächter M, Selmar D (2015) New insights explain that drought stress enhances the quality of spice and medicinal plants: potential applications. Agron Sustain Dev 35, 121-131.
Ma Y, Yuan L, Wu B et al. (2012) Genome-wide identification and characterization of novel genes involved in terpenoid biosynthesis in Salvia miltiorrhiza. J Exp Bot 63, 2809-2823.
Mohammadabadi MR, Jafari AHD, Bordbar F (2017) Molecular analysis of CIB4 gene and protein in Kermani sheep. Brazil J Med Biol Res 50, e6177.
Mohammadabadi MR, Tohidinejad F 2017. Charachteristics determination of Rheb gene and protein in Raini Cashmere goat. Iranian Journal of Applied Animal Science 7 (2), 289-295
Morshedloo MR, Craker LE, Salami A et al. (2017) Effect of prolonged water stress on essential oil content, compositions and gene expression patterns of mono-and sesquiterpene synthesis in two Oregano (Origanum vulgare L.) subspecies. Plant Physiol Biochem 111, 119-128.
Pateraki I, Kanellis AK (2010) Stress and developmental responses of terpenoid biosynthetic genes in Cistus creticus subsp. Creticus. Plant Cell Rep 29, 629-641.
Radwan AM. The impact of drought stress on monoterpene biosynthesis in sage (Salvia officinalis): Dehydrins and monoterpene synthases as molecular markers. PhD thesis, .Technische Universität Carolo-Wilhelmina zu Braunschweig. pp. 2-29
Rahimi Y, Taleei A, Ranjbar M (2017) Changes in the expression of key genes involved in the biosynthesis of menthol and menthofuran in Mentha piperita L. under drought stress. Acta Physiol Plant 39, 203.
Rohmer M, (2003) Mevalonate-independent methylerythritol phosphate pathway for isoprenoid biosynthesis. Elucidation and distribution. Pure and Applied Chemistry 75, 375-388.
Salehi M, Hejazi SMH, Tabaei R (2015) Genetic Differentiation of two Dracocephalum (Lamiaceae) species and populations in Iran by Polyacrylamide Gel Electrophoresis. Biological Forum – An international journal 7, 300.
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nature protocols 3, 1101.
Shariat A, Karimzadeh G, Assareh MH, Hadian J (2018) Metabolite profiling and molecular responses in a drought-tolerant savory, Satureja rechingeri exposed to water deficit. 3 Biotech 8, 477.
Soleymani F, Taheri H, Shafeinia AR (2017) Relative expression of genes of menthol biosynthesis pathway in peppermint (Mentha piperita L.) after chitosan, gibberellic acid and methyl jasmonate treatments. Russ J Plant physiol 64, 59-66.
Tohidi nezhad F, Mohammadabadi MR, Esmailizadeh AK, Najmi Noori A (2015) Comparison of different levels of Rheb gene expression in different tissues of Raini Cashmir goat. Agric Biotechnol J 6, 35-50 (in Persian).
Wei H, Movahedi A, Xu C et al. (2019) Overexpression of PtDXS Enhances Stress Resistance in Poplars. Int J Mol Sci 20, 1669.
Weissenborn DL, Denbow CJ, Laine M et al. (1995) HMG‐CoA reductase and terpenoid phytoalexins: Molecular specialization with in a complex pathway. Physiol Plant 93, 393-400.
Yazaki K, Arimura GI, Ohnishi T (2017) Hidden’terpenoids in plants: their biosynthesis, localization and ecological roles. Plant Cell Physiol 58, 1615-1621.
Agricultural Biotechnology Journal
Volume 11, Issue 3
Volume 11, No 3
October 2019
Pages 1-18

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