Expression analysis of biosynthetic genes of thymol and carvacrol in different tissues of thyme (Thymus vulgaris)

Authors

1 Sc., Department of Agriculture, College of Agriculture, University of Kurdistan, Sanandaj, I.R. Iran

2 Assist. Prof., College of Agriculture, University of Kurdistan, Sanandaj, I.R. Iran.

Abstract

Thymus vulgaris is one of the most important medicinal plants with more than 20 types of known important chemical compositions. A gene named 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) acts as an important control point, as this is the first committed step in the MEP pathway. γ-terpinene is a key component in essential oil of thyme which constituted up to 30% of the essential oil in the species and is biosynthesized via γ-terpinene as a penultimate precursor. Different cytochrome P450 including CYP71D178 and CYP71D182 are involved in the biosynthesis of thymol and carvacrol in thyme and oregano via the γ-terpinene as an intermediate. The aim of the present work was to study the expression patterns of genes involved in thymol and carvacrol biosynthesis including DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes in flowers and leaves of thyme. Gene expression analysis of DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes were carried out by semi-quantitative RT-PCR technique. The relative gene expression of flowers compared with leaves for DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes were 1.7, 2.7, 1.9 and 2 fold higher, respectively and the gene expression intensity varied for each gene. Expression of aforementioned genes in glandular trichomes were high, hence the higher gene expression in flowers might be related to higher density of glandular trichomes. Based on these results, higher content of essential oils in flowers compared to leaves is most likely further related to the higher expression of related genes in flowers rather than leaves.

Keywords

Main Subjects

References

-      Anonymous. 2008. http://drnelson.uthsc.edu/ P450stats.Feb2008.htm
-      Anonymous. 2009. http://drnelson.uthsc.ed /P450. Statistics.Aug 2009.pdf.
-      Behnia, M., Haghighi, A., Komeylizadeh, H., Tabaei, S.J.S. and Abadi, A., 2008. Inhibitory effects of Iranian Thymus vulgaris extracts on in vitro growth of Entamoeba histolytica. The Korean Journal of Parasitology, 46: 153-156.
-      Braga, P.C., Culici, M., Alfieri, M. and Dal Sasso, M., 2008. Thymol inhibits Candida albicans biofilm formation and mature biofilm. International Journal of Antimicrobial Agents, 31: 472-477.
-      Burke, C.C., Wildung, M.R. and Croteau, R., 1999. Geranyl diphosphate synthase: cloning, expression, and characterization of this prenyltransferase as a heterodimer. Proceedings of the National Academy of Sciences, 96: 13062-13067.
-      Burlat, V., Ou32din, A., Courtois, M., Rideau, M. and St‐Pierre, B., 2004. Co‐expression of three MEP pathway genes and geraniol 10‐hydroxylase in internal phloem parenchyma of Catharanthus roseus implicates multicellular translocation of intermediates during the biosynthesis of monoterpene indole alkaloids and isoprenoid‐derived primary metabolites. The Plant Journal, 38:131-141.
-      Crocoll, C., 2011. Biosynthesis of the phenolic monoterpenes, thymol and carvacrol, by terpene synthases and cytochrome P450s in oregano and thyme, PhD Thesis, Friedrich-Schiller-Universitat.
-      Crocoll, C., Asbach, J., Novak, J., Gershenzon, J. and Degenhardt, J., 2010. Terpene synthases of oregano (Origanum vulgare L.) and their roles in the pathway and regulation of terpene biosynthesis. Plant Molecular Biology, 73: 587-603.
-      Das, R. and Laederach, A., 2004. GelQuant user's manual. Stanford University, USA.
-      Davis, E.M. and Croteau, R., 2000. Cyclization enzymes in the biosynthesis of monoterpenes, sesquiterpenes, and diterpenes, Biosynthesis. Springer, pp. 53-95.
-      Elena-Rossello, J., 1981. Cytotaxonomic and evolutionary studies in Thymus (Labiatae); Relationships of the members of section Thymus jalas (1), Anales del Jardín Botánico de Madrid, pp. 51-60.
-      Fahn, A., 1988. Secretory tissues in vascular plants. New Phytologist, 108: 229-257.
-      Gershenzon, J., Maffei, M. and Croteau, R., 1989. Biochemical and histochemical localization of monoterpene biosynthesis in the glandular trichomes of spearmint (Mentha spicata). Plant Physiology, 89: 1351-1357.
-      Grausgruber-Gröger, S., Schmiderer, C., Steinborn, R. and Novak, J., 2012. Seasonal influence on gene expression of monoterpene synthases in Salvia officinalis (Lamiaceae). Journal of Plant Physiology. 169: 353-359.
-      Grosso, C., Figueiredo, A.C., Burillo, J., Mainar, A.M., Urieta, J.S., Barroso, J.G., Coelho, J.A. and Palavra, A.M., 2010. Composition and antioxidant activity of Thymus vulgaris volatiles: comparison between supercritical fluid extraction and hydrodistillation. Journal of Separation Science, 33: 2211-2218.
-      He, L., Zhao, S. and Hu. Z.,  2012. A novel cytochrome P450 gene from Catharanthus roseus cell line C 20 hi: cloning and characterization of expression." Acta Pharmaceutica Sinica B, 2: 250-255.
-      Horwath, A.B., Grayer, R.J., Keith-Lucas, D.M. and Simmonds, M.S., 2008. Chemical characterisation of wild populations of Thymus from different climatic regions in southeast Spain. Biochemical Systematics and Ecology, 36: 117-133.
-      Hummelbrunner, L.A. and Isman, M.B., 2001. Acute, sublethal, antifeedant, and synergistic effects of monoterpenoid essential oil compounds on the tobacco cutworm, Spodoptera litura (Lep., Noctuidae). Journal of Agricultural and Food Chemistry, 49: 715-720.
-      Isman, M.B., 2000. Plant essential oils for pest and disease management. Crop Protection, 19: 603-608.
-      Julliard, J.H., 1992. Biosynthesis of the pyridoxal ring (vitamin B6) in higher plant chloroplasts and its relationship with the biosynthesis of the thiazole ring (vitamin B1). Comptes rendus de l'Académie des sciences. Série 3, Sciences de la vie, 314: 285-290.
-      Julliard, J.H. and Douce, R., 1991. Biosynthesis of the thiazole moiety of thiamin (vitamin B1) in higher plant chloroplasts. Proceedings of the National Academy of Sciences, 88: 2042-2045.
-      Lambert, E., Faizal, A. and Geelen, D., 2011. Modulation of triterpene saponin production: in vitro cultures, elicitation, and metabolic engineering. Applied Biochemistry and Biotechnology, 164: 220-237.
-      Lopez‐Pujol, J., Bosch, M., Simon, J. and Blanche, C., 2004. Allozyme diversity in the tetraploid endemic Thymus loscosii (Lamiaceae). Annals of Botany, 93: 323-332.
-      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. Proceedings of the National Academy of Sciences, 98: 8915-8920.
-      Mahmoud, S.S. and Croteau, R.B., 2003. Menthofuran regulates essential oil biosynthesis in pepper mint by controlling a downstream monoterpene reductase. Proceedings of the National Academy of Sciences, 100: 14481-14486.
-      Majdi, M., Liu, Q., Karimzadeh, Gh., Malboobi, M.A., Beekwilder, J., Cankar, K., Vos, R.D., Todorović, S., Simonović, A. and Bouwmeester, H., 2011. Biosynthesis and localization of parthenolide in glandular trichomes of feverfew (Tanacetum parthenium). Phytochemistry, 72: 1739-1750.
-      Majdi, M., Karimzadeh, Gh. and Malboobi. M.A., 2014. Spatial and developmental expression of key genes of terpene biosynthesis in Tanacetum parthenium. Biologia Plantarum, 58: 379-384
-      Matarese, F., Cuzzola, A., Scalabrelli, G. and D’Onofrio, C., 2014. Expression of terpene synthase genes associated with the formation of volatiles in different organs of Vitis vinifera. Phytochemistry, 105: 12-24.
-      Nagegowda, D.A., 2010. Plant volatile terpenoid metabolism: Biosynthetic genes, transcriptional regulation and subcellular compartmentation. Febs Letters, 584: 2965-2973.
-      Nieuwenhuizen, N.J., Wang, M.Y., Matich, A.J., Green, S.A., Chen, X., Yauk, Y.K., Beuning, L.L., Nagegowda, D.A., Dudareva, N. and Atkinson, R.G., 2009. Two terpene synthases are responsible for the major sesquiterpenes emitted from the flowers of kiwifruit (Actinidia deliciosa). Journal of Experimental Botany, 60: 3203-3219.
-      Ozguven, M. and Tansi, S., 1998. Drug yield and essential oil of Thymus vulgaris L. as in influenced by ecological and ontogenetical variation. Turkish Journal of Agriculture and Forestry, 22: 537-542.
-      Rodríguez-Concepción, M., Ahumada, I., Diez-Juez, E., Sauret-Güeto, S., Lois, L.M., Gallego, F., Carretero-Paulet, L., Campos, N. and Boronat, A., 2001. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase and plastid isoprenoid biosynthesis during tomato fruit ripening. Plant Journal, 27:213-222.
-      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. Biochemical Journal, 295: 517-524.
-      Sedy, K. and Koschier, E., 2003. Bioactivity of carvacrol and thymol against Frankliniella occidentalis and Thrips tabaci. Journal of Applied Entomology, 127: 313-316.
-      Turner, G., Gershenzon, J., Nielson, E.E., Froehlich, J.E. and Croteau, R., 1999. Limonene synthase, the enzyme responsible for monoterpene biosynthesis in peppermint, is localized to leucoplasts of oil gland secretory cells. Plant Physiology, 120: 879-886.
-      Turner, G.W., Gershenzon, J. and Croteau, R.B., 2000. Development of peltate glandular trichomes of peppermint. Plant Physiology, 124: 665-680.
-      Ultee, A., Bennik, M. and Moezelaar, R., 2002. The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus. Applied and Environmental Microbiology,  68: 1561-1568.
-      Van Schie, C.C., Ament, K., Schmidt, A., Lange, T., Haring, M.A. and Schuurink, R.C., 2007. Geranyl diphosphate synthase is required for biosynthesis of gibberellins. The Plant Journal, 52: 752-762.
-      Vernet, P., Gouyon, R. and Valdeyron, G., 1986. Genetic control of the oil content in Thymus vulgaris L: a case of polymorphism in a biosynthetic chain. Genetica, 69: 227-231.

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