Subscribe to RSS
DOI: 10.1055/a-1809-7862
Studies of the Major Gene Expression and Related Metabolites in Cannabinoids Biosynthesis Pathway Influenced by Ascorbic Acid
Abstract
Cannabis sativa L. is an annual dioecious plant that belongs to the Cannabaceae family and is essential for different pharmaceutical and nutritional properties. The most important and prevalent cannabinoids in cannabis are cannabidiol and delta-9-tetrahydrocannabinol. The application of elicitors is an effective method to improve secondary metabolite production, leading to a whole spectrum of molecular, genetic, and physiological modifications. Therefore, the expression changes of four key genes (THCAS, CBDAS, PT, and OLS) of the cannabinoids pathway along with the delta-9-tetrahydrocannabinol and cannabidiol metabolites fluctuation were surveyed following the application of ascorbic acid as an elicitor. Cannabis was sprayed immediately before flowering with ascorbic acid. Treated and untreated (control) plants were sampled in different time courses for real-time PCR and HPLC experiments. Results showed significant increases in THCAS, CBDAS, PT, and OLS expression after ascorbic acid treatments. The results of metabolite quantification also indicated that secondary metabolites, especially delta-9-tetrahydrocannabinol and cannabidiol, increased after the ascorbic acid application. This study contributes to the growing body of knowledge of the functions of key genes in the cannabinoids pathway to the engineering of cannabis for improving the production of delta-9-tetrahydrocannabinol and cannabidiol metabolites in this plant.
Key words
Cannabis sativa - Cannabaceae - ascorbic acid - cannabidiol - delta-9-tetrahydrocannabinol - gene expression - metabolitesPublication History
Received: 17 October 2021
Received: 06 February 2022
Accepted: 25 March 2022
Article published online:
30 May 2022
© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).
Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany
-
References
- 1 Ainsworth C. Boys and girls come out to play: the molecular biology of dioecious plants. Ann Bot 2000; 86: 211-221
- 2 Russo EB, Jiang HE, Li X, Sutton A, Carboni A, Del Bianco F, Mandolino G, Potter DJ, Zhao YX, Bera S, Zhang YB. Phytochemical and genetic analyses of ancient cannabis from Central Asia. J Exp Bot 2008; 59: 4171-4182
- 3 Small E. The Role of Agriculture in Supplying Nutritional, Medicinal, and Recreational Cannabis Products. Planta Medica International Open 2018; 5: S3-S4
- 4 ElSohly MA, Slade D. Chemical constituents of marijuana: the complex mixture of natural cannabinoids. Life Sci 2005; 78: 539-548
- 5 Clark PA, Capuzzi K, Fick C. Medical marijuana: medical necessity versus political agenda. Med Sci Monit 2011; 17: RA249
- 6 Chandra S, Lata H, Khan IA, ElSohly MA. Temperature response of photosynthesis in different drug and fiber varieties of Cannabis sativa L. Physiol Mol Biol Plants 2011; 17: 297-303
- 7 Gaoni Y, Mechoulam R. Isolation, structure, and partial synthesis of an active constituent of hashish. J Am Chem Soc 1964; 86: 1646-1647
- 8 Watson SJ, Benson JA, Joy JE. Marijuana and medicine: assessing the science base: a summary of the 1999 Institute of Medicine report. Arch Gen Psychiatry 2000; 57: 547-552
- 9 Moldzio R, Unterberger A, Krewenka C, Kranner B, Radad K. Neuroprotective Effects of Delta-9-Tetrahydrocannabinol against FeSO4-and H2O2-Induced Cell Damage on Dopaminergic Neurons in Primary Mesencephalic Cell Culture. Planta Medica International Open 2021; 8: e88-e95
- 10 Fellermeier M, Eisenreich W, Bacher A, Zenk MH. Biosynthesis of cannabinoids: incorporation experiments with 13C-labeled glucoses. Eur J Biochem 2001; 268: 1596-1604
- 11 Raharjo TJ, Verpoorte R. Methods for the analysis of cannabinoids in biological materials: a review. Phytochem Anal 2004; 15: 79-94
- 12 Shoyama Y, Hirano H, Nishioka I. Biosynthesis of propyl cannabinoid acid and its biosynthetic relationship with pentyl and methyl cannabinoid acids. Phytochemistry 1984; 23: 1909-1912
- 13 Taura F, Morimoto S, Shoyama Y. Purification and characterization of cannabidiolic-acid synthase from Cannabis sativa L.: biochemical analysis of a novel enzyme that catalyzes the oxidocyclization of cannabigerolic acid to cannabidiolic acid. J Biol Chem 1996; 271: 17411-17416
- 14 Morimoto S, Komatsu K, Taura F, Shoyama Y. Purification and characterization of cannabichromenic acid synthase from Cannabis sativa . Phytochemistry 1998; 49: 1525-1529
- 15 Fischedick JT, Glas R, Hazekamp A, Verpoorte R. A qualitative and quantitative HPTLC densitometry method for the analysis of cannabinoids in Cannabis sativa L. Phytochem Anal 2009; 20: 421-426
- 16 Stout JM, Boubakir Z, Ambrose SJ, Purves RW, Page JE. The hexanoyl-CoA precursor for cannabinoid biosynthesis is formed by an acyl-activating enzyme in Cannabis sativa trichomes. Plant J 2012; 71: 353-365
- 17 Taura F, Tanaka S, Taguchi C, Fukamizu T, Tanaka H, Shoyama Y, Morimoto S. Characterization of olivetol synthase, a polyketide synthase putatively involved in cannabinoid biosynthetic pathway. FEBS lett 2009; 583: 2061-2066
- 18 Luo X, Reiter MA, d’Espaux L, Wong J, Denby CM, Lechner A, Zhang Y, Grzybowski AT, Harth S, Lin W, Lee H. Complete biosynthesis of cannabinoids and their unnatural analogues in yeast. Nature 2019; 567: 123-126
- 19 Lanyon VS, Turner JC, Mahlberg PG. Quantitative analysis of cannabinoids in the secretory product from capitate-stalked glands of Cannabis sativa L.(Cannabaceae). Bot Gaz 1981; 142: 316-319
- 20 Dayanandan P, Kaufman PB. Trichomes of Cannabis sativa L.(Cannabaceae). Am J Bot 1976; 63: 578-591
- 21 Petrovska BB. Historical review of medicinal plants’ usage. Pharmacogn Rev 2012; 6: 1-5
- 22 Govindaraju S, Arulselvi PI. Effect of cytokinin combined elicitors (l-phenylalanine, salicylic acid and chitosan) on in vitro propagation, secondary metabolites and molecular characterization of medicinal herb– Coleus aromaticus Benth (L). J Saudi Soc Agric Sci 2018; 17: 435-544
- 23 O’Brien ER, Read E, Deyholos M, Nelson L. Effects of Nitric oxide Producing Bacteria Azospirillum brasilense on Microbial Composition and Secondary Metabolite Profile of Cannabis. Planta Medica International Open 2018; 5 CAN06P
- 24 Alsoufi AS, Pączkowski C, Szakiel A, Długosz M. Effect of jasmonic acid and chitosan on triterpenoid production in Calendula officinalis hairy root cultures. Phytochem Lett 2019; 31: 5-11
- 25 Shi GR, Cai QS, Liu QQ, Wu L. Salicylic acid-mediated alleviation of cadmium toxicity in hemp plants in relation to cadmium uptake, photosynthesis, and antioxidant enzymes. Acta Physiol Plant 2009; 31: 969-977
- 26 Arrigoni O, De Tullio MC. The role of ascorbic acid in cell metabolism: between gene-directed functions and unpredictable chemical reactions. J Plant Physiol 2000; 157: 481-488
- 27 Conklin PL. Recent advances in the role and biosynthesis of ascorbic acid in plants. Plant Cell Environ 2001; 24: 383-394
- 28 Burkey KO, Eason G. Ozone tolerance in snap bean is associated with elevated ascorbic acid in the leaf apoplast. Physiol Plant 2002; 114: 387-394
- 29 Cook D, Rimando AM, Clemente TE, Schröder J, Dayan FE, Nanayakkara ND, Pan Z, Noonan BP, Fishbein M, Abe I, Duke SO. Alkylresorcinol synthases expressed in Sorghum bicolor root hairs play an essential role in the biosynthesis of the allelopathic benzoquinone sorgoleone. Plant Cell 2010; 22: 867-887
- 30 Gagne SJ, Stout JM, Liu E, Boubakir Z, Clark SM, Page JE. Identification of olivetolic acid cyclase from Cannabis sativa reveals a unique catalytic route to plant polyketides. PNAS 2012; 109: 12811-12816
- 31 Jalali S, Salami SA, Sharifi M, Sohrabi S. Signaling compounds elicit expression of key genes in cannabinoid pathway and related metabolites in cannabis. Ind Crops Prod 2019; 133: 105-110
- 32 Schachtsiek J, Warzecha H, Kayser O, Stehle F. Current perspectives on biotechnological cannabinoid production in plants. Planta Med 2018; 84: 214-220
- 33 Schmelz S, Naismith JH. Adenylate-forming enzymes. Curr Opin Struct Biol 2009; 19: 666-671
- 34 Hu Y, Gai Y, Yin L, Wang X, Feng C, Feng L, Li D, Jiang XN, Wang DC. Crystal structures of a Populus tomentosa 4-coumarate:CoA ligase shed light on its enzymatic mechanisms. Plant Cell 2010; 22: 3093-3104
- 35 Facchini PJ, Johnson AG, Poupart J, De Luca V. Uncoupled defense gene expression and antimicrobial alkaloid accumulation in elicited opium poppy cell cultures. Plant Physiol 1996; 111: 687-697
- 36 Melchior F, Hohmann F, Schwer B, Kindl H. Induction of stilbene synthase by Botrytis cinerea in cultured grapevine cells. Planta 1991; 183: 307-314
- 37 Rolfs CH, Fritzemeier KH, Kindl H. Cultured cells of Arachis hypogaea susceptible to induction of stilbene synthase (resveratrol-forming). Plant Cell Rep 1981; 1: 83-85
- 38 Kakani VG, Reddy KR, Zhao D, Mohammed AR. Effects of ultraviolet-B radiation on cotton (Gossypium hirsutum L.) morphology and anatomy. Ann Bot 2003; 91: 817-826
- 39 Caldwell MM, Bornman JF, Ballaré CL, Flint SD, Kulandaivelu G. Terrestrial ecosystems, increased solar ultraviolet radiation, and interactions with other climate change factors. Photochem Photobiol Sci 2007; 6: 252-266
- 40 Potter DJ, Duncombe P. The effect of electrical lighting power and irradiance on indoor-grown cannabis potency and yield. J Forensic Sci 2012; 57: 618-622
- 41 Chandra S, Lata H, Khan IA, Elsohly MA. Photosynthetic response of Cannabis sativa L. to variations in photosynthetic photon flux densities, temperature and CO2 conditions. Physiol Mol Biol Plants 2008; 14: 299-306
- 42 Mansouri H, Asrar Z, Amarowicz R. The response of terpenoids to exogenous gibberellic acid in Cannabis sativa L. at vegetative stage. Acta Physiol Plant 2011; 33: 1085-1091
- 43 Złotek U, Świeca M, Jakubczyk A. Effect of abiotic elicitation on main health-promoting compounds, antioxidant activity and commercial quality of butter lettuce (Lactuca sativa L.). Food Chem 2014; 148: 253-260
- 44 Pérez-Balibrea S, Moreno DA, García-Viguera C. Improving the phytochemical composition of broccoli sprouts by elicitation. Food Chem 2011; 129: 35-44
- 45 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 2001; 25: 402-408