Effect of different concentrations of KH2PO4 on the expression of UGT85C2 and KO genes and some biochemical traits of Stevia Rebaudiana bertoni

Document Type : Research Paper

Authors

1 Assistant Professor, Department of Agriculture, Payam Noor University, Iran

2 M.Sc. of Plant Breeding, Department of Agriculture, Payam Noor University, Iran

3 Associate Professor, Department of Agriculture, Payam Noor University, Iran

Abstract

Stevia rebaudiana bertoni is a natural sweetener that is sweeter than sugar; however it does not have any calorie and can be used for diabetic patients. The sweet taste of stevia is because of the steviol glycosides (SGs) accumulated in the leaves. Rebaudioside A-F and Stevioside are the main compounds of stevia. KO and UGT85C2 genes are two important genes involved in biosynthesis pathway of steviol and steviolmonoside that are precursors of other SGs. Due to the very low germination rate of stevia seeds, tissue culture is the fastest way for propagation of it. Therefore, optimizing the composition of the culture medium is necessary to increase of pharmaceutical ingredients in stevia. This experiment was conducted to study the effect of different concentrations of KH2PO4 (0, 4.25, 8.5, 17, 34 µM) on the expression of KO and UGT85C2 genes using semi- quantitative RT-PCR technique as well as soluble sugar content and antioxidant activity of stevia. Analysis of variance showed a significant difference between the treatments (p≤0.01) for all measured parameters. Mean comparison using LSD (p≤0.05) showed that UGT85C2 and KO genes possessed the highest expression level in 4.25 and 17 µM KH2PO4respectively. Also, the stevia showed the highest content of soluble sugars and antioxidant activity in 34 µM KH2PO4. According to our results it seems that total steviol glycosides increased after adding KH2PO4into the culture medium, but its effect on different glycosides was not the same and probably changed the ratio of different SGs in this plant.

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Main Subjects


Aladakatti YR, Palled YB, chetti MB, Halikatti SI, Alagundagi SC, Patil PL, patil1 VC, janawade AD (2012) Effect of nitrogen, phosphorus and potassium levels on growth and yield of stevia (Stevia rebaudiana Bertoni.). Karnataka J. Agr. Sci. 25(1): 25-29.
Alavi Matin SM, Rahnama A, Meskarbashee M (2016).  Effect of potassium supply on the activity of some antioxidant enzymes of two durum wheat cultivars under salt stress. The Plant Production, 38(4): 1-12. (In Pertsian)
Behroozi P, Baghizadeh A, Saei A, Kharazmi S (2017) Quantitative Analysis of Uridine Diphosphate Glycosyl transferase UGT85C2, UGT74G1 and UGT76G1 Genes Expression in Stevia rebaudiana under Different Irrigations. Russian J. Plant Physiol. 64: 67-72.
Brandle JE, Starratt AN, Gijzen M (1998) Stevia rebaudiana: Its agricultural, biological, and chemical properties. Can. J. Plant Sci. 78: 527-536.
Cakmak I (2005) The role of potassium in alleviating detrimental effects of abiotic stresses in plants. J. Plant Nutr. Sc. 168: 521-530.  
Curry LL, Ashley R (2008) Subchronic toxicity of rebaudioside A. Food Chem. Toxicol. 46: S11-S20.
Devi BSR, Kim YJ, Selvi SK, Gayathri S, Altanzul K, Parvin S, Yang DU, Lee OR, Lee S, Yang DC (2012) Influence of potassium nitrate on antioxidant level and secondary metabolite genes under cold stress in Panax ginseng. Russian J. Plant Physiol. 59: 318-325.
Dubios MK, Gilles A, Hamilton JK, Roberts PA, Smith F (1956) Colorometric method for determination of sugars and related substrate. Annu. Chem. 28: 350-356.
Fakhrul RH, Norrizah JS, Jaapar SS, Noor Anilizawatima S (2014) The Effect of Potassium Concentrations on the Growth and Development of Stevia rebaudiana (Bertoni) and Production of Stevioside and Rebaudioside A. American-Eurasian J. Sustainable Agric. 8: 42-51.
Gluria P, Yadav SK (2013) Agrobacterium mediated transient gene silencing (AMTS) in Stevia rebaudiana: insights into steviol glycoside biosynthesis pathway, PLOS One, 8(9): e74731.
Gupta E, Purwar SH, Sundaram SH, Rai, G (2013) Nutritional and therapeutic values of Stevia rebaudiana: A review. J. Med. Plants Res. 7: 3343-3353
Hanson JR, White AF (1968) Studies in terpenoid biosynthesis–ii. the biosynthesis of steviol. Phytochemistry, 7: 595- 597
Ibrahim IA, Nasr MI, Mohammedm BR, El-Zefzafi MM (2008) Nutrient factors effecting in vitro cultivation of stevia rebaudiana. Sugar Technol. 10: 248-253.
Inugraha M, Maghfoer D, Widaryanto E. (2014) Response of Stevia (Stevia rebaudiana Bertoni M) to Nitrogen and Potassium Fertiliza­tion. Int. J. Agric. Vet. Sci.; 10 (1): 47-55.
Kahrizi D, Ghari SM Ghaheri M, Fallah F, Ghorbani T, Beheshti Ale Agha A, Kazemi E, Ansarypour Z, (2017) Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions.  Cell Mol Biol. 63(7): 107-111.
Karimi M, Ahmadi A, Hashemi J, Abbasi A, Tavarini S, Guglielminetti L (2015) The effect of soil moisture depletion on Stevia (Stevia rebaudiana Bertoni) grown in greenhouse conditions: growth, steviol glycosides content, soluble sugars and total antioxidant capacity, Sci. Hortic. 183: 93-99.
Kumar H, Kaul K, Bajpai-Gupta S, Kumar Kaul V, Kumar S (2012) A comprehensive analysis of fifteen genes of steviol glycosides biosynthesis pathway in Stevia rebaudiana (Bertoni). Gene, 492: 276-284.
Luwanska A, Perz A, Mankowska G, Wielgus K (2015) Application of in vitro stevia (Stevia rebaudiana Bertoni) cultures in obtaining steviol glycoside rich material. Herba Pol. 61(1): 50-63.
Malekoti MJ, Shahabi A, Bazargan K (2016) Potassium in agriculture "The role of potassium in the production of healthy agricultural products" Publishing amount. 331 p.
Marshner H, (1995) Mineral nutrition of higher plants. Academic Press, London.
Mohamed A, Ceunen S, Geuns  J, Ende W, Ley M (2011) UDP-dependent glycosyltransferases involved in the biosynthesis of steviol glycosides. J. Plant Physiol. 168: 1136-1141.
Moini A, Kahrizi D (2003) Plant tissue culture (Persian Translated “Plant Tissue Culture Practice: Taji A., Williams, R., Dodd R., and William, A.”), Basij Student Organization Publications.
Molyneux p (2004) The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin J. Sci. Technol. 26: 211-219.
Moradi-Peynevandi K, Sharifi M, Behmanesh M, (2014) Effects of methyl jasmonate, on stevioside and rebaudioside A content and expression of the ent-Kaurenoic acid 13-hydroxylase gene in Stevia rebaudiana Bert. in vitro. Iranian J. Plant Biol. 21: 99-110 (In Persian)
Richman AS, Gijzen M, Starratt AN, Yang Z, Brandle JE (1999) Diterpene synthesis in Stevia rebaudiana: Recruitment and up-regulation of key enzymes from the gibberellin biosynthetic pathway. Plant J. 19(4): 411-421.
Saber-Hamyshegy p, Torang A, Moballeghi M, Dehpouri A, Saber-Hamyshegy Z (2012) The Effect of different levels of nitrogen and potassium on morphological and chemical characteristics of Stevia rebaudiana Bertoni. New Agric. Findings, 7: 127-135. (In Persian)
Sathyanarayana BN (2007) Plant Tissue Culture: Practices and New Experimental Protocols. I. K. International Pvt Ltd, Botany, Experi­mental, 316 pages.
Tavakoli Hasanaklou N, Ebadi A, Davari M, Tavakoli Hasanaklou H (2016) Effect of potassium and nitrogen on the wheat resistance against fusarium head blight. Cereal Res. 6(2): 159-171 (In Persian)
Tisdale SL, Nelson WL, beaton JD, Havlein JL (1993) Soil fertility and fertilizer. 5th. Eds.Ma cmillan, Pub. Co. New York.
Yadav S, Guleria P (2012) Steviol glycosides from stevia: biosynthesis pathway review and their application in foods and medicine. Crit. Rev. Food Sci. Nutr. 52: 988-998.