Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni

Danial Kahrizi; Matin Ghaheri; Zahra Yari; Khirollah Yari; Sohbat Bahraminejad

doi:10.14715/cmb/2018.64.2.11

Issue

Vol. 64 No. 2: New biotechnological aspects in Stevia rebaudiana Bertoni

Issue Published : February 11, 2018

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Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni

https://doi.org/10.14715/cmb/2018.64.2.11

Danial Kahrizi

Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

Matin Ghaheri

Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

Zahra Yari

Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

Khirollah Yari

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

Sohbat Bahraminejad

Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

Corresponding Author(s) : Matin Ghaheri

matingh92@gmail.com

Cellular and Molecular Biology, Vol. 64 No. 2: New biotechnological aspects in Stevia rebaudiana Bertoni
Article Published : February 10, 2018

Abstract

Stevia rebaudiana Bertoni is one of two species that contains steviol glycosides. Among steviol glycosides that extracted from leaves, stevioside and rebaudioside A are the two major and the sweetest glycosides that are about 200-300 times sweeter than sucrose with zero calories. The best method for stevia propagation is tissue culture. So, for investigation of nutrients in medium, we studied the effect of different concentrations of MS media (MS, 0.5 MS, 0.25 MS, 0 MS) on morphological traits, UGT74G1 and UGT76G1 genes expression and accumulation of steviol glycosides in stevia leaves. The best growth rate (0.472 mm/d) has occurred in plants grown in MS media. Also, the highest gene expression of UGT74G1 gene (1.000 Total lab unit) was seen under MS treatment. However, the highest expression level of UGT76G1 gene (1.701 Total lab unit) was observed at plants grown in 0 MS. The highest amount of both Stevioside and Rebaudioside A (14.23 and 8.12, respectively) were accumulated in plants under MS treatment. Obviously, dilution of MS media associated with decreasing in both expression of the intended genes and accumulation of steviol glycosides.

Keywords

Stevia rebaudiana Bertoni MS media Steviol glycoside Semi-quantitative RT-PCR HPLC.

Kahrizi, D., Ghaheri, M., Yari, Z., Yari, K., & Bahraminejad, S. (2018). Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cellular and Molecular Biology, 64(2), 23–27. https://doi.org/10.14715/cmb/2018.64.2.11

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References

Kinghorn AD, Soejarto DD, Nanayakkara NPD, Compadre CM, Makapugay HC, Hovanec-Brown JM, et al. A phytochemical screening procedure for sweet ent-kaurene glycosides in the genus Stevia. J Nat Prod 1984; 47:439-444.
Ohtani K, Aikawa Y, Kasai R, Chou WH, Yamasaki K, Tanaka O. Minor diterpene glycosides from sweet leaves of Rubus sauvissimus. Phytochemistry 1992; 31:155-1559.
Brandle JE, Rosa N. Heritability for yield, leaf: stem ratio and stevioside content estimated from a landrace cultivar of Stevia rebaudiana. Can J Plant Sci 1992; 72:1263–1266.
Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist (online first) 2015; art. 151415.
Mondaca RL, Gálvez AV, Bravo L, Hen KA. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehen¬sive review on the biochemical, nutritional and functional aspects. J Food Chem 2012; 132:1121-1132.
Soufi S, D'Urso G, Pizza C, Rezgui S, Bettaieb T, Montoro. Steviol glycosides targeted analysis in leaves of Stevia rebaudiana (Bertoni) from plants cultivated under chilling stress conditions. J Food Chem 2016; 190:572–580
Debnath M. Propagation and antimicrobial activity of an ende¬mic medicinal plant Stevia rebaudiana. J Med Plant Res 2008; 2:45–51.
Anbazhagan M, Kalpana M, Rajendran R, Natarajan V, Dhana¬vel D. In vitro production of Stevia rebaudiana Bertoni. Emir J food agric 2010; 22:216-222.
Totté N, Charon L, Rohmer M, Compernolle F, Baboeuf I, Geuns JMC. Biosynthesis of the diterpenoid steviol, an ent-kaurene derivative from Stevia rebaudiana Bertoni, via the methylerythritol phosphate pathway. Tetrahedron Lett 2000; 41:6407–6410.
Brandle J, Telmer P. Steviol glycoside biosynthesis. Phytochemistry 2007; 68:1855-1863.
Richman A, Swanson A, Humphrey T, Chapman R, McGarvey B, Pocs R, et al. Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana. Plant J 2005; 41:56-67.
Pandey H, Pandey P, Pandey Sh, Singh S, Banerjee S. Meeting the challenge of stevioside production in the hairy roots of Stevia rebaudiana by probing the underlying process. Plant Cell Tiss Organ Cult 2016; 126:511–52.
Ma L, Shi Y. Effects of potassium fertilizer on physiological and biochemical index of Stevia rebaudiana Bertoni. J Energy Procedia 2011; 5:581-586.
Yadav P, Kumari P, Arya A, Tripathi S, Kumar S. Effect of ni¬trogen sources on rooting of in vitro culture of Stevia rebaudiana Bertoni. J Biotechnol 2013; 4:41-46.
Kamran Khan M, Misra P, Sharma P, Shukla PK, Ramteke PW. Effect of adenine sulphate on in vitro mass propagation of Stevia rebaudiana Bertoni. J Med Plants Res 2014; 8(13):543-549.
Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an Anti-diabetic Plant - Stevia rebaudiana Bertoni, (Natural Citation as Sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA J Biol Sci 2012; 1(3):37-42.
Inugraha M, Maghfoer D, Widaryanto E. Response of Stevia (Stevia rebaudiana Bertoni M) To Nitrogen and Potassium Fertilization. Int J Agric Vet Sci 2014; 10 (1):47-55.
Nepovim A, Vanek T. In vitro propagation of Stevia rebaudina plants using multiple shoot culture. Planta Med 1998; 64:775-776.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15:473–479.
Marone M, Mozzetti S, Ritis DD, Pierelli L, Scambia G. Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 2001; 3(1):19-25.
Jaitak V, Singh BB, Kaul VK. An efficient microwave-assisted extraction process of stevioside and rebaudioside-A from Stevia rebaudiana (Bertoni). Phytochem. Anal 2009; 20:240–245.
George EF, de Klerk GJ. In Plant Propagation by Tissue Culture. The Background (eds George EF, Hall MA, de Klerk GJ) 3rd edn. Springer, Dordrecht, The Netherlands, 2008, pp. 65–113
Sathyanarayana BN. Plant Tissue Culture: Practices and New Experimental Protocols. I. K. International Pvt Ltd. Botany, Experimental 2007, pp. 316.
Tavares AR, Kanashiro S, Ribeiro RCS, Gonçalves AN, Jocys T. Effect of phosphorus on in vitro growth and development of bromeliad aechmea blanchetiana. Acta Hort (ISHS) 2015; 1083:241-248.

References

Kinghorn AD, Soejarto DD, Nanayakkara NPD, Compadre CM, Makapugay HC, Hovanec-Brown JM, et al. A phytochemical screening procedure for sweet ent-kaurene glycosides in the genus Stevia. J Nat Prod 1984; 47:439-444.

Ohtani K, Aikawa Y, Kasai R, Chou WH, Yamasaki K, Tanaka O. Minor diterpene glycosides from sweet leaves of Rubus sauvissimus. Phytochemistry 1992; 31:155-1559.

Brandle JE, Rosa N. Heritability for yield, leaf: stem ratio and stevioside content estimated from a landrace cultivar of Stevia rebaudiana. Can J Plant Sci 1992; 72:1263–1266.

Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist (online first) 2015; art. 151415.

Mondaca RL, Gálvez AV, Bravo L, Hen KA. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehen¬sive review on the biochemical, nutritional and functional aspects. J Food Chem 2012; 132:1121-1132.

Soufi S, D'Urso G, Pizza C, Rezgui S, Bettaieb T, Montoro. Steviol glycosides targeted analysis in leaves of Stevia rebaudiana (Bertoni) from plants cultivated under chilling stress conditions. J Food Chem 2016; 190:572–580

Debnath M. Propagation and antimicrobial activity of an ende¬mic medicinal plant Stevia rebaudiana. J Med Plant Res 2008; 2:45–51.

Anbazhagan M, Kalpana M, Rajendran R, Natarajan V, Dhana¬vel D. In vitro production of Stevia rebaudiana Bertoni. Emir J food agric 2010; 22:216-222.

Totté N, Charon L, Rohmer M, Compernolle F, Baboeuf I, Geuns JMC. Biosynthesis of the diterpenoid steviol, an ent-kaurene derivative from Stevia rebaudiana Bertoni, via the methylerythritol phosphate pathway. Tetrahedron Lett 2000; 41:6407–6410.

Brandle J, Telmer P. Steviol glycoside biosynthesis. Phytochemistry 2007; 68:1855-1863.

Richman A, Swanson A, Humphrey T, Chapman R, McGarvey B, Pocs R, et al. Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana. Plant J 2005; 41:56-67.

Pandey H, Pandey P, Pandey Sh, Singh S, Banerjee S. Meeting the challenge of stevioside production in the hairy roots of Stevia rebaudiana by probing the underlying process. Plant Cell Tiss Organ Cult 2016; 126:511–52.

Ma L, Shi Y. Effects of potassium fertilizer on physiological and biochemical index of Stevia rebaudiana Bertoni. J Energy Procedia 2011; 5:581-586.

Yadav P, Kumari P, Arya A, Tripathi S, Kumar S. Effect of ni¬trogen sources on rooting of in vitro culture of Stevia rebaudiana Bertoni. J Biotechnol 2013; 4:41-46.

Kamran Khan M, Misra P, Sharma P, Shukla PK, Ramteke PW. Effect of adenine sulphate on in vitro mass propagation of Stevia rebaudiana Bertoni. J Med Plants Res 2014; 8(13):543-549.

Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an Anti-diabetic Plant - Stevia rebaudiana Bertoni, (Natural Citation as Sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA J Biol Sci 2012; 1(3):37-42.

Inugraha M, Maghfoer D, Widaryanto E. Response of Stevia (Stevia rebaudiana Bertoni M) To Nitrogen and Potassium Fertilization. Int J Agric Vet Sci 2014; 10 (1):47-55.

Nepovim A, Vanek T. In vitro propagation of Stevia rebaudina plants using multiple shoot culture. Planta Med 1998; 64:775-776.

Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15:473–479.

Marone M, Mozzetti S, Ritis DD, Pierelli L, Scambia G. Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 2001; 3(1):19-25.

Jaitak V, Singh BB, Kaul VK. An efficient microwave-assisted extraction process of stevioside and rebaudioside-A from Stevia rebaudiana (Bertoni). Phytochem. Anal 2009; 20:240–245.

George EF, de Klerk GJ. In Plant Propagation by Tissue Culture. The Background (eds George EF, Hall MA, de Klerk GJ) 3rd edn. Springer, Dordrecht, The Netherlands, 2008, pp. 65–113

Sathyanarayana BN. Plant Tissue Culture: Practices and New Experimental Protocols. I. K. International Pvt Ltd. Botany, Experimental 2007, pp. 316.

Tavares AR, Kanashiro S, Ribeiro RCS, Gonçalves AN, Jocys T. Effect of phosphorus on in vitro growth and development of bromeliad aechmea blanchetiana. Acta Hort (ISHS) 2015; 1083:241-248.