Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni

Fatemeh Esmaeili; Matin Ghaheri; Danial Kahrizi; Mohsen Mansouri; Seyed Mehdi Safavi; Tayebeh Ghorbani; Sarre Muhammadi; Elham Rahmanian; Siavash Vaziri

doi:10.14715/cmb/2018.64.2.1

Issue

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

Issue Published : February 11, 2018

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Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni

https://doi.org/10.14715/cmb/2018.64.2.1

Fatemeh Esmaeili

Department of Medicinal Plants, Institute of Higher Education, Jahad-e-Daneshgahi, Kermanshah Unit, Iran

Matin Ghaheri

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

Danial Kahrizi

Fertility and Infertility Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran

Mohsen Mansouri

Department of Persian Medicine, Medicine Faculty, Aja University of Medical Sciences, Tehran, Iran

Seyed Mehdi Safavi

Department of Persian Medicine, Medicine Faculty, Aja University of Medical Sciences, Tehran, Iran

Tayebeh Ghorbani

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

Sarre Muhammadi

Department of Cardiology, Kermanshah University of Medical Sciences, Kermanshah, Iran

Elham Rahmanian

Department of Sexual Medicine; and The Rhazes Center for Research in Family Health and Sexual Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

Siavash Vaziri

Department of Infectious Diseases, Kermanshah University of Medical Sciences, 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 the most important biologically sourced and low-calorie sweeteners that contains a lots of Steviol glycosides. Tissue culture is the best method for propagation of stevia and micro nutrients can affect both morphological traits and steviol glycosides production. In the present study, we investigated the effect of different concentrations of glutamine (10, 20, 30 and 40 g/l) on expression of UGT74G1 and UGT76G1 genes and stevioside and rebaudioside A accumulation in the leaves of stevia under in vitro conditions. The highest level of expression for UGT74G1 (1.000 Total lab unit) was seen at plants grown in MS media without glutamine and the highest gene expression level for UGT76G1 (1.321 Total lab unit) was observed at plants grown in 2% glutamine. Based on HPLC results, the highest amount of stevioside (22.74) was accumulated in plants which were under 3% glutamine treatment and the lowest production level of stevioside (16.19) was resulted under MS (0 glutamine) medium. The highest rebaudioside A (12.19) accumulation was observed under 2% glutamine treatment and the lowest accumulation of rebaudioside A (8.41) was seen at plants grown in MS medium.

Keywords

Stevia rebaudiana Bertoni Tissue Culture Glutamine Semi quantitative RT- PCR HPLC.

Esmaeili, F., Ghaheri, M., Kahrizi, D., Mansouri, M., Safavi, S. M., Ghorbani, T., Muhammadi, S., Rahmanian, E., & Vaziri, S. (2018). Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cellular and Molecular Biology, 64(2), 1–5. https://doi.org/10.14715/cmb/2018.64.2.1

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References

Katayama O, Sumida T, Hayashi H, Mitsuhashi H. The practical application of Stevia and research and development data, I.S.U. Company, Japan, 1976; 747 pp.
Mitsuhashi H, Ueno J, Sumita T, Studies on the cultivation of Stevia rebaudiana Bertoni, Determination of stevioside, Yakugaku Zasshi, 1975; 95: 127-130
Lewis WH. Early uses of Stevia rebaudiana (Asteraceae) leaves as a sweetener in Paraguay, Econ Bot 1992; 46: 336-337.
Lee JI, Kang KK, Lee EU. Studies on new sweetening resource plant stevia (Stevia rebaudiana Bert.) in Korea. I, Effects of transplanting date shifting by cutting and seeding dates on agronomic characteristics and dry leaf yields of stevia, Res Rep ORD 1979; 21: 171-179.
Fors A, A new character in the sweetener scenario, Sugar J. 1995; 58: 30.
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.
Singh S, Garg V, Yadav D, Beg MN, Sharma N. In-vitro antioxidative and antibacterial activities of various parts of stevia rebaudiana (Bertoni). Int J Pharm Pharm Sci 2012; 4(3): 468-473.
Prakash I, Markosyan A, Bunders C. Development of Next Generation Stevia Sweetener: Rebaudioside M. Foods 2014; (3): 162-175
Mondaca RL, Glvez AV, Bravo LZ, Hen KA. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehensive review on the biochemical, nutritional and functional aspects. 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.
Brandle J, Telmer P. Steviol glycoside biosynthesis. Phytochemistry 2007; 68: 1855-1863.
Richman A, Swanson A, Humphrey T, Chapman R, McGarvey B, Pocs R, Brandle J. Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana. Plant J, 2005; 41: 56-67.
Raina R, Bhandari SK, Chand R, Sharma Y. Strategies to improve poor seed germination in Stevia rebaudiana, a low calorie sweetener. J Med Plants Res 2013; 7: 1793-1799.
Savita S, Sheela K, Sunanda S, Shankar A, Ramakrishna P. Stevia rebaudiana–a functional component for food industry. J Hum Ecol 2004; 15: 261–264.
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: 543-549
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15: 473–479.
Basu A, Sethi U, Mukherjee SP. Regulation of cell proliferation and morphogeneis by amino acids in brassica tissue cultures and its correlation with threonine deaminase. Plant Cell Rep 1989; 8: 333-335.
Wang Y, Ruemmela BA, Chandlee JM, Sullivan WM, Knapp JE, Kausch AP. Embryogenic callus induction and plant regeneratio media for bentgrasses and annual blue grass. In Vitro Cell. Dev. Biol. Plant 2002; 38: 460-467.
Kopertekh LG, Stribnaya LA. Plant regeneration from wheat leaf explants. Russ. J. Plant Physiol 2003; 50: 365-368.
Abdullah R, Alizah Z, Wee WH, Leaw CL, Yeap CB.
Immature embryo: A useful tool for oil palm (Elaies guineensis Jacq.) genetic transformation studies. Elect. J Biotechnol 2005; 8: 25-34.
Li M, Villemur R, Hulley PJ, Silflow CD, Grontt JS, Snustad DP. Differential expression of sixglutamine synthase genes in Zea mays. Plant Mol. Biol. 1993; 23: 229-329.
Higashi K, Kamada H, Harada H. The effect of reduced nitrogenous compound sugar in that glutamine synthase activity is involved in the development of somatic embryo carrot. Plant Cell Tiss. Org. Cult 1996; 45: 109-114.
Das A, Mandal N. Enhanced Development of Embryogenic Callus in Stevia rebaudiana Bert. by Additive and Amino acids. Biotechnol. J. 2010; 9: 368-372.
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.

References

Katayama O, Sumida T, Hayashi H, Mitsuhashi H. The practical application of Stevia and research and development data, I.S.U. Company, Japan, 1976; 747 pp.

Mitsuhashi H, Ueno J, Sumita T, Studies on the cultivation of Stevia rebaudiana Bertoni, Determination of stevioside, Yakugaku Zasshi, 1975; 95: 127-130

Lewis WH. Early uses of Stevia rebaudiana (Asteraceae) leaves as a sweetener in Paraguay, Econ Bot 1992; 46: 336-337.

Lee JI, Kang KK, Lee EU. Studies on new sweetening resource plant stevia (Stevia rebaudiana Bert.) in Korea. I, Effects of transplanting date shifting by cutting and seeding dates on agronomic characteristics and dry leaf yields of stevia, Res Rep ORD 1979; 21: 171-179.

Fors A, A new character in the sweetener scenario, Sugar J. 1995; 58: 30.

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.

Singh S, Garg V, Yadav D, Beg MN, Sharma N. In-vitro antioxidative and antibacterial activities of various parts of stevia rebaudiana (Bertoni). Int J Pharm Pharm Sci 2012; 4(3): 468-473.

Prakash I, Markosyan A, Bunders C. Development of Next Generation Stevia Sweetener: Rebaudioside M. Foods 2014; (3): 162-175

Mondaca RL, Glvez AV, Bravo LZ, Hen KA. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehensive review on the biochemical, nutritional and functional aspects. 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.

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

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

Raina R, Bhandari SK, Chand R, Sharma Y. Strategies to improve poor seed germination in Stevia rebaudiana, a low calorie sweetener. J Med Plants Res 2013; 7: 1793-1799.

Savita S, Sheela K, Sunanda S, Shankar A, Ramakrishna P. Stevia rebaudiana–a functional component for food industry. J Hum Ecol 2004; 15: 261–264.

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: 543-549

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

Basu A, Sethi U, Mukherjee SP. Regulation of cell proliferation and morphogeneis by amino acids in brassica tissue cultures and its correlation with threonine deaminase. Plant Cell Rep 1989; 8: 333-335.

Wang Y, Ruemmela BA, Chandlee JM, Sullivan WM, Knapp JE, Kausch AP. Embryogenic callus induction and plant regeneratio media for bentgrasses and annual blue grass. In Vitro Cell. Dev. Biol. Plant 2002; 38: 460-467.

Kopertekh LG, Stribnaya LA. Plant regeneration from wheat leaf explants. Russ. J. Plant Physiol 2003; 50: 365-368.

Abdullah R, Alizah Z, Wee WH, Leaw CL, Yeap CB.

Immature embryo: A useful tool for oil palm (Elaies guineensis Jacq.) genetic transformation studies. Elect. J Biotechnol 2005; 8: 25-34.

Li M, Villemur R, Hulley PJ, Silflow CD, Grontt JS, Snustad DP. Differential expression of sixglutamine synthase genes in Zea mays. Plant Mol. Biol. 1993; 23: 229-329.

Higashi K, Kamada H, Harada H. The effect of reduced nitrogenous compound sugar in that glutamine synthase activity is involved in the development of somatic embryo carrot. Plant Cell Tiss. Org. Cult 1996; 45: 109-114.

Das A, Mandal N. Enhanced Development of Embryogenic Callus in Stevia rebaudiana Bert. by Additive and Amino acids. Biotechnol. J. 2010; 9: 368-372.

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.

Author biographies is not available.