Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression

T. Ghorbani; D. Kahrizi; M. Saeidi; I. Arji

doi:10.14715/cmb/2017.63.8.8

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Vol. 63 No. 8: Issue 8

Issue Published : September 4, 2017

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Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression

https://doi.org/10.14715/cmb/2017.63.8.8

T. Ghorbani

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

D. Kahrizi

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

M. Saeidi

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

I. Arji

Horticultural Crop Research Department, Kermanshah Agricultural and Natural Resources Research and Education Center, AREEO, Kermanshah, Iran

Corresponding Author(s) : D. Kahrizi

dkahrizi@yahoo.com

Cellular and Molecular Biology, Vol. 63 No. 8: Issue 8
Article Published : August 30, 2017

Abstract

Stevia rebaudiana (Bert.) Bertoni is known as sweet plant which it contains a high level of steviol glycosides in the leaves. This plant has been used from centuries ago as a sweetener for tea. One of the most important steviol glycosides is stevioside that is attractive for diabetic persons. Tissue culture is the only rapid process for the mass propagation of stevia. One of the most important factors in the medium is sucrose that is a necessary for plant growth. In the present study, we use nodal segments of the stem as explants in mediums with different sucrose concentration (50 mM, 100mM and 150mM). Several morphological traits were measured in a 28 day period. Results analysis showed a significant variation between treatments. The highest growth rate, rooting and leaf production was obtained in medium with 100mM sucrose. The correlation between measured traits was significant at the 0.01 level. To investigation of UGT74G1, UGT76G1, UGT85C2 and KS genes expression that are involved in the synthesis of SGs, RT- PCR was done with the housekeeping gene of as internal control. There were significant differences between all media. The results showed thatsucrose 100 mM containing media was more desirable than others for expression of UGT76G1 and UGT85C2 genes. Whereas, the best medium for expression of UGT74G1 was sucrose 150 mM and sucrose 50 mM for KS gene. Totally, it seems that sucrose at a concentration of 100 mMprovides the best condition for stevia growth and steviol glycosides production.

Keywords

Stevia rebaudiana Bertoni Tissue culture Sucrose Gene expression Semi-quantitative RT- PCR.

Ghorbani, T., Kahrizi, D., Saeidi, M., & Arji, I. (2017). Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cellular and Molecular Biology, 63(8), 33–37. https://doi.org/10.14715/cmb/2017.63.8.8

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References

Madan S, Ahmad S, Singh G, Kohli K, Kumar Y, Singh R, Garg M. Stevia rebaudiana (Bert.) Bertoni-a review. Indian J Nat Prod Resour. 2010; 1: 267-286.
Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an Anti diabetic Plant-Stevia rebaudiana Bertoni,(Natural Sweetener) in Hadoti Region of South-East Rajasthan. India ISCA J Biol Sci. 2012; 1: 37-42.
Ahmed MB, Salahin M, Karim R, Razvy MA, Hannan MM, Sultana R, Hossain M, Islam R. An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana Bertoni.) in Bangladesh American-Eurasian. J Sci Res 2007; 2: 121-125.
Brandle J, Telmer P. Steviol glycoside biosynthesis. Phytochem 2007; 68: 1855-1863.
Khan SA, Rahman L, Verma R, Shanker K. Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiol Plant 2016; 38: 1-12.
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.
Guleria P, Kumar V, Yadav SK. Effect of sucrose on steviol glycoside biosynthesis pathway in Stevia rebaudiana. Asian J Plant Sci 2011; 10: 401-409.
Koch K. Carbohydrate-modulated gene expression in plants. Annu Rev Plant Biol 1996; 47: 509-540.
Loreti E, De Bellis L, Alpi A, Perata P. Why and how do plant cells sense sugars? Ann Bot 2001; 88: 803-812.
Smeekens S. Sugar-induced signal transduction in plants. Annu Rev Plant Biol 2000; 51: 49-81.
Ramesh. K, Singh V, Megeji NW. Cultivation of Stevia [Stevia rebaudiana (Bert.) Bertoni]. Adv Agron 2006; 89: 137-177.
Nepovim A, Vanek T. In vitro propagation of Stevia rebaudina plants using multiple shoot culture. Planta Med 1998; 64: 775-776.
Mazinga K, Mario G, Baboy L, Useni S, Nyembo K, Kasongo L, Van K. Effets des sources de carbone (sucrose, glucose), et des doses de saccharose sur l'induction de la rhizogénèse chez l'hybride FHIA-01 (AAAB) du bananier en culture in vitro. J Appl Biosci 2015; 73: 5991-6001.
Gauchan D. Effect of different sugars on shoot regeneration of maize (Zea mays L.). KUSET.2012; 8: 119-124.
Nambiar N, Tee C, Maziah M. Effects of organic additives and different carbohydrate sources on proliferation of protocormlike bodies in'Dendrobium'Alya Pink plant. Omics J 2012; 5: 10-18.
Preethi D, Sridhar T, Naidu C. Carbohydrate concentration influences on in vitro plant regeneration in Stevia rebaudiana. J Phytol 2011; 3: 61-64.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 1962; 1: 473-497.
Marone M, Mozzetti S, De Ritis D, 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: 19-25.
Chivers JE, Gong W, King EM, Seybold J, Mak JC, Donnelly LE, Holden NS, Newton R. Analysis of the dissociated steroid RU24858 does not exclude a role for inducible genes in the anti-inflammatory actions of glucocorticoids. Mol Pharmacol 2006; 70: 2084-2095.
Ismaili A, Mohammadi PP. Effect of genotype, induction medium, carbohydrate source, and polyethylene glycol on embryogenesis in maize (Zea mays L.) anther culture. Acta Physiol Plant 2016; 38, 1-8.
Rhee Y, Hwang KR, Cho S, Lee M, Kil EJ, Choi S, Hahn BS, Kim D, Auh CK, Lee S. Expression analysis of D-type cyclin in potato (Solanum tuberosum L.) under different culture conditions. Acta Physiol Plant 2016; 38: 1-12.
Bondarev N, Reshetnyak O, Nosov A. Peculiarities of diterpenoid steviol glycoside production in vitro cultures of Stevia rebaudiana Bertoni. Plant Sci 2001; 161: 155-163.
Esmaeili F, Kahrizi D, Mansouri M, Yari K, Kazemi N, Ghaheri M. Cell dedifferentiation in Stevia rebauiana as a pharmaceutical and medicinal plant. J Rep Pharma Sci 2016; 5(1): 12-17.
Akbari F, Arminian A, Kahrizi D, Fazeli A. Effect of nitrogen sources on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Cell Mol Biol 2017; 63(2): 107-111.
Ghaheri M, Kahrizi D, Bahrami Gh.. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist 2017 (online first): art. 151415.

References

Madan S, Ahmad S, Singh G, Kohli K, Kumar Y, Singh R, Garg M. Stevia rebaudiana (Bert.) Bertoni-a review. Indian J Nat Prod Resour. 2010; 1: 267-286.

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

Ahmed MB, Salahin M, Karim R, Razvy MA, Hannan MM, Sultana R, Hossain M, Islam R. An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana Bertoni.) in Bangladesh American-Eurasian. J Sci Res 2007; 2: 121-125.

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

Khan SA, Rahman L, Verma R, Shanker K. Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiol Plant 2016; 38: 1-12.

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.

Guleria P, Kumar V, Yadav SK. Effect of sucrose on steviol glycoside biosynthesis pathway in Stevia rebaudiana. Asian J Plant Sci 2011; 10: 401-409.

Koch K. Carbohydrate-modulated gene expression in plants. Annu Rev Plant Biol 1996; 47: 509-540.

Loreti E, De Bellis L, Alpi A, Perata P. Why and how do plant cells sense sugars? Ann Bot 2001; 88: 803-812.

Smeekens S. Sugar-induced signal transduction in plants. Annu Rev Plant Biol 2000; 51: 49-81.

Ramesh. K, Singh V, Megeji NW. Cultivation of Stevia [Stevia rebaudiana (Bert.) Bertoni]. Adv Agron 2006; 89: 137-177.

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

Mazinga K, Mario G, Baboy L, Useni S, Nyembo K, Kasongo L, Van K. Effets des sources de carbone (sucrose, glucose), et des doses de saccharose sur l'induction de la rhizogénèse chez l'hybride FHIA-01 (AAAB) du bananier en culture in vitro. J Appl Biosci 2015; 73: 5991-6001.

Gauchan D. Effect of different sugars on shoot regeneration of maize (Zea mays L.). KUSET.2012; 8: 119-124.

Nambiar N, Tee C, Maziah M. Effects of organic additives and different carbohydrate sources on proliferation of protocormlike bodies in'Dendrobium'Alya Pink plant. Omics J 2012; 5: 10-18.

Preethi D, Sridhar T, Naidu C. Carbohydrate concentration influences on in vitro plant regeneration in Stevia rebaudiana. J Phytol 2011; 3: 61-64.

Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 1962; 1: 473-497.

Marone M, Mozzetti S, De Ritis D, 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: 19-25.

Chivers JE, Gong W, King EM, Seybold J, Mak JC, Donnelly LE, Holden NS, Newton R. Analysis of the dissociated steroid RU24858 does not exclude a role for inducible genes in the anti-inflammatory actions of glucocorticoids. Mol Pharmacol 2006; 70: 2084-2095.

Ismaili A, Mohammadi PP. Effect of genotype, induction medium, carbohydrate source, and polyethylene glycol on embryogenesis in maize (Zea mays L.) anther culture. Acta Physiol Plant 2016; 38, 1-8.

Rhee Y, Hwang KR, Cho S, Lee M, Kil EJ, Choi S, Hahn BS, Kim D, Auh CK, Lee S. Expression analysis of D-type cyclin in potato (Solanum tuberosum L.) under different culture conditions. Acta Physiol Plant 2016; 38: 1-12.

Bondarev N, Reshetnyak O, Nosov A. Peculiarities of diterpenoid steviol glycoside production in vitro cultures of Stevia rebaudiana Bertoni. Plant Sci 2001; 161: 155-163.

Esmaeili F, Kahrizi D, Mansouri M, Yari K, Kazemi N, Ghaheri M. Cell dedifferentiation in Stevia rebauiana as a pharmaceutical and medicinal plant. J Rep Pharma Sci 2016; 5(1): 12-17.

Akbari F, Arminian A, Kahrizi D, Fazeli A. Effect of nitrogen sources on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Cell Mol Biol 2017; 63(2): 107-111.

Ghaheri M, Kahrizi D, Bahrami Gh.. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian Biologist 2017 (online first): art. 151415.

Author biographies is not available.