Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions

D. Kahrizi; S. M. Ghari; M. Ghaheri; F. Fallah; T. Ghorbani; A. Beheshti Ale Agha; E. Kazemi; Z. Ansarypour

doi:10.14715/cmb/2017.63.7.18

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Vol. 63 No. 7: Issue 7

Issue Published : August 22, 2017

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Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions

https://doi.org/10.14715/cmb/2017.63.7.18

D. Kahrizi

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

S. M. Ghari

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

M. Ghaheri

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

F. Fallah

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

T. Ghorbani

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

A. Beheshti Ale Agha

Department of Soil Science, Faculty of Agriculture, Razi University, Kermanshah, Iran

E. Kazemi

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

Z. Ansarypour

Isfahan University of Technology, College of Agriculture, Department of Biotechnology, Isfahan, Iran

Corresponding Author(s) : D. Kahrizi

dkahrizi@yahoo.com

Cellular and Molecular Biology, Vol. 63 No. 7: Issue 7
Article Published : August 15, 2017

Abstract

Stevia rebaudiana is one of the most important biologically sourced and low-calorie sweeteners Bertoni that has a lot of steviol glycosides. Tissue culture is the best for propagation of stevia and micro nutrients can affect both morphological traits and steviol glycosides production. Therefore, the effect of different concentrations of KH2PO4on stevia growth factors and gene expression had been studied by tissue culture methods, RT-PCR and HPLC. According the results, bud numbers had increased significantly in MS + 0.034 mMKH2PO4 media and the highest measured length was seen in plants grown under MS + 0.034 mM KH2PO4 treatment. Also, the highest growth rate (1.396 mm/d) was observed in MS + 0.034 mMKH2PO4.The best concentration of KH2PO4 for expression of UGT74G1 was 0.00425mMand the best one for UGT76G1 expression was 0.017mM. Interestingly, the best media for both stevioside and rebaudioside A accumulation was 0.017mM KH2PO4containing media. There was positive correlation between the best media for gene expression and the best one for steviol glycosides production.

Keywords

Stevia rebaudiana Bertoni Tissue Culture KH2PO4 RT- PCR HPLC UGT74G1 UGT76G1.

Kahrizi, D., Ghari, S. M., 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. Cellular and Molecular Biology, 63(7), 107–111. https://doi.org/10.14715/cmb/2017.63.7.18

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References

Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian biologist 2017; 11(2): on-first (online first)
Esmaeilia F, Kahrizi D, Mansouri M, Yari Kh, Kazemi N, Ghaheri M. Cell Dedifferentiation in Stevia rebauiana as a Pharmaceutical and Medicinal Plant. J. Rep. Pharm. Sci. 2016; 5(1), 12-17.
Chalapathi MV and Thimmegowda S. Natural non-calorie sweetener stevia (Stevia rebaudiana Bertoni): A future crop of India. Crop Res. Hisar 1997; 14(2): 347-350.
Liu J and Li SFY. Separation and determination of stevia sweeteners by capillary electrophoresis and high performance liquid chromatography. J. Liquid Chromatography 1995; 18(9): 1703-1719.
Jitendra M, Monika S, Ratan SD, Priyanka G, Priyanka S, Kiran DJ. Micropropagation of an antidiabetic plant - Stevia rebaudiana Bertoni, (Natural Sweetener) in Hadoti Region of South-East Rajas¬than, India. ISCA. J Biol Sci 2012; 1: 37-42.
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.
Verma R, Gena DD, Lal Jat B. In vitro propagation of Stevia re¬baudiana Bertoni (a sweeting plant). J Pharm Res 2016; 5: 666-685.
Brandle J, Telmer P. Steviol glycoside biosynthesis. Phytochemistry 2007; 68: 1855-1863.
Richman A, Swanson A, Humphrey T, Chapman R, Mc Garvey 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.
Abdul Razak UNA, Ong CB, Yu TS, Lau LK. In vitro Micropropagation of Stevia rebaudiana Bertoni in Malaysia. Braz Arch Biol Technol 2014; 1: 23-28.
Anbazhagan M, Kalpana M, Rajendran R, Natarajan V, Dhanavel D. In vitro production of Stevia rebaudiana Bertoni. Emir J food agric 2010; 22: 216-222.
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.
Mukundan U, Sivaram L. In vitro culture studies on Stevia rebaudiana. J In Vitro Cell Dev Biol 2003; 5: 520-523.
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
Fronza D and Folegatti MV. Water consumption of the Stevia (Stevia rebaudiana Bert.) crop estimated through microlysimiter. J Sci. agric. 2003; 60(3): 595-599.
Geuns JMC. Molecules of interest stevioside. J Phytochem. 2003; 6: 913-921.
Sathyanarayana BN. Plant Tissue Culture: Practices and New Experimental Protocols. I. K. International Pvt Ltd, Botany, Experimental 2007; 316 pages.
George Edwin F, Hall Michael A, De Klerk GJ (Eds.). Plant Propagation by Tissue Culture. Plant Sci. 2008; 1: 65–113.
Tavares AR, Kanashiro S, Ribeiro RCS, Gonçalves AN, Jocys T. Effect of phosphorus on in vitro growth and development of bromeliad Achmea blanchetlana. Acta Hort. 2015; 241-248
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.

References

Ghaheri M, Kahrizi D, Bahrami Gh. Effect of mannitol on some morphological characteristics of in vitro stevia rebaudiana Bertoni. Biharian biologist 2017; 11(2): on-first (online first)

Esmaeilia F, Kahrizi D, Mansouri M, Yari Kh, Kazemi N, Ghaheri M. Cell Dedifferentiation in Stevia rebauiana as a Pharmaceutical and Medicinal Plant. J. Rep. Pharm. Sci. 2016; 5(1), 12-17.

Chalapathi MV and Thimmegowda S. Natural non-calorie sweetener stevia (Stevia rebaudiana Bertoni): A future crop of India. Crop Res. Hisar 1997; 14(2): 347-350.

Liu J and Li SFY. Separation and determination of stevia sweeteners by capillary electrophoresis and high performance liquid chromatography. J. Liquid Chromatography 1995; 18(9): 1703-1719.

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

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.

Verma R, Gena DD, Lal Jat B. In vitro propagation of Stevia re¬baudiana Bertoni (a sweeting plant). J Pharm Res 2016; 5: 666-685.

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

Richman A, Swanson A, Humphrey T, Chapman R, Mc Garvey 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.

Abdul Razak UNA, Ong CB, Yu TS, Lau LK. In vitro Micropropagation of Stevia rebaudiana Bertoni in Malaysia. Braz Arch Biol Technol 2014; 1: 23-28.

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

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.

Mukundan U, Sivaram L. In vitro culture studies on Stevia rebaudiana. J In Vitro Cell Dev Biol 2003; 5: 520-523.

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

Fronza D and Folegatti MV. Water consumption of the Stevia (Stevia rebaudiana Bert.) crop estimated through microlysimiter. J Sci. agric. 2003; 60(3): 595-599.

Geuns JMC. Molecules of interest stevioside. J Phytochem. 2003; 6: 913-921.

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

George Edwin F, Hall Michael A, De Klerk GJ (Eds.). Plant Propagation by Tissue Culture. Plant Sci. 2008; 1: 65–113.

Tavares AR, Kanashiro S, Ribeiro RCS, Gonçalves AN, Jocys T. Effect of phosphorus on in vitro growth and development of bromeliad Achmea blanchetlana. Acta Hort. 2015; 241-248

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.

Author biographies is not available.