Issue

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

The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.

Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions

https://doi.org/10.14715/cmb/2018.64.2.3
Fariba Akbari
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran
Ali Arminian
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran
Danial Kahrizi
author
Arash Fazeli
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran
Matin Ghaheri
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Razi University, Kermanshah, Iran

Corresponding Author(s) : Ali Arminian

a.arminian@ilam.ac.ir

Cellular and Molecular Biology, Vol. 64 No. 2: New biotechnological aspects in Stevia rebaudiana Bertoni

Abstract

Stevia rebaudiana (Bertoni) is a non-caloric sweetener belonging to Asteraceae family. Stevia compounds such as steviol glycosides (SGs) are 200 times sweeter than sugar. Stevioside and rebaudioside A are the two major steviol glycosides. Nitrogen is an essential element for plant growth and development. In this study the effects of nitrogen influenced by different concentrations of NH4NO3 (0, 825 and 1650 mg/l) and KNO3 (0, 950 and 1900 mg/l) is examined in MS medium. To analysis the UGT74G1 and UGT76G1 genes expression, involved in the synthesis of SGs, RT-qPCR technique was performed. Data showed that there were significant differences between all media. The shoot length, seedlings dry weight and leaf fresh weight of stevia increased with applying NH4NO3 along with KNO3. The highest expression of UGT74G1 gene, was observed in plantlets grown on MS medium with 0 mg/l NH4NO3 and 950 mg/l KNO3 (1.291 total lab unit) but the highest expression of UGT76G1 gene, was observed in plantlets grown on MS medium added by 1650 mg/l NH4NO3 +950 mg/l KNO3 (1.08 total lab unit). Moreover, the lowest value of UGT74G1 gene expression were revealed in MS medium added by 1650 mg/l NH4NO3 +0 mg/l KNO3 (0.80 total lab unit) and the lowest values of UGT76G1 gene expression seen in MS medium with 0 mg/l NH4NO3 +950 mg/l KNO3 (0.85 total lab unit) concentrations. The results of this study could be valuable in stevia breeding programs through glycosides biosynthesis pathways.

Keywords

KNO3 NH4NO3 Stevia rebaudiana (Bertoni) Tissue culture UGT74G1 UGT76G1.
Akbari, F., Arminian, A., Kahrizi, D., Fazeli, A., & Ghaheri, M. (2018). Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions. Cellular and Molecular Biology, 64(2), 11–16. https://doi.org/10.14715/cmb/2018.64.2.3
Download Citation
Endnote/Zotero/Mendeley (RIS)
BibTeX
References
  1. Philippe, R.N., M.D. Mey, and J. Anderson, Biotechnological production of natural zero-calorie sweeteners. Current Opinions in Biotechnology, 2014. 26: p. 155–161.
  2. Akbari, F., et al., Effect of nitrogen sources on some morphological characteristics of in vitro Stevia rebaudiana (Bertoni) Cellular and Molecular Biology 2016. 63: p. 2.
  3. Dacomi, E., et al., Evaluation of antioxidant activity of some plant extracts and their application in biscuit. Journal of Food Chemistry, 2005. 90: p. 118-121.
  4. Geuns, J.M.C., Stevioside. Phytochemistry 2003. 64: p. 913–921.
  5. Ghahery, M., D. Kahrizi, and G. Bahrami, Effect of mannitol on some morphological characteristics of in vitro Stevia rebaudiana (Bertoni) . Journal of Biharian Biologist, 2015. (online first): art. 151415. .
  6. Hajihashemi, S. and A.A. Ehsanpour, Antioxidant response of Stevia rebaudiana (Bertoni) To polyethylene glycol and paclobutrazol treatments under in vitro culture. Applied Biochemistry and Biotechnology, 2014. 172: p. 4038–4052.
  7. Madan, S., et al., Stevia rebaudiana (Bertoni) - A review. Indian Journal of Natural Production Resources, 2010. 1(3): p. 267- 286.
  8. Mohamed, R. and A.D. Alhady, Micropropagation of Stevia rebaudiana (Bertoni) A New Sweetening Crop in Egypt. Global Journal of Biotechnology and Biochemistry, 2011. 6(4): p. 178-182.
  9. Ojha, A., V.N. Sharma, and V. Sharma, An efficient protocol for in vitro clonal propagation of natural sweetener plant Stevia rebaudiana (Bertoni) African Journal of Plant Science, 2010. 4(8): p. 319-321.
  10. Richman, A.S., et al., Diterpene synthesis in Stevia rebaudiana: recruitment and up-regulation of key enzymes from the gibberellin biosynthetic pathway. The Plant Journal, 1999. 19: p. 411–421.
  11. Kinghorn, A.D., Stevia: the genus Stevia. 2002: Taylor and Francis. 202.
  12. Saifi, M., et al., In silico analysis and expression profiling of miRNAs targeting genes of steviol glycosides biosynthetic pathway and their relationship with steviol glycosides content in different tissues of Stevia rebaudiana. Plant Physiology and Biochemistry, 2015. 94: p. 57-64.
  13. Kumar, H., et al., A comprehensive analysis of ï¬fteen genes of steviol glycosides biosynthesis pathway in Stevia rebaudiana (Bertoni). Gene, 2012a. 492: p. 276-274.
  14. Ahmad Khan, S., et al., Physical and chemical mutagenesis in Stevia rebaudiana: variant generation with higher UGT expression and glycosidic profile but with low photosynthetic capabilities. Acta Physiolia Plantarum, 2016. 38: p. 4.
  15. Brandle, J.E. and P.G. Telmer, Steviol glycoside biosynthesis. Phytochemistry 2007. 68: p. 1855–1863.
  16. Guleria, P. and S. Kumar Yadav, Identification of miR414 and Expression Analysis of Conserved miRNAs from Stevia rebaudiana. Genomics Proteomics Bioinformatics, 2011. 9(6): p. 211-217.
  17. Hajihashemi, S. and A.A. Ehsanpour, Influence of Exogenously Applied Paclobutrazol on Some Physiological Traits and Growth of Stevia rebaudiana under in Vitro Drought Stress. Biologia 2013. 68: p. 414-420.
  18. Kumar, H., K. Singh, and S. Kumar, 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Stevia rebaudiana (Bertoni) is a functional gene. Molecular Biology Reproduction, 2012b. 39(12): p. 10971-8.
  19. Inugraha, M., D. Maghfoer, and E. Widaryanto, Response of Stevia rebaudiana (Bertoni) to nitrogen and potassium fertilization. International Journal of Agriculture and Veterinary Science, 2014. 10 (1): p. 47-55.
  20. Mubarak, M., et al., Improving sweet leaf Stevia rebaudiana var. (Bertoni) ) resistance to bialaphos herbicide via bar gene transfer Plant Omics Journal, 2015. 8(3): p. 232-237.
  21. Ali Awad, A.S., Cytogenetic studies on Stevia rebaudiana produced by tissue culture and affected by gamma rays and drought in National Center for Radiation Research and Technology. Botany Department. 2009, Ain Shams University University College for Women (Arts, Science and Education)
  22. Bespalhok-Filho, J.C. and K. Hattori, Embryogenic callus formation and logical studies from Stevia rebaudiana Bertoni floret explants. Revista Brasileira de-Fisiologia-Vegetal, 1997. 9(3): p. 185-188.
  23. Bespalhok-Filho, J.C., L.A. Vieira, and J.M. Hashimoto, Factors influencing the in vitro micropropagation of axillary shoots of Stevia rebaudiana Bert. . Revista Brasileira de- Fisiologia-Vegetal, 1992. 4(1): p. 59-61.
  24. Chen, S.Y., Studies on tissue culture of Hevea brasiliensis I. Role of osmotic concentration carbohydrate and pH value in induction of callus growth in plumule tissue from Hevea seeding. Journal of Research Inst. Malaya, 1993. 199: p. 272-276.
  25. Swanson, S.M., G.B. Mahady, and C.W.W. Beecher, Stevioside biosynsis by callus, root, and rooted – shoot culture in vitro. Plant Cell Tissue and Organ Culture, 1992. 28(2): p. 151-157.
  26. Tamura, Y., et al., Clonal propagation of Stevia rebaudiana Bertoni by stem – tip culture. Plant Cell Reports, 1984. 3: p. 183- 185.
  27. Wei, S., et al., The role of nitrogen in leaf senescence of summer maize and analysis of underlying mechanisms using comparative proteomics. Plant Science, 2015. 233: p. 72-81.
  28. Jitendra, M., et al., Micropropagation of an anti-diabetic plant - Stevia rebaudiana (Bertoni) , (Natural Citation as Sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA Journal of Biological Scisnce, 2012. 1(3): p. 37-42.
  29. Kamran Khan, M., et al., Effect of adenine sulphate on in vitro mass propagation of Stevia rebaudiana (Bertoni) . Journal of Medicinal Plants Research, 2014. 8(13): p. 543-549.
  30. Krishna, H., et al., Somaclonal variations and their applications in horticultural crops improvement. Biotechnology, 2016. 6(1): p. 54-72.
  31. Aladakatti, Y.R., et al., Effect of nitrogen, phosphorus and potassium levels on growth and yield of stevia (Stevia rebaudiana Bertoni). Journal of Agricultural Science, 2012. 25: p. 25-29.
  32. Ibrahim, I.A., et al., Nutrient factors affecting in vitro cultivation of Stevia rebaudiana. Journal of Sugar Technology, 2008. 10: p. 248-253.
  33. Kumar, S., et al., Effect of nitrogen on micropropagation of Stevia rebaudiana (Bertoni) Journal of Developmental Research, 2013. 8: p. 23-29.
  34. Yadav, P., et al., Effect of nitrogen sources on rooting of in vitro culture of Stevia rebaudiana (Bertoni) Journal of Biotechnology, 2013. 4: p. 41-46.
  35. Murashige, T. and F. Skoog, A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 1962. 15: p. 473–479.
  36. Pitekelabou, R., A. Aí¯dam, and K. Kokou, Influence of various carbohydrates on the in vitro micropropagation of Nauclea diderrichii (De Wild &T. Durand) Merrill, an endangered forest species in Togo. African Journal of Biotechnology, 2015. 14: p. 1283-1289.
  37. Marone, M., et al., Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biological Proced., 2001. 3(1): p. 19-25.
  38. Nakamura, S. and Y. Tamura, Variation in the main glycosides of Stevia rebaudiana Bertoni. Japanese Journal of Tropical Agriculture, 1985. 29(2): p. 109-116.
  39. Luitel, A., In vitro stevioside production and molecular characterization of callus and cell suspension system of stevia rebaudiana (Bertoni), in 2015 FIP Congress in Düsseldorf (Germany). 2015, © FIP - International Pharmaceutical Federation (www.fip.org): Germany.
  40. Pandey, H., et al., Meeting the challenge of stevioside production in the hairy roots of Stevia rebaudiana by probing the underlying process. Plant Cell, Tissue, and Organ Culture, 2016. 126: p. 511–512.
  41. Kishore, G., et al., Recombinant Production of Steviol Glycosides. WO Patent World Intellectual Property Organization. . 2011.
  42. Tamada, T., Effects of nitrogen sources on growth and leaf nutrient concentrations of ‘Tifblue'rabbiteye blueberry under water culture. Small Fruits Review, 2004. 3(1-2): p. 149-158.
  43. Evans, J.M., Genetic and environmental control of steviol glycoside biosynthesis in Stevia rebaudiana 2014, Michigan State University.
  44. Shock, C., Rebaudi's stevia: natural noncaloric sweetners. . 1982: California Agriculture. p. 4–5.
  45. Barbet-Massin, C., et al., Nitrogen limitation alters biomass production but enhances steviol glycoside concentration in Stevia rebaudiana bertoni. PLOS one, 2015. DOI:10.1371/journal.pone.0133067: p. 1-16.
  46. Pal, P.K., R. Prasad, and V. Pathania, Effect of decapitation and nutrient applications on shoot branching, yield, and accumulation of secondary metabolites in leaves of Stevia rebaudiana Bertoni. . Journal of Plant Physiology, 2013. 170: p. 1526–1535.
  47. Koch, K., Carbohydrate-modulated gene expression in plants. Annual Review of Plant Biology, 1996. 47: p. 509-540.
Read More

Author Biography

Danial Kahrizi, author
agronomy and plant breeding
Crossref
Scopus
Google Scholar
Europe PMC
Download this PDF file
PDF
Statistic
Read Counter : 1911 Download : 597