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Study of gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni under various mannitol concentrations

  • Matin Ghaheri
  • Danial Kahrizi
  • Gholamreza Bahrami
  • Hamid-Reza Mohammadi-Motlagh
Original Article
  • 15 Downloads

Abstract

Stevia rebaudiana produces sweet steviol glycosides that are 300 times sweeter than sugar and have the beneficial effects on human health including anti-hyperglycaemic. Tissue culture is the best method with high efficacy to propagate stevia. Abiotic stress has an impact on steviol glycoside contents in stevia. Therefore, we investigated the effect of mannitol on the expression of four genes involved in the biosynthesis of stevia including UGT74G1, UGT76G1, kaurene oxidase and kaurene synthase genes and steviol glycosides accumulation in stevia under in vitro conditions. The highest expression of UGT76G1 gene occurred in the plants grown under 20 g/l mannitol. While for the kaurene synthase gene, the highest amount of gene expression was observed at 40 g/l mannitol. The results were different about kaurene oxidase gene. As the highest and lowest gene expression were seen in 50 and 30 g/l mannitol conditions respectively. There were the same results for UGT74G1 that means the most appropriate and also the most inopportune treatment for the gene expression were same as the condition for the kaurene oxidase gene. Compared with control, adding mannitol to media in all concentrations increases the expression of UGT76G1 gene. Estimation of steviol glycosides contents under different treatments of mannitol carried out by HPLC. According to the results, the highest amount of stevioside was produced under 20 g/l mannitol treatment. However, rebaudioside A was accumulated in its maximum amounts under 30 g/l mannitol. It can be concluded that adding mannitol to media in the certain concentration increases steviol glycoside contents in the stevia.

Keywords

Stevia rebaudiana Bertoni Real Time PCR Rebaudioside A Stevioside HPLC 

Abbreviations

PCR

Polymerase chain reaction

RNA

Ribonucleic acid

HPLC

High-performance liquid chromatography

Ct

Cycle threshold

DXS

Deoxyxyulose-5-phosphate synthase

DXR

Deoxyxyulose-5-phosphate reductoisomerase

CMS

4-Diphosphocytidyl-2-C-methyl-d-erythritol synthase

CMK

4-Diphosphocytidyl-2-C-methyl-d-erythritol kinase

MCS

4-Diphosphocytidyl-2-C-methyl-d-erythritol 2,4-cyclodiphosphate synthase

HDS

1-Hydroxy-2-methyl-2(E)-butenyl 4-diphosphate synthase

HDR

1-Hydroxy-2-methyl-2(E)-butenyl 4-diphosphate reductase

GGDPS

Geranylgeranyl diphosphate synthase

CPS

Copalyl diphosphate synthase

KS

Kaurene synthase

KO

Kaurene oxidase

KAH

Kaurenoic acid 13-hydroxylase

UGT85C2

UDP-glycosyltransferase 85C2

UGT74G1

UDP-glycosyltransferase 74G1

UGT76G1

UDP-glycosyltransferase 76G1

Notes

Acknowledgements

I would like to thank Dr. Tayebeh Ghorbani for her kindly helps in doing this research. Thanks to Zagros Bioidea Co., Razi University Incubator for all supports.

Author Contributions

MG: Execution research project, Data analysis, Manuscript preparation. DK: Experimental design, Data analysis, Manuscript preparation. GB: Data analysis, proofreading of the article. HMM: Experimental design, Data analysis, proofreading of the article.

Compliance with ethical standards

Conflict of interest

Matin Ghaheri, Danial Kahrizi, Gholamreza Bahrami, and Hamidreza Mohammadi Motlagh declares that they have no conflict of interest.

Ethical approval

This study was approved by the Ethics Committee of Razi University, Kermanshah, Iran. All participants signed the informed consent form, in line with the principles of the Helsinki II declarations.

Supplementary material

11033_2018_4250_MOESM1_ESM.docx (3.1 mb)
Supplementary material 1 (DOCX 3172 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Agronomy and Plant Breeding, Faculty of AgricultureRazi UniversityKermanshahIran
  2. 2.Medical Biology Research CenterKermanshah University of Medical SciencesKermanshahIran
  3. 3.School of PharmacyKermanshah University of Medical SciencesKermanshahIran

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