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Comparative Study
. 2019 Jun 12;9(1):8559.
doi: 10.1038/s41598-019-44989-4.

Comparison of uridine diphosphate-glycosyltransferase UGT76G1 genes from some varieties of Stevia rebaudiana Bertoni

Nader R Abdelsalam  1 , William A Botros  2 , Ahmed E Khaled  3 , Mohamed A Ghonema  4 , Shimaa G Hussein  4 , Hayssam M Ali  5   6 , Mohamed S Elshikh  5
Affiliations
Comparative Study

Comparison of uridine diphosphate-glycosyltransferase UGT76G1 genes from some varieties of Stevia rebaudiana Bertoni

Nader R Abdelsalam et al. Sci Rep. .

Abstract

Stevia leaves contain various components, such as flavonoids, labdanes, chlorophylls, sterols, triterpenoids, mono-disaccharides, organic acids and inorganic salts. Stevia is known to accumulate diterpenoid steviol glycosides, which are approximately 300 times sweeter than regular sugar. Stevioside and rebaudioside A are the main diterpenic glycosides in stevia. Steviol glycosides are the secondary metabolites responsible for the sweetness of stevia. The main objectives of the present study were to determine the concentrations of diterpenic glycosides (stevioside and rebaudioside A) in three stevia varieties (Stevia rebaudiana) via the HPLC-UV technique and to amplify the UGT76G1 gene by PCR using gene-specific primers. The expression levels of the UGT76G1 gene were determined in the three stevia varieties. The PCR products were sequenced and analyzed, and the nucleotide sequences of the UGT76G1 gene were submitted to GenBank and assigned to the following three varieties: Egy1 (MH087463), China1 (MH087464) and Sponti (MH087465). Cluster analysis was used to separate the three varieties into two major clusters based on their phylogenetic relationship. In addition, chemical analysis was carried out to evaluate stevioside and rebaudioside A. The present study concluded that Egy1 and Sponti are closely related varieties as they fall in the same cluster, while China1 forms a separate cluster. Bioprospecting studies could be useful for selection of superior ecotypes of Stevia rebaudiana.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
HPLC of stevia varieties Egy1 (A), China1 (B)and Sponti (C).
Figure 2
Figure 2
PCR products of the UGT76G1 gene amplified from Egy1, China1 and Sponti.
Figure 3
Figure 3
Nucleotide sequencing of the UGT76G1 gene from stevia plants of different varieties: (A) Egy1 (519 bp); (B) China1 (634); (C) Sponti (549).
Figure 4
Figure 4
Multiple DNA sequence alignments of UGT76G1 genes with the genes from three different varieties of stevia. Completely conserved residues across all the aligned sequences are marked with an asterisk (*) below. Absent nucleotides are indicated by dashes (−).
Figure 5
Figure 5
Multiple DNA sequence alignments of the UGT76G1 genes with the genes from three different varieties of stevia available GenBank. Completely conserved residues across all the aligned sequences are marked with an asterisk (*) below. Absent nucleotides are indicated by dashes (−).
Figure 6
Figure 6
Dendrogram illustrating the phylogenetic relationship of different varieties based on DNA nucleotide sequencing of the UGT76G1 gene.
Figure 7
Figure 7
Dendrogram illustrating the phylogenetic relationship of different varieties based on DNA nucleotide sequencing of the UGT76G1 gene and comparison with the same gene listed in GenBank.
Figure 8
Figure 8
Quantitative estimation of UGT76G1 gene expression in stevia leaves.
See this image and copyright information in PMC

References

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