scholarly journals Characterizing glycosyltransferases by a combination of sequencing platforms applied to the leaf tissues of Stevia rebaudiana

2020 ◽  
Author(s):  
Shaoshan Zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong Chen ◽  
Renfeng Xiao ◽  
...  

Abstract Background: Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides.Results: Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a more complete and correct full-length transcriptome of S. rebaudiana . Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained, including SrUGT85C2 , SrUGT74G1 , SrUGT76G1 and one SrUGT91D2, which have already been reported to be involved in the glucosylation of steviol glycosides, illustrating the reliability of our results.Conclusion: Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and forty-four candidate SrUGTs involved in the glycosylation of steviol glucosides were obtained. Moreover, the complete and correct full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana .

2020 ◽  
Author(s):  
shaoshan zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong chen ◽  
Renfeng xiao ◽  
...  

Abstract Background: Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides. Results: Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a more complete and correct full-length transcriptome of S. rebaudiana. Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained, including SrUGT85C2, SrUGT74G1, SrUGT76G1 and one SrUGT91D2, which have already been reported to be involved in the glucosylation of steviol glycosides, illustrating the reliability of our results.Conclusion: Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and forty-four candidate SrUGTs involved in the glycosylation of steviol glucosides were obtained. Moreover, the complete and correct full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana.


BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Shaoshan Zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong Chen ◽  
Renfeng Xiao ◽  
...  

Abstract Background Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides. Results Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a full-length transcriptome of S. rebaudiana. Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained, including SrUGT85C2, SrUGT74G1, SrUGT76G1 and SrUGT91D2, which have already been reported to be involved in the glucosylation of steviol glycosides, illustrating the reliability of our results. Conclusion Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and forty-four candidate SrUGTs involved in the glycosylation of steviol glucosides were obtained. Moreover, the full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana.


2020 ◽  
Author(s):  
shaoshan zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong chen ◽  
Renfeng xiao ◽  
...  

Abstract Background: Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides. Results: Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a full-length transcriptome of S. rebaudiana. Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained, including SrUGT85C2, SrUGT74G1, SrUGT76G1 and one SrUGT91D2, which have already been reported to be involved in the glucosylation of steviol glycosides, illustrating the reliability of our results.Conclusion: Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and forty-four candidate SrUGTs involved in the glycosylation of steviol glucosides were obtained. Moreover, the full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana.


2020 ◽  
Author(s):  
shaoshan zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong chen ◽  
Renfeng xiao ◽  
...  

Abstract Background: Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides. Results: Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a full-length transcriptome of S. rebaudiana. Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained, including SrUGT85C2, SrUGT74G1, SrUGT76G1 and SrUGT91D2, which have already been reported to be involved in the glucosylation of steviol glycosides, illustrating the reliability of our results.Conclusion: Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and forty-four candidate SrUGTs involved in the glycosylation of steviol glucosides were obtained. Moreover, the full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana.


2020 ◽  
Author(s):  
Shaoshan Zhang ◽  
Qiong Liu ◽  
Chengcheng Lyu ◽  
Jinsong Chen ◽  
Renfeng Xiao ◽  
...  

Abstract Background: Stevia rebaudiana (Bertoni) is considered one of the most valuable plants because of the steviol glycosides (SGs) that can be extracted from its leaves. Glycosyltransferases (GTs), which can transfer sugar moieties from activated sugar donors onto saccharide and nonsaccharide acceptors, are widely distributed in the genome of S. rebaudiana and play important roles in the synthesis of steviol glycosides. Results: Six stevia genotypes with significantly different concentrations of SGs were obtained by induction through various mutagenic methods, and the contents of seven glycosides (stevioboside, Reb B, ST, Reb A, Reb F, Reb D and Reb M) in their leaves were considerably different. Then, NGS and single-molecule real-time (SMRT) sequencing were combined to analyse leaf tissue from these six different genotypes to generate a more complete and correct full-length transcriptome of S. rebaudiana. Two phylogenetic trees of glycosyltransferases (SrUGTs) were constructed by the neighbour-joining method and successfully predicted the functions of SrUGTs involved in SG biosynthesis. With further insight into glycosyltransferases (SrUGTs) involved in SG biosynthesis, the weighted gene co-expression network analysis (WGCNA) method was used to characterize the relationships between SrUGTs and SGs, and forty-four potential SrUGTs were finally obtained. Of these potential SrUGTs, twenty-seven have complete ORFs and have not been verified to date, and enzyme assays were subsequently performed, but none of them had activity towards SGs. In addition, SrUGT (SrUGT88B1-1) could utilize UDP-glucose as a sugar donor to glycosylate isoquercetin to form three products containing one, two, and three glucoses, respectively.Conclusion: Combined with the results obtained by previous studies and those of this work, we systematically characterized glycosyltransferases in S. rebaudiana and confirmed that four enzymes (SrUGT85C2, SrUGT74G1, SrUGT76G1 and SrUGT91D2) are primarily involved in the glycosylation of steviol glucosides. Moreover, the complete and correct full-length transcriptome obtained in this study will provide valuable support for further research investigating S. rebaudiana.


Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 4090
Author(s):  
Morteza Sheikhalipour ◽  
Behrooz Esmaielpour ◽  
Gholamreza Gohari ◽  
Maryam Haghighi ◽  
Hessam Jafari ◽  
...  

High salt levels are one of the significant and major limiting factors on crop yield and productivity. Out of the available attempts made against high salt levels, engineered nanoparticles (NPs) have been widely employed and considered as effective strategies in this regard. Of these NPs, titanium dioxide nanoparticles (TiO2 NPs) and selenium functionalized using chitosan nanoparticles (Cs–Se NPs) were applied for a quite number of plants, but their potential roles for alleviating the adverse effects of salinity on stevia remains unclear. Stevia (Stevia rebaudiana Bertoni) is one of the reputed medicinal plants due to their diterpenoid steviol glycosides (stevioside and rebaudioside A). For this reason, the current study was designed to investigate the potential of TiO2 NPs (0, 100 and 200 mg L−1) and Cs–Se NPs (0, 10 and 20 mg L−1) to alleviate salt stress (0, 50 and 100 mM NaCl) in stevia. The findings of the study revealed that salinity decreased the growth and photosynthetic traits but resulted in substantial cell damage through increasing H2O2 and MDA content, as well as electrolyte leakage (EL). However, the application of TiO2 NPs (100 mg L−1) and Cs–Se NPs (20 mg L−1) increased the growth, photosynthetic performance and activity of antioxidant enzymes, and decreased the contents of H2O2, MDA and EL under the saline conditions. In addition to the enhanced growth and physiological performance of the plant, the essential oil content was also increased with the treatments of TiO2 (100 mg L−1) and Cs–Se NPs (20 mg L−1). In addition, the tested NPs treatments increased the concentration of stevioside (in the non-saline condition and under salinity stress) and rebaudioside A (under the salinity conditions) in stevia plants. Overall, the current findings suggest that especially 100 mg L−1 TiO2 NPs and 20 mg L−1 Cs–Se could be considered as promising agents in combating high levels of salinity in the case of stevia.


2021 ◽  
Vol 11 (8) ◽  
pp. 3634
Author(s):  
Teresa Leszczyńska ◽  
Barbara Piekło ◽  
Aneta Kopeć ◽  
Benno F. Zimmermann

This study compares the content of basic nutrients (proteins, fats, digestible carbohydrates, dietary fiber and ash), steviol glycosides, selected antioxidants (vitamin C, total polyphenols) and antioxidant activity in dried leaves of Stevia rebaudiana Bertoni cultivated in Poland, Paraguay and Brazil and available in the direct sale. The basic chemical composition was determined by standard AOAC (Association of Official Analytical Chemists) methods. Content of steviol glycosides was determined by the UHPLC-UV chromatographic method. Total polyphenols content was expressed as gallic acid equivalent (GAE) and catechins equivalent (CE). Antioxidant activity was measured as ABTS●+ free radical scavenging activity. Dried leaves of S. rebaudiana grown in Poland had significantly higher contents of dietary fiber, and lower protein and ash content, compared to those derived from Paraguay and Brazil. The former had, however, considerably higher contents of total steviol glycosides, stevioside and rebaudioside D, compared to the remaining two plants. In the Paraguay-derived dried leaves, the content of rebaudioside A, C, E and rubusoside was found to be significantly lower. Dried leaves of S. rebaudiana Bertoni, cultivated in Poland, contained substantially more vitamin C and a similar content of total polyphenols, compared to those from Brazil and Paraguay. The examined material from Brazil and Paraguay plantations showed similar antioxidant activity, while that obtained from Polish cultivation was characterized by a significantly lower value of this parameter.


Author(s):  
Ma. Cristina Vazquez-Hernandez ◽  
Aurora Alvarado-Mariana ◽  
Humberto Aguirre-Becerra ◽  
Genaro Martin Soto-Zarazua ◽  
Ana Angelica Feregrino-Perez ◽  
...  

Processes ◽  
2018 ◽  
Vol 6 (8) ◽  
pp. 105 ◽  
Author(s):  
Monica Gallo ◽  
Andrea Formato ◽  
Gaetano Formato ◽  
Daniele Naviglio

Abstract: Stevia rebaudiana Bertoni is a perennial shrub belonging to the Asteraceae family. The leaves contain a mixture of steviol glycosides with extraordinary sweetening properties, among which the most important are stevioside and rebaudioside A. These components have a high sweetening power, which is about 300 times that of sucrose, and a negligible calorie content. However, their extraction and purification are not easy. In this paper, the extraction technique under cyclic pressure, known as rapid solid-liquid dynamic extraction (RSLDE), was compared using a Naviglio extractor (NE) with conventional maceration. The aim was to identify an efficient and economically viable method for obtaining high amounts of steviol glycosides in a short time. Furthermore, a numerical model was set up for the solid-liquid extraction process of value-added compounds from natural sources. Several parameters must be evaluated in relation to the characteristics of the parts of the plant subjected to extraction. Therefore, since diffusion and osmosis are highly dependent on temperature, it is necessary to control the temperature of the extraction system. On the other hand, the final aim of this work was to provide a scientific and quantitative basis for RSLDE. Therefore, the results obtained from stevia extracts using the corresponding mathematical model allowed hypothesizing the application of this model to the extraction processes of other vegetable matrices.


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