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

Saifi, Monica; Nasrullah, Nazima; Ahmad, Malik M.; Ali, Athar; Khan, J. A.; Abdin, M. Z.

doi:10.1016/j.plaphy.2015.05.009

Cited by 25 publications

(12 citation statements)

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“…Among all the miRNAs analysed for their expression levels, we found the two (miR319g and miRStv_11) which showed the expression patterns similar to that of their target mRNAs. On the other hand, the other miRNAs were found to have expression pattern opposite to their target mRNAs [28]. Thus, it became quite interesting to understand how these two miRNAs (miR319g and miRStv_11) are involved in regulation of steviol glycosides biosynthetic pathway genes.…”

Section: Discussionmentioning

confidence: 99%

“…β-actin was used as an internal control. Reaction mixture and cycling conditions were same as described in our previous report [28]. For amplification of precursor miRNAs and target mRNAs, RT-PCR was carried out using two step RT-PCR kit (Qiagen) with precursor miRNA and mRNA specific forward and reverse primers, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Doing so would significantly increase the quality of the stevia sweetener, not to mention the potential to be used on a wider scale. In our previous study [28], we for the first time identified miRNAs associated with genes of steviol gycosides biosynthetic pathway in S. rebaudiana. Among all the miRNAsanalysed for their expression levels, we found the two miRNAs (miR319g and miRStv_11) which showed the expression patterns similar to their target mRNAs.…”

Section: Introductionmentioning

confidence: 99%

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Co-expression of anti-miR319g and miRStv_11 lead to enhanced steviol glycosides content in Stevia rebaudiana

et al. 2019

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Background miRNAs are major regulators of gene expression and have proven their role in understanding the genetic regulation of biosynthetic pathways. Stevioside and rebaudioside-A, the two most abundant and sweetest compounds found in leaf extract of Stevia rebaudiana , have been used for many years in treatment of diabetes. It has been found that the crude extract is more potent than the purified extract. Stevioside, being accumulated in higher concentration, imparts licorice like aftertaste. Thus, in order to make the sweetener more potent and palatable, there is a need to increase the intrinsic concentration of steviol glycosides and to alter the ratio of rebaudioside-A to stevioside. Doing so would significantly increase the quality of the sweeteners, and the potential to be used on a wider scale. To do so, in previous report, miRNAs associated with genes of steviol glycosides biosynthetic pathway were identified in S. rebaudiana. In continuation to that in this study, the two miRNAs (miR319g and miRStv_11) targeting key genes of steviol glycosides biosynthetic pathway were modulated and their impact was evaluated on steviol glycosides contents. Results The over-expression results showed that miRStv_11 induced, while miR319g had repressive action on its target genes. The knock-down constructs for miR319g and miRStv_11 were then prepared and it was demonstrated that the expression of anti-miR319g produced inhibitory effect on its target miRNA, resulting in enhanced expression of its target genes. On the other hand, anti-miRStv_11 resulted in down-regulation of miRStv_11 and its target gene. Further miRStv_11 and anti-miR319gwere co-expressed which resulted in significant increase in stevioside (24.5%) and rebaudioside-A (51%) contents. Conclusion In conclusion, the role of miR319g and miRStv_11 was successfully validated in steviol gycosides biosynthetic pathway gene regulation and their effect on steviol gycosides contents. In this study, we found the positively correlated miRNA-mRNA interaction network in plants, where miRStv_11 enhanced the expression of KAH gene. miRNAs knock-down was also successfully achieved using antisense precursors. Overall, this study thus reveals more complex nature and fundamental importance of miRNAs in biosynthetic pathway related gene networks and hence, these miRNAs can be successfully employed to enhance the ratio of rebaudioside-A to stevioside, thus enhancing the sweetening indices of this plant and making it more palatable. Electronic supplementary material The online version of this article (10.1186/s12870-019-1871-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Co-expression of anti-miR319g and miRStv_11 lead to enhanced steviol glycosides content in Stevia rebaudiana

et al. 2019

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“…Similarly, miR-4995 was predicted to target mRNA of an enzyme 3-deoxy-7-phosphoheptulonate synthase, which is involved in the picroside biosynthetic pathway in a medicinal herb P. kurroa (Vashisht et al, 2015). In addition, Saifi et al (2015), have mined and validated 11 miRNAs which are involved in steviol glycoside biosynthetic pathway (Table 1) in Stevia and established the relationship pattern with the expression levels of their target mRNAs as well as steviol glycoside contents. Using NGS technology, several miRNAs involved in the sesquiterpene biosynthesis pathway have been mapped and validated in X. strumarium .…”

Section: Role Of Mirnas In Terpenoid Biosynthesismentioning

confidence: 99%

Contemporary Understanding of miRNA-Based Regulation of Secondary Metabolites Biosynthesis in Plants

et al. 2017

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Plant's secondary metabolites such as flavonoids, terpenoids, and alkaloids etc. are known for their role in the defense against various insects-pests of plants and for medicinal benefits in human. Due to the immense biological importance of these phytochemicals, understanding the regulation of their biosynthetic pathway is crucial. In the recent past, advancement in the molecular technologies has enabled us to better understand the proteins, enzymes, genes, etc. involved in the biosynthetic pathway of the secondary metabolites. miRNAs are magical, tiny, non-coding ribonucleotides that function as critical regulators of gene expression in eukaryotes. Despite the accumulated knowledge of the miRNA-mediated regulation of several processes, the involvement of miRNAs in regulating secondary plant product biosynthesis is still poorly understood. Here, we summarize the recent progress made in the area of identification and characterizations of miRNAs involved in regulating the biosynthesis of secondary metabolites in plants and discuss the future perspectives for designing the viable strategies for their targeted manipulation.

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“…miR8154 and miR5298b increase taxol, phenylpropanoid, and flavonoid biosynthesis in subcultured Taxus cells [56]. In silico analysis indicate that miRstv_7* target ugt76g1, KAH, KO, for steviol glycoside biosynthesis [47]. In Arabidopsis thaliana, miR826 and miR5090 share the target AOP2, which encodes a 2-oxoglutarate-dependent dioxygenase that is involved in glucosinolate biosynthesis [48].…”

Section: Other Secondary Metabolitesmentioning

confidence: 99%

MicroRNAs Associated with Secondary Metabolites Production

Vargas-Hernández¹,

Vázquez-Marrufo²,

Agustín³

et al. 2019

Plant Physiological Aspects of Phenolic Compounds

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MicroRNAs (miRNAs) are noncoding RNAs that play an important role in the regulation of the genetic expression in animals and plants by targeting mRNAs for cleavage or translational repression. Several miRNAs regulate the plant development, the metabolism, and the responses to biotic and abiotic stresses. Characterization of an miRNA has helped to show its role in fine tuning the mechanisms of posttranscriptional gene regulation. Although there is a lot of information related to miRNA regulation of some processes, the role of miRNA involved in the regulation of biosynthesis of secondary plant product is still poorly understood. In this chapter, we summarize the identification and characterization of miRNAs that participate in the regulation of the biosynthesis of secondary metabolites in plants and their use in the strategies to manipulate a controlled manipulation.

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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

Cited by 25 publications

References 30 publications

Co-expression of anti-miR319g and miRStv_11 lead to enhanced steviol glycosides content in Stevia rebaudiana

Co-expression of anti-miR319g and miRStv_11 lead to enhanced steviol glycosides content in Stevia rebaudiana

Contemporary Understanding of miRNA-Based Regulation of Secondary Metabolites Biosynthesis in Plants

MicroRNAs Associated with Secondary Metabolites Production

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