Identification of a 301 bp promoter core region of the SrUGT91D2 gene from Stevia rebaudiana that contributes to hormone and abiotic stress inducibility
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View abstract on PubMed
Summary
This summary is machine-generated.The promoter of the UDP-glucuronosyltransferase 91D2 (SrUGT91D2) gene was analyzed, revealing a key 26 bp insertion that enhances its activity. This finding provides insights into steviol glycoside biosynthesis regulation.
Area Of Science
- Plant Molecular Biology
- Biochemistry
- Metabolic Engineering
Background
- The UDP-glucuronosyltransferase 91D2 (SrUGT91D2) gene is essential for steviol glycoside (SG) biosynthesis, catalyzing the formation of key glucosidic bonds.
- It plays a critical role in synthesizing rebaudioside M (RM) and rebaudioside D (RD), important steviol glycosides.
- Understanding the regulation of SrUGT91D2 gene expression through its promoter is crucial, but its functional analysis remained unexplored.
Purpose Of The Study
- To isolate and characterize the promoter of the SrUGT91D2 gene (pSrUGT91D2) from different Stevia rebaudiana lines.
- To investigate the functional activity of pSrUGT91D2 and its variants in regulating gene expression.
- To identify regulatory elements within the promoter, particularly a 26 bp insertion, and their role in gene transcription.
Main Methods
- Isolation of pSrUGT91D2 from six S. rebaudiana lines.
- Multiple sequence alignment to identify sequence variations, including a 26 bp insertion/deletion (inDel).
- Bioinformatic analysis to predict cis-regulatory elements (CREs).
- Construction of plant expression vectors with the reporter gene β-glucuronidase (GUS) fused to full-length and truncated promoter fragments.
- Transient transformation in tobacco leaves to assess promoter activity via GUS staining and enzyme assays.
- Response analysis to exogenous hormones (ABA, IAA) and light.
Main Results
- A 26 bp inDel fragment was identified in the pSrUGT91D2 of several lines (pSrUGT91D2-B1188 type) but absent in line 023 (pSrUGT91D2-023 type).
- Bioinformatics predicted numerous CREs responsive to various stimuli like ABA, light, and hormones.
- Both full-length and truncated pSrUGT91D2 fragments demonstrated transcriptional activity in tobacco.
- A 301 bp fragment containing the inDel (P8-pSrUGT91D2-B1188) showed enhanced GUS gene expression.
- The inDel region was found essential for the promoter's transcriptional activity and responsiveness to ABA, IAA, and light.
Conclusions
- The study elucidates the regulatory mechanisms of the SrUGT91D2 promoter, highlighting the role of a specific inDel region.
- Findings provide a theoretical foundation for further research on the interaction between pSrUGT91D2 CREs and transcription factors.
- This enhances understanding of steviol glycoside biosynthesis regulation at the gene expression level.
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