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Isolation and functional analysis of SrMYB1, a direct transcriptional repressor of SrUGT76G1 in Stevia rebaudiana

ZHANG Ting, ZHANG Yong-xia, SUN Yu-ming, XU Xiao-yang, WANG Yin-jie, CHONG Xin-ran, YANG Yong-heng and YUAN Hai-yan
2023, 22 (4): 1058-1067.   DOI: 10.1016/j.jia.2023.03.001
Abstract212)      PDF in ScienceDirect      

SrUGT76G1, the most well-studied diterpene glycosyltransferase in Stevia rebaudiana, is key to the biosynthesis of economically important steviol glycosides (SGs).  However, the molecular regulatory mechanism of SrUGT76G1 has rarely been explored.  In this study, we identified a MYB transcription factor, SrMYB1, using a yeast one-hybrid screening assay.  SrMYB1 belongs to the typical R2R3-type MYB protein and is specifically localized in the nucleus with strong transactivation activity.  The transcript of SrMYB1 is predominantly accumulated in flowers, but is also present at a lower level in leaves.  Yeast one-hybrid and electrophoretic mobility shift assays verified that SrMYB1 binds directly to the MYB binding sites in the F4-3 fragment (+50–(–141)) of the SrUGT76G1 promoter.  Furthermore, we found that SrMYB1 could significantly repress the expression of SrUGT76G1 in both epidermal cells of tobacco leaves and stevia callus.  Taken together, our results demonstrate that SrMYB1 is an essential upstream regulator of SrUGT76G1 and provide novel insight into the regulatory network for the SGs metabolic pathway in S. rebaudiana.

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Potassium deficiency inhibits steviol glycosides synthesis by limiting leaf sugar metabolism in stevia (Stevia rebaudiana Bertoni) plants
SUN Yu-ming, HUANG Xiao-lei, ZHANG Ting, YANG Yong-heng, CHENG Xiao-fang, XU Xiao-yang, YUAN Hai-yan
2021, 20 (11): 2932-2943.   DOI: 10.1016/S2095-3119(20)63472-4
Abstract134)      PDF in ScienceDirect      
The steviol glycosides (SGs) in stevia (Stevia rebaudiana Bertoni) leaves are becoming increasingly valuable due to its high sweetness but low calorific value, which is driving the development of stevia commercial cultivation.  Optimizing fertilization management can effectively increase SGs productivity, but knowledge on the relationship between potassium (K) fertilization and SGs production is still lacking.  In this study, pot experiments were conducted in order to investigate the effect of K deficiency on SGs synthesis in stevia leaves, as well as the underlying mechanisms.  Our results showed that when compared with standard K fertilization, K deficiency treatment has no significant effect on the biomass of stevia plant grown in a given soil with high K contents.  However, K deficiency critically decreased leaf SGs contents as well as the expression of SGs synthesis-related genes.  The contents of different sugar components decreased and the activities of sugar metabolism-related enzymes were inhibited under the K deficiency condition.  Moreover, spraying sucrose on the leaves of stevia seedlings diminished the inhibitory effect caused by K deficiency.  Our results also revealed the significant positive correlations between sucrose, glucose and SGs contents.  Overall, our results suggest that K deficiency would suppress the synthesis of SGs in stevia leaves, and this effect may be mediated by the leaf sugar metabolism.  Our findings provide new insights into the improvement of SGs production potential. 
 
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