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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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Changes in speciation, mobility and bioavailability of Cd, Cr and As during the transformation process of pig manure by black soldier fly larvae (Hermetia illucens)
WANG Xiao-bo, WU Nan, CAI Rui-jie, GENG Wei-na, XU Xiao-yan
2021, 20 (5): 1157-1166.   DOI: 10.1016/S2095-3119(20)63333-0
Abstract132)      PDF in ScienceDirect      
The black soldier fly larvae (BSFL) (Hermetia illucens) are a good candidate for poultry and livestock manure treatment.  The harvested insect bodies and feces can be used as animal feed and organic fertilizer.  However, heavy metals have a negative impact on the transformation process of the manure by BSFL.  Here we evaluated the effects of Cd, Cr and As in pig manure on the growth of BSFL, as well as the mobility and changes in speciation of the metals during the transformation process.  The results showed that As significantly reduced the weight of BSFL, but Cr and Cd had no significant effects on BSFL growth.  The bioaccumulation factors (BAFs) at various concentrations of Cd (2.8–3.7) were significantly higher than those of Cr (0.44–0.62) and As (0.43–0.45).  The heavy metals in pig manure were mainly transferred to BSFL feces, and the distribution percentages of Cd, Cr and As in the BSFL feces were 52.6–62.0%, 90.3–94.2% and 93.0–93.3%, respectively.  Cd concentrations in BSFL feces were significantly lower than those in the pig manure, while there were no significant differences in the concentrations of Cr and As between BSFL feces and pig manure (except for the treatment with the addition of 150 mg Cr kg–1).  The metal speciation (weak acid soluble, reducible, oxidizable and residual fractions) in BSFL feces obviously changed when compared with pig manure.  In BSFL feces, the reducible proportion of Cr decreased while the oxidizable proportion increased.  Cd mainly existed in the weak acid soluble and reducible states in pig manure and BSFL feces.  For As, the proportions of weak acid soluble and reducible states decreased in BSFL feces.  After the BSFL transformation process, the bioavailable fraction of Cr decreased by 17.3–23.1%, but those of Cd and As did not change significantly, except for As in the CK group.  These findings contribute to our understanding of the roles of BSFL during the biotransformation process of livestock manure as well as the safe utilization of transformed products.
 
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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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