Enhanced production of phenolic acids in Salvia miltiorrhiza hairy root cultures by combing the RNAi-mediated silencing of chalcone synthase gene with salicylic acid treatment
“…In this study, we detected 4 differentially expressed CHS genes that were all upregulated (Table 1 ), and metabolic analysis showed that the abundance of naringenin chalcone, which is primarily catalyzed by CHS, was also higher in JIZI 2 than in JIZI 1 (Table S1 ). Zhang et al reported that the silenced CHS gene enhanced the phenolic acid content and decreased the accumulation of flavonoids in Salvia miltiorrhiz a 32 . Matoušek et al reported that naringenin chalcone is a precursor of prenylflavonoids and plays an important role in the biosynthesis pathways of flavonoid 33 .…”
Perilla frutescens (L.) is an important medicinal and edible plant in China with nutritional and medical uses. The extract from leaves of Perilla frutescens contains flavonoids and volatile oils, which are mainly used in traditional Chinese medicine. In this study, we analyzed the transcriptomic and metabolomic data of the leaves of two Perilla frutescens varieties: JIZI 1 and JIZI 2. A total of 9277 differentially expressed genes and 223 flavonoid metabolites were identified in these varieties. Chrysoeriol, apigenin, malvidin, cyanidin, kaempferol, and their derivatives were abundant in the leaves of Perilla frutescens, which were more than 70% of total flavonoid contents. A total of 77 unigenes encoding 15 enzymes were identified as candidate genes involved in flavonoid biosynthesis in the leaves of Perilla frutescens. High expression of the CHS gene enhances the accumulation of flavonoids in the leaves of Perilla frutescens. Our results provide valuable information on the flavonoid metabolites and candidate genes involved in the flavonoid biosynthesis pathways in the leaves of Perilla frutescens.
“…In this study, we detected 4 differentially expressed CHS genes that were all upregulated (Table 1 ), and metabolic analysis showed that the abundance of naringenin chalcone, which is primarily catalyzed by CHS, was also higher in JIZI 2 than in JIZI 1 (Table S1 ). Zhang et al reported that the silenced CHS gene enhanced the phenolic acid content and decreased the accumulation of flavonoids in Salvia miltiorrhiz a 32 . Matoušek et al reported that naringenin chalcone is a precursor of prenylflavonoids and plays an important role in the biosynthesis pathways of flavonoid 33 .…”
Perilla frutescens (L.) is an important medicinal and edible plant in China with nutritional and medical uses. The extract from leaves of Perilla frutescens contains flavonoids and volatile oils, which are mainly used in traditional Chinese medicine. In this study, we analyzed the transcriptomic and metabolomic data of the leaves of two Perilla frutescens varieties: JIZI 1 and JIZI 2. A total of 9277 differentially expressed genes and 223 flavonoid metabolites were identified in these varieties. Chrysoeriol, apigenin, malvidin, cyanidin, kaempferol, and their derivatives were abundant in the leaves of Perilla frutescens, which were more than 70% of total flavonoid contents. A total of 77 unigenes encoding 15 enzymes were identified as candidate genes involved in flavonoid biosynthesis in the leaves of Perilla frutescens. High expression of the CHS gene enhances the accumulation of flavonoids in the leaves of Perilla frutescens. Our results provide valuable information on the flavonoid metabolites and candidate genes involved in the flavonoid biosynthesis pathways in the leaves of Perilla frutescens.
“…S3), so they were placed on a medium containing only 400 mg/L Cef. Hairy root lines were established using rapidly growing root lines that were free from bacterial contamination [30]. Fig.…”
Section: Methodsmentioning
confidence: 99%
“…Elicitors, which are defined as signaling compounds, can enhance or induce the biosynthesis of metabolites by activating pathways that respond to external stress [28, 29]. As with other in vitro culture systems, the accumulation of target metabolites can be readily increased by adding elicitors to hairy root cultures [30–32]. Previous studies have documented the effect of endogenous hormones and elicitors, such as salicylic acid (SA), methyl jasmonate (MJ), and yeast extract (YE), on tanshinone metabolism in hairy root cultures of S. miltiorrhiza [11, 22, 33].…”
Background
Tanshinones are diterpenoid compounds that are used to treat cardiovascular diseases. As current extraction methods for tanshinones are inefficient, there is a pressing need to improve the production of these bioactive compounds to meet increasing demand.
Results
Overexpression of
SmMDS
(2-
c
-methyl-
d
-erythritol 2,4-cyclodiphosphate synthase, a tanshinone biosynthesis gene) in transgenic
Salvia miltiorrhiza
hairy roots significantly increased the tanshinone yield compared to the control, and total tanshinone content in
SmMDS
-overexpressing lines increased after elicitor treatment. Total tanshinones increased to 2.5, 2.3, and 3.2 mg/g DW (dry weight) following treatment with Ag
+
, YE (yeast extract), and MJ (methyl jasmonate), respectively, compared with the non-induced transgenic line (1.7 mg/g DW). Also, qRT-PCR analysis showed that the expression levels of two pathway genes was positively correlated with increased accumulation of tanshinone.
Conclusions
Our study provides an effective strategy for increasing the content of tanshinones and other natural compounds using a combination of genetic engineering and elicitor treatment.
Electronic supplementary material
The online version of this article (10.1186/s13007-019-0439-3) contains supplementary material, which is available to authorized users.
“…In response, many studies have since focused on increasing this species phenolic acid contents by either overexpressing or suppressing its transcription factors or key enzyme genes within the relevant metabolic pathways. For example, the overexpression of SmTAT , SmC4H , SmHPPR , AtPAP1 , AtEDT1 , and SmPAP1 induced a substantial accumulation of phenolic acids in the transgenic S. miltiorrhiza ( Zhang Y. et al, 2010 ; Xiao et al, 2011 ; Hao et al, 2016 ; Liu Y. et al, 2016 ), whereas the downregulation of SmMYB39 , SmHPPD , SmCCR1 , or SmCHS all increased the content of phenolic acids ( Xiao et al, 2011 ; Wang et al, 2012 ; Zhang et al, 2013 ; Zhang et al, 2015 ).…”
Subclass III members of the sucrose non-fermenting-1-related protein kinase 2 (SnRK2) play essential roles in both the abscisic acid signaling and abiotic stress responses of plants by phosphorylating the downstream ABA-responsive element (ABRE)-binding proteins (AREB/ABFs). This comprehensive study investigated the function of new candidate genes, namely SmSnRK2.3, SmSnRK2.6, and SmAREB1, with a view to breeding novel varieties of Salvia miltiorrhiza with improved stress tolerance stresses and more content of bioactive ingredients. Exogenous ABA strongly induced the expression of these genes. PlantCARE predicted several hormones and stress response cis-elements in their promoters. SmSnRK2.6 and SmAREB1 showed the highest expression levels in the leaves of S. miltiorrhiza seedlings, while SmSnRK2.3 exhibited a steady expression in their roots, stems, and leaves. A subcellular localization assay revealed that both SmSnRK2.3 and SmSnRK2.6 were located in the cell membrane, cytoplasm, and nucleus, whereas SmAREB1 was exclusive to the nucleus. Overexpressing SmSnRK2.3 did not significantly promote the accumulation of rosmarinic acid (RA) and salvianolic acid B (Sal B) in the transgenic S. miltiorrhiza hairy roots. However, overexpressing SmSnRK2.6 and SmAREB1 increased the contents of RA and Sal B, and regulated the expression levels of structural genes participating in the phenolic acid-branched and side-branched pathways, including SmPAL1, SmC4H, Sm4CL1, SmTAT, SmHPPR, SmRAS, SmCHS, SmCCR, SmCOMT, and SmHPPD. Furthermore, SmSnRK2.3 and SmSnRK2.6 interacted physically with SmAREB1. In summary, our results indicate that SmSnRK2.6 is involved in stress responses and can regulate structural gene transcripts to promote greater metabolic flux to the phenolic acid-branched pathway, via its interaction with SmAREB1, a transcription factor. In this way, SmSnRK2.6 contributes to the positive regulation of phenolic acids in S. miltiorrhiza hairy roots.
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