Mediator subunit MED8 interacts with heat shock transcription factor HSF3 to promote fucoxanthin synthesis in the diatom Phaeodactylum tricornutum.

Fucoxanthin, a natural carotenoid that has substantial pharmaceutical value due to its anticancer, antioxidant, antiobesity, and antidiabetic properties, is biosynthesized from glyceraldehyde-3-phosphate (G3P) via a series of enzymatic reactions. However, our understanding of the transcriptional mechanisms involved in fucoxanthin biosynthesis remains limited. Using reverse genetics, the med8 mutant was identified based on its phenotype of reduced fucoxanthin content, and the biological functions of MED8 in fucoxanthin synthesis were characterized using approaches such as gene expression, protein subcellular localization, protein-protein interaction and chromatin immunoprecipitation assay. Gene-editing mutants of MED8 exhibited decreased fucoxanthin content as well as reduced expression levels of six key genes involved in fucoxanthin synthesis, namely DXS, PSY1, ZDS-like, CRTISO5, ZEP1, and ZEP3, when compared to the wild-type (WT) strain. Furthermore, we showed that MED8 interacts with HSF3, and genetic analysis revealed their shared involvement in the genetic pathway governing fucoxanthin synthesis. Additionally, HSF3 was required for MED8 association with the promoters of the six fucoxanthin synthesis genes. In conclusion, MED8 and HSF3 are involved in fucoxanthin synthesis by modulating the expression of the fucoxanthin synthesis genes. Our results increase the understanding of the molecular regulation mechanisms underlying fucoxanthin synthesis in the diatom P. tricornutum.

Jasmonate regulates the FAMA/mediator complex subunit 8-THIOGLUCOSIDE GLUCOHYDROLASE 1 cascade and myrosinase activity.

Myrosinases are ß-thioglucoside glucosidases that are unique to the Brassicales order. These enzymes hydrolyze glucosinolates to produce compounds that have direct antibiotic effects or that function as signaling molecules in the plant immune system, protecting plants from pathogens and insect pests. However, the effects of jasmonic acid (JA), a plant hormone that is crucial for plant disease resistance, on myrosinase activity remain unclear. Here, we systematically studied the effects of JA on myrosinase activity and explored the associated internal transcriptional regulation mechanisms. Exogenous application of JA significantly increased myrosinase activity, while the inhibition of endogenous JA biosynthesis and signaling reduced myrosinase activity. In addition, some myrosinase genes in Arabidopsis (Arabidopsis thaliana) were upregulated by JA. Further genetic and biochemical evidence showed that transcription factor FAMA interacted with a series of JASMONATE ZIM-DOMAIN proteins and affected JA-mediated myrosinase activity. However, among the JA-upregulated myrosinase genes, only THIOGLUCOSIDE GLUCOHYDROLASE 1 (TGG1) was positively regulated by FAMA. Further biochemical analysis showed that FAMA bound to the TGG1 promoter to directly mediate TGG1 expression in conjunction with Mediator complex subunit 8 (MED8). Together, our results provide evidence that JA acts as an important signal upstream of the FAMA/MED8-TGG1 pathway to positively regulate myrosinase activity in Arabidopsis.