Association of BrERF72 with methyl jasmonate-induced leaf senescence of Chinese flowering cabbage through activating JA biosynthesis-related genes

The ethylene response factor(ERF)and phytohormone jasmonate(JA)are reported to function in leaf senescence.The involvement of ERF in JA-mediated leaf senescence,however,needs to be elucidated.In the present work,we demonstrate a Chinese flowering cabbage ERF transcription factor(TF),BrERF72,that is associated with JA-promoted leaf senescence.Exogenous application of methyl jasmonate(MeJA)-accelerated leaf senescence of Chinese flowering cabbage,evidenced by the data that MeJA treatment led to the stronger reduction in the maximum quantum yield(Fv/Fm),photosynthetic electron transport rate(ETR),and total chlorophyll content,while significant induction in the expression of several senescence-associated genes(SAGs)including BrSAG12,BrSAG19,and chlorophyll catabolic genes(CCGs)BrPAO1,BrNYC1,BrPPH1,and BrSGR1.Increases in levels of endogenous JA and transcripts of JA biosynthetic genes BrLOX4,BrAOC3,and BrOPR3 were also found after MeJA treatment.BrERF72 was a MeJA-inducible,nucleus-localized protein,and possessed trans-activation ability.Transient overexpression of BrERF72 in tobacco leaves also promoted leaf senescence.More importantly,further experiments revealed that BrERF72 directly activated expression of BrLOX4,BrAOC3,and BrOPR3 through binding to their promoters via the GCC or DRE/CRT cis-element.Together,the novel JA-ERF association reported in our study uncovers a new insight into the transcriptional regulation of JA production mediated by ERF during JA-promoted leaf senescence in Chinese flowering cabbage.

Mutagenesis of SlNAC4 by CRISPR/Cas9 alters gene expression and softening of ripening tomato fruit

Softening is one of the key fruit quality traits,which results from the selective expression of cell wall metabolism genes during ripening.The identification of transcription factors(TFs)that regulate fruit softening is an important field in order to understand and control fruit softening.In tomato,NAC(NAM,ATAF,and CUC)TFs members have been demonstrated to be involved in fruit ripening regulation,including NAC-NOR(nonripening),NOR-like1,SlNAC4,SlNAC1.Here,we generated slnac4 mutant knockout(CR-SlNAC4)tomato plant by a clustered regularly interspaced short palindromic repeats genomic targeting system(CRISPR/Cas9)and SlNAC4 overexpressing(OE-SlNAC4)plant.In addition to confirming the previously reported results that SlNAC4 positively regulates fruit ripening,we found that SlNAC4 has a strong effect on tomato fruit softening.Compared with the control fruit,fruit softening was inhibited in slnac4 fruit and conversely was accelerated in OE-SlNAC4 tomato fruit.Through RNA-sequencing(RNA-seq)analysis,we found that expression levels of SlEXP1(expansin)and SlCEL2(endo-β-1,4 glucanase)genes involved in cell wall metabolism were significantly different in WT(wild type)/slnac4 and WT/OE-SlNAC4 fruit.Further study showed that these genes contained a NAC TF binding domain in their promoter regions.In vitro electrophoretic mobility shift assays(EMSA)and dual-luciferase reporter assays(DLR)demonstrated that these two genes were the direct targets of SlNAC4 binding and transactivation.The results enriched the function of SlNAC4 and provided a new dimension in understanding the regulation of tomato fruit softening.

Emerging role of molecular diagnosis and personalized therapy for hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is a prevalent malignancy worldwide,ranking as the sixth most common malignancy and the third leading cause of cancer-related mortality.Late diagnosis,limited management options,and its complex etiology contribute to the poor prognosis and high mortality rates.Recent advances in understanding the molecular mechanisms of HCC and innovations in high-throughput sequencing technologies have led to the development of molecular diagnostics and personalized therapies for this challenging malignancy.This review provides a comprehensive overview of research on the molecular diagnosis and individualized treatment for HCC.We highlight key advances and potential future directions and discuss the application of next-generation sequencing technologies to identify and characterize genetic and epigenetic alterations in HCC patients.These technologies may aid in the selection of targeted therapies,prediction of treatment response,and monitoring disease progression.Furthermore,we explore the role of liquid biopsy in HCC diagnosis,prognosis prediction,and treatment monitoring,focusing on circulating tumor cells,circulating tumor DNA,and extracellular vesicles.We also explore the evolving landscape of personalized therapy for HCC,including targeted therapies against key oncogenic signaling pathways,immune checkpoint inhibitors,tumor-agnostic therapies,and innovative cellbased therapies.We discuss the challenges and opportunities that lie ahead in the quest to improve HCC patient outcomes through the integration of molecular diagnostics and individualized precision therapies.We emphasize the need for multi-interdisciplinary collaboration,refinement of predictive and prognostic biomarkers,and the development of more effective combination strategies for HCC management in the new area of precision medicine.