Identification, characterization, and expression profiling of the putative U-box E3 ubiquitin ligase gene family in Sorghum bicolor.

The U-box family is one of the main E3 ubiquitin ligase families in plants. The U-box family has been characterized in several species. However, genome-wide gene identification and expression profiling of the U-box family in response to abiotic stress in Sorghum bicolor remain unclear. In this study, we broadly identified 68 U-box genes in the sorghum genome, including 2 CHIP genes, and 1 typical UFD2 (Ub fusion degradation 2) gene. The U-box gene family was divided into eight subclasses based on homology and conserved domain characteristics. Evolutionary analysis identified 14, 66, and 82 U-box collinear gene pairs in sorghum compared with arabidopsis, rice, and maize, respectively, and a unique tandem repeat pair (SbPUB26/SbPUB27) is present in the sorghum genome. Gene Ontology (GO) enrichment analysis showed that U-box proteins were mainly related to ubiquitination and modification, and various stress responses. Comprehensive analysis of promoters, expression profiling, and gene co-regulation networks also revealed that many sorghum U-box genes may be correlated with multiple stress responses. In summary, our results showed that sorghum contains 68 U-box genes, which may be involved in multiple abiotic stress responses. The findings will support future gene functional studies related to ubiquitination in sorghum.

Characterization of histone deacetylases and their roles in response to abiotic and PAMPs stresses in Sorghum bicolor.

BACKGROUND: Histone deacetylases (HDACs) play an important role in the regulation of gene expression, which is indispensable in plant growth, development, and responses to environmental stresses. In Arabidopsis and rice, the molecular functions of HDACs have been well-described. However, systematic analysis of the HDAC gene family and gene expression in response to biotic and abiotic stresses has not been reported for sorghum. RESULTS: We conducted a systematic analysis of the sorghum HDAC gene family and identified 19 SbHDACs mainly distributed on eight chromosomes. Phylogenetic tree analysis of SbHDACs showed that the gene family was divided into three subfamilies: RPD3/HDA1, SIR2, and HD2. Tissue-specific expression results showed that SbHDACs displayed different expression patterns in different tissues, indicating that these genes may perform different functions in growth and development. The expression pattern of SbHDACs under different stresses (high and low temperature, drought, osmotic and salt) and pathogen-associated molecular model (PAMPs) elf18, chitin, and flg22) indicated that SbHDAC genes may participate in adversity responses and biological stress defenses. Overexpression of SbHDA1, SbHDA3, SbHDT2 and SbSRT2 in Escherichia coli promoted the growth of recombinant cells under abiotic stress. Interestingly, we also showed that the sorghum acetylation level was enhanced when plants were under cold, heat, drought, osmotic and salt stresses. The findings will help us to understand the HDAC gene family in sorghum, and illuminate the molecular mechanism of the responses to abiotic and biotic stresses. CONCLUSION: We have identified and classified 19 HDAC genes in sorghum. Our data provides insights into the evolution of the HDAC gene family and further support the hypothesis that these genes are important for the plant responses to abiotic and biotic stresses.

Cytochrome P450 Superfamily: Evolutionary and Functional Divergence in Sorghum (Sorghum bicolor) Stress Resistance.

Cytochrome P450 (CYP) genes encode enzymes that catalyze various growth-, development-, and stress-related reactions. Sorghum (Sorghum bicolor) is a type of C4 plant and an important cash crop. However, systematic identification and analysis of functional differentiation and evolution of CYP genes have not been carried out in this species. In the present study, we revealed that the sorghum genome contains 351 CYP genes, which can be divided into nine classes. These genes are from ancestors and repeated segments, rather than tandem repeats. Based on collinearity results, a large number of CYPs were extended before cotyledon differentiation, during the emergence of Gramineae, suggesting that genomewide duplication events and stress adaptation processes were important for the expansion of CYP genes. Their gene structure and motifs contain conserved regions and include various changes and loci. The expression characteristics and functional annotation of CYP genes indicated tissue specificity and selective expression. Overall, we identified all CYP genes in the sorghum genome and preliminarily explored their naming, structure, evolution, expression, and functional differentiation. The results advanced our understanding of plant gene family evolution and functional differentiation.

Histone deacetylase SbHDT701 in Sorghum bicolor reveals functions in response to stress factors by enhancing acetylation.

Histone acetylation plays important roles in eukaryotic chromatin modification and gene expression regulation. Acetylation levels are modulated by histone deacetylases (HDACs), which function as key epigenetic factors that regulate gene expression in response to various stresses. HDT701, a member of the HD2 subfamily of HDACs, plays crucial roles in plant responses to abiotic stress and pathogen infection. Here, we analysed the expression pattern of SbHDT701 in sorghum. Real-time fluorescence quantitative PCR (RT-qPCR) results showed that expression of SbHDT701 was tissue-specific, and up-regulated under drought (d-mannitol) and salt (NaCl) stresses. We also determined the optimal expression conditions for SbHDT701 protein accumulation, and successfully expressed and purified SbHDT701 protein. Besides, overexpression of SbHDT701 in could promote the growth of recombinant cells under abiotic stress. SbHDT701 expression in Escherichia coli also increased acetylation modification levels following treatment with 750 mM NaCl, and 100 mM or 300 mM d-mannitol. In summary, the sorghum HDAC SbHDT701 mediates stress responses by enhancing acetylation modification levels.

Identification of the constituents and metabolites in rat plasma after oral administration of HuanglianShangqing pills by ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

An ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry method was established to detect and identify the chemical constituents and metabolites of HuanglianShangqing pill (HLSQ) in vitro and in vivo. A total of 96 compounds were characterized in HLSQ extracts. In vivo, 45 prototype components and 53 metabolites of HLSQ were detected in rat plasma and tentatively identified, three of which are new. The novel metabolic pathways of rhein and hesperetin were revealed. The bioactive compounds of HLSQ undergo phase I metabolic routes of hydrogenation, hydroxylation, hydrolysis, demethylation and phase II metabolic routes of glucuronide, sulfation, acetylation and methylation. Among these, glucuronide conjugation is the main metabolic pathway of the active compounds of HLSQ in rat plasma.