Flower abscission is an important developmental process that can significantly reduce the yield of horticultural plants. We previously reported that SmMYB113 is a key transcription factor promoting anthocyanin biosynthesis and improve fruit quality. However, the overexpression of SmMYB113 in eggplant increased flower drop rate and reduced fruit yield. Here, we elucidate the regulatory mechanisms of SmMYB113 on flower abscission in eggplant. RNA-seq analysis indicated that the regulation of flower abscission by SmMYB113 was associated with altered expression of genes related to ethylene biosynthesis and signal transduction, including ethylene biosynthetic genes SmACS1, SmACS8 and SmACO4. Then, the ethylene content in flowers and the function of ethephon (ETH, which promotes fruit ripening) and 1-Methylcyclopropene (1-MCP, which acts as an ethylene perception inhibitor) were analyzed, which revealed that SmMYB113 directly regulates ethylene-dependent flower abscission. Yeast one-hybrid and dual-luciferase assays revealed that SmMYB113 could directly bind to the promoters of SmACS1, SmACS8, and SmACO4 to activate their expression. Through construction of a yeast two-hybrid (Y2H) screening library, the protein SmERF38 was found to interact with SmMYB113, and verified by Y2H, bimolecular fluorescence complementation (BiFC), and luciferase complementation assay. Furthermore, dual-luciferase assays showed that SmERF38 enhanced the role of SmMYB113 on the promoters of SmACS1. Our results provided new insight into the molecular mechanism of flower abscission in eggplant.

Eggplant (Solanum melongena L.) is a highly nutritious vegetable. Here, the molecular mechanism of color formation in eggplants was determined using six eggplant cultivars with different peel colors and two SmMYB113-overexpressing transgenic eggplants with a purple peel and pulp. Significant differentially expressed genes (DEGs) were identified by RNA-sequencing analysis using the following criteria: log2(sample1/sample2) ≥ 0.75 and q-value ≤ 0.05. Two analytical strategies were used to identify genes related to the different peel color according to the peel color, flavonoids content, delphinidins/flavonoids ratio, and the content of anthocyanins. Finally, 27 novel genes were identified to be related to the color difference among eggplant peels and 32 novel genes were identified to be related to anthocyanin biosynthesis and regulated by SmMYB113. Venn analysis revealed that SmCytb5, SmGST, SmMATE, SmASAT3, and SmF3'5'M were shared among both sets of novel genes. Transient expression assay in tobacco suggested that these five genes were not sufficient for inducing anthocyanin biosynthesis alone, but they play important roles in anthocyanin accumulation in eggplant peels. Yeast one-hybrid, electrophoretic mobility shift assay and dual-luciferase assays indicated that the expression of the five genes could be directly activated by SmMYB113 protein. Finally, a regulatory model for the mechanism of color formation in eggplant was proposed. Overall, the results of this study provide useful information that enhances our understanding of the molecular mechanism underlying the different color formation in eggplant.

To understand the color formation mechanism in eggplant (Solanum melongena L.) peel, a metabolomic analysis was performed in six cultivars with different peel colors. A total of 167 flavonoids, including 16 anthocyanins, were identified based on a UPLC-MS/MS approach. Further analysis revealed that the delphinidins/flavonoids ratio was consistent with the purple coloration of eggplant peels, and SmF3 ' 5 ' H expression level was consistent with the delphinidin 3-O-glucoside and delphinidin 3-O-rutinoside contents, the main anthocyanins in the purple-peels eggplant cultivars identified in this study. SmMYB113 overexpression promoted anthocyanins accumulation in eggplant peels and pulps. Metabolomic analysis revealed that delphinidins were still the main anthocyanins class in the peels and pulps of SmMYB113-OE4, but most anthocyanins were glycosylated at the 5-position of the B-ring. Our results provide new insights into the anthocyanin composition of eggplant peels and demonstrate the importance of SmMYB113 in stimulating anthocyanin biosynthesis in eggplant fruits.

The basic helix-loop-helix (bHLH) superfamily is considered the second largest transcription factor (TF) family. It plays regulatory roles in the developmental processes of plants and in their defense responses. In recent years, many bHLH superfamily genes have been identified and characterized in herbaceous and woody plants. However, the comprehensive genomic and functional analyses of these genes in eggplant (Solanum melongena L.) have not been reported. In this study, 121 bHLH TFs were identified in the recently released eggplant genome. The phylogeny, gene structure and conserved motifs of the SmbHLH gene were comprehensively studied. Subsequently, the phylogenetic relationship between the bHLH of eggplant and the bHLH of other species was analyzed, and the proteins were classified into 17 subfamilies. Among these protein sequences, 16 subgroups were clustered into the functional clades of Arabidopsis. Two candidate genes (SmbHLH1, SmbHLH117) that may be involved in anthocyanin biosynthesis were screened. The tissue specificity or differential expression of the bHLH genes in different tissues and under various light and temperature conditions suggested the differential regulation of tissue development and metabolism. This study not only provides a solid foundation for the functional dissection of the eggplant bHLH gene family but may also be useful for the future synthesis of anthocyanins in eggplant.

Plant phospholipase D (PLD), which can hydrolyze membrane phospholipids to produce phosphatidic acid (PA), a secondary signaling molecule, has been proposed to function in diverse plant stress responses. Both PLD and PA play key roles in plant growth, development, and cellular processes. PLD was suggested to mediate the regulation of stomatal movements by abscisic acid (ABA) as a response to water deficit. In this research, we characterized the roles of the cucumber phospholipase D alpha gene (CsPLDα, GenBank accession number EF363796) in the growth and tolerance of transgenic tobacco (Nicotiana tabacum) to drought stress.

The R2R3MYB proteins comprise one of the largest families of transcription factors and play regulatory roles in developmental processes and defense responses in plants. However, limited comprehensive genomic and functional analyses of these genes in eggplant (Solanum melongena L.) have been performed. In this study, 73 R2R3MYB genes were identified in the recently released eggplant genome and further classified into 20 subgroups based on the phylogenetic analysis of their complete protein sequences. Phylogenetic comparisons of this superfamily’s members among eggplant, tomato, Arabidopsis, grape, rice, poplar, soybean, cucumber and apple revealed that the putative functions of some of the eggplant R2R3MYB proteins were clustered into the Arabidopsis functional clades. The tissue specificity or differential expression of SmR2R3MYBs in different tissues suggested the differential regulation of tissue development as well as metabolism. The transcript abundance analysis under abiotic stress identified a group of R2R3MYB genes that responded to one or more treatments, suggesting that SmR2R3MYBs played major roles in the plants response to abiotic stress and were involved in signal transduction pathways. Genes related to anthocyanin synthesis were also identified. This study not only provides a solid foundation for the functional dissection of the eggplant R2R3MYB gene family, but may also be useful for the future synthesis of anthocyanins in eggplant.