Lettuce is an important leafy vegetable that represents a significant dietary source of antioxidants and bioactive compounds. However, the levels of metabolites in different lettuce cultivars are poorly characterized. In this study, we used combined GC × GC-TOF/MS and UPLC-IMS-QTOF/MS to detect and relatively quantify metabolites in 30 lettuce cultivars representing large genetic diversity. Comparison with online databases, the published literature, standards as well using collision cross-section values enabled putative identification of 171 metabolites. Sixteen of these 171 metabolites (including phenolic acid derivatives, glycosylated flavonoids, and one iridoid) were present at significantly different levels in leaf and head type lettuces, which suggested the significant metabolomic variations between the leaf and head types of lettuce are related to secondary metabolism. A combination of the results and metabolic network analysis techniques suggested that leaf and head type lettuces contain not only different levels of metabolites but also have significant variations in the corresponding associated metabolic networks. The novel lettuce metabolite library and novel non-targeted metabolomics strategy devised in this study could be used to further characterize metabolic variations between lettuce cultivars or other plants. Moreover, the findings of this study provide important insight into metabolic adaptations due to natural and human selection, which could stimulate further research to potentially improve lettuce quality, yield, and nutritional value.

Cucumber (Cucumis sativus L.) is a typical monoecious vegetable with individual male and female flowers, and has been used as a model plant for sex determination. It is well known that sex differentiation of cucumber can be regulated by phytohormones, such as gibberellic acid (GA) and ethylene. The molecular mechanism of female sex expression modulated by ethylene has been widely understood, but how GA controls male sex expression remains elusive. In hermaphroditic Arabidopsis and rice, GA can regulate stamen and anther development via the transcriptional regulation of GAMYB. Here we characterized a GAMYB homologue CsGAMYB1 in cucumber. We found that CsGAMYB1 is predominantly expressed in male flower buds, where its expression is upregulated by GA₃ treatment. CsGAMYB1 protein is localized in the nucleus. CsGAMYB1 can partially rescue stamen development and fertility phenotypes of an Arabidopsis myb33 myb65 double mutant. However, constitutive overexpression of CsGAMYB1 in wild-type Arabidopsis resulted in male sterility, which mimics the effect of GA overdose in flower development. Knockdown of CsGAMYB1 in cucumber decreases the ratio of nodes with male and female flowers, and ethylene is not involved in this process. Our data suggest that CsGAMYB1 regulates sex expression of cucumber via an ethylene-independent pathway.

The cucumber (Cucumis sativus L.) fruit is covered with bloom trichomes and warts (comprised of spines and tubercules), which have an important impact on the commercial value of the crop. However, little is known about the regulatory mechanism underlying their formation. Here, we reported that the cucumber WD-repeat homolog, CsTTG1, which is localized in the nucleus and cytomembrane, plays an important role in the formation of cucumber fruit bloom trichomes and warts. Functional characterization of CsTTG1 revealed that it is mainly expressed in the epidermis of cucumber fruit and that its overexpression in cucumber alters the density of fruit bloom trichomes and spines, thereby promoting the warty (Wty) fruit trait. Conversely, silencing CsTTG1 expression inhibits the initiation of fruit spines. Molecular and genetic analyses showed that CsTTG1 acts in parallel to Mict/CsGL1, a key trichome formation factor, to regulate the initiation of fruit trichomes, including fruit bloom trichomes and spines, and that the further differentiation of fruit spines and formation of tubercules regulated by CsTTG1 is dependent on Mict. Using yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) assay, we determined that CsTTG1 directly interacts with Mict. Collectively, our results indicate that CsTTG1 is an important component of the molecular network that regulates fruit bloom trichome and wart formation in cucumber.

Summary: The fruit epidermal features such as the size of tubercules are important fruit quality traits for cucumber production. But the mechanisms underlying tubercule formation remain elusive. Here, tubercule size locus CsTS1 was identified by map‐based cloning and was found to encode an oleosin protein. Allelic variation was identified in the promoter region of CsTS1, resulting in low expression of CsTS1 in all 22 different small‐warty or nonwarty cucumber lines. High CsTS1 expression levels were closely correlated with increased fruit tubercule size among 44 different cucumber lines. Transgenic complementation and RNAi‐mediated gene silencing of CsTS1 in transgenic cucumber plants demonstrated that CsTS1 positively regulates the development of tubercules. CsTS1 is highly expressed in the peel at fruit tubercule forming and enlargement stage. Auxin content and expression of three auxin signalling pathway genes were altered in the 35S:CsTS1 and CsTS1‐RNAi fruit tubercules, a result that was supported by comparing the cell size of the control and transgenic fruit tubercules. CsTu, a C₂H₂ zinc finger domain transcription factor that regulates tubercule initiation, binds directly to the CsTS1 promoter and promotes its expression. Taken together, our results reveal a novel mechanism in which the CsTu‐TS1 complex promotes fruit tubercule formation in cucumber.