Integration of transcriptome and metabolome reveals regulatory mechanisms of volatile flavor formation during tomato fruit ripening

Tomato is an important economic crop all over the world. Volatile flavors in tomato fruit are key factors influencing consumer liking and commercial quality. However, the regulatory mechanism controlling the volatile flavors of tomatoes is still not clear. Here, we integrated the metabolome and transcriptome of the volatile flavors in tomato fruit to explore the regulatory mechanism of volatile flavor formation, using wild and cultivated tomatoes with significant differences in flavors. A total of 35 volatile flavor compounds were identified, based on the solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). The content of the volatiles, affecting fruit flavor, significantly increased in the transition from breaker to red ripe fruit stage. Moreover, the total content of the volatiles in wild tomatoes was much higher than that in the cultivated tomatoes. The content variations of all volatile flavors were clustered into 10 groups by hierarchical cluster and Pearson correlation analysis (PCC). The fruit transcriptome was also patterned into 10 groups, with significant variations both from the mature green to breaker fruit stage and from the breaker to red ripe fruit stage. Combining the metabolome and the transcriptome of the same developmental stage of fruits by co-expression analysis, we found that the expression level of 1 182 genes was highly correlated with the content of volatile flavor compounds, thereby constructing two regulatory pathways of important volatile flavors. One pathway is tetrahydrothiazolidine N-hydroxylase (SlTNH1)-dependent, which is regulated by two transcription factors (TFs) from the bHLH and AP2/ERF families, controlling the synthesis of 2-isobutylthiazole in amino acid metabolism. The other is lipoxygenase (SlLOX)-dependent, which is regulated by one TF from the HD-Zip family, controlling the synthesis of hexanal and (Z)-2-heptenal in fatty acid metabolism. Dual-luciferase assay confirmed the binding of bHLH and AP2/ERF to their structural genes. The findings of this study provide new insights into volatile flavor formation in tomato fruit, which can be useful for tomato flavor improvement.