Silencing of SlGH3-2 in tomato alters auxin and ethylene levels during fruit ripening and cause reduced lycopene accumulation in the transgenic fruits. While auxin's role during fleshy fruit ripening is widely acknowledged to be important, the physiological functions of several ripening-induced genes, especially those involved in the maintenance of cellular auxin homeostasis, largely remain under-explored. In the present study, the updated inventory shows that 24 members constitute the Gretchen-Hagen 3 (GH3) gene family in tomato. Their characterization using an expression profiling approach revealed that SlGH3-2, a member of the group II IAA-amido synthetase, is strongly induced at the commencement of fruit ripening. Phylogenetic analysis and homology modeling revealed that SlGH3-2 is the closest homolog of pepper CcGH3 and grapevine VvGH3-1. Expression profiling revealed that the mRNA level of SlGH3-2 is inhibited in ripening mutants such as ripening-inhibitor (rin) and Never-ripe (Nr). Whereas both auxin and ethylene were found to act as positive regulators of its transcript accumulation. The fruits of 35S::SlGH3-2 RNAi lines exhibited prolonged shelf-life. Both ethylene production and lycopene accumulation were affected in the fruits of SlGH3-2 silenced lines. These observations were corroborated by the altered expression of key ethylene and carotenoid biosynthesis genes such as ACS2 and PSY1, respectively, in the RNAi lines. Additionally, the SlGH3-2 silenced line fruits had higher IAA and IBA levels at the ripening stages, and showed increased transcript accumulation of several known auxin-induced SlIAA and SlGH3 genes. Altogether, the present study suggests that SlGH3-2 influences fruit ripening in tomato via modulating ethylene and auxin crosstalk, especially during the early phase.
Keywords: Auxin homeostasis; Fruit ripening; GH3; Lycopene; Shelf-life; Tomato; Transgenic.