Abstract
We previously reported that sound wave treatment (1 kHz) delays fruit ripening in tomato (Solanum lycopersicum), affecting the expression of ethylene biosynthesis-related genes encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthases (ACS) and ACC oxidases (ACO). In this study, we investigated the activity of the transcription factors RIN and HB-1, which function in the ethylene biosynthetic pathway, in response to sound treatment. To investigate whether RIN and HB-1 directly activate the transcription of ACS and ACO, we performed transcriptional activation analysis in Arabidopsis thaliana leaf protoplasts, transiently expressing RIN or HB-1 and using reporter constructs with promoters of the tomato ACS and ACO genes. Activation of the endogenous AtACS and AtACO genes was also measured by qPCR. The RIN- and HB-1-induced expression of these genes decreased, but the HB-1-induced expression of some genes increased after sound treatment. To confirm these results, we performed transient assays in Nicotiana tabacum, which produced results similar to those observed in Arabidopsis. The major ethylene biosynthesis-related genes harbor a CArG-box as a RIN-binding motif. These findings indicate that RIN and HB-1 affect the expression of ethylene biosynthesis-related genes in response to sound treatment, and they suggest that RIN may regulate the ethylene biosynthesis-related genes by binding to their CArG-boxes.
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