Dynamic changes of endogenous H2S generation during responding to developmental and environmental signals in Solanum lycopersicum L.

Abstract

Increasing studies have confirmed the functions of hydrogen sulfide (H2S) as a gas signal molecule (gasotransmitter) in a variety of physiological activities in plants. However, the dynamic changes of endogenous H2S generation, especially the response speed and degree of H2S-generation-associated members, to both developmental and environmental cues in Solanum lycopersicum L. are still poorly understood. Here, we identified and charactered the H2S-generation-associated enzymes in tomato, including O-acetylserine (mercaptan) lyases (OAS-TL), L/D-Cysteine desulfhydrases (L/D-CDes) and nitrogen fixation S (NIFS)-like enzymes, possessing LCD-like activity. Analysis of cis-regulatory elements (CREs) showed that the promoter regions of these H2S-generation-associated genes contained lots of light-, hormone- and stresses-responsive CREs, and transcriptional factors (TFs) binding sites, suggesting that the regulatory mechanism of endogenous H2S generation might involve TFs mediated transcriptional regulation or/and interacting with phytohormones signals. Levels of endogenous H2S exhibited diversity in different organs and developmental stages in tomato, and the top and second highest level of H2S in flowers and ripened fruits hinted the significance of H2S signal in tomato reproduction and fruit development. Furthermore, the expression of SlOAS4, SlLCD, SlDCD2, SlNFS1, and SlNFS2 enhanced continuously during fruit ripening, and the SlOAS4 expression exhibited the most significant up-regulation. The SlDCD enzymatic activity responded more quickly to fruit development cues, while the SlLCD remained at a higher level almost throughout the whole fruit ripening. During environmental stimulus, it was found that the transcriptions of SlDCD2 and SlOAS2 responded faster and stronger to heat stress, while SlOAS4 and SlLCD were activated more quickly and significantly by drought stress. Both SlLCD and SlDCD enzymatic activities could be enhanced by drought and heat stress to varying degrees, and it seemed that the SlLCD was activated to a greater extent and remained at the higher activity for a longer period. With the extension of exogenous phytohormones (ABA, SA, GA, JA, ACC, NAA) treatment, the SlLCD activity and the endogenous H2S content increased in a specific time-dependent manner, and ABA, SA and NAA had more rapid effects on SlLCD activity. By widely explored the dynamic responses of endogenous H2S generation, our study would provide some references for understanding the regulatory mechanism upstream of H2S signaling in tomato growth developments and environmental adaptations.