Characterization and Transcriptome Analysis Reveal Abnormal Pollen Germination in Cytoplasmic Male Sterile Tomato.

Cytoplasmic male sterility (CMS) is a plant trait wherein plants cannot develop normal male organs because of the mitochondrial genes. Although the mitochondrial gene has been identified as the CMS-causing gene in tomatoes, its function remains unclear. In this study, we characterized the sterile male phenotypes and analyzed the CMS pollen transcriptome. Microscopic and calcium imaging analyses revealed that CMS pollen exhibited abnormal germination from multiple apertures, accompanied by elevated calcium concentrations and vesicle accumulation, which are typically observed in pollen tube tips. RNA-Seq analysis revealed 440 differentially expressed genes, including four () genes that were highly expressed in the pollen. PME activity was significantly reduced in CMS pollen, suggesting its association with abnormal pollen germination. ATP and reactive oxygen species (ROS) levels, which are key mediators of mitochondrial retrograde signaling (MRS), remained unchanged in CMS pollen, and the expression of the mitochondrial stress marker was not elevated. These findings suggest that triggers an alternative MRS pathway independent of ATP or ROS, potentially leading to upregulation and abnormal pollen germination. Our results reveal a previously unrecognized mechanism of CMS-induced male sterility in tomatoes involving nuclear gene regulation through unconventional mitochondrial signaling.