Tomato pollen tube integrity is critical for pollen function at high temperature and is controlled by RALF signaling.
Reproductive success at high temperature (HT) depends on the ability of pollen to germinate a pollen tube that delivers sperm for double fertilization. We recently found that Solanum lycopersicum (tomato) cultivars capable of setting fruit at HT produce pollen tubes that continue to grow under heat stress. Our present goal is to define the molecular basis of thermotolerant pollen tube growth using quantitative live imaging, transcriptomic and proteomic profiling, and genetic analysis. We found that pollen tube integrity - the ability to maintain an intact tip during germination and elongation - was the pollen performance parameter that distinguished thermotolerant cultivars. We identified the components of the Rapid Alkalinization Factor (RALF) pollen tube integrity signal transduction pathway in tomato and found that RALF peptides repress germination at control temperature and that signaling is antagonized by HT. Loss of a single RALF peptide changes pollen tube cell wall pectin distribution and enhances pollen tube integrity at HT in thermotolerant Heinz, revealing that RALF signaling regulates thermotolerant pollen tube growth and establishing a molecular framework for improving reproductive heat-resilience in crop plants.