Pathogenic bacteria can trigger active defense responses in higher plants, leading to hypersensitive programmed cell death (PCD) to against those bacteria. However, related research on seaweeds is very limited. Pseudoalteromonas piscicida X-8 (PpX-8) has been identified as the pathogen that causes bleaching disease in commercially farmed Saccharina japonica. In this study, using an inoculation assay and microscopic observations, we found that the proportion of bleaching tissue pieces inoculated with PpX-8 extracellular compounds was significantly higher (p < 0.05) than that inoculated with heated extracellular compounds, indicating that the virulence factors of PpX-8 exist in extracellular compounds and they are heat-sensitive. Using TEM, we observed typical morphological characteristics of PCD after inoculation with extracellular compounds, including chloroplast shrinkage, cytoplasmic vacuolation, and intact mitochondrial structures. Moreover, we detected biochemical characteristics of PCD, such as 3′-OH ends resulting from DNA cleavage and caspase-3-like enzymatic activity, using a TUNEL assay and fluorescence staining. Therefore, PpX-8 extracellular compounds can induce PCD, thus triggering active defense responses in S. japonica. These results indicate that seaweeds and higher plants are conservative in their active defense responses against pathogenic bacteria. The results of this study lay the foundation for further investigation of the virulence mechanisms of PpX-8.
Keywords: Pseudoalteromonas piscicida X-8; Saccharina japonica; active defense responses; bleaching disease; programmed cell death.