D-Lactic acid secreted by Chlorella fusca primes pattern-triggered immunity against Pseudomonas syringae in Arabidopsis.

Biological control agents including microbes and their products have been studied as sustainable crop protection strategies. Although aquatic microalgae have been recently introduced as a biological control agent, the underlying molecular mechanisms are largely unknown. The aim of the present study was to investigate the molecular mechanisms underlying biological control by microalgae Chlorella fusca. Foliar application of C. fusca elicits induced resistance in Arabidopsis thaliana against Pseudomonas syringae pv. tomato DC3000 that activates plant immunity rather than direct antagonism. To understand the basis of C. fusca-triggered induced resistance at the transcriptional level, we conducted RNA sequencing (RNA-seq) analysis. RNA-seq data showed that upon pathogen inoculation, C. fusca treatment primed the expression of cysteine-rich receptor-like kinases (CRKs), WRKY transcription factor genes, and salicylic acid and jasmonic acid signaling-related genes. Intriguingly, the application of C. fusca primed pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), characterized by reactive oxygen species (ROS) burst and callose deposition, upon flagellin 22 treatment. The attempts to find C. fusca determinants allowed us to identify D-lactic acid secreted in the supernatant of C. fusca as a defense priming agent. This is the first report of the mechanism of innate immune activation by aquatic microalgae Chlorella in higher plants.