Tryptophan specialized metabolism and ER body-resident myrosinases modulate root microbiota assembly

Abstract

Indole glucosinolates (IGs) are tryptophan (Trp)-derived sulfur-containing specialized metabolites that play a crucial role in plant-microbe interactions in plants of the order Brassicales, including Arabidopsis thaliana. Despite the growing body of evidence implicating IG biosynthetic pathways in root-microbiota interactions, how myrosinases, the enzymes that convert inert IGs into bioactive intermediate/terminal products, contribute to this process remains unknown. Here, we describe the roles of the PYK10 and BGLU21 myrosinases in root-microbiota assembly partly via metabolites secreted from roots into the rhizosphere. PYK10 and BGLU21 localize to the endoplasmic reticulum (ER) body, an ER-derived organelle observed in plants of the family Brassicaceae. We investigated the root microbiota structure of mutants defective in the Trp metabolic (cyp79b2b3 and myb34/51/122) and ER body (nai1 and pyk10bglu21) pathways and found that these factors together contribute to the assembly of root microbiota. Microbial community composition in soils as well as in bacterial synthetic communities (SynComs) treated with root exudates axenically collected from pyk10bglu21 and cyp79b2b3 differed significantly from those treated with exudates derived from wild-type plants, pointing to a direct role of root-exuded compounds. We also show that growth of the pyk10bglu21 and cyp79b2b3 mutants was severely inhibited by fungal endophytes isolated from healthy A. thaliana plants. Overall, our findings demonstrate that root ER body-resident myrosinases influencing the secretion of Trp-derived specialized metabolites represent a lineage-specific innovation that evolved in Brassicaceae to regulate root microbiota structure. SignificanceER bodies were first identified in roots of Brassicaceae plants more than 50 years ago, but their physiological functions have remained uncharacterized. A series of previous studies have suggested their possible role in root-microbe interactions. Here, we provide clear experimental evidence showing a role for ER bodies in root-microbiota interactions, which overlaps with that of root-exuded Trp-derived metabolites. Our findings delineate a plant lineage-specific innovation involving intracellular compartments and metabolic enzymes that evolved to regulate plant-microbe interactions at the root-soil interface.