Differentiation of Xanthomonas oryzae pv. oryzae in vitro and during rice leaf infection

Highlights- Xoo produces filamentous morphology, which is transient and yields pleomorphic progenies in vitro - In planta, initial attachment of rod-shaped Xoo is detected at xylem pits - The Xoo infection front migrates basipetally in the vascular bundle and progresses laterally from major to minor veins via transverse veins - Xoo breaks out of the xylem vessels and enter the neighboring xylem parenchyma - Xoo assumes filamentous morphology that can traverse from the xylem across the bundle sheath into mesophyll tissue - Mobility in xylem vessels depends predominantly on rod-shaped Xoo, while infection of mesophyll tissue at later stages appears to be linked to filamentous morphology SummaryXanthomonas oryzae pv. oryzae (Xoo) is classified as a xylem pathogen responsible for bacterial blight of rice causing substantial yield losses in Asia and Africa. Xoo virulence depends on the ability to trigger SWEET sucrose efflux transporters in the xylem parenchyma (XP) by injection of transcription activation like effectors (TALe) into host cells, likely to access host-derived sucrose. To establish infection, Xoo must overcome physical barriers, immune responses and the hydraulic xylem flow. To gain insights into the colonization process, we used translational SWEET11a-GUS reporter lines, scanning electron microscopy, and confocal laser scanning microscopy of Xoo tagged with a fluorescent protein. We found that Xoo can differentiate in vitro into filamentous forms. We mapped the infection route of Xoo along the vasculature, identified distinct spatiotemporal phases of Xoo colonization marked by rod-shaped and, notably, filamentous Xoo cells. Rod-shaped Xoo were found to attach to xylem pits during basipetal progression of the infection. Notably, we found that at later infection stages, Xoo could enter the XP. Strikingly, Xoo adopted a filamentous phenotype that traversed bundle sheath cells and entered mesophyll cells. Chlorosis and necrosis of leaves is thus likely not just due to blockage of xylem flow, but to direct tissue damage. Filamentation had been reported as important for virulence of human pathogens e.g. Yersinia pestis, uropathogenic E. coli and Shigella and had been associated to sugar utilization in Bacillus subtilis. We thus hypothesize that Xoo differentiation during host colonization is critical for virulence. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/680524v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@f66d8borg.highwire.dtl.DTLVardef@91ca66org.highwire.dtl.DTLVardef@17f2846org.highwire.dtl.DTLVardef@12d46e7_HPS_FORMAT_FIGEXP M_FIG C_FIG