Type 2 Nep1-Like Proteins from the Biocontrol Oomycete Suppress Infection in Solanaceous Plants.

As a non-pathogenic oomycete, the biocontrol agent is able to control plant diseases through direct mycoparasite activity and boosting plant immune responses. Several elicitors have been found to activate plant immunity as microbe-associated molecular patterns (MAMPs). Necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are a group of MAMPs widely distributed in eukaryotic and prokaryotic plant pathogens. However, little is known about their distribution and functions in and its sister species . Here, we identified a total of 25 NLPs from (PyolNLPs) and (PypeNLPs). Meanwhile, we found that PyolNLPs/PypeNLPs genes cluster in two chromosomal segments, and our analysis suggests that they expand by duplication and share a common origin totally different from that of pathogenic oomycetes. Nine PyolNLPs/PypeNLPs induced necrosis in   by agroinfiltration. Eight partially purified PyolNLPs/PypeNLPs were tested for their potential biocontrol activity. PyolNLP5 and PyolNLP7 showed necrosis-inducing activity in via direct protein infiltration. At sufficient concentrations, they both significantly reduced disease severity and suppressed the in planta growth of in solanaceous plants including (tobacco), (tomato) and (pepper). Our assays suggest that the suppression effect of PyolNLP5 and PyolNLP7 is irrelevant to reactive oxygen species (ROS) accumulation. Instead, they induce the expression of antimicrobial plant defensin genes, and the induction depends on their conserved nlp24-like peptide pattern. This work demonstrates the biocontrol role of two NLPs for solanaceous plants, which uncovers a novel approach of utilizing NLPs to develop bioactive formulae for oomycete pathogen control with no ROS-caused injury to plants.