Synthetic peptides bioactive against phytopathogens have lower impact on some beneficial bacteria: An assessment of peptides biosafety in agriculture

The emergence of bacterial resistance and the increasing restrictions on the use of agrochemicals are boosting the search for novel, sustainable antibiotics. Antimicrobial peptides (AMPs) arise as a new generation of antibiotics due to their effectiveness at low doses and biocompatibility. We compared the antimicrobial activity of four promising AMPs (CA-M, BP100, RW-BP100, and 3.1) against a collection of notorious phytopathogens, and quantified their impact on plant beneficial bacteria. Plant growth promoters (PGP) and biological control agents (BCA) were also included to study the feasibility of integrating AMPs with bio-based strategies to mitigate diseases impacts and promote crop production. Flow cytometry and fluorescence microscopy revealed that the AMPs' effects on the membrane integrity of both gram-negative and gram-positive strains were time- and concentration-dependent. Bacterial strains were separated into three groups of susceptibility to the AMPs. Group 1 was represented by the most sensitive, gram-negative phytopathogenic belonging to Xanthomonadales and Pseudomonadales and the gram-positive C. michiganensis subsp. michiganensis. Group 2 encompassed bacteria showing intermediate susceptibility, namely P. carotovorum subsp. carotovorum, P. cerasi, both phytopathogens, as well as the plant growth promoters P. fluorescens and P. putida. Finaly, Group 3 was represented by the bacteria with the lowest susceptibility to AMPs. It included beneficial bacteria (B. zhangzhouensis, B. subtilis, B. safensis, P. azotoformans), a phytopathogen (R. solanacearum), and a strain reported as able to act as both (P. aeruginosa). This work demonstrates that the minimum inhibitory concentrations (MICs) needed to act against the beneficial Bacillus and Pseudomonas strains were higher than those needed to produce bacteriostatic or bactericidal effects on the phytopathogens tested, hence supporting that these AMPs might be environmentally safe antibiotics with low likeliness of disrupting the beneficial microbial communities. The possibility of mixing these AMPs with BCA/PGP, in a combined biocontrol strategy, is also discussed.