Abstract
The situation of drug resistance has become more complicated due to the scarcity of plant resistance genes, and overcoming this challenge is imperative. Isatis indigotica has been used for the treatment of wounds, viral infections, and inflammation for centuries. Antimicrobial peptides (AMPs) are found in all classes of life ranging from prokaryotes to eukaryotes. To identify AMPs, I. indigotica was explored using a novel, sensitive, and high-throughput Bacillus subtilis screening system. We found that IiR515 and IiR915 exhibited significant antimicrobial activities against a variety of bacterial (Xanthomonas oryzae, Ralstonia solanacearum, Clavibacter michiganensis, and C. fangii) and fungal (Phytophthora capsici and Botrytis cinerea) pathogens. Scanning electron microscope and cytometric analysis revealed the possible mechanism of these peptides, which was to target and disrupt the bacterial cell membrane. This model was also supported by membrane fluidity and electrical potential analyses. Hemolytic activity assays revealed that these peptides may act as a potential source for clinical medicine development. In conclusion, the plant-derived novel AMPs IiR515 and IiR915 are effective biocontrol agents and can be used as raw materials in the drug discovery field.
Highlights
Drug resistance in pathogenic microbes is an emerging challenge in crop production and human health care [1,2]
A drawback of this strategy is that strong Antimicrobial peptides (AMPs) will kill B. subtilis cells too rapidly to detect the clones, and only AMPs with weak killing effects are preferred
B. subtilis has a good secretory system, which may reduce the toxicity of strong AMPs
Summary
Drug resistance in pathogenic microbes is an emerging challenge in crop production and human health care [1,2]. Multiresistant bacteria have many strategies that threaten the health of animals and plants [3], such as beta-lactamases that are enzymes synthesized by bacteria to break host resistance [4]. This situation has been provoked by the recent attention paid to identifying new antimicrobial genes against emerging bacterial resistance [5]. The Gram-positive bacterial cell envelope is composed of an outer cell wall (a thick peptidoglycan layer and a polysaccharide coat) and an inner cytoplasmic membrane To achieve their bactericidal activities, AMPs interact with the cell wall or cytomembrane, resulting in membrane interruption and cell lysis [12]. In most of these cases, AMPs are reported to cause disruptions in cell wall or cell membrane integrity, perforation, deformation, and increased water ion and molecular flow across the membrane, which causes microbial death [13]
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