Paenibacillus polymyxaproduces fusaricidin-type antifungal antibiotics active againstLeptosphaeria maculans, the causative agent of blackleg disease of canola

Abstract

A bacterial isolate capable of inhibiting the growth of Leptosphaeria maculans (Desmaz.) Ces. & De Not., the causative agent of blackleg disease of canola (Brassica napus L. and Brassica rapa L.), was identified as a potential biological control agent. This environmental isolate was determined to be Paenibacillus polymyxa based on its (i) biochemical and growth characteristics and (ii) 16S rRNA sequence similarity, and was given the strain designation PKB1. Antifungal peptides were produced by P. polymyxa PKB1 around the onset of sporulation, with optimal production on potato dextrose broth. The antifungal peptides were extracted from P. polymyxa PKB1 cells and (or) spores using methanol and were purified using size exclusion and reverse-phase chromatography. Characterization of the antifungal peptides was done using amino acid compositional analysis, Edman degradation sequencing from partially hydrolyzed material, and a variety of mass spectrometric methods. The purified antifungal material was found to be a mixture of related peptides of molecular masses 883, 897, 948, and 961 Da, with the most likely structure of the 897-Da component determined to be a cyclic depsipeptide with an unusual 15-guanidino-3-hydroxypentadecanoic acid moiety bound to a free amino group. These compounds are therefore members of the fusaricidin group of cyclic depsipeptides.Key words: cyclic depsipeptide, biocontrol agent, allo-threonine.