Fitness of Isogenic Colony Morphology Variants of Pseudomonas aeruginosa in Murine Airway Infection

Chronic lung infections with Pseudomonas aeruginosa are associated with the diversification of the persisting clone into niche specialists and morphotypes, a phenomenon called ‘dissociative behaviour’. To explore the potential of P. aeruginosa to change its morphotype by single step loss-of–function mutagenesis, a signature-tagged mini-Tn5 plasposon library of the cystic fibrosis airway isolate TBCF10839 was screened for colony morphology variants under nine different conditions in vitro. Transposon insertion into 1% of the genome changed colony morphology into eight discernable morphotypes. Half of the 55 targets encode features of primary or secondary metabolism whereby quinolone production was frequently affected. In the other half the transposon had inserted into genes of the functional categories transport, regulation or motility/chemotaxis. To mimic dissociative behaviour of isogenic strains in lungs, pools of 25 colony morphology variants were tested for competitive fitness in an acute murine airway infection model. Six of the 55 mutants either grew better or worse in vivo than in vitro, respectively. Metabolic proficiency of the colony morphology variant was a key determinant for survival in murine airways. The most common morphotype of self-destructive autolysis did unexpectedly not impair fitness. Transposon insertions into homologous genes of strain PAO1 did not reproduce the TBCF10839 mutant morphotypes for 16 of 19 examined loci pointing to an important role of the genetic background on colony morphology. Depending on the chosen P. aeruginosa strain, functional genome scans will explore other areas of the evolutionary landscape. Based on our discordant findings of mutant phenotypes in P. aeruginosa strains PAO1, PA14 and TBCF10839, we conclude that the current focus on few reference strains may miss modes of niche adaptation and dissociative behaviour that are relevant for the microevolution of complex traits in the wild

Management of Diseases Caused by Pectobacterium and Dickeya Species

Management of soft rot Pectobacteriaceae (SRP) is a challenge as there are no control agents available and no effective resistance present in commercial cultivars. In addition, many species of SRP have a broad host range and spread via rotten plant material takes place readily. In this chapter, the possibilities for disease management are outlined. Management is mainly based on seed certification to limit the risks of using infected planting material, and on hygiene and cultivation practices that reduce cross-contamination within and between seed lots. Balanced nutrition also supports the suppressiveness of crops against SRP. Experimental data show that inoculum in seed tubers can be reduced by thermotherapy and the use of biocides. Under controlled conditions, application of seed potatoes with biocontrol agents has showed promising results but few data are present on the efficacy of biocontrol in the field. Resistance in wild Solanum species against SRP has been found but to date no genes have been transferred to cultivars. However, new breeding technologies, such as CRISPR/CAS 9 and the use of true potato seed (TPS), will give us new perspectives on the generation of resistant cultivars