Being spontaneous has its costs! Characterization of spontaneous phage □D5-resistant mutants ofDickeya solanistrain IPO 2222

Abstract

Lytic bacteriophages able to infect and kill Dickeya spp. can be readily isolated from virtually all Dickeya spp.-containing environments, yet little is known about the selective pressure those viruses exert on their hosts. Here, we identified two spontaneous D. solani IPO 2222 mutants (0.8% of all obtained mutants), DsR34 and DsR207, resistant to infection caused by lytic phage vB_Dsol_D5 (ΦD5) that expressed a reduced ability to macerate potato tuber tissues compared to the wild-type, phage-susceptible D. solani IPO 2222 strain. Genome sequencing revealed that genes encoding: secretion protein HlyD (mutant DsR34) and elongation factor Tu (EF-Tu) (mutant DsR207) were altered in these strains. Both mutations impacted the proteomes of cells grown in both rich and minimal media, including the abundance of the cell envelope and transmembrane transport-associated proteins. Furthermore, features essential for the ecological success of these mutants in a plant environment, including their ability to use various carbon and nitrogen sources, produce plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta were assessed. Compared to the wild-type strain, D. solani strain IPO 2222, mutants DsR34 and DsR207 had a reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants. The implications of the ΦD5 resistance on driving traits affecting the ecological performance of D. solani are discussed.