Conjugal Transfer from Streptococcus lactis ME2 of Plasmids Encoding Phage Resistance, Nisin Resistance and Lactose-fermenting Ability: Evidence for a High-frequency Conjugative Plasmid Responsible for Abortive Infection of Virulent Bacteriophage

Abstract

S. lactis ME2 exhibits at least 3 mechanism which confer resistance to virulent bacteriophage. These include plasmid-induced interference with phage adsorption, host-controlled restriction and modification activities, and a heat-sensitive mechanism which suppresses development of virulent phage. Conjugal mating experiments were done with S. lactis ME2 to determine if phage-defense mechanisms present in this strain could be mobilized, associated with plasmid DNA elements and phenotypically characterized in transconjugants. Agar-surface matings of S. lactis ME2 with S. lactis LM0230 demonstrated that lactose-fermenting ability (Lac+) was transferred in a conjugation-like process at frequencies of 10-6 per donor cell and was associated with a 40 MDal (megadalton) plasmid designated pTR1040. Resistance to nisin (Nisr) was acquired or lost simultaneously with Lac+, indicating that pTR1040 carried determinants for both phenotypes. Lac+ Nisr transconjugants that carried a 30 MDal plasmid (pTR2030) exhibited a heat-sensitive phage-defence mechanism (Hsp+) which limited the burst size and plaque size of phage c2 without altering the efficiency of plaquing (eop) or the level of adsorption. The ability of phage c2 to initiate plaquing at an eop of 1.0 indicated that DNA injection and early viral gene expression are not affected in the Hsp+ transconjugants. The Hsp+ phenotype may result from plasmid-induced abortive infection of phage dependent on the presence of pTR2030. Hsp+ transconjugants carrying pTR2030 also promoted high-frequency conjugal transfer of Lac+ Nisr associated with pTR1040 (> 10-1/donor cell). Thus, Hsp+ and determinants for conjugal transfer ability (Tra+) are located on pTR2030.