Spontaneous mutations in gas vesicle genes of Planktothrix spp. affect gas vesicle production and critical pressure.

Wild-type strains of the cyanobacterium Planktothrix rubescens have a cluster of gas vesicle (gvp) genes with repeats of alternating gvpA and gvpC. The gvpC occurs in three length variants, all with the same 3'-sequence, OmegaC. Spontaneous non-buoyant mutants had lost some of the alternating gvpAC copies and their gvpC genes had a novel 3'-end sequence, PsiC; additional gvpC genes terminating in this sequence were also found in the wild-type and representatives of other GV genotypes. Alleles of gvpC terminating in PsiC occurred only at the downstream ends of the gvpAC clusters investigated; all other gvpCs terminated in OmegaC. Mutants of strains with the GV3 genotype produced only 30-50% of the gas vesicles present in the wild-type; their gas vesicles had lower mean critical pressures (0.70-0.78 MPa) than those in the wild-type (1.05-1.10 MPa).

The sequence of the major gas vesicle protein, GvpA, influences the width and strength of halobacterial gas vesicles.

Transformation experiments with Haloferax volcanii show that the amino acid sequence of the gas vesicle protein GvpA influences the morphology and strength of gas vesicles produced by halophilic archaea. A modified expression vector containing p-gvpA was used to complement a Vac(-) strain of Hfx. volcanii that harboured the entire p-vac region (from Halobacterium salinarum PHH1) except for p-gvpA. Replacement of p-gvpA with mc-gvpA (from Haloferax mediterranei) led to the synthesis of gas vesicles that were narrower and stronger. Other gene replacements (using c-gvpA from Hbt. salinarum or mutated p-gvpA sequences) led to a significant but smaller increase in gas vesicle strength, and less marked effects on gas vesicle morphology.

Growth competition between Halobacterium salinarium strain PHH1 and mutants affected in gas vesicle synthesis.

To investigate the role of the buoyancy provided by gas vesicles in the facultative anaerobe Halobacterium salinarium PHH1, the growth of a gas-vacuolate (Gv+) strain in competition with two gas-vesicle-defective (Gvdef) mutants was examined. The Gv+ strain synthesized gas vesicles throughout its growth cycle, and floated up to form a thick surface scum during the exponential growth phase in static culture. Mutant Gvdef1 produced significantly fewer gas vesicles than the Gv+ strain in corresponding stages of growth, although in late stationary phase a small proportion of cells floated up to the surface of static cultures. Mutant Gvdef2 had a much lower gas vesicle content in shaken culture and produced negligible amounts of gas vesicles in static culture. The Gv+ and the two Gvdef strains grew equally well in shaken cultures, but in static cultures, where steep vertical gradients of oxygen concentration were established, Gvdef1 was outgrown by the Gv+ strain. Gvdef2 outcompeted the Gv+ strain in shallow static cultures, perhaps because Gvdef2 carried a smaller protein burden, which offset the benefits of buoyancy. This selection for Gvdef2 was lost in deeper static cultures, although it could be restored by aerating static cultures from below. The results support the hypothesis that the role of buoyancy in halobacteria is to maintain cells at the more aerated surface of brine pools.