Pseudomonas syringae Responds to the Environment on Leaves by Cell Size Reduction

Abstract

The length and volume of cells of the plant-pathogenic bacterium Pseudomonas syringae strain B728a were measured in vitro and with time after inoculation on bean leaf surfaces to assess both the effect of nutrient availability on the cell size of P. syringae and, by inference, the variability in nutrient availability in the leaf surface habitat. Cells of P. syringae harboring a green fluorescent protein marker gene were visualized by epifluorescence microscopy after recovery from leaves or culture and their size was estimated by analysis of captured digital images. The average cell length of bacteria grown on leaves was significantly smaller than that of cultured cells, and approached that of cells starved in phosphate buffer for 24 h. The average length of cells originally grown on King's medium B decreased from ≈2.5 to ≈1.2 μm by 7 days after inoculation on plants. Some decrease in cell size occurred during growth of cells on leaves and continued for up to 13 days after cell multiplication ceased. Although cultured cells exhibited a normal size distribution, the size of cells recovered from bean plants at various times after inoculation was strongly right-hand skewed and was described by a log-normal distribution. The skewness of the size distribution tended to increase with time after inoculation. The reduced cell size of P. syringae B728a on plants was readily reversible when recovered cells were grown in culture. Direct in situ measurements of cell sizes on leaves confirmed that most cells of P. syringae respond to the leaf environment by reducing their size. The spatial heterogeneity of cell sizes observed on leaves suggest that nutrient availability is quite variable on the leaf surface environment.