Figure 3.

Rhizosphere fitness of mutant strains in competition with KT2440RTn7-Sm. The knocked-out open reading frame in the mutant strains is indicated by the locus name. Proportion of mutant (grey) and wild type (white), which was 50% ± 2% in the initial inocula, is plotted after 12 days of colonization. Data are the averages and standard error for six plants. KT2440RTn7-Sm, a streptomycin resistant derivative of KT2440R (see Materials and methods), was used as the wild-type strain in the experiments. KT2440RTn7-Sm and KT2440R are equally competitive in root colonization (not shown). Sm resistance marker of the wild-type bacteria allowed their specific selection against the mutants, which were kanamycin resistant derivatives of KT2440R. Statistical analysis was carried out using SPSS software (version 12.0.1 for Windows, SPSS Inc., Chicago, IL, USA). The linear model univariate analysis of variance rendered significant differences for the mutants shown in the figure (P value < 0.05) in comparison with the wild type. Seed adhesion rate was similar for mutants and KT2440R (0.5%), with the exception of PP0110 and PP1477 (0.1%). The growth of the mutants under laboratory conditions (rich and defined medium) was indistinguishable from that of KT2440RTn7-Sm. The transcriptional organization of mutated genes is shown in the bottom. The space between the 3' end of PP1476 and the 5' end of PP1477 is 7 bp. Translational coupling between PP3281 and PP3280 (8 bp) was observed. PP3279 is probably in an independent operon; however, PP3279 and PP3281 code for enzymes in the same degradative pathway [28]. Translational coupling between PP4987 and PP4988 was also observed (8 bp). Arrows indicate direction of transcription. Transposon insertion is indicated by inverted triangles.