Growth of <i>Methylobacterium</i>-inoculated barley.

<p>The data are presented as mean ± standard deviation, and analyzed with one-way ANOVA and Dunnett’s test. Statistical significance was indicated with</p><p>* (<i>p</i> < 0.05),</p><p>** (<i>p</i> < 0.01),</p><p>*** (<i>p</i> < 0.001),</p><p>**** (<i>p</i> < 0.0001).</p><p>Growth of <i>Methylobacterium</i>-inoculated barley.</p

Result of DGGE analysis for <i>R. japonicum</i> liquid culture samples.

<p>Identification was done at EZtaxon site (version 2.1). The closest relatives are listed.</p

Identification and composition of <i>Methylobacterium</i> species isolated from rice plants subjected to <i>Methylobacterium</i> inoculation.

<p>Total number of <i>Methylobacterium</i> isolates is shown in parentheses. Identification was performed by WC-MS and 16S RNA sequencing, and details are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129509#pone.0129509.g001" target="_blank">Fig 1</a>.</p

Effect of strain 22A application on other bryophytes.

<p>Strain 22A was applied onto protonemata of <i>H. microphyllum</i>, <i>T. microphylla</i>, and <i>Bryum</i> sp., and the plants were grown for 38 days on Y medium. The plants that received no bacterial cells served as controls. Grid, 5 mm.</p

Growth promotion of seed plants by added <i>Methylobacterium</i> species.

<p>Data presented as mean ± SD. Fresh mass indicates that of entire plants.</p><p>nt: not tested.</p>*<p>, p<0.05 and.</p>**<p>, p<0.01 (Student's T-test).</p

Growth promotion of <i>R. japonicum</i> by <i>Methylobacterium</i> isolates.

<p>A, Protonemata growth on Y medium in the presence of bacterial isolates (35 days of cultivation). Grid, 5 mm. B, Quantification of protonemata growth. White bars, 22 days; light gray bars, 35 days; and gray bars, 63 days. Growth was evaluated by measuring the area of protonemata proliferation, regarded as ellipses. Error bars, standard deviation (n = 8). C, Strain 22A growing with protonemata on Y medium. Grid, 5 mm. D, Induction of gametophyte formation by application of bacterial isolates. A suspension of <i>R. japonicum</i> protonemata was inoculated on Florialite, to which 50 µL of water (control), kinetin (3 mg/L), or a bacterial suspension (11A, 21C and 22A) was applied. Images were taken after 60 days of cultivation.</p

MSP dendrogram based on WC-MS analysis of the isolates from leaves of inoculated rice seed.

<p>The isolates were named with the treated strain name and isolate numbers. The inoculated strains are colored in red. The representatives selected from each cluster are colored in blue with the closest type strain name and percentage identity of 16S RNA gene in parentheses. Isolate 22A-9 was pink-pigmented fungus and was identified by ITS region sequencing, as described previously [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129509#pone.0129509.ref019" target="_blank">19</a>].</p

MSP dendrogram based on WC-MS analysis of the isolates from barley of different cultivars.

<p>The representatives are shown in blue with the closest species name and percentage identity of the 16S RNA gene in parentheses. Strains shown in gray were not pink-pigmented.</p

Maximum-likelihood phylogenetic tree of <i>Methylobacterium</i> isolates and related taxa, based on 16S rRNA gene sequences.

<p>Isolates from rice are colored in green and those from barley are in blue. Numbers in parentheses indicate isolates belonging to the same species, estimated by WC-MS analysis. For isolates from rice, the inoculation effect is not taken into account in the figure. <i>M aquaticum</i> strain 22A is taken as a representative strain for the <i>M</i>. <i>platani/aquaticum</i> cluster shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129509#pone.0129509.g001" target="_blank">Fig 1</a>. Bootstrap percentages based on 1000 replicates are shown if greater than 80%. <i>Microvirga flocculans</i> TFB (AB098515) was used as an outgroup. Bar, 0.1 changes per nucleotide position.</p

Detection of methanol emitted from <i>R. japonicum.</i>

<p>A. Control observation with <i>P. pastoris</i> PPY12 (pYA005 and pYA006) grown on YNB and YNM for 2 days. B. YNB-grown cells were applied onto Y medium, Y+0.5% methanol (YM); protonemata were grown on Y medium and non-sterile gametophytes were inoculated on Y medium. Left panel, fluorescence; right panel, differential interference contrast images. Bar, 20 µm.</p