Phylogenetic analysis of the 14-3-3 family proteins from soybean, Arabidopsis and rice.

<p>The maximum likelihood phylogenetic tree was constructed by using MEGA5.0 based on the full-length amino acid method with 1000 bootstrap replicates.</p

Chromosomal localization and synteny analysis of soybean 14-3-3 family genes.

<p>The chromosomal localization of the 14-3-3 family genes was determined by using the MapInspect software. For synteny analysis, the synteny blocks of the soybean genome were downloaded from the Plant Genome Duplication Database (PGDD, <a href="http://chibba.agtec.uga.edu/duplication/" target="_blank">http://chibba.agtec.uga.edu/duplication/</a>). Duplicated gene pairs were connected by light blue lines for group I, dark blue lines for group II, green lines for group III, and red lines for group IV.</p

Ectopic expression of <i>GsCML27</i> in <i>Arabidopsis</i> influences plant by modifying cellular ionic content.

<p><b>A</b>. Growth performance of WT and ectopic expression seedlings on 1/2 MS medium with 50, 100, 150 mM KCl. Seedlings with open and green leaves was recorded 7 days after stratification. <b>B</b>. Growth performance of WT and ectopic expression seedlings on 1/2 MS medium with 10, 20, 30 mM LiCl. Seedlings with open and green leaves was recorded 7 days after stratification. Data shown represent the means (±SE) of three independent experiments. *P < 0.05 and **P < 0.01 by Student’s t-test.</p

Increased ABA sensitivity of <i>GsGF14o</i> transgenic lines during the seed germination and seedling growth stages.

<p>(A) The growth performance of WT and <i>GsGF14o</i> OX seedlings under ABA stress. Bars are 1cm. (B) Seed germination rates of WT and <i>GsGF14o</i> OX lines. (C) Percentage of seedlings with open and green leaves. (D) Percentage of seedlings with four leaves. (E) Phenotypes of WT and OX seedlings. Bars are 1cm. (F) Primary roots of WT and OX seedlings. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01 by Student’s <i>t</i> test.</p

<i>GsCML27</i> ectopic expression altered expression patterns of a set of osmotic stresses signal related genes.

<p>Expression patterns of osmotic stress related genes. Relative expression levels were determined by quantitative RT-PCR using <i>ACTIN2</i> as an internal control. All of the values represent the means of three fully independent biological replicates; error bars indicate the SD. *P < 0.05 and **P < 0.01 by Student’s t-test.</p

Phylogenetic analysis and exon-intron structures of soybean 14-3-3 family members.

<p>(A) The phylogenetic tree of soybean 14-3-3 family proteins. <b>(</b>B) The exon-intron structures of soybean 14-3-3 family genes. Yellow and blue boxes represented exons and lines represented introns.</p

Ectopic expression of <i>GsCML27</i> in <i>Arabidopsis</i> enhanced bicarbonate tolerance, but decreased salt and osmotic tolerance.

<p><b>A</b>. Semi-quantitative RT-PCR and quantitative RT-PCR analysis of <i>GsCML27</i> expressions in WT and three ectopic expression lines. <b>B</b>. Growth performance of WT and ectopic expression seedlings on 1/2 MS medium without or with 10 mM NaHCO₃, 125 mM NaCl or 300 mM mannitol. Photographs were taken 7 days after stratification. <b>C</b>. Seed germination rates of WT and ectopic expression lines under stresses. Germination was recorded daily up to 6 days. <b>D</b>. Seedlings with open and green leaves was recorded 7 days after stratification. All of the values represent the means of three fully independent biological replicates; error bars indicate the SD. *P < 0.05 and **P < 0.01 by Student’s t-test.</p

Sequence analysis of <i>GsCML27</i>.

<p><b>A</b>. Multiple sequence alignment of the full-length amino acid sequences of <i>GsCML27</i> with homologous <i>CMLs</i> from <i>Arabidopsis</i>. Sequences were aligned using MEGA 5.0. <b>B</b>. The phylogenetic relationships between <i>GsCML27</i> and homologous protein kinases with the conserved amino acid sequences. The phylogenetic tree was constructed using MEGA 5.0.</p

<i>GsGF14o</i> overexpression promoted ABA induced stomata closure but not proline accumulation.

<p>(A) Stomata from WT and GsGF14o OX lines in response to ABA stress. Bars are 20μm. (B) Presented photos to show stomata from WT and OX lines. Bars are 10μm. (C) Percentage of different stomata in WT and OX lines. (D) Stomata aperture of WT and OX lines. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01 by Student’s t test. (E) Proline content of WT and OX lines. Different letters indicated statistical differences among means by Duncan's Multiple Range Test (<i>P</i> < 0.05).</p

Segmental duplication analyses of soybean 14-3-3 family genes in Group I (A), Group II (B), Group III (C), and Group IV (D).

<p>The segmental duplication of soybean 14-3-3 genes was obtained based on the locus search at PGDD website.</p