Table 1.
Foxtail millet ALDH genes and superfamilies.
Figure 1.
Phylogenetic analysis of foxtail millet and other plant ALDHs.
Phylogenetic tree was constructed with ALDH protein sequences from S.bicolor (Sb), Z.mays (Zm), O.sativa (Os), S.italica (Si), and A. thaliana (At). Members of respective ALDH families are depicted in a specific background color.
Figure 2.
Distribution and synteny of ALDH genes on foxtail millet chromosomes and synteny analysis of ALDH genes between foxtail millet and rice.
(A) Chromosomes 1–9 are depicted as horizontal gray bars. ALDH genes are indicated by vertical green line. Colored bars denote syntenic of the foxtail millet genome. (B) Foxtail millet and rice chromosomes are depicted as horizontal black and blue bars, respectively. Foxtail millet and rice ALDH gene are indicated by vertical black lines, Colored bars denote syntenic regions between foxtail millet and rice chromosomes. A twisted colored bar indicates that syntenic regions are in opposite orientations. SiALDH genes don't located in the syntenc blocks between rice and foxtail millet chromosomes were marked by red oval.
Figure 3.
Phylogenetic analysis and exon-intron structures of foxtail millet and rice ALDH genes.
Numbers above or below branches of the tree indicate bootstrap values. Coding exons, represented by ashy, were drawn to scale. Dashed lines connecting two exons represent introns. Members of respective ALDH families are depicted in a specific background color.
Figure 4.
Semi-quantitative RT-PCR results of 20 foxtail millet ALDH genes in root, stem, and leaf.
(A), (B), (C), (D) and (E): The RT-PCR products, generated with 20 SiALDHs and actin (AF288226.1) gene specific primers, were electrophoresed in a 1.5%agarose gel and densitometric analysis of the corresponding band to SiALDHs. The bars represent the mean ± SD of the results from three separate experiments. One-way ANOVA analysis of variance showed significant differences between group means (P<0.05). Tukey's multiple comparison test showed significant differences between the means of groups depicted by the different letters on the bars (P<0.05). Actin mRNA (AF288226.1) was used as an internal control.
Figure 5.
QRT-PCR analysis of the 20 foxtail millet ALDH genes.
Time course expression analysis of the 20 foxtail millet genes under various stresses. (A) Cold at 4°C; (B) 250 mM NaCl; (C) 20%PEG-6000; (D) 200 µM H2O2; (E) 100 µM ABA; (F) Heat at 42°C. Actin mRNA (AF288226.1) was used as an internal control. The bars represent the mean± SD of the results from three separate experiments.
Figure 6.
The viabilities of the strains were detected on different LB plated with additional 500/L NaCl (B), 800 mmol/L NaCl and normal LB plates (A). Strains were Control: strain harboring plasmid pET-28(a), pET-ALDHs: strain harboring different SiALDH proteins. 1.5 microliters of the serially diluted E. coli cells cultured for 16 h at 16°C with 0.5 mmol/L IPTG were dotted onto LB agar plates containing NaCl at the marked concentration.