Expression levels of eleven genes in leaf tissues of <i>Suaeda salsa</i> from control, Hg²⁺ (20 µg L⁻¹), salinity (NaCl, 500 mM) and Hg²⁺+salinity-treated groups after exposure for 30 days.

<p>Data (<i>n</i> = 6) were expressed as mean ± standard deviation. Significant difference between control and exposed groups was tested by one-way analysis of variance with Tukey’s test and indicated by *(<i>P</i><0.05) and **(<i>P</i><0.01).</p

One representative 1-dimensional 500 MHz ¹H NMR spectrum of metabolite extracts of the leaf tissue from <i>S.</i><i>salsa</i> in control group using extraction solvent system of methanol/water (1/1).

<p>Metabolite assignments: (1) branched chain amino acids: leucine, isoleucine and valine, (2) ethanol, (3) threonine, (4) alanine, (5) acetate, (6) glutamate, (7) glutamine, (8) succinate, (9) malate, (10) aspartate, (11) asparagine, (12) malonate, (13) choline, (14) phosphocholine, (15) betaine, (16) glycine, (17) fructose, (18) glucose, (19) unknown (5.39 ppm) and (20) fumarate.</p

The activities (U mg⁻¹ protein) of SOD (A), POD (B) and CAT (C) in leaf tissues from control, Hg²⁺ (20 µg L⁻¹), salinity (NaCl, 500) and Hg²⁺+salinity-exposed <i>S.</i><i>salsa</i> samples (<i>n</i> = 6).

<p>Data were expressed as mean ± standard deviation. Significant difference between control and exposed groups was tested by one-way analysis of variance with Tukey’s test and indicated by *(<i>P</i><0.05) and **(<i>P</i><0.01).</p

Number of protein spots from <i>S.</i><i>salsa</i> samples (A) and the differentially expressed proteins in <i>S. salsa</i> from Hg²⁺ (20 µg L⁻¹), salinity (NaCl, 500 mM) and Hg²⁺+salinity (B).

<p>Number of protein spots from <i>S.</i><i>salsa</i> samples (A) and the differentially expressed proteins in <i>S. salsa</i> from Hg²⁺ (20 µg L⁻¹), salinity (NaCl, 500 mM) and Hg²⁺+salinity (B).</p

Metabolite concentrations (µmol g⁻¹ wet tissue) in leaf tissues of <i>S. salsa</i> exposed to Hg²⁺ (20 µg L⁻¹), salinity (NaCl, 500 mM) and combined Hg²⁺ and salinity.

<p>Values are presented as mean ± standard deviation (<i>n</i> = 6).</p>a<p>Statistical significances (<i>P</i><0.05,^*and <i>P</i><0.01,^**) between control and heavy metal-exposed <i>S. salsa</i> samples were determined by one-way ANOVA.</p>b<p>s = singlet, d = doublet, dd = double doublet, t = triplet, m = multiplet, ABX = complex multiplet involving 2 protons (A and B) and a heavy atom (X).</p

The list of primers used for the determination of internal control and quantification of gene expressions by qRT-PCR.

<p>The list of primers used for the determination of internal control and quantification of gene expressions by qRT-PCR.</p

Schematic presentation of molecular responsive-mechanisms in <i>S.</i><i>salsa</i> to Hg²⁺ co-exposed without (A) or with (B) the environmentally relevant salinity (NaCl, 500 mM) according to Kyoto Encyclopedia of Genes and Genomes (http://www.genome.jp/kegg/) and Uniprot (http://www.uniprot.org/).

<p>The identified proteins and metabolites were shown by marking the protein names in red (up-regulated) or blue (down-regulated).</p

The average length (A) and fresh weight (B) of <i>S.</i><i>salsa</i> seedlings from control, Hg²⁺−, salinity- and Hg²⁺+salinity-treated groups.

<p>The average length (A) and fresh weight (B) of <i>S.</i><i>salsa</i> seedlings from control, Hg²⁺−, salinity- and Hg²⁺+salinity-treated groups.</p

Data_Sheet_1_Plant resistance to tomato yellow leaf curl virus is enhanced by Bacillus amyloliquefaciens Ba13 through modulation of RNA interference.docx

IntroductionTomato yellow leaf curl virus (TYLCV), which is a typical member of the genus Begomovirus, causes severe crop yield losses worldwide. RNA interference (RNAi) is an important antiviral defense mechanism in plants, but whether plant beneficial microbes used as biocontrol agents would modulate RNAi in defense against TYLCV remains unclear.MethodsHere, we employed whole-transcriptome, bisulfite, and small RNA sequencing to decipher the possible role of Bacillus amyloliquefaciens Ba13 as a bacterial biocontrol agent against TYLCV in RNAi modulation.ResultsPotted tomato plants were exposed to whiteflies for natural viral infection 14 days after bacterial inoculation. Compared with non-inoculated controls, the abundance of TYLCV gene in the leaves of inoculated plants decreased by 70.1% at 28 days post-infection, which mirrored the pattern observed for plant disease index. The expression of the ARGONAUTE family genes (e.g., AGO3, AGO4, AGO5, and AGO7) involved in antiviral defense markedly increased by 2.44–6.73-fold following bacterial inoculation. The methylation level at CpG site 228 (in the open reading frame region of the RNA interference suppressing gene AV2) and site 461 (in the open reading frame regions of AV1 and AV2) was 183.1 and 63.0% higher in inoculated plants than in non-inoculated controls, respectively. The abundances of 10 small interfering RNAs matched to the TYLCV genome were all reduced in inoculated plants, accompanied by enhancement of photosystem and auxin response pathways.DiscussionThe results indicate that the application of Ba. amyloliquefaciens Ba13 enhances plant resistance to TYLCV through RNAi modulation by upregulating RNAi-related gene expression and enhancing viral genome methylation.</p