qRT-PCR validation of a selection of genes.

<p>The table shows the relative expression levels, obtained using mRNA-Seq or qRT-PCR on an independent sample, comparing systemic tissue of <i>H. oryzae</i> infected plants with uninfected plants at time point 3 dpi.</p><p>qRT-PCR validation of a selection of genes.</p

Visualization of transcriptome data of shoots of rice infected with rice root nematode <i>Hirschmanniella oryzae</i>.

<p>(A) Parametric Analysis of Gene Set Enrichment of the differentially expressed genes in the shoots of infected versus uninfected plants. The Z-scores of the significantly enriched secondary level GO terms are shown. (B) Mapman visualization of the expression profiles of genes involved in the general metabolism of the rice plant. The visualization shows the observed differential expression patterns, based on the Log₂ fold changes of mRNA levels, in shoots of infected versus uninfected control plants. Red dots indicate that the gene is upregulated in infected plants versus the corresponding healthy control plants, while blue indicates downregulation.</p

Differentially expressed genes (FDR<0.05), when comparing systemic tissue of <i>H. oryzae</i> infected plants with uninfected plants at both time points (3 and 7 dpi).

<p>The table shows the rice locus number, the annotation (MSU7.0), the Log₂ of the fold-change (FC) of infected versus uninfected shoot tissue and the FDR-value. Novel transcriptionally active regions received a gene ID starting with RNSR. FDR: false discovery rate. RNSR: Root Nematode Systemic Respons.</p><p>Differentially expressed genes (FDR<0.05), when comparing systemic tissue of <i>H. oryzae</i> infected plants with uninfected plants at both time points (3 and 7 dpi).</p

Overview of the obtained sequencing data and mapping of these sequences onto the rice genome.

<p>Overview of the obtained sequencing data and mapping of these sequences onto the rice genome.</p

Primers used in PCR for cloning and real-time quantitative PCR.

<p>Primers used in PCR for cloning and real-time quantitative PCR.</p

Overview of identified genes associated to RISC complex in <i>C. puncticollis.</i> (FS) frame shift; (RF) reading frame.

<p>Overview of identified genes associated to RISC complex in <i>C. puncticollis.</i> (FS) frame shift; (RF) reading frame.</p

Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.

<p>Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.</p

Inhibition of <i>laccase2</i> expression in second-instar larvae of <i>Cylas puncticollis</i> at 1, 3, 5 and 10 days after injection with dsRNA targeting <i>laccase2</i> at 0.2 µg/mg of body weight.

<p>Injection with dsRNA targeting <i>gfp</i> was used as a control. As internal controls, ribosomal protein L32 and Actin were used. Values are based on two repetitions of three biological samples and expressed as mean ± SEM. Each sample contains 5 pooled insects. The p-values were calculated by unpaired t-test.</p

Overview of identified genes related to the RNAi pathways in <i>C. puncticollis</i>.

<p>(FS) frame shift; (RF) reading frame.</p><p>Overview of identified genes related to the RNAi pathways in <i>C. puncticollis</i>.</p

Image_2_The Globodera pallida SPRYSEC Effector GpSPRY-414-2 That Suppresses Plant Defenses Targets a Regulatory Component of the Dynamic Microtubule Network.JPEG

<p>The white potato cyst nematode, Globodera pallida, is an obligate biotrophic pathogen of a limited number of Solanaceous plants. Like other plant pathogens, G. pallida deploys effectors into its host that manipulate the plant to the benefit of the nematode. Genome analysis has led to the identification of large numbers of candidate effectors from this nematode, including the cyst nematode-specific SPRYSEC proteins. These are a secreted subset of a hugely expanded gene family encoding SPRY domain-containing proteins, many of which remain to be characterized. We investigated the function of one of these SPRYSEC effector candidates, GpSPRY-414-2. Expression of the gene encoding GpSPRY-414-2 is restricted to the dorsal pharyngeal gland cell and reducing its expression in G. pallida infective second stage juveniles using RNA interference causes a reduction in parasitic success on potato. Transient expression assays in Nicotiana benthamiana indicated that GpSPRY-414-2 disrupts plant defenses. It specifically suppresses effector-triggered immunity (ETI) induced by co-expression of the Gpa2 resistance gene and its cognate avirulence factor RBP-1. It also causes a reduction in the production of reactive oxygen species triggered by exposure of plants to the bacterial flagellin epitope flg22. Yeast two-hybrid screening identified a potato cytoplasmic linker protein (CLIP)-associated protein (StCLASP) as a host target of GpSPRY-414-2. The two proteins co-localize in planta at the microtubules. CLASPs are members of a conserved class of microtubule-associated proteins that contribute to microtubule stability and growth. However, disruption of the microtubule network does not prevent suppression of ETI by GpSPRY-414-2 nor the interaction of the effector with its host target. Besides, GpSPRY-414-2 stabilizes its target while effector dimerization and the formation of high molecular weight protein complexes including GpSPRY-414-2 are prompted in the presence of the StCLASP. These data indicate that the nematode effector GpSPRY-414-2 targets the microtubules to facilitate infection.</p