9 research outputs found
Heatmap of the variance stabilization transformed data (vsd) of ds<i>Pcsut6</i>-injected <i>vs</i>. ds<i>gfp</i>-injected samples.
<p>Samples derived from glandular tissue. For further explanation see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084461#pone-0084461-g008" target="_blank">Figure 8</a>.</p
Heatmap of the variance stabilization transformed data (vsd) of ds<i>Pcsut2</i>-injected <i>versus</i> ds<i>gfp</i>-injected samples.
<p>Samples derived from glandular tissue. For further explanation see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084461#pone-0084461-g008" target="_blank">Figure 8</a>.</p
Number of assembled transcripts and average length after assembly and reassembly showing the usefulness of reassembling.
<p>Number of assembled transcripts and average length after assembly and reassembly showing the usefulness of reassembling.</p
Heatmap of the variance stabilization transformed data (vsd) of ds<i>Pcsut1</i>-injected <i>vs</i>. ds<i>gfp</i>-injected samples.
<p>Samples derived from glandular tissue. For this, the transcript counts of the sugar transporters of each sample after dsRNA-injection have been normalized to the effective library size and the variance over all samples has been stabilized by applying the DESeq package. For each heatmap, the 30 most abundant sugar transporter transcripts are shown. Ds<i>gfp</i>-injected samples are the same in each heatmap.</p
Putative Sugar Transporters of the Mustard Leaf Beetle <i>Phaedon cochleariae</i>: Their Phylogeny and Role for Nutrient Supply in Larval Defensive Glands
<div><p>Background</p><p>Phytophagous insects have emerged successfully on the planet also because of the development of diverse and often astonishing defensive strategies against their enemies. The larvae of the mustard leaf beetle <i>Phaedon cochleariae</i>, for example, secrete deterrents from specialized defensive glands on their back. The secretion process involves ATP-binding cassette transporters. Therefore, sugar as one of the major energy sources to fuel the ATP synthesis for the cellular metabolism and transport processes, has to be present in the defensive glands. However, the role of sugar transporters for the production of defensive secretions was not addressed until now.</p><p>Results</p><p>To identify sugar transporters in <i>P. cochleariae</i>, a transcript catalogue was created by Illumina sequencing of cDNA libraries. A total of 68,667 transcripts were identified and 68 proteins were annotated as either members of the solute carrier 2 (SLC2) family or trehalose transporters. Phylogenetic analyses revealed an extension of the mammalian GLUT6/8 class in insects as well as one group of transporters exhibiting distinctive conserved motifs only present in the insect order Coleoptera. RNA-seq data of samples derived from the defensive glands revealed six transcripts encoding sugar transporters with more than 3,000 counts. Two of them are exclusively expressed in the glandular tissue. Reduction in secretions production was accomplished by silencing two of four selected transporters. RNA-seq experiments of transporter-silenced larvae showed the down-regulation of the silenced transporter but concurrently the up-regulation of other SLC2 transporters suggesting an adaptive system to maintain sugar homeostasis in the defensive glands.</p><p>Conclusion</p><p>We provide the first comprehensive phylogenetic study of the SLC2 family in a phytophagous beetle species. RNAi and RNA-seq experiments underline the importance of SLC2 transporters in defensive glands to achieve a chemical defense for successful competitive interaction in natural ecosystems.</p></div
Differential expression analysis using DESeq package.
<p>baseMeanA: mean of normalized counts value of ds<i>gfp</i>-injected samples. baseMeanB: mean of normalized counts values of dsRNA-<i>gfp</i>-injected, dsRNA-<i>Pcsut1</i>-injected, dsRNA-<i>Pcsut2</i>-injected, dsRNA-<i>Pcsut6</i>-injected samples. Fold-change: baseMeanA compared to baseMeanB. Log<sub>2</sub>fold-change: logarithm (to base 2) of fold-change values. Pval: p-value for the statistical significance of this change. Padj: p-value adjusted for multiple testing with Benjamini-Hochberg procedure which controls false discovery rate.</p
Characterized trehalose as well as glucose/fructose transporters.
<p>They are listed with their description, accession number and organism. Those transporters were added to <i>P. cochleariae</i>’s chosen sequences to calculate phylogenetic trees.</p
The phylogenetic tree of the nine sequences derived from <i>P. cochleariae</i> belonging to the purple branch (see <b>Figure 3</b> and <b>4</b>, Figure S4) and homologous sequences derived from the whole tree of life, especially from <i>Dendroctonus ponderosae</i> (Dc) as well as from <i>Tribolium castaneum</i> (Tc), was calculated using RaxML.
<p>Indicated in purple, it can be seen that the beetles’ sequences build up a separate branch.</p
Distribution of mRNA levels of putative SLC2 transporters in various tissues of juvenile <i>P. cochleariae</i> by using quantitative real-time PCR.
<p>Distribution of mRNA levels of putative SLC2 transporters in various tissues of juvenile <i>P. cochleariae</i> by using quantitative real-time PCR.</p