Expression patterns of four candidate reference genes in head samples of foragers and nurses.

<p>Seasonal expression patterns, indicated by quantification cycle (C_q) values of four candidate reference genes over a yearlong cycle, are represented for the foragers and nurses by the closed black circle and open white circle, respectively (A-D). C_q values of each gene over the year in foragers and nurses were statistically analyzed with repeated-measures ANOVA (A-D). Average and standard error of C_q values of each gene for forager head (For) and nurse head (Nur) were calculated from the data of three colonies for 12 months (E). Integrated expression level of each gene between foragers and nurses were statistically analyzed with Student’s t-test (E).</p

Optimal number of reference genes for normalization calculated by geNorm.

<p>Pairwise variations between the normalization factors (NF_n and NF_n+1) were separately analyzed to determine the optimal number of reference genes for normalization in forager head and nurse head samples (A). In order to compare the target gene expression between forager and nurse heads, the optimal number of reference genes for normalization was analyzed using combined data from forager and nurse (B).</p

Description, accession number of the reference gene, size and GC percentage of PCR products, qRT-PCR efficiency, and correlation coefficient (R²) of the selected reference genes.

<p>Description, accession number of the reference gene, size and GC percentage of PCR products, qRT-PCR efficiency, and correlation coefficient (R²) of the selected reference genes.</p

Validation of quantitative real-time PCR reference genes for the determination of seasonal and labor-specific gene expression profiles in the head of Western honey bee, <i>Apis mellifera</i>

<div><p>Honey bee is not only considered an important pollinator in agriculture, but is also widely used as a model insect in biological sciences, thanks to its highly evolved sociality, specialization of labor division, and flexibility of colony management. For an intensive investigation of the seasonal and labor-dependent expression patterns of its genes, accurate quantification of the target gene transcription level is a fundamental step. To date, quantitative real-time PCR (qRT-PCR) has been widely used for rapid quantification of gene transcripts, with reliable reference gene(s) for normalization. To this end, in an attempt to search for reliable reference genes, the amplification efficiencies of six candidate reference genes (<i>rp49</i>, <i>rpL32</i>, <i>rpS18</i>, <i>tbp</i>, <i>tub</i>, and <i>gapdh</i>) were determined. Subsequently, four genes (<i>rpL32</i>, <i>rpS18</i>, <i>tbp</i>, and <i>gapdh</i>) with PCR efficiencies of 90% to 110% were evaluated for their expression stabilities with three programs (geNorm, NormFinder, and BestKeeper) and used for normalization of seasonal expression patterns of target genes in the forager and nurse heads. Although the three programs revealed slightly different results, two genes, <i>rpS18</i> and <i>gapdh</i>, were suggested to be the optimal reference genes for qRT-PCR-based determination of seasonal and labor-specific gene expression profiles. Furthermore, the combined use of these two genes yielded a more accurate normalization, compared with the use of a single gene in the head of honey bee. The validated reference genes can be widely used for quantification of target gene expression in honey bee head although it is still remained to be elucidated the expression levels of the selected reference genes in specific tissues in head.</p></div

Average expression stability values of the five reference genes in forager, nurse, and integration of two tasks calculated by geNorm and NormFinder.

<p>Average expression stability values of the five reference genes in forager, nurse, and integration of two tasks calculated by geNorm and NormFinder.</p

Gene expression stability values of the five reference genes in forager, nurse, and integration of two tasks calculated by BestKeeper.

<p>Gene expression stability values of the five reference genes in forager, nurse, and integration of two tasks calculated by BestKeeper.</p

Phylogenetic tree of <i>Tetranychus</i> mites based on mtCOI partial sequences.

<p>Two <i>Oligonychus spp</i>. were used as outgroups. A maximum likelihood test was conducted and a bootstrap value over 50% is shown. The Korean populations are designated with blue circles.</p

Phenotypic- and Genotypic-Resistance Detection for Adaptive Resistance Management in <i>Tetranychus urticae</i> Koch

<div><p>Rapid resistance detection is necessary for the adaptive management of acaricide-resistant populations of <i>Tetranychus urticae</i>. Detection of phenotypic and genotypic resistance was conducted by employing residual contact vial bioassay (RCV) and quantitative sequencing (QS) methods, respectively. RCV was useful for detecting the acaricide resistance levels of <i>T</i>. <i>urticae</i>, particularly for on-site resistance detection; however, it was only applicable for rapid-acting acaricides (12 out of 19 tested acaricides). QS was effective for determining the frequencies of resistance alleles on a population basis, which corresponded to 12 nonsynonymous point mutations associated with target-site resistance to five types of acaricides [organophosphates (monocrotophos, pirimiphos-methyl, dimethoate and chlorpyrifos), pyrethroids (fenpropathrin and bifenthrin), abamectin, bifenazate and etoxazole]. Most field-collected mites exhibited high levels of multiple resistance, as determined by RCV and QS data, suggesting the seriousness of their current acaricide resistance status in rose cultivation areas in Korea. The correlation analyses revealed moderate to high levels of positive relationships between the resistance allele frequencies and the actual resistance levels in only five of the acaricides evaluated, which limits the general application of allele frequency as a direct indicator for estimating actual resistance levels. Nevertheless, the resistance allele frequency data alone allowed for the evaluation of the genetic resistance potential and background of test mite populations. The combined use of RCV and QS provides basic information on resistance levels, which is essential for choosing appropriate acaricides for the management of resistant <i>T</i>. <i>urticae</i>.</p></div

Toxicity parameter by residual contact vial bioassay to PyriF strains in <i>Tetranychus urticae</i>.

<p>^a The grouping was based on the mode of action classification by Insecticide Resistance Action Committee (IRAC) (<a href="http://www.irac-online.org/documents/moa-structures-poster-english/?ext=pdf" target="_blank">http://www.irac-online.org/documents/moa-structures-poster-english/?ext=pdf</a>)</p><p>^b ND represents ‘not determined’</p><p>^c Determined by leaf dipping methods based on emergence rate. The unit of toxicity levels were ppm.</p><p>Toxicity parameter by residual contact vial bioassay to PyriF strains in <i>Tetranychus urticae</i>.</p

Anti-bacterial effects of components from <i>Sanguisorba officinalis</i> L. on <i>Vibrio vulnificus</i> and their soluble epoxide hydrolase inhibitory activity

<p><i>Sanguisorba officinalis</i> L. is a traditional herbal medicine, which is prevailingly applied to cure hemorrhoids, wounds and ulcers in Eastern Asian countries. The purpose of this study was to investigate the antibacterial and soluble epoxide hydrolase (sEH) inhibitory effects of the extracts and components from <i>S. officinalis</i>. The methanol extract was divided into ethyl acetate (EtOAc), <i>n</i>-butanol (<i>n</i>-BuOH), and water layers. In our screening procedure, the EtOAc and <i>n</i>-BuOH extracts and compounds (<b>1</b>–<b>2</b>) remarkably suppressed the growth of <i>V. vulnificus</i> in a dose-dependent manner. In addition, the EtOAc extract and compound <b>1</b> exhibited significant inhibitory effect on the <i>V</i>. <i>vulnificus</i> induced cytotoxicity on HeLa cells. Furthermore, compound <b>4</b> displayed an inhibition against sEH with an IC₅₀ value of 7.0 ± 0.5 μM. A kinetic analysis demonstrated that the inhibitory effect of compound <b>4</b> was a mixed type, with an inhibitory constant (<i>K</i>_<i>i</i>) 0.22 ± 0.0 μM.</p