Fatty acids differ significantly in castes of the Formosan subterranean termite (Isoptera: Rhinotermitidae)

© The Authors 2015. Identification and quantitation of fatty acids (FAs) in nymphs, alates, workers, presoldiers, and soldiers of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were determined by gas chromatography- mass spectrometry, showing quantitative and qualitative differences among groups. Total FAs content of nymphs and alate females was about 1.5-fold higher than alate males, about 2-fold higher than workers, 6-fold higher than presoldiers, and 12-fold higher than soldiers. Overall differences in total FAs content were due to oleic acid (C18:1), stearic acid (C18:0), linoleic acid (C18:2), and palmitic acid (C16:0). Soldiers contained two unique FAs among the castes - lignoceric (C24:0) and hexacosanoic acid (C26:0). Nymphs had the highest ratio between triacylglycerols and phospholipids probably for energy storage in alate development. Four branched FAs - 13-methyl myristic, 14-methyl pentadecenoic, 15-methyl palmitic, and 14-methyl palmitic - and three oddnumbered FAs - pentadecanoic (C15:0), heptadecanoic (C17:0), and heptadecenoic (C17:1) - were found in nymphs, alates, workers, presoldiers, and soldiers. Interestingly, all FAs were distributed in different percentages both in triglycerides and phospholipids of the different developmental stages and castes, indicating a function both for energy storage and membrane components. Five different 2-hydroxy FAs - 2-OH C16:0, 2-OH C18:0, 2-OH C20:0, 2-OH C22:0, and 2-OH C24:0 - were identified, the latter only in soldiers. Total 2-hydroxy FAs content in soldiers was significantly higher than that in other groups (6.01-7.9-fold vs. presoldiers and 41.7- 132.6-fold vs. nymphs, alates, and workers), and the quantity in presoldiers was significantly higher than in nymphs, alates, and workers, with no difference among nymphs, alates, and workers

Influence of Rearing Temperature on Fatty Acid Metabolism in the Pea Aphid, Acyrthosiphon Pisum

Entomolog

Diagnostic Molecular Markers for Phosphine Resistance in US Populations of Tribolium castaneum and Rhyzopertha dominica

Citation: Chen, Z., Schlipalius, D., Opit, G., Subramanyam, B., & Phillips, T. W. (2015). Diagnostic Molecular Markers for Phosphine Resistance in US Populations of Tribolium castaneum and Rhyzopertha dominica. Plos One, 10(3), 14. doi:10.1371/journal.pone.0121343Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T. castaneum and R. dominica with strong resistance was identified as P45S in T. castaneum and P49S in R. dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T. castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population

浅析中药学与民族药学对木香的临床应用异同

Costusroot is commonly used in traditional medicine,Mongolian medicine,Tibetan medicine,Dai medicine and Uighur medicine. We discuss similarities and differences in clinical applications of costusroot in traditional medicine,Mongolian medicine,Tibetan medicine,Dai medicine and Uighur medicine, from aspects of its property and flavor,functions and indications ,and clinical applications. For sake of retaining their own features, the authors intended to integrate the essences of the two, broaden its clinical applications, make costusroot to play its role fully and fully understand the natural medicinal plants ,which is more conducive to modernize the traditional medicine.木香是中、蒙、藏、傣以及维医药学的常用药。本文从木香的性味、功效主治及临床应用等方面，比较探讨木香在中医药学、蒙医药学、藏医药学、傣医药学及维医药学中的临床应用异同，希冀在保留各自特色的基础上，相互借鉴，拓宽临床应用，使之发挥其更全面的作用，同时全面认识自然界药用植物，更有利于促进中药现代化

Genes related to mitochondrial functions are differentially expressed in phosphine-resistant and -susceptible Tribolium castaneum

Background: Phosphine is a valuable fumigant to control pest populations in stored grains and grain products. However, recent studies indicate a substantial increase in phosphine resistance in stored product pests worldwide.Results: To understand the molecular bases of phosphine resistance in insects, we used RNA-Seq to compare gene expression in phosphine-resistant and susceptible laboratory populations of the red flour beetle, Tribolium castaneum. Each population was evaluated as either phosphine-exposed or no phosphine (untreated controls) in triplicate biological replicates (12 samples total). Pairwise analysis indicated there were eight genes differentially expressed between susceptible and resistant insects not exposed to phosphine (i.e., basal expression) or those exposed to phopshine (>8-fold expression and 90 % C.I.). However, 214 genes were differentially expressed among all four treatment groups at a statistically significant level (ANOVA, p < 0.05). Increased expression of 44 cytochrome P450 genes was found in resistant vs. susceptible insects, and phosphine exposure resulted in additional increases of 21 of these genes, five of which were significant among all treatment groups (p < 0.05). Expression of two genes encoding anti-diruetic peptide was 2- to 8-fold reduced in phosphine-resistant insects, and when exposed to phosphine, expression was further reduced 36- to 500-fold compared to susceptible. Phosphine-resistant insects also displayed differential expression of cuticle, carbohydrate, protease, transporter, and many mitochondrial genes, among others. Gene ontology terms associated with mitochondrial functions (oxidation biological processes, monooxygenase and catalytic molecular functions, and iron, heme, and tetrapyyrole binding) were enriched in the significantly differentially expressed dataset. Sequence polymorphism was found in transcripts encoding a known phosphine resistance gene, dihydrolipoamide dehydrogenase, in both susceptible and resistant insects. Phosphine-resistant adults also were resistant to knockdown by the pyrethroid deltamethrin, likely due to the increased cytochrome P450 expression.Conclusions: Overall, genes associated with the mitochondria were differentially expressed in resistant insects, and these differences may contribute to a reduction in overall metabolism and energy production and/or compensation in resistant insects. These data provide the first gene expression data on the response of phosphine-resistant and -susceptible insects to phosphine exposure, and demonstrate that RNA-Seq is a valuable tool to examine differences in insects that respond differentially to environmental stimuli.Peer reviewedEntomology and Plant Patholog

Fatty Acids Differ Significantly in Castes of the Formosan Subterranean Termite (Isoptera: Rhinotermitidae)

© The Authors 2015. Identification and quantitation of fatty acids (FAs) in nymphs, alates, workers, presoldiers, and soldiers of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were determined by gas chromatography- mass spectrometry, showing quantitative and qualitative differences among groups. Total FAs content of nymphs and alate females was about 1.5-fold higher than alate males, about 2-fold higher than workers, 6-fold higher than presoldiers, and 12-fold higher than soldiers. Overall differences in total FAs content were due to oleic acid (C18:1), stearic acid (C18:0), linoleic acid (C18:2), and palmitic acid (C16:0). Soldiers contained two unique FAs among the castes - lignoceric (C24:0) and hexacosanoic acid (C26:0). Nymphs had the highest ratio between triacylglycerols and phospholipids probably for energy storage in alate development. Four branched FAs - 13-methyl myristic, 14-methyl pentadecenoic, 15-methyl palmitic, and 14-methyl palmitic - and three oddnumbered FAs - pentadecanoic (C15:0), heptadecanoic (C17:0), and heptadecenoic (C17:1) - were found in nymphs, alates, workers, presoldiers, and soldiers. Interestingly, all FAs were distributed in different percentages both in triglycerides and phospholipids of the different developmental stages and castes, indicating a function both for energy storage and membrane components. Five different 2-hydroxy FAs - 2-OH C16:0, 2-OH C18:0, 2-OH C20:0, 2-OH C22:0, and 2-OH C24:0 - were identified, the latter only in soldiers. Total 2-hydroxy FAs content in soldiers was significantly higher than that in other groups (6.01-7.9-fold vs. presoldiers and 41.7- 132.6-fold vs. nymphs, alates, and workers), and the quantity in presoldiers was significantly higher than in nymphs, alates, and workers, with no difference among nymphs, alates, and workers

Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica

Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T.castaneum and R.dominica with strong resistance was identified as P45S in T.castaneum and P49S in R.dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T.castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population

Frequency of phosphine resistance phenotypes in seven Kansas population of <i>T</i>. <i>castaneum</i>.

<p>Phenotypes were evaluated using discriminating dose bioassays for weak resistance (30 ppm phosphine for 20 hrs) and strong resistance (180 ppm for 20 hrs). The percent surviving the bioassay for a given population is equivalent to the percent resistant reported here. Also shown are the CAPS results for the frequency of the strong resistance allele in each population.</p

Deduced amino acid sequences of DLD from <i>T</i>. <i>castaneum</i> and <i>R</i>. <i>dominica</i> by ClustalW2.

<p>Only the portions of the amino acid sequences with relevant mutations in both species are shown. <b>Fig 2A:</b> Sequences from susceptible and resistant strains of <i>T</i>. <i>castaneum</i> from the USA aligned against Australian strains. <b>Fig 2B:</b> Sequences from susceptible and resistant strains of <i>R</i>. <i>dominica</i> from the USA aligned against Australian strains. Differences between the phosphine-susceptible and phosphine-resistant sequences are boxed. The asterisk denotes the P45S phosphine-resistance mutations described in the <i>T</i>. <i>castaneum</i> population TcOK-G. The triangle denotes the P49S phosphine-resistance mutations described in the text. The squares denote all the other phosphine-resistance mutations described only in Australian strains.</p