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In this study, the cDNAs of five cytochromes P450 genes (named CYP345P1, CYP358B1, CYP4FD2, CYP4CD2, and CYP6JN1) contained open reading frames from 1,500 to 1,554 nucleotides that encoded 499 to 517 amino acids were cloned from the psocid Liposcelis entomophila. They are characterized by predicted molecular weights from 57.67 to 59.64 kDa and theoretical isoelectric points of 5.57-9.07. Quantitative real-time PCR analysis showed these five genes were expressed at all tested developmental stages and higher expressions were observed in adults. CYP358B1 was expressed at higher levels in egg and adult compared to the larval stages. mRNA abundances of five genes were detected in both sexes and were relatively more abundant in adult females than in adult males. Synergism bioassay showed that the synergic ratio was 2.20 and 2.45 when insects were treated with the mixture of deltamethrin or malathion with the synergist piperonyl butoxide (PBO). Because PBO induces cytochrome P450s in some insects, this suggested to us that cytochromes P450 might participate in detoxification of these insecticides. The transcripts of the five cytochromes P450 genes in adult psocids could be induced to the highest level at 12 h after the exposure to malathion. After exposure to deltamethrin, CYP358B1 reached maximum expression at 24 h. The maximum expression of the other four genes occurred at 36 h. Treatments with the carbamate propoxur did not influence transcription of the cytochromes P450 gene. The induction profiles suggested that these five cytochrome P450 genes may be associated with deltamethrin and malathion metabolism in psocids., (© 2016 Wiley Periodicals, Inc.)

Fertility life table provides a comprehensive description of arthropod population dynamics by the estimation of parameters about arthropod population growth potential. It can also clarify the sublethal effects of chemicals on insects. Ethacrynic acid (EA), an inhibitor of glutathione S-transferases, is a diuretic compound that has been confirmed to modulate drug resistance in organisms. In this study, the effects of EA on growth and development of Liposcelis bostrychophila Badonnel were investigated in the laboratory to explore the potential possibilities of EA as an active agent to manage insecticide-resistant psocids. The treatment of psocids was obtained by feeding on the routine diet containing 3% EA for three successive generations, and psocids on routine diet served as control. The results indicated that EA possessed some negative effects on the life-table parameters of the psocid in F1 and F2 generations. The addition of EA to diet stunted psocids growth by lengthening development time and increasing mortality with a greater effect in the F2 generation. In the third generation of psocids on EA diet, it seemed there was a return to normal. Psocid fitness was influenced by addition of EA to standard diet. Using rm values, the fitness for EA diet in F1, F2, and F3 compared with the counterpart of routine diet was calculated as 0.80, 0.74, and 0.87, respectively., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: The genus Liposcelis (Psocoptera: Troctomorpha) has more than 120 species with a worldwide distribution and they pose a risk for global food security. The organization of mitochondrial (mt) genomes varies between the two species of booklice investigated in the genus Liposcelis. Liposcelis decolor has its mt genes on a single chromosome, like most other insects; L. bostrychophila, however, has a multipartite mt genome with genes on two chromosomes., Results: To understand how multipartite mt genome organization evolved in the genus Liposcelis, we sequenced the mt genomes of L. entomophila and L. paeta in this study. We found that these two species of booklice also have multipartite mt genomes, like L. bostrychophila, with the mt genes we identified on two chromosomes. Numerous pseudo mt genes and non-coding regions were found in the mt genomes of these two booklice, and account for 30% and 10% respectively of the entire length we sequenced. In L. bostrychophila, the mt genes are distributed approximately equally between the two chromosomes. In L. entomophila and L. paeta, however, one mt chromosome has most of the genes we identified whereas the other chromosome has largely pseudogenes and non-coding regions. L. entomophila and L. paeta differ substantially from each other and from L. bostrychophila in gene content and gene arrangement in their mt chromosomes., Conclusions: Our results indicate unusually fast evolution in mt genome organization in the booklice of the genus Liposcelis, and reveal different patterns of mt genome fragmentation among L. bostrychophila, L. entomophila and L. paeta.

Booklice in the genus Liposcelis are pests of stored grain products. They pose a considerable economic threat to global food security and safety. To date, the complete mitochondrial genome has only been determined for a single booklouse species Liposcelis bostrychophila. Unlike most bilateral animals, which have their 37 mt genes on one circular chromosome, ≈15 kb in size, the mt genome of L. bostrychophila has two circular chromosomes, 8 and 8.5 kb in size. Here, we report the mt genome of another booklouse, Liposcelis decolor. The mt genome of L. decolor has the typical mt chromosome of bilateral animals, 14,405 bp long with 37 genes (13 PCGs, 22 tRNAs and 2 rRNAs). However, the arrangement of these genes in L. decolor differs substantially from that observed in L. bostrychophila and other insects. With the exception of atp8-atp6, L. decolor differs from L. bostrychophila in the arrangement of all of the other 35 genes. The variation in the mt genome organization and mt gene arrangement between the two Liposcelis species is unprecedented for closely related animals in the same genus. Furthermore, our results indicate that the two-chromosome mt genome organization observed in L. bostrychophila likely evolved recently after L. bostrychophila and L. decolor split from their most recent common ancestor.

Background: Recent studies indicate that infestations of psocids pose a new risk for global food security. Among the psocids species, Liposcelis bostrychophila Badonnel has gained recognition in importance because of its parthenogenic reproduction, rapid adaptation, and increased worldwide distribution. To date, the molecular data available for L. bostrychophila is largely limited to genes identified through homology. Also, no transcriptome data relevant to psocids infection is available., Methodology and Principal Findings: In this study, we generated de novo assembly of L. bostrychophila transcriptome performed through the short read sequencing technology (Illumina). In a single run, we obtained more than 51 million sequencing reads that were assembled into 60,012 unigenes (mean size = 711 bp) by Trinity. The transcriptome sequences from different developmental stages of L. bostrychophila including egg, nymph and adult were annotated with non-redundant (Nr) protein database, gene ontology (GO), cluster of orthologous groups of proteins (COG), and KEGG orthology (KO). The analysis revealed three major enzyme families involved in insecticide metabolism as differentially expressed in the L. bostrychophila transcriptome. A total of 49 P450-, 31 GST- and 21 CES-specific genes representing the three enzyme families were identified. Besides, 16 transcripts were identified to contain target site sequences of resistance genes. Furthermore, we profiled gene expression patterns upon insecticide (malathion and deltamethrin) exposure using the tag-based digital gene expression (DGE) method., Conclusion: The L. bostrychophila transcriptome and DGE data provide gene expression data that would further our understanding of molecular mechanisms in psocids. In particular, the findings of this investigation will facilitate identification of genes involved in insecticide resistance and designing of new compounds for control of psocids.

Background: As a major stored-product pest insect, Liposcelis entomophila has developed high levels of resistance to various insecticides in grain storage systems. However, the molecular mechanisms underlying resistance and environmental stress have not been characterized. To date, there is a lack of genomic information for this species. Therefore, studies aimed at profiling the L. entomophila transcriptome would provide a better understanding of the biological functions at the molecular levels., Methodology/principal Findings: We applied Illumina sequencing technology to sequence the transcriptome of L. entomophila. A total of 54,406,328 clean reads were obtained and that de novo assembled into 54,220 unigenes, with an average length of 571 bp. Through a similarity search, 33,404 (61.61%) unigenes were matched to known proteins in the NCBI non-redundant (Nr) protein database. These unigenes were further functionally annotated with gene ontology (GO), cluster of orthologous groups of proteins (COG), and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. A large number of genes potentially involved in insecticide resistance were manually curated, including 68 putative cytochrome P450 genes, 37 putative glutathione S-transferase (GST) genes, 19 putative carboxyl/cholinesterase (CCE) genes, and other 126 transcripts to contain target site sequences or encoding detoxification genes representing eight types of resistance enzymes. Furthermore, to gain insight into the molecular basis of the L. entomophila toward thermal stresses, 25 heat shock protein (Hsp) genes were identified. In addition, 1,100 SSRs and 57,757 SNPs were detected and 231 pairs of SSR primes were designed for investigating the genetic diversity in future., Conclusions/significance: We developed a comprehensive transcriptomic database for L. entomophila. These sequences and putative molecular markers would further promote our understanding of the molecular mechanisms underlying insecticide resistance or environmental stress, and will facilitate studies on population genetics for psocids, as well as providing useful information for functional genomic research in the future.

Alternative splicing greatly contributes to the structural and functional diversity of voltage-gated sodium channels (VGSCs) by generating various isoforms with unique functional and pharmacological properties. Here, we identified a new optional exon 23 located in the linker between domains II and III, and four mutually exclusive exons (exons 27A, 27B, 27C, and 27D) in domains IIIS3 and IIIS4 of the sodium channel of Liposcelis bostrychophila (termed as LbVGSC). This suggested that more alternative splicing phenomena remained to be discovered in VGSCs. Inclusion of exon 27C might lead to generation of non-functional isoforms. Meanwhile, identification of three alternative exons (exons 11, 13A, and 13B), which were located in the linker between domains II and III, indicated that abundant splicing events occurred in the DSC1 ortholog channel of L. bostrychophila (termed as LbSC1). Exons 13A and 13B were generated by intron retention, and the presence of exon 13B relied on the inclusion of exon 13A. Exon 13B was specifically expressed in the embryonic stage and contained an in-frame stop codon, inclusion of which led to generation of truncated proteins with only the first two domains. Additionally, several co-occurring RNA editing events were identified in LbSC1. Furthermore, remarkable similarity between the structure and expression patterns of LbVGSC and LbSC1 were discovered, and a closer evolutionary relationship between VGSCs and DSC1 orthologs was verified. Taken together, the data provided abundant molecular information on VGSC and DSC1 orthologs in L. bostrychophila, a representative Psocoptera storage pest, and insights into the alternative splicing of these two channels.

The cDNAs of three novel P450 genes, CYP4CB1, CYP4CC1, and CYP4CD1 (GenBank accessions EU979550, EU979549, and EU979551, respectively), were sequenced and characterized from the psocid Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae). CYP4CB1, CYP4CC1, and CYP4CD1 contain open reading frames of 1533, 1512, and 1536 nucleotides that encode 511,504, and 512 amino acid residues, respectively. The putative proteins of CYP4CB1, CYP4CC1, and CYP4CD1 show predicted molecular mass of 59.65, 58.87, and 59.71 kDa, with a theoretical isoelectric point of 6.59, 8.80, and 8.84, respectively. The N-terminal transmembrane domain was only found in CYP4CB1 suggested it is a typical microsomal P450. Phylogenetic analysis showed a close relationship of CYP4CB1, CYP4CC1, and CYP4CD1 with CYP4AW1, CYP4L4, and CYP4E2. Quantitative real-time reverse transcriptase-polymerase chain reaction indicated that these three genes were expressed at all tested developmental stages. In addition, the highest expression occurred in the adult stage, which suggested that these three P450 genes may play important roles in adulthood. The transcripts of CYP4CB1 and CYP4CC1 in adult psocids could be induced to the highest level at 36 and 24 h after the exposure to deltamethrin and paraoxon-methyl (50% lethal concentration [LC50]), respectively, whereas CYP4CD1 remained unchanged. CYP4CD1 transcripts, however, increased rapidly at 8 h after aldicarb (LC50) induction and reached the peak at 36 h. The induction profiles of the three P450 genes suggested that CYP4CB1 and CYP4CC1 are possibly associated with deltamethrin and paraoxonmethyl metabolism in psocids, whereas CYP4CD1 is probably involved in aldicarb metabolism. However, our assumption needs to be further verified by recombinant protein expression of these proteins as well as RNA interference of these genes.

Booklice (order Psocoptera) in the genus Liposcelis are major pests to stored grains worldwide and are closely related to parasitic lice (order Phthiraptera). We sequenced the mitochondrial (mt) genome of Liposcelis bostrychophila and found that the typical single mt chromosome of bilateral animals has fragmented into and been replaced by two medium-sized chromosomes in this booklouse; each of these chromosomes has about half of the genes of the typical mt chromosome of bilateral animals. These mt chromosomes are 8,530 bp (mt chromosome I) and 7,933 bp (mt chromosome II) in size. Intriguingly, mt chromosome I is twice as abundant as chromosome II. It appears that the selection pressure for compact mt genomes in bilateral animals favors small mt chromosomes when small mt chromosomes co-exist with the typical large mt chromosomes. Thus, small mt chromosomes may have selective advantages over large mt chromosomes in bilateral animals. Phylogenetic analyses of mt genome sequences of Psocodea (i.e. Psocoptera plus Phthiraptera) indicate that: 1) the order Psocoptera (booklice and barklice) is paraphyletic; and 2) the order Phthiraptera (the parasitic lice) is monophyletic. Within parasitic lice, however, the suborder Ischnocera is paraphyletic; this differs from the traditional view that each suborder of parasitic lice is monophyletic.

Background: The psocids Liposcelis bostrychophila and L. entomophila (Psocoptera: Liposcelididae) are found throughout the world and are often associated with humans, food stores and habitations. These insects have developed high levels of resistance to various insecticides in grain storage systems. However, the population genetic structure and gene flow of psocids has not been well categorized, which is helpful to plan appropriate strategies for the control of these pests., Methodology/principal Findings: The two species were sampled from 15 localities in China and analyzed for polymorphisms at the mitochondrial DNA (Cytb) and ITS (ITS1-5.8S-ITS2) regions. In total, 177 individual L. bostrychophila and 272 individual L. entomophila were analysed. Both Cytb and ITS sequences showed high genetic diversity for the two species with haplotype diversities ranged from 0.154 ± 0.126 to 1.000 ± 0.045, and significant population differentiation (mean F(ST) = 0.358 for L. bostrychophila; mean F(ST) = 0.336 for L. entomophila) was also detected among populations investigated. A Mantel test indicated that for both species there was no evidence for isolation-by-distance (IBD). The neutrality test and mismatch distribution statistics revealed that the two species might have undergone population expansions in the past., Conclusion: Both L. bostrychophila and L. entomophila displayed high genetic diversity and widespread population genetic differentiation within and between populations. The significant population differentiation detected for both psocids may be mainly due to other factors, such as genetic drift, inbreeding or control practices, and less by geographic distance since an IBD effect was not found.

Psocids (Psocoptera: Liposcelididae: Liposcelis spp.) are major pests of stored grain and commonly occur on a wide range of stored products. Increasingly, the genus of Liposcelis has gained recognition of their importance due to their feeding on stored grains, contaminating food, and agricultural commodities as well as transmitting harmful microorganisms, including fungi and bacteria. Psocids have close morphological similarities and often commix occur at the same ecosystems. Therefore, a first step necessary to further implement population studies is the accurate identification of species, based on molecular methods. In this study, we determined nucleotide sequences of the nuclear rDNA internal transcribed spacer (ITS)1-5.8S-ITS2 region in 100 individuals of six Liposcelis species (including Liposcelis bostrychophila Badonnel, Liposcelis entomophila (Enderlein), Liposcelis decolor (Pearman), Liposcelis tricolor Badonnel, Liposcelis paeta Pearman, and Liposcelis yunnaniensis Li & Li) from 16 locations of China. We evaluated the suitability of this marker for phylogenetic inference study in the Liposcelis species. We also developed a molecular identification method for six Liposcelis species based on ITS2 sequence. Results demonstrate that ITS1-5.8S-ITS2 sequences are a useful tool for the population genetic study and phylogeny estimation of Liposcelis species. The results of this study indicate that the ITS2 sequences can be a reliable tool for species discrimination of the six species of psocids tested here. In addition, the multiplex method described proved reliable when tested across different geographical populations.

We investigated the effects often constant temperatures (20.0, 22.5, 25.0, 27.5, 30.0, 32.5, 35.0, 37.5, 39.0, and 41.0 degrees C) on the development, survival, and reproduction of the psocid Liposcelis yunnaniensis Li & Li (Psocoptera: Liposcelididae). At 39.0 and 41.0 degrees C, none of individuals could develop successfully or reproduce. From 20 to 37.5 degrees C, the development period from egg to adult ranged from 64.3 d at 20 degrees C to 16.1 d at 35 degrees C. The lower developmental threshold for egg, nymph, and combined immature stages were estimated at 15.08, 15.13, and 14.77 degrees C, respectively. After emergence the females went through a preoviposition period that ranged from 18.5 d at 22.5 degrees C to 3.11 d at 35 degrees C, whereas it was 16.3 d at 20 degrees C. Liposcelis yunnaniensis produced most eggs at 35 degrees C and the fewest at 22.5 degrees C. The population reared at 35 degrees C had the highest intrinsic rate of increase, shorter mean generation time, and shortest population doubling time compared with other temperatures. According to Weibull frequency distribution, L. yunnaniensis reared at all the temperatures had type III survivorship curves (c < 1.0). Based on life-table parameter estimations, we suggest that optimum range of temperatures for this species is from 25 to 37.5 degrees C. These data give us useful information on population biology of L. yunnaniensis and can be used to better manage this species.

Acetylcholinesterace (AChE) is known to be the major target for organophophate and carbamate insecticides and biomolecular changes to AChE have been demonstrated to be an important mechanism for insecticide resistance in many insect species. In this study, AChE from three field populations of Liposcelis entomophila (Enderlein) (Psocoptera: Liposcelididae) was purified by affinity chromatography and subsequently characterized by its Michaelis-Menten kinetics to determine if detectable changes to AChE have occurred. Bioassays revealed that the potential resistance threat of psocids in Sichuan Province (GH) was greater than either Hubei Province (WH) or Chongqing Municipality (BB). Compared to the other two populations, the WH population possessed the highest specific activity of purified AChE. Kinetic analyses indicated that the purified AChE from GH population expressed a significantly lower affinity to the substrate and a higher catalytic activity toward acetylthiocholine iodide (ATChI) (i.e., higher K(m) and V(max) values) than BB and WH populations. In vitro studies of AChE suggest that five inhibitors (aldicarb, eserine, BW284C51, omethoate, and propoxur) all possess strong inhibitory effects with eserine having the strongest inhibitory effect against purified AChE. According to bimolecular rate constants (k(i)), the purified AChE from GH population was least sensitive to all inhibitors except for omethoate. The differences in AChE among the three populations may be partially attributed to the differences in pesticide application and control practices for psocids among the three locations., (© 2010 Wiley Periodicals, Inc.)

The development, survival, and reproduction of Liposcelis paeta Pearman (Psocoptera: Liposcelididae), an important insect pest of stored products, were evaluated at nine constant temperatures (20-40 degrees C). Few insects could complete the development or reproduce at 20 or 40 degrees C. Between 22.5 and 37.5 degrees C, the developmental period from egg to adult varied from 45.4 d at 22.5 degrees C to 11.5 d at 37.5 degrees C. The lower temperature developmental thresholds were estimated at 18.1-21.9 degrees C, and the upper temperature thresholds were 40.4-42.0 degrees C. The percentage of survival from egg to adult was 52.79% at 32.5 degrees C and 18.79% at 22.5 degrees C. After emergence, the adult had a short preoviposition period that ranged from 6.3 d at 22.5 degrees C to 0.8 d at 37.5 degrees C. L. paeta produced the most eggs at 27.5 degrees C and the fewest at 37.5 degrees C. The population reared at 32.5 degrees C had the highest intrinsic rate of increase compared with the other temperatures. Based on our data, the optimal range of temperature for L. paeta population growth was 30-35 degrees C. These data give us better understanding of L. paeta population dynamics, and they can be used to develop effective management strategies for this psocid.

Enzymes that possess glutathione S-transferase (GST) activity were purified to homogeneity by glutathione-agarose affinity chromatography from three field populations of Liposcelis paeta (Pearman). These populations were collected from Nanyang city of Henan Province (NY), Wuzhou (WZ) and Hezhou (HZ) cities of Guangxi Province, China, and had different susceptibilities to dichlorvos [LC(50)s of the NY (281.48 mg/m(2)), the WZ (285.07 mg/m(2)), and the HZ (243.52 mg/m(2)), respectively]. The specific activities of purified enzymes from these three populations increased 32.24-, 99.81-, and 42.52-fold, respectively. Kinetic analyses showed that the catalytic activity of purified GST from NY population towards GSH was much higher than the others, while WZ population reached the highest in V(max) (CDNB). SDS-polyacrylamide electrophoresis revealed that the purified GST had two subunits with a molecular mass of 23.31 and 20.43 kDa for NY, 53.14 and 20.13 kDa for WZ, and 50.79 and 19.42 kDa for HZ, respectively. The in vitro inhibition studies of GSTs indicated that three kinds of insecticides (chlorpyrifos, carbosulfan, and cypermethrin) and five metallic ions (Zn(2+), Ba(2+), Ca(2+), Hg(2+), Mn(2+), and Mg(2+)) all possessed inhibitory effects on purified GST, and ethacrynic acid (EA, a specific inhibitor of GST) expressed inhibitory effects. In the bioassay, three populations of L. paeta had different susceptibilities to different insecticides, even after they were reared on diets consisting of 25% EA. The GST activities of L. paeta from different areas also showed different temperature and pH stabilities. The differences in GST among the three populations may be attributed partially to the differences in control practices for psocids between Henan and Guangxi Provinces. (c) 2009 Wiley Periodicals, Inc.

Endosymbiotic bacteria that potentially influence reproduction and other fitness-related traits of their hosts are widespread in arthropods, and their appeal to researchers' interest is still increasing. In this study, the effects of removal of Cardinium infection on development, survival, and reproduction of Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae) were investigated in the laboratory. The Cardinium-free strain was obtained by the removal of Cardinium infection by using 1% rifampicin treatment on the Cardinium-infected strain (control) for 4 wk, and no Cardinium gene product was detected in this strain throughout the experiment. The results showed that the removal of Cardinium infection had negative effects on fitness of L. bostrychophila. Compared with the control strain, the Cardinium-free strain (both in first [F1] and second [F2] generation) had a similar developmental time, reduced survivorship of immature stages, as well as reduced fecundity, which resulted in much smaller r(m) values. Using r(m) values, the fitness for Cardinium-free F1 and F2 relative to the control was calculated as 0.81 and 0.74, respectively. We concluded that the use of antibiotics combined with heat treatment might be a good control measure for L. bostrychophila.