Ecological Interactions of Predatory Mites, Cheyletus eruditus (Schrank) (Trombidiformes: Cheyletidae) and Cheyletus malaccensis Oudemans, and Prey, Liposcelis decolor (Pearman) (Psocodea: Liposcelididae), under Different Thermo-Hygrometric Regimes.

Predator-prey interactions are linked through trophic relationships, and individual population dynamics are a function of multiple interactions among many ecological factors. The present study considered the efficacy of the predatory mites Cheyletus eruditus (Schrank) (Trombidiformes: Cheyletidae) and Cheyletus malaccensis Oudemans to manage Liposcelis decolor (Pearman) (Psocodea: Liposcelididae). Prey population suppression and progeny replacement efficiency of the predators were assessed under different predator-prey ratios (0:20, 1:20, 2:20, 4:20, and 10:20), temperatures (20, 24, 28, and 32 °C), and relative humidities (RH) (63, 75, and 85%) over 40 days under laboratory conditions of 0:24 (L:D) photoperiod. Suppression of L. decolor population when C. eruditus-related predator-to-prey ratios of 1:20, 2:20, 4:20, and 10:20 were used was ~61.7, 79.7, 85.1, and 87.5%, respectively, relative to the Control ratio (0:20). In the case of C. malaccensis, suppression of 70, 82.1, 92.9, and 96.5%, respectively, was achieved. Although the low 63% RH limited efficacy of these cheyletid mites, both predatory mites caused pest population suppression of ~67.1-97.2% and increased their progeny by ~96.7-844.4% fold for the predator-prey ratios of 1:20, 2:20, 4:20, and 10:20, temperatures of 20, 24, 28, and 32 °C, and RH levels of 63, 75, and 85%. The levels of psocid population suppression achieved indicate the potential of both predatory mites for psocid management.

New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a.

BACKGROUND: Khapra beetle (Trogoderma granarium Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for T. granarium based on recombinase polymerase amplification (RPA) and the CRISPR/Cas12a system. RESULTS: We designed and selected the first khapra beetle-specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a/CrRNA = 100 nM/500 nM). With only a 37 °C-heat-source and a blue light torch, RPA and CRISPR/CAS12a-based visualization assays can be completed within 40 min to differentiate between khapra beetle and nine similar Dermestidae species. After DNA extraction using a kit (4-5 h) or a simple method (5 min), the specific amplicons were obtained after a 15 min RPA reaction at 37 °C, followed by a 15 min color reaction under 37 °C in dark conditions using a CRISPR/CAS12a system and a fluorescent probe (5'-FAM/3'-BHQ1 labeled). This method is ingenious to low levels of DNA (10-1 ng µL-1 ) and meets the sensitivity requirements for detecting a single khapra beetle's egg (≈0.7 mm). CONCLUSION: Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37 °C), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management, and provides value as a reference for monitoring and identification of other pests. © 2023 Society of Chemical Industry.

An advanced approach for rapid visual identification of Liposcelis bostrychophila (Psocoptera: Liposcelididae) based on CRISPR/Cas12a combined with RPA.

Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae) is a booklouse pest that is a threat to commodity storage security worldwide. Accurate and sensitive methods of L. bostrychophila on-site identification are essential prerequisites for its effective management. Evidence suggests that L. bostrychophila contains 3 intraspecific biotypes that are morphologically indistinguishable but can be discriminated at the level of mitochondrial genome organization and sequences. The traditional molecular identification methods, such as DNA barcoding and PCR-RFLP, are instrumentally demanding and time-consuming, limiting the application of the identification in the field. Therefore, this study developed a new CRISPR/Cas12a-based visual nucleic acid system based on the mitochondrial gene coding for NADH dehydrogenase subunit 2 (nad2), combined with recombinase polymerase amplification (RPA) to accurately identify L. bostrychophila from 4 other common stored-product booklice, and also differentiate 3 biotypes of this species at the same time. The entire identification process could be completed at 37 °C within 20 min with high sensitivity. The system could stably detect at least 1 ng/µl of DNA template. The green fluorescence signal produced by the trans-cleaving of the single-stranded DNA reporter could be observed by the naked eye under blue light. Additionally, the suggested system combined with the crude DNA extraction method to extract DNA rapidly, enabled identification of all developmental stages of L. bostrychophila. With crude DNA, this novel diagnostic system successfully identified an unknown booklouse by holding the reaction tubes in the hand, thus can be considered as an accurate, rapid, highly sensitive, and instrument-flexible method for on-site visual identification of L. bostrychophila.

Numerical responses of the predatory mites, Cheyletus eruditus (Trombidiformes: Cheyletidae) and Cheyletus malaccensis, to Liposcelis decolor (Psocodea: Liposcelididae).

Predatory mites display diverse ecological mechanisms to suppress pest population density below certain thresholds known to cause economic loss. The current study explored the numerical responses of the predatory mites, Cheyletus eruditus (Schrank) (Trombidiformes: Cheyletidae) and Cheyletus malaccensis Oudemans, to Liposcelis decolor (Pearman) (Psocodea: Liposcelididae). The numerical responses of these 2 cheyletid mites to nymphs, adult males, and adult females of L. decolor were determined under laboratory conditions at 24 ± 1 °C, 85 ± 5 RH, and 0:24 (L:D) photoperiod. Oviposition rate, oviposition efficiency, and efficiency of conversion of ingested (ECI) food resources were the key numerical response parameters assessed. The present study revealed a general trend of a strong negative and positive correlation between oviposition rates and increase in prey densities (number of prey per 16.98 cm2) for C. eruditus and C. malaccensis, respectively. The oviposition efficiency was mostly similar for both predatory mites and was inversely related to prey density. Generally, ECI (%) decreased considerably with increasing prey density across different prey types for both predators, however, C. malaccensis was more efficient than C. eruditus in utilizing prey biomass. Given the relatively weak numerical responses, we recommended further assessment of these predatory mites before recommending their use for managing stored-product insect pests in the United States.

Evaluation of Hermetic Storage Bags for the Preservation of Yellow Maize in Poultry Farms in Dormaa Ahenkro, Ghana.

Using low-quality maize, resulting from insect pests and fungal attack, for formulating feed reduces chicken performance. This study evaluated the effectiveness of hermetic storage bags to keep insect pest and mycotoxin levels in check in yellow maize. The study was conducted in storehouses at three poultry farms in Dormaa Ahenkro, Bono Region, Ghana. The experiment was set up in a Randomized Complete Block Design with ZeroFly® Hermetic (ZFH), Purdue Improved Crop Storage (PICS), and Polypropylene (PP) bags as treatments. In each treatment, twelve 50 kg samples of untreated maize were each put in 100 kg capacity bags. Two bags in each treatment were destructively sampled monthly for 6 months. The number of insects was significantly higher in the PP bag (161.00 ± 4.25), compared to the PICS and ZFH bags: 7.00 ± 0.29 and 4.50 ± 0.76, respectively. The PICS and ZFH bags had less insect damage and lower weight loss than the PP bags. Aflatoxin and fumonisin levels were below the recommended safe thresholds of 15 ppb and 4 ppm, respectively, in all the bags. With the exception of ash, proximate analyses were higher for all variables in the PICS and ZFH bags. The study showed that PICS and ZFH bags conserved maize quality better than the PP bag.

A chromosome-level genome of the booklouse, Liposcelis brunnea, provides insight into louse evolution and environmental stress adaptation.

BACKGROUND: Booklice (psocids) in the genus Liposcelis (Psocoptera: Liposcelididae) are a group of important storage pests, found in libraries, grain storages, and food-processing facilities. Booklice are able to survive under heat treatment and typically possess high resistance to common fumigant insecticides, hence posing a threat to storage security worldwide. RESULTS: We assembled the genome of the booklouse, L. brunnea, the first genome reported in Psocoptera, using PacBio long-read sequencing, Illumina sequencing, and chromatin conformation capture (Hi-C) methods. After assembly, polishing, haplotype purging, and Hi-C scaffolding, we obtained 9 linkage groups (174.1 Mb in total) ranging from 12.1 Mb to 27.6 Mb (N50: 19.7 Mb), with the BUSCO completeness at 98.9%. In total, 15,543 genes were predicted by the Maker pipeline. Gene family analyses indicated the sensing-related gene families (OBP and OR) and the resistance-related gene families (ABC, EST, GST, UGT, and P450) expanded significantly in L. brunnea compared with those of their closest relatives (2 parasitic lice). Based on transcriptomic analysis, we found that the CYP4 subfamily from the P450 gene family functioned during phosphine fumigation; HSP genes, particularly those from the HSP70 subfamily, were upregulated significantly under high temperatures. CONCLUSIONS: We present a chromosome-level genome assembly of L. brunnea, the first genome reported for the order Psocoptera. Our analyses provide new insights into the gene family evolution of the louse clade and the transcriptomic responses of booklice to environmental stresses.

Fragmentation in mitochondrial genomes in relation to elevated sequence divergence and extreme rearrangements.

BACKGROUND: A single circular mitochondrial (mt) genome is a common feature across most metazoans. The mt-genome includes protein-coding genes involved in oxidative phosphorylation, as well as RNAs necessary for translation of mt-RNAs, whose order and number are highly conserved across animal clades, with few known exceptions of alternative mt-gene order or mt-genome architectures. One such exception consists of the fragmented mitochondrial genome, a type of genome architecture where mt-genes are split across two or more mt-chromosomes. However, the origins of mt-genome fragmentation and its effects on mt-genome evolution are unknown. Here, we investigate these origin and potential mechanisms underlying mt-genome fragmentation, focusing on a genus of booklice, Liposcelis, which exhibits elevated sequence divergence, frequent rearrangement of mt-gene order, and fragmentation of the mt genome, and compare them to other Metazoan clades. RESULTS: We found this genus Liposcelis exhibits very low conservation of mt-gene order across species, relative to other metazoans. Levels of gene order rearrangement were, however, unrelated to whether or not mt-genomes were fragmented or intact, suggesting mitochondrial genome fragmentation is not affecting mt-gene order directly. We further investigated possible mechanisms underpinning these patterns and revealed very high conservation of non-coding sequences at the edges of multiple recombination regions across populations of one particular Liposcelis species, supportive of a hypothesis that mt-fragmentation arises from recombination errors between mt-genome copies. We propose these errors may arise as a consequence of a heightened mutation rate in clades exhibiting mt-fragmentation. Consistent with this, we observed a striking pattern across three Metazoan phyla (Arthropoda, Nematoda, Cnidaria) characterised by members exhibiting high levels of mt-gene order rearrangement and cases of mt-fragmentation, whereby the mt-genomes of species more closely related to species with fragmented mt-genomes diverge more rapidly despite experiencing strong purifying selection. CONCLUSIONS: We showed that contrary to expectations, mt-genome fragmentation is not correlated with the increase in mt-genome rearrangements. Furthermore, we present evidence that fragmentation of the mt-genome may be part of a general relaxation of a natural selection on the mt-genome, thus providing new insights into the origins of mt-genome fragmentation and evolution.

Effects of Dehumidification on the Survivorship of Four Psocid Species.

Psocids are damaging stored-product pests. In this study, eggs and early-instar nymphs, adults, and all life stages of Liposcelis entomophila, L. decolor, L. bostrychophila, and L. paeta were subjected to 43, 50, or 75% (Control) relative humidity (RH) for 2, 4, 6, 8, 10, 12, 14, or 16 d at 30.0°C. All adults of these species died within 8 d at both 43 and 50% RH, except for L. bostrychophila, which required 12 d at 50% RH for 100% mortality to occur. For all life stages and eggs and early-instar nymphs, maximum survival times (times to 100% mortality) at 43 or 50% RH for L. entomophila, L. decolor, L. bostrychophila, and L. paeta, were 8 and 10 d, 8 and 12 d, 12 and 14 d, and 12 and 16 d, respectively. During this study, numbers of nymphs and adults of all species 14 d after the RH treatments increased within the 75% RH Control arenas. Different species and life stages responded differently to 43 and 50% RH, as time to kill all stages of the four psocid species was 8-12 and 10-16 d, respectively. Results indicate that using a specific RH environment may be effective in psocid management.

Categories of Resistance to Sugarcane Aphid (Hemiptera: Aphididae) Among Sorghum Genotypes.

The sugarcane aphid Melanaphis sacchari (Zehnter) (Hemiptera: Aphididae) has emerged as a potential threat to sorghum (Sorghum bicolor (L.) Moench) production in the United States. Since the late summer of 2013, finding and advancing M. sacchari-resistant germplasm has been a priority for all stakeholders involved. We evaluated 23 sorghum genotypes for resistance to the sugarcane aphid by testing for tolerance, and antixenosis. In addition, nine sorghum germplasm were evaluated for the expression of antibiosis. Free-choice and no-choice tests were conducted to explore the functional categories of resistance. Levels of resistance to M. sacchari were compared with the known resistant 'TX 2783' and the susceptible 'KS 585'. Sorghum entries AG1201, AG1301, W844-E, and DKS 37-07 were identified as expressing tolerance, antibiosis, and antixenosis, while H13073 expressed antibiosis and GW1489 expressed both tolerance and antibiosis. These resistant sorghums identified during this study will have a significant impact on reducing economic damage from the sugarcane aphid infestations.

The Highly Divergent Mitochondrial Genomes Indicate That the Booklouse, Liposcelis bostrychophila (Psocoptera: Liposcelididae) Is a Cryptic Species.

The booklouse, Liposcelis bostrychophila is an important storage pest worldwide. The mitochondrial (mt) genome of an asexual strain (Beibei, China) of the L. bostrychophila comprises two chromosomes; each chromosome contains approximate half of the 37 genes typically found in bilateral animals. The mt genomes of two sexual strains of L. bostrychophila, however, comprise five and seven chromosomes, respectively; each chromosome contains one to six genes. To understand mt genome evolution in L. bostrychophila, and whether L. bostrychophila is a cryptic species, we sequenced the mt genomes of six strains of asexual L. bostrychophila collected from different locations in China, Croatia, and the United States. The mt genomes of all six asexual strains of L. bostrychophila have two chromosomes. Phylogenetic analysis of mt genome sequences divided nine strains of L. bostrychophila into four groups. Each group has a distinct mt genome organization and substantial sequence divergence (48.7-87.4%) from other groups. Furthermore, the seven asexual strains of L. bostrychophila, including the published Beibei strain, are more closely related to two other species of booklice, L. paeta and L. sculptilimacula, than to the sexual strains of L. bostrychophila Our results revealed highly divergent mt genomes in the booklouse, L. bostrychophila, and indicate that L. bostrychophila is a cryptic species.

Molecular Identification of ten species of stored-product psocids through microarray method based on ITS2 rDNA.

Stored-product psocids (Psocoptera: Liposcelididae) are cosmopolitan storage pests that can damage stored products and cause serious economic loss. However, because of the body size (~1 mm) of eggs, nymphs, and adults, morphological identification of most stored-product psocids is difficult and hampers effective identification. In this study, 10 economically important stored-product Liposcelis spp. psocids (Liposcelis brunnea, L. entomophila, L. decolor, L. pearmani, L. rufa, L.mendax, L. bostrychophila, L. corrodens, L. paeta, and L. tricolor) were collected from 25 geographic locations in 3 countries (China, Czech Republic, and the United States). Ten species-specific probes for identifying these 10 psocid species were designed based on ITS2 sequences. The microarray method and reaction system were optimized. Specificity of each of the ten probes was tested, and all probes were found suitable for use in identification of the respective10 Liposcelis spp. psocids at 66 °C. This method was also used to identify an unknown psocid species collected in Taian, China. This work has contributed to the development of a molecular identification method for stored-product psocids, and can provide technical support not only to facilitate identification of intercepted samples in relation to plant quarantine, but also for use in insect pest monitoring.

DNA barcoding, species-specific PCR and real-time PCR techniques for the identification of six Tribolium pests of stored products.

Flour beetles of the genus Tribolium Macleay (Coleoptera: Tenebrionidae) are important stored product pests in China and worldwide. They are often found or are intercepted in grain depots, flour mills, and entry-exit ports, etc. Traditionally, Tribolium species are identified according to the morphological characteristics of the adult. However, it is almost impossible to rapidly identify adult fragments and non-adult stages based on external morphological characteristics. Molecular techniques for the rapid and accurate identification of Tribolium species are required, particularly for pest monitoring and the quarantine of stored products pests. Here, we establish DNA barcoding, species-specific PCR, and real-time PCR techniques for the identification of six stored-product pest Tribolium species including T. castaneum, T. confusum, T. destructor, T. madens, T. freemani and T. brevicornis. We detected the mitochondrial DNA cytochrome oxidase subunit I (COI) barcodes for Tribolium from 18 geographic populations and 101 individuals, built a Tribolium DNA barcode library, and designed species-specific primers and TaqMan probes for the above six Tribolium species. The three techniques were applied to identify Tribolium collected from stored samples and samples captured from quarantine ports. The results demonstrated that three techniques were all able to identify the six species of Tribolium both rapidly and accurately.

The establishment of species-specific primers for the molecular identification of ten stored-product psocids based on ITS2 rDNA.

Psocids are important stored product pests found worldwide that can be spread through grain trade. Most stored-product psocids, including eggs, nymphs, and adults, are very small (~1 mm) and difficult to identify morphologically. Here, we collected 10 economically important stored-product Liposcelis spp. psocids (L. bostrychophila, L. entomophila, L. decolor, L. paeta, L. brunnea, L. corrodens, L. mendax, L. rufa, L. pearmani, and L. tricolor) from 35 geographical locations in 5 countries (China, Czech Republic, Denmark, Germany, and the United States). The ITS2 rDNA gene was extracted and sequenced. The interspecific genetic distance of the stored-product psocids was significantly higher than the intraspecific genetic distance according to the barcoding gap analysis. Ten pairs of species-specific primers based on the ITS2 rDNA were developed for psocid identification. The sensitivity estimation indicated that the species-specific primers could correctly amplify the target ITS2 gene and successfully identify psocids at 1.0 ng/mL. Additionally, these species-specific primers could quantify specificity and identify 10 stored-product psocids; this approach could also be used to accurately identify other stored-product psocids. This work provides a practical approach for the precise examination of 10 stored-product psocid species and also contributes to the development of an identification method using ITS2 rDNA.

Effectiveness of Sulfuryl Fluoride Fumigation for the Control of Phosphine-Resistant Grain Insects Infesting Stored Wheat.

A field experiment was conducted in eight 13.6-MT steel bins containing 6.8 MT each of wheat to assess efficacy of sulfuryl fluoride or SF fumigant to control phosphine-resistant and susceptible Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst). Approximately 400 adults of each type of beetle were added to each bin. Additionally, muslin bags containing immature stages and adults, with their respective diets, were also placed in bins. Four bins were fumigated with SF and others were untreated control bins. The SF dosages in treated bins ranged from 1,196­1,467 mg-h/liter. Mortality of adults in each bag was assessed 5 d postfumigation; diet minus adults was incubated in a jar, and number of adults counted after 8 wk. No significant change occurred in number of insect-damaged kernels in SF-treated bins. In trier samples from SF-treated bins, R. dominica numbers declined from 24 prefumigation to 0 at 3- and 6-wk postfumigation; T. castaneum numbers were unchanged. In WBII traps from SF-treated bins, numbers R. dominica and T. castaneum declined from 25 and 33, respectively, prefumigation to 0 or near 0 at 3- and 6-wk postfumigation. Mortalities of resistant and susceptible adult R. dominica, and adult and large larvae of T. castaneum in SF-treated bags was 100%. For all four types of beetles, adult numbers in jars associated with SF-treated bins were 0 or near 0. Results show SF is effective against all life stages of phosphine-resistant R. dominica and T. castaneum, and can be used for phosphine resistance management.

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.

Array of Synthetic Oligonucleotides to Generate Unique Multi-Target Artificial Positive Controls and Molecular Probe-Based Discrimination of Liposcelis Species.

Several species of the genus Liposcelis are common insect pests that cause serious qualitative and quantitative losses to various stored grains and processed grain products. They also can contaminate foods, transmit pathogenic microorganisms and cause allergies in humans. The common occurrence of multi-species infestations and the fact that it is difficult to identify and discriminate Liposcelis spp. make accurate, rapid detection and discriminatory tools absolutely necessary for confirmation of their identity. In this study, PCR primers and probes specific to different Liposcelis spp. were designed based on nucleotide sequences of the cytochrome oxidase 1 (CO1) gene. Primer sets ObsCo13F/13R, PeaCo15F/14R, BosCO7F/7R, BruCo5F/5R, and DecCo11F/11R were used to specifically detect Liposcelis obscura Broadhead, Liposcelis pearmani Lienhard, Liposcelis bostrychophila Badonnel, Liposcelis brunnea Motschulsky and Liposcelis decolor (Pearman) in multiplex endpoint PCRs, which amplified products of 438-, 351-, 191-, 140-, and 87-bp, respectively. In multiplex TaqMan qPCR assays, orange, yellow, red, crimson and green channels corresponding to reporter dyes 6-ROXN, HEX, Cy5, Quasar705 and 6-FAM specifically detected L. obscura, L. brunnea, L. bostrychophila, L. pearmani and L. decolor, respectively. All developed primer and probe sets allowed specific amplification of corresponding targeted Liposcelis species. The development of multiplex endpoint PCR and multiplex TaqMan qPCR will greatly facilitate psocid identification and their management. The use of APCs will streamline and standardize PCR assays. APC will also provide the opportunity to have all positive controls in a single tube, which reduces maintenance cost and labor, but increases the accuracy and reliability of the assays. These novel methods from our study will have applications in pest management, biosecurity, quarantine, food safety, and routine diagnostics.

Morphological and molecular characterization of a sexually reproducing colony of the booklouse Liposcelis bostrychophila (Psocodea: Liposcelididae) found in Arizona.

The booklouse, Liposcelis bostrychophila, is a worldwide pest of stored products. For decades, only thelytokous parthenogenetic reproduction was documented in L. bostrychophila. Male L. bostrychophila were first found in Hawaii in 2002. In 2009, a sexual strain was found in Arizona. We examined the morphology of both males and females of the Arizona strain and compared the Arizona sexual strain with the Hawaii sexual strain and the parthenogenetic strains of L. bostrychophila. The sexual and parthenogenetic strains show some differences in eye morphology. To examine the relationship between sexual and asexual lineages, we sequenced the mitochondrial 12S and 16S ribosomal RNA genes of males and females from the Arizona strain. Phylogenetic analyses of L. bostrychophila individuals revealed that: 1) the sexually reproducing colony found in Arizona contains two closely related mitochondrial DNA haplotypes--one present in only females and the other in both males and females; and 2) the Arizona sexual strain was most closely related to a parthenogenetic strain in Illinois. We detected Rickettsia in all of the parthenogenetic individuals we checked but not in any Arizona sexual individuals. Further evidence is required to establish whether the presence of Rickettsia is linked to asexual reproduction in Liposcelis.

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.

Maternal transmission, sex ratio distortion, and mitochondria.

In virtually all multicellular eukaryotes, mitochondria are transmitted exclusively through one parent, usually the mother. In this short review, we discuss some of the major consequences of uniparental transmission of mitochondria, including deleterious effects in males and selection for increased transmission through females. Many of these consequences, particularly sex ratio distortion, have well-studied parallels in other maternally transmitted genetic elements, such as bacterial endosymbionts of arthropods. We also discuss the consequences of linkage between mitochondria and other maternally transmitted genetic elements, including the role of cytonuclear incompatibilities in maintaining polymorphism. Finally, as a case study, we discuss a recently discovered maternally transmitted sex ratio distortion in an insect that is associated with extraordinarily divergent mitochondria.

Efficacies of spinosad and a combination of chlorpyrifos-methyl and deltamethrin against phosphine-resistant Rhyzopertha dominica (Coleoptera: Bostrichidae) and Tribolium castaneum (Coleoptera: Tenebrionidae) on wheat.

Highly phosphine-resistant populations of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) have recently been found in Oklahoma grain storage facilities. These findings necessitate development of a phosphine resistance management strategy to ensure continued effective use of phosphine. Therefore, we investigated the efficacies of two grain insecticides, namely, spinosad applied at label rate of 1 ppm and a mixture of chlorpyrifos-methyl and deltamethrin applied at label rates of 3 and 0.5 ppm, respectively, against highly phosphine-resistant R. dominica and T. castaneum. Adult mortality and progeny production suppression of spinosad- or chlorpyrifos-methyl + deltamethrin mixture-treated wheat that had been stored for 2, 84, 168, 252, and 336 d posttreatment were assessed. We found that both spinosad and chlorpyrifos-methyl + deltamethrin were effective against phosphine-resistant R. dominica and caused 83-100% mortality and also caused total progeny production suppression for all storage periods. Spinosad was not effective against phosphine-resistant T. castaneum; the highest mortality observed was only 3% for all the storage periods. Chlorpyrifos-methyl + deltamethrin was effective against phosphine-resistant T. castaneum only in treated wheat stored for 2 and 84 d, where it caused 93-99% mortality. However, chlorpyrifos-methyl + deltamethrin was effective and achieved total suppression of progeny production in T. castaneum for all the storage periods. Spinosad was not as effective as chlorpyrifos-methyl + deltamethrin mixture at suppressing progeny production of phosphine-resistant T. castaneum. These two insecticides can be used in a phosphine resistance management strategy for R. dominica and T. castaneum in the United States.