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A single degenerated primer significantly improves COX1 barcoding performance in soil nematode community profiling

Soil Ecology Letters 2024年第2期6卷 31-44页

作者： Yincai Ren Dorota L. Porazinska Quanhui Ma Shuhan Liu Hongmei Li Xue Qing Department of Plant Pathology Nanjing Agricultural UniversityNanjing210095China Department of entomology and nematology University of FloridaGainesvilleFL32611USA Institute of Grassland Science Key Laboratory of Vegetation Ecology of the Ministry of EducationJilin Songnen Grassland Ecosystem National Observation and Research StationNortheast Normal UniversityChangchun130024China

A new COX1 primer for soil nematode metabarcoding was designed,and this primer outperforms other commonly used COX1 primer pairs in species recovery and quantity of PCR products.•The lack of reference database is the main reason that led to the low species recovery in COX1 metabarcoding.•We expanded current NCBI database by adding 51 newly generated COX1 reference *** nematodes play important roles in soil ecosystems and often serve as bioindicators of soil *** identification of soil nematodes is often difficult due to their limited diagnostic characters and high phenotypic *** barcoding and metabarcoding techniques are promising but lack universal primers,especially for mitochondrial COX1 *** this study a degenerated COX1 forward primer COIFGED was *** primer pair(COIFGED/JB5GED)outperforms other four commonly used COX1 primer pairs in species recovery and quantity of polymerase chain reaction(PCR)*** metabarcoding analysis,the reads obtained from the new primer pair had the highest sequencing saturation threshold and amplicon sequence variant(ASV)diversity in comparison to other COX1 as well as 18S rRNA *** annotation of ASVs suggested the new primer pair initially recovered 9 and 6 out of 25 genera from mock communities,respectively,outperformed other COX1 primers,but underperformed the widely used 18S NF1/18Sr2b primers(16 out of 25 genera).By supplementing the COX1 database with our reference sequences,we recovered an additional 6 mock community species bringing the tally closer to that obtained with 18S *** summary,our newly designed COX1 primers significantly improved species recovery and thus can be supplementary or alternative to the conventional 18S metabarcoding.

关键词： degenerated primers DNA metabarcoding mitochondrial cytochrome oxidase c subunit I gene phylogeny ribosomal RNA gene soil nematodes

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The effectiveness of eyespots and masquerade in protecting artificial prey across ontogenetic and seasonal shifts

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Current Zoology 2022年第4期68卷 451-458页

作者： Elizabeth G Postema Department of entomology and nematology University of CaliforniaDavisCA 95616USA

When constraints on antipredator coloration shift over the course of development,it can be advantageous for animals to adopt different color strategies for each life *** caterpillars in the genus Papilio exhibit unique ontogenetic color sequences:for example,early instars that masquerade as bird feces,with later instars possessing eyespots.I hypothesize that larvae abandon feces masquerade in favor of eyespots due to ontogenetic changes in signaler *** ontogenetic pattern also occurs within broader seasonal shifts in background color and predator activity.I conducted predation experiments with artificial prey to determine how potential signaling constraints(specifically size and season)shape predation risk,and consequently the expression of ontogenetic color change in Papilio ***,both predation and background greenness declined significantly from July to September,though there was little evidence that these patterns impacted the effectiveness of either color *** size and color strongly affected the attack rate of avian predators:attacks increased with prey size regardless of color,and eyespotted prey were attacked more than masquerading prey *** results may reflect a key size-mediated tradeoff between conspicuousness and intimidation in eyespotted prey,and raise questions about how interwoven aspects of behavior and signal environment might maintain the prevalence of large,eyespotted larvae in nature.

关键词： ontogenetic color change eyespots masquerade predator-prey dynamics season

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Cold acclimation increases Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) survival during exposure to freezing temperatures

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Insect Science 2022年第2期29卷 531-538页

作者： Xavier Martini Kathi Malfa Dara Stockton Monique JRivera Department of entomology and nematology North Florida Research and Education CenterQuincyFloridaUSA USDA-ARS U.S.Pacific Basin Agricultural Research CenterHiloHawaiiUSA Department of entomology University of CaliforniaRiversideCaliforniaUSA

The Asian citrus psyllid,Diaphorina citri,is the vector of the pathogenic bacteria that causes Huanglongbing,the most devasting disease affecting citrus *** cultivation of citrus tends to expand northward,the tolerance of psyllids to freezing temperatures needs to be *** mortality of *** to cold temperature has been previously studied,much less is known regarding the acclimation potential of psyllids to cold *** first evaluated cold resistance of *** depending on color morph and ***,we compared mortality of *** at−4°C for 10 hours between unacclimated psyllids,psyllids that were gradually acclimated to cold over 1 or 2 weeks,and psyllids intermittently acclimated also for 1 or 2 *** did not find major differences in cold resistance between color morphs and *** found that after 1 week of gradual acclimation,the survival of psyllids at−4°C increased to 86%,compared to 20%in the control *** did not increase after the second week of gradual *** intermittent acclimation improved survival compared to the control group,it was less effective than gradual acclimation with a survival at 30%and 70%after 1 and 2 weeks of acclimation,respectively,although this difference with gradual acclimation was not significant at week *** data show that gradual cold acclimation allows *** to survive brief periods below *** may serve as a mechanism responsible for increasing northern establishment of this pest.

关键词： cold acclimation cold hardy citrus cold resistance citrus greening Diaphorina citri overwintering

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Anopheles gambiae strain(Ag55)cultured cells originated from Anopheles coluzzii and are phagocytic with hemocyte-like gene expression

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Insect Science 2022年第5期29卷 1346-1360页

作者： Ruchir Mishra Gang Hua Ujwal R.Bagal Donald E.Champagne Michael J.Adang Department of entomology College of Agricultural and Environmental SciencesUniversity of GeorgiaAthensGeorgiaUSA Department of entomology and nematology University of FloridaGainesvilleFloridaUSA Institute of Bioinformatics University of GeorgiaAthensGeorgiaUSA Department of Biochemistry and Molecular Biology University of GeorgiaAthensGeorgiaUSA

Anopheles gambiae and Anopheles coluzzii are closely related species that are predominant vectors of malaria in ***,*** form M was renamed *** and we now conclude on the basis of a diagnostic PCR-restriction fragment length polymorphism assay that Ag55 cells were derived from *** established an Ag55 cell transcriptome,and KEGG pathway analysis showed that Ag55 cells are enriched in phagosome pathway *** Ag55 transcriptome has an abundance of specific transcripts characteristic of mosquito *** *** bioparticle uptake experiments visualized by fluorescence microscopy and confocal microscopy and quantified by flow cytometry establish the phagocytic competence of Ag55 *** from this investigation of Ag55 cell properties will guide researchers in the use and engineering of the Ag55 cell line to better enable investigations of Plasmodium,other microbes,and insecticidal toxins.

关键词： Anopheles coluzzii transcriptome Anopheles gambiae confocal microscopy flow-cytometry FPKM hemocyte-like phagocytosis

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Potential of termite-based biomass pre-treatment strategies for use in bioethanol production

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Insect Science 2010年第3期17卷 166-174页

作者： Michael E. Scharf Drion G. Boucias entomology and nematology Department University of Florida Gainesville Florida USA

When considering the current state of the biorefinery industry, it is readily apparent that industrial cellulose and hemicellulose digestion processes are relatively advanced, whereas enzymatic pre-treatment strategies for biomass delignification and cellulose solubilization are not well developed. The need for efficient biomass pre-treatment strategies presents a significant opportunity for researchers studying lignocellulose digestion in termites and other insects. With an emphasis on industrial biomass pre-treatment, this review provides an overview of：（i） industrial biorefining operations （feedstocks, processing, and economics）; （ii） recent findings from termite research that have revealed candidate enzymes; and （iii） research needs and opportunities for consideration by entomologists working in this area. With respect to research findings, recently identified candidate lignases （laccases, catalases, peroxidases, esterases）, other potentially important detoxification enzymes （cytochrome P450, superoxide dismutase）, and phenolic acid esterases （carboxylesterases） that may assist in hemicellulose solubilization are overviewed. Regarding research needs and opportunities, several approaches for identification of candidate pre-treatment enzymes from upstream, symbiont-free gut regions are also described.

关键词： bioenergy bioethanol biorefining Isoptera lignocellulose renewable energy

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Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA

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Journal of Pharmaceutical Analysis 2023年第9期13卷 1013-1023页

作者： Huiyi Yang Meng Qi Qiyi He Sung Hee Hwang Jun Yang Mark McCoy Christophe Morisseau Suqing Zhao Bruce D.Hammock Department of Pharmaceutical Engineering School of Biomedical and Pharmaceutical SciencesGuangdong University of TechnologyGuangzhouChina Department of entomology and nematology and UCD Comprehensive Cancer Center University of CaliforniaDavisCA 95616USA Langfang Normal University LangfangHebei065000China

To ensure proper dosage of a drug,analytical quantification of it in biofluid is *** chromatography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and ***,it requires expensive instrumentation and is not appropriate for bedside *** soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human *** optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and *** assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate *** addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory *** results indicate that nanobody-based ELISA methods can efficiently analyze drug like *** methods could be easily implemented by the bedside,in the field in remote areas or in veterinary *** work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and *** of nanobody based pharmaceutical assays are discussed.

关键词： Nanobody Immunoassay Soluble epoxide hydrolase inhibitors Metabolites Small molecules

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Transcytosis of Junonia coenia densovirus VP4 across the gut epithelium of Spodoptera frugiperda(Lepidoptera:Noctuidae)

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Insect Science 2020年第1期27卷 22-32页

作者： Mariah Kemmerer Bryony C.Bonning Department of entomology Iowa State UniversityAmesIowaUSA Department of entomology and nematology University of FloridaGainesvilleFloridaUSA

The Junonia coenia densovirus rapidly traverses the gut epithelium of the host lepidopteran without replicating in the gut *** ability of this virus to transcytose across the gut epithelium is of interest for the potential use of virus structural proteins as delivery vehicles for insecticidal peptides that act within the insect hemocoel,rather than in the *** this study,we used fall armyworm,Spodoptera frugiperda to examine the binding of the virus to brush border membrane vesicle proteins by two-dimensional ligand blot *** also assessed the rate of flux of the primary viral structural protein,VP4 fused to eGFP with a proline-rich linker(VP4-P-eGFP)through the gut epithelium ex vivo in an Ussing *** mechanisms involved with transcytosis of VP4-P-eGFP were assessed by use of *** serum albumin(BSA)and eGFP were used as positive and negative control proteins,*** contrast to BSA,which binds to multiple proteins on the brush border membrane,VP4-P-eGFP binding was specific to a protein of high molecular *** flux was significantly higher for VP4-P-eGFP after 2 h than for albumin or eGFP,with rapid transcytosis of VP4-P-eGFP within the first 30 *** contrast to BSA which transcytosed following clathrin-mediated endocytosis,the movement of VP4-P-eGFP was vesicle-mediated but *** specificity of binding combined with the efficiency of transport across the gut epithelium suggest that VP4 will provide a useful carrier for insecticidal peptides active within the hemocoel of key lepidopteran pests including ***.

关键词： BSA gut epithelium JcDNV Lepidoptera transcytosis virus

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Characterization of the voltage-gated sodium channel of the Asian citrus psyllid, Diaphorina citri

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Insect Science 2017年第1期24卷 47-59页

作者： Bin Liu Monique R. Coy Jin-Jun Wang Lukasz L. Stelinski entomology and nematology Department Citrus Research and Education Center University of Florida Florida USA Key Laboratory of entomology and Pest Control Engineering College of Plant Protection Southwest University Chongqing China

The Asian citrus psyllid, Diaphorina citri Kuwayama （Hemiptera： Liviidae）, is an important insect pest of citrus. It is the vector of ＇Candidatus＇ Liberibacter asiaticus, a phloem-limited bacterium that infects citrus, resulting in the disease Huanglongbing （HLB）. Disease management relies heavily on suppression of D. cirri populations with insecticides, including pyrethroids. In recent annual surveys to monitor insecticide resistance, reduced susceptibility to fenpropathrin was identified in several field populations olD. citri. The primary target ofpyrethroids is the voltage-gated sodium channel （VGSC）. The VGSC is prone to target-site insensitivity because of mutations that either reduce pyrethroid binding and/or alter gating kinetics. These mutations, known as knockdown resistance or kdr, have been reported in a wide diversity of arthropod species. Alternative splicing, in combination with kdr mutations, has been also associated with reduced pyrethroid efficacy. Here we report the molecular characterization of the VGSC in D. citri along with a survey of alternative splicing across developmental stages of this species. Previous studies demonstrated that D. citri has an exquisite enzymatic arsenal to detoxify insecticides resulting in reduced efficacy. The results from the current investigation demonstrate that target-site insensitivity is also a potential basis for insecticide resistance to pyrethroids in D. citri. The VGSC sequence and its molecular characterization should facilitate early elucidation of the underlying cause of an established case of resistance to pyrethroids. This is the first characterization of a VGSC from a hemipteran to this level of detail, with the majority of the previous studies on dipterans and lepidopterans.

关键词： citrusgreening diseasemanagement knockdownresistance(kdr) target-site insensitivity

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Slow-growth high-mortality： A meta-analysis for insects

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Insect Science 2018年第2期25卷 337-351页

作者： Ke-Wei Chen1 and Yigen Chen2 College of Agriculture South China Agricultural University Guangzhou China Department of entomology and nematology University of California One Shields Avenue Davis California USA

The slow growth-high mortality hypothesis （SG-HG） predicts that slower growing herbivores suffer greater mortality due to a prolonged window of vulnerability. Given diverse plant-herbivore-natural enemy systems resulting from different feeding ecologies of herbivores and natural enemies, this hypothesis might not always be applicable to all systems. This is evidenced by mixed support from empirical data. In this study, a meta- analysis of the SG-HM hypothesis for insects was conducted, aiming to find conditions that favor or reject SG-HM. The analysis revealed significant within- and between-group heterogeneity for almost a11 explanatory variables and overall did not support SG-HM. In this analysis, SG-HM was supported when any of the following 5 conditions was met：（1） host food consisted of artificial diet; （2） herbivore growth was measured as larval mass; （3） herbivores were generalists; （4） no or multiple species of natural enemies were involved in the study; and （5） parasitoids （i.e., parasitic insects） involved in the study were gregarious. SG-HM was rejected when any of the following 5 conditions was met：（1） herbivores were from the order Hymentoptera; （2） parasitoids from more than 1 order caused herbivore mortality; （2） parasitoids were specialists; （3） parasitoids were solitary; （4） parasitoids were idiobionts or koinobionts; and （5） single species of natural enemy caused mortality of specialist herbivores. All known studies investigated herbivore mortality for a short period of their life cycle. Researchers are encouraged to monitor herbivore mortality during the entire window of susceptibility or life cycle using life tables. Studies involving multiple mortality factors （i.e., both biotic and abiotic） or multiple natural enemy species are also encouraged since herbivores in nature face a multitude of risks during the entire life cycle. More comprehensive studies may increase our understanding of factors influencing the relationships between herbivore gr

关键词： feeding ecology natural enemy parasitoid predator tritrophic interactions

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Systemic RNAi in western corn rootworm, Diabrotica virgifera virgifera, does not involve transitive pathways

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Insect Science 2018年第1期25卷 45-56页

作者： Huarong Li Andrew J. Bowling Premchand Gandra Murugesan Rangasamy Heather E. Pence Robert E. McEwan Chitvan Khajuria Blair D. Siegfried Kenneth E. Narva Dow AgroSciences LLC 9330 Zionsville Rd. Indianapolis Indiana USA Department of entomology University of Nebraska Lincoln Nebraska USA Monsanto Company Chesterfield Missouri USA entomology and nematology Department University of Florida Gainesville Florida USA

Western corn rootworm （WCR, Diabrotica virgifera virgifera LeConte） is highly sensitive to orally delivered double-stranded RNA （dsRNA）. RNAi in WCR is systemic and spreads throughout the insect body. This raises the question whether transitive RNAi is a mechanism that functions in WCR to amplify the RNAi response via production of secondary siRNA. Secondary siRNA production is achieved through RNA-dependent RNA polymerase （RdRP） activity in other eukaryotic organisms, but RdRP has not been identified in WCR and any other insects. This study visualized the spread of the RNAi- mediated knockdown of Dr v-ATPase C mRNA throughout the WCR gut and other tissues using high-sensitivity branched DNA in situ hybridization. Furthermore, we did not detect either secondary siRNA production or transitive RNAi in WCR through siRNA sequence profile analysis. Nucleotide mismatched sequences introduced into either the sense or antisense strand of v-ATPase C dsRNAs were maintained in siRNAs derived from WCR fed with the mismatched dsRNAs in a strand specific manner. The distribution of all siRNAs was restricted to within the original target sequence regions, which may indicate the lack of new dsRNA synthesis leading to production of secondary siRNA. Thus, the systemic spread of RNAi in WCR may be derived from the original dsRNA molecules taken up from the gut lumen. These results indicate that the initial dsRNA dose is important for a lethal systemic RNAi response in WCR and have implications in developing effective dsRNA traits to control WCR and in resistance management to prolong the durability of RNAi trait technology.

关键词： corn rootworm Diabrotica secondary siRNA production systemic RNAi transitive RNAi

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