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Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

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Insect Science 2019年第6期26卷 964-972页

作者： San-Yuan Ma guy smagghe Qing-You Xia State Key Laboratory of Silkworm Genome Biology Southwest University ChongqingChina Chongqing Engineering and Technology Research Center for Novel Silk Materials Southwest University Chongqing China Department of Crop Protection Faculty of Bioscience Engineering Ghent University Ghent Belgium

In recent years, research in life sciences has been remarkably revolutionized owing to the establishment, development and application of genome editing technologies. Genome editing has not only accelerated fundamental research but has also shown promising applications in agricultural breeding and therapy. In particular, the clustered, regularly interspaced, short palindromic repeat (CRISPR) technology has become an indispensable tool in molecular biology owing to its high efficacy and simplicity. Genome editing tools have also been established in silkworm (Bombyx mori), a model organism of Lepidoptera insects with high economic importance. This has remarkably improved the level and scope of silkworm research and could reveal new mechanisms or targets in basic entomology and pest management studies. In this review, we summarize the progress and potential of genome editing in silkworm and its applications in functional genomic studies for generating novel genetic materials.

关键词： CRISPR genome editing silkworm TALEN ZFN

Discovery of a widespread presence bunyavirus that may have symbiont-like relationships with different species of aphids

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Insect Science 2022年第4期29卷 1120-1134页

作者： Xin An Wei Zhang Chao Ye guy smagghe Jin-Jun Wang Jinzhi Niu Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina International Joint Laboratory of China–Belgium on Sustainable Crop Pest Control Academy of Agricultural SciencesSouthwest UniversityChongqingChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Aphids are important agricultural pests,vectors of many plant viruses and have sophisticated relationships with symbiotic *** asymptomatic RNA viruses have been reported in aphids due to the application of RNA-seq,but aphid-virus interactions remain *** is the most abundant RNA virus order,which can infect mammals,arthropods,and ***,many bunyaviruses have specific hosts,such as ***,we discovered 18 viruses from 10 aphid species by ***,a widespread presence bunyavirus,Aphid bunyavirus 1(ABV-1),was determined to have a wide host range,infecting and replicating in all 10 tested aphid ***-1 may be transmitted horizontally during feeding on plant leaves and vertically through *** a comparison of the physiological parameters of ABV-1high and ABV-1low strains of pea aphid,higher ABV-1 titers reduced the total nymphal duration and induced the ***,viral titer significantly affected the lipid and protein contents in pea *** summary,we proposed that ABV-1 may have stable symbiont-like relationships with aphids,and these observations may provide a new direction for studying bunyaviruses in aphids and establishing a model for virus-aphid interactions.

关键词： insec-specific virus pea aphid Phasmaviridae RNA virus RNAi

Comparative effects of insecticides with different mechanisms of action on Chrysoperla externa （Neuroptera＂ Chrysopidae）＂ Lethal, sublethal and dose-response effects

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Insect Science 2013年第6期20卷 743-752页

作者： Moises Joao Zotti Anderson Dionel Grutzmacher Isac Heres Lopes guy smagghe UGent-Crop Protection Ghent Oost-Vlaanderen Belgium UFPel - Phytosanitary Pelotas RS Brazil

The comprehensive knowledge that the delayed systemic and reproduction side effects can be even more deleterious than acute toxicity, has caused a shift in focus toward sublethal effects assessment on physiology and behavior of beneficial insects. In this study, we assessed the risks posed by some insecticides with different mode of action through lethal and delayed systemic sublethal effects on the pupation, adult emergence, and repro- duction of the chrysopid Chrysoperla externa （Hagen, 1861; Neuroptera： Chrysopidae）, an important predator in pest biological control. The maximum field recommended dose （MFRD） and twice （2xMFRD） for chlorantraniliprole, tebufenozide, and pyriproxyfen were harmless to C. externa. In contrast, all the tested chitin synthesis inhibitors （CSIs） were highly detrimental to the predator, despite of their lack of acute lethal toxicity. There- fore, the safety assumed by using IGRs toward beneficial insects is not valid for chrysopids. Dose-response data showed that although all CSIs have a similar mechanism of action, the relative extent of toxicity may differ （novaluron 〉 lufenuron 〉 teflubenzuron）. For CSIs, the delayed systemic effects became obvious at adult emergence, where the predicted no observable effect dose （NOED） was 1/2 048 of the MFRD for novaluron （0.085 ng/insect）, and 1/256 of the MFRD for both lufenuron （0.25 ng/insect） and teflubenzuron （0.6 ng/insect）. Finally, this work emphasized the significance of performing toxicity risk assessments with an adequate posttreatment period to avoid underestimating the toxicities of insecticides, as the acute lethal toxicity assays may not provide accurate information regarding the long-range effects of hazardous compounds.

关键词： Chrysoperla externa dose-response effects ecotoxicology IGRs insecti-cide risk assessment topical application

Ferritin acts as a target site for the snowdrop lectin （GNA） in the midgut of the cotton leafworm Spodoptera littoralis

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Insect Science 2008年第6期15卷 513-519页

作者： Amin Sadeghi guy smagghe Paul Proost Els J.M. Van Damme Laboratory of Agrozoology Department of Crop Protection Laboratory of Biochemistry and Glycobiology Department of Molecular Biotechnology Faculty of Bioscience Engineering Ghent University Ghent Rega Institute Laboratory of Molecular Immunology Kathofieke Universiteit Leuven Minderbroedersstraat 10 B-3000 Leuven Belgium

The snowdrop lectin GNA （Galanthus nivalis agglutinin） has been shown to possess insecticidal activity to a range of economically important insect pests. However, the precise mechanism of insecticidal action of GNA against insects remains unknown. In this investigation, we attempted to purify and identify receptor（s） responsible for binding of GNA in the larval midgut of a major lepidopteran pest （the cotton leafworm, Spodoptera littoralis） to better understand its mode of action. Therefore, cytoplasmic as well as membrane proteins from 800 larval midguts were chromatographed on a column with immobilized GNA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the proteins eluted from the GNA column followed by sequencing of the GNA-binding proteins and BLAST analyses revealed that the N-terminal sequences of a 24 kDa polypeptide purified from the cytoplasmic and membrane protein fraction revealed sequence similarity to sequences encoding heavy chain homologs of ferritin from Manduca sexta （76% sequence identity）, Calpodes ethlius （80% sequence identity） and Bombyx mori （61% sequence identity）. Furthermore, the N-terminal sequence of a 31 kDa polypeptide from the membrane protein fraction showed sequence similarity to a light chain homolog of ferritin from Manduca sexta （88% sequence identity）.

关键词： ferritin Galanthus nivalis agglutinin GNA Lepidoptera midgut receptors Spodoptera littoralis

Identification and immune analysis of antimicrobial peptides from the cigarette beetle(Lasioderma serricorne)

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Insect Science 2024年第4期31卷 1121-1134页

作者： Yue Zhang Yi Yan guy smagghe Hong Yang Ren-huai Dai Wen-jia Yang Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region Institute of EntomologyGuizhou UniversityGuiyangChina Guizhou Provincial Key Laboratory for Rare Animal and Economic Insect of the Mountainous Region College of Biology and Environmental EngineeringGuiyang UniversityGuiyangChina

Antimicrobial peptides(AMPs)in insects are endogenous peptides that are effector components of the innate defense system of the *** may serve as antimicrobial agents because of their small molecular weight and broad-spectrum antimicrobial *** this study,we performed transcriptome analysis of cigarette beetle(Lasioderma serricorne)larvae,parasitized by the ectoparasitic wasp,Anisopteromalus *** AMP genes were significantly upregulated following *** parasitism,postulating the hypothesis that the parasitization enhanced the host's resistance against pathogenic microorganisms through the regulation of host AMP ***,3 AMP genes(LsDef1,LsDef2,and LsCole)were significantly upregulated and we studied their immune function in *** challenge and functional analysis showed that LsCole was responsible for the immune response against Gram-negative and Gram-positive bacteria,while LsDef1 and LsDef2 were involved in insect defense against Gram-positive *** recombinant LsCole exhibited antimicrobial activities against the Gram-negative bacterium Escherichia coli and the Gram-positive bacterium Staphylococcus ***2 showed an antibacterial effect against *** and LsDef2 exhibited antibiofilm activity against *** 2 AMPs disrupted cell membranes and caused leakage of *** cell *** results indicated that the 3 AMPs in *** are involved in the innate immunity of this pest *** AMPs may have potential as antimicrobial agents for bacterial infection ***,data are discussed in relation to new control strategies with greater biosafety against pest insects with use of microbial biocontrol agents in combination with RNA interference against the insect's defensive AMP genes.

关键词： antimicrobial activity antimicrobial peptides innate immune response Lasioderma serricorne RNA interference

Molting process revealed by the detailed expression profiles of RXR1/RXR2 and mining the associated genes in a spider mite, Panonychus citri

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Insect Science 2022年第2期29卷 430-442页

作者： Gang Li Xun-Yan Liu guy smagghe Jin-Zhi Niu Jin-Jun Wang Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionInternational Joint Laboratory of China-Belgium on Sustainable Crop Pest ControlAcademy of Agricultural SciencesSouthwest UniversityChongqingChina Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions Institute of EntomologyGuizhou UniversityGuiyangChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Spider mites have one ecdysone receptor(EcR)and multiple retinoid X receptors(RXRs).However,the function of these RXRs in spider mite development is ***,we screened the expression dynamics of two PcRXR isoforms at 4 h intervals in the deutonymphal stage of Panonychus *** results showed that PcEcR had an expression pattern similar to that of *** PcRXR1,its expression remained at a certain high level,when there was a decrease of both PcEcR and *** situ hybridization showed that PcRXR2 was detected in the central nervous mass,while the ecdysteroid biosynthesis gene PcSpo was mainly expressed at the edge of the central nervous ***-based silencing of PcRXR1 or PcRXR2 showed the same phenotype as in mites with that of silencing ***,RNA-seq was used to mine the genes associated with the expression dynamics of PcRXR1 or PcRXR2,which revealed that the heterodimer of EcR-RXR2 in spider mites might be linked with the cell autophagy and tissue remodeling during apolysis,and RXR1 might be linked with new epicuticle and exocuticle secretion during *** together,these results increase our understanding of the regulation mechanism of ecdysteroid signal pathway in spider mite development.

关键词： ecdysone receptor(EcR) in situ hybridization retinoid X receptors RNA interference RNA-seq

A different gut microbial community between larvae and adults of a wild bumblebee nest （Bombus pascuorum）

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Insect Science 2018年第1期25卷 66-74页

作者： Anneleen Parmentier Ivan Meeus Filip Van Nieuwerburgh Dieter Deforce Peter Vandamme guy smagghe Laboratory of Agrozoology Department of Crop Protection Faculty of Bioscience Engineering Ghent University Laboratory of Pharmaceutical Biotechnology Faculty of Pharmaceutical Sciences Ghent University Laboratory of Microbiology Department of Biochemistryand Microbiology Faculty of Sciences Ghent University Ghent Belgium

Although the gut microbial communities in adult bumblebees and their as- sociated functionalities are widely studied, descriptive data on the larval gut microbiota are still limited. The gut microbiota of a fully sampled wild Bombus pascuorum nest has been characterized, using the multiplexed Illumina MiSeq 16S ribosomal RNA ampli- con sequencing technique. The nesters and foragers inhabiting the same nest showed the typical core bacterial sequences and only marginal differences in their characterized gut microbiota. The gut microbial communities within the adult and larval specimens differed strongly, as the typical core gut bacteria in the adult bumblebees are absent in the larval bumblebees. The bacterial communities within the larval gut are dominated by bacterial phylotypes of Enterobacteriaceae and Lactobacillaceae, supplemented with genera be- longing to Corynebacteriales and Bacillales. The function of this larval gut microbiota, being different from the adult, remains to be determined.

关键词： bumblebee gut Illumina sequencing larvae microbiota nest sample size

Colony contact contributes to the diversity of gut bacteria in bumblebees （Bombus terrestris）

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Insect Science 2017年第2期24卷 270-277页

作者： Annelies Billiet Ivan Meeus Filip Van Nieuwerburgh Dieter Deforce Felix Wackers guy smagghe Laboratory of Agrozoology Department of Crop Protection Faculty of Bioscience Engineering Ghent University B-9000 Ghent Laboratory of Pharmaceutical Biotechnology Faculty of Pharmaceutical Sciences Ghent University B-9000 Ghent BiobestNV B-2260 Westerlo Belgium

Social bees, like honeybees and bumblebees, have a close contact with nest mates of different developmental stages and generations. This could enhance bacterial transfer between nest mates and offers opportunities for direct transfer of symbionts from one generation to the next, resulting in a stable host specific gut microbiota. Gut sym- bionts of honeybees and bumblebees have been suggested to contribute in digestion and protection against parasites and pathogens. Here we studied the impact of contact with the bumblebee colony on the colonization potential of the bacterial families （i.e., Neisseri- aceae, Orbaceae, Lactobacillaceae and Bifidobacteriaceae） occurring in the gut of adult bumblebees （Bombus terrestris）. Bacterial profiles of the gut microbiota of B. terrestris were determined based on the hypervariable V4 region of the 16S rRNA using paired-end Illumina sequencing. In our experiments, we created different groups in which we grad- ually reduced the contact with nest mates and hive material. We made 3 observations：（i） reducing the contact between the colony and the bumblebee during adult life resulted in a significant drop in the relative abundance ofLactobacillus bombicola and Lactobacillus bombi; （ii） Bifidobacteriaceae required contact with nest mates to colonize the gut of B. terrestris and a significant lower bacterial diversity was observed in bumblebees that were completely excluded from colony contact during the adult life; （iii） Snodgrassella and Gilliamella were able to colonize the gut of the adult bumblebee without any direct contact with nest mates in the adult life stage. These results indicate the impact of the colony life on the diversity of the characteristic bumblebee gut bacteria.

关键词： bacterial transfer bumblebee gut Illumina sequencing microbiota social

RNAi of the N-glycosylation-related genes confirms their importance in insect development and α-1,6-fucosyltransferase plays a role in the ecdysis event for the hemimetabolous pest insect Nilaparvata lugens

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Insect Science 2022年第1期29卷 91-99页

作者： Qun Yang Kristof De Schutter Pengyu Chen Els J.M.Van Damme guy smagghe Laboratory of Agrozoology Department of Plants and CropsFaculty of Bioscience EngineeringGhent UniversityCoupure links 653Ghent9000Belgium Laboratory of Biochemistry and Glycobiology Department of BiotechnologyFaculty of Bioscience EngineeringGhent UniversityCoupure links 653Ghent9000Belgium

Recently N-glycosylation was found to be required for the postembryonic development and metamorphosis of the holometabolous beetle Tribolium castaneum. However, the role of N-glycosylation in the development of hemimetabolous insects is unknown. To further elucidate the role of N-glycosylation in the development of insects, a functional characterization of the N-glycosylation-related genes (NGRGs) was performed in a model insect for hemimetabolous development, namely the brown planthopper Nilaparvata lugens. In this project, we report the effects of RNAi-mediated silencing of 15 NGRGs on the postembryonic development of N. lugens. Two major observations were made. First, interruption of the early steps of N-glycan processing led to a lethal phenotype during the transition from nymph to adult as was observed in T. castaneum. Second, we report here on an essential function for the α-1,6-fucosyl transferase in the ecdysis event of N. lugens between nymphal instars, since gene-silencing by RNAi led to failure of ecdysis and subsequent mortality of the treated insect.

关键词： development glycosylation Nilaparvata lugens N-glycan postembryonic RNA interference