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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 unknown.Here,we screened the expression dynamics of two PcRXR isoforms at 4 h intervals in the deutonymphal stage of Panonychus citri.The results showed that PcEcR had an expression pattern similar to that of PcRXR2.For PcRXR1,its expression remained at a certain high level,when there was a decrease of both PcEcR and PcRXR2.In 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 mass.RNAi-based silencing of PcRXR1 or PcRXR2 showed the same phenotype as in mites with that of silencing PcEcR.Furthermore,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 ecdysis.Taken 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

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Characterization of carotenoid biosynthetic pathway genes in the pea aphid(Acyrthosiphon pisum)revealed by heterologous complementation and RNA interference assays

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Insect Science 2022年第3期29卷 645-656页

作者： Bi-Yue Ding Xiu-Cheng Xie Feng Shang Guy Smagghe Jin-Zhi Niu Jin-Jun Wang Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina International Joint laboratory of China-Belgium on Sustainable Crop pest ControlAcademy of Agricultural SciencesSouthwest UniversityChongqingChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Carotenoids are involved in many essential physiological functions and are produced from geranylgeranyl pyrophosphate through synthase,desaturase,and cyclase activities.In the pea aphid(Acyrthosiphon pisum),the duplication of carotenoid biosynthetic genes,including carotenoid synthases/cyclases(ApCscA-C)and desaturases(ApCdeA-D),through horizontal gene transfer from fungi has been detected,and ApCdeB has known dehydrogenation functions.However,whether other genes contribute to aphid carotenoid biosynthesis,and its specific regulatory pathway,remains unclear.In the current study,functional analyses of seven genes were performed using heterologous complementation and RNA interference assays.The bifunctional enzymes ApCscA-C were responsible for the synthase of phytoene,and ApCscC may also have a cyclase activity.ApCdeA,ApCdeC,and ApCdeD had diverse dehydrogenation functions.ApCdeA catalyzed the enzymatic conversion of phytoene to neurosporene(three-step product),ApCdeC catalyzed the enzymatic conversion of phytoene to ζ-carotene(two-step product),and ApCdeD catalyzed the enzymatic conversion of phytoene to lycopene(four-step product).Silencing of ApCscs reduced the expression levels of ApCdes,and silencing these carotenoid biosynthetic genes reduced the α-,β-,and γ-carotene levels,as well as the total carotenoid level.The results suggest that these genes were activated and led to carotenoid biosynthesis in the pea aphid.

关键词： carotenoid biosynthesis heterologous complementary pea aphid RNA interference

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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 microorganisms.Abundant asymptomatic RNA viruses have been reported in aphids due to the application of RNA-seq,but aphid-virus interactions remain unclear.Bunyavirales is the most abundant RNA virus order,which can infect mammals,arthropods,and plants.However,many bunyaviruses have specific hosts,such as insects.Here,we discovered 18 viruses from 10 aphid species by RNA-seq.Importantly,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 species.ABV-1 may be transmitted horizontally during feeding on plant leaves and vertically through reproduction.In 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 reproduction.Moreover,viral titer significantly affected the lipid and protein contents in pea aphids.In 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

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Performance and Enzyme Activity of Beet Armyworm Spodoptera exigua (Hübner) (Lepidoptera:Noctuidae) Under Various Nutritional Conditions

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Agricultural Sciences in China 2011年第5期10卷 737-746页

作者： Helen Hull Sanders Chongqing Key Laboratory of Entomology and Pest Control Engineering Southwest University

We explored the influence of the artificial diets with different protein and glucose contents on larval development,fecundity and enzyme activities of the beet armyworm Spodoptera exigua （Hübner）.Our results suggested that development,fecundity and population growth index of beet armyworm increased with increased nutritional content in the artificial diets;however,when the yeast to cellulose content ratio reached 46.8：53.2,the fecundity and population growth had reached a maximum and additional protein did not yield additional growth.Additionally,3rd instar beet armyworm larvae fed on different artificial diets had increased enzymatic activities of acetylcholinesterase （AChE） with the increased nutritional content,but carboxylesterase （CarE） activities did not significantly change under variation in the nutritional content.

关键词： beet armyworm nutritional variation acetylcholinesterase carboxylesterase protein carbohydrate enzymeactivity

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Prey life stages modulate effects of predation stress on prey lifespan,development,and reproduction in mites

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Insect Science 2023年第3期30卷 844-856页

作者： Xiaoying Weil Guangyun Li Zhi-Qiang Zhang School of Biological Sciences The University of AucklandAucklandNew Zealand Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina Manaaki Whenua-Landcare Research AucklandNewZealand

The non-consumptive effects of predator-induced stress can influence a variety of life-history traits.Many previous studies focused only on short-term effects such as development and reproductive rates.Recent studies have showed that long-term predation stress(given during the whole life of the prey)and short-term predation stress(provided during the immature stage of the prey)could generate completely opposite results:the former could decrease lifespan,whereas the later could increase lifespan.However,it is still unclear whether the advantage is because of the short duration of exposure or the early stage of life during which exposure was exerted.Thus,in this study,the prey(Tyrophagus putrescentiae)was exposed to predation stress from the predator(Neoseiulus cucumeris)during different life stages(larva,protonymph,tritonymph,first 5 d of oviposition,the full lifespan or none of the above).The results showed that the predation stress supplied during larval and protonymphal stage delayed development,reduced fecundity and prolonged lifespan of the prey,while the stress given during tritonymphal stage only reduced lifespan slightly and the stress given during the first 5 d of oviposition did not change lifespan but reduced fecundity.This study indicated that the effects of predation stress are dependent on prey life stage and the predation stress experienced in the early life stages is important to lifespan modulation.

关键词： early life experience lifespan non-consumptive effects predation stress tyrophagus

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CRISPR-mediated mutagenesis of the odorant receptor co-receptor(Orco)gene disrupts olfaction-mediated behaviors in Bactrocera dorsalis

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Insect Science 2022年第5期29卷 1275-1286页

作者： Li Xu Hong-Bo Jiang Kai-Yue Tang Ying Yan Marc F.Schetelig Jin-Jun Wang 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 Insect Biotechnology in Plant Protection Justus-Liebig-University GiessenInstitute for Insect BiotechnologyGiessenGermany

Olfaction plays an essential role in insect behavior such as host location,foraging,mating,and oviposition.The odorant receptor co-receptor(Orco)is an obligatory odorant receptor and indispensable in odor perception.Here,we characterized the Orco gene from the oriental fruit fly,Bactrocera dorsalis(Hendel),a notorious agriculture pest.The olfactory deficiency mutants were generated by editing the BdorOrco gene using the CRISPR/Cas9 system.Electroantennograms(EAG)and olfactory preference assays confirmed that BdorOrco^(−/−)mutant flies had reduced perception of methyl eugenol,β-caryophyllene,and ethyl acetate.Oviposition bioassays showed that the eggs laid by BdorOrco^(−/−)females mediated by benzothiazole and 1-octen-3-ol were significantly decreased.In addition,BdorOrco^(−/−)mutant flies took a significantly longer time to locate the food source compared with wild type(WT)flies.Altogether,our data indicated that Orco is essential for multiple physiological processes in B.dorsalis,and it expands our understanding of the function of insect Orco.

关键词： CRISPR/Cas9 foraging olfaction oriental fruit fly oviposition

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Identification of the key chitinase genes in Tetranychus cinnabarinus (Boisduval) based on the expression and sequence characteristic analysis

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Journal of Integrative Agriculture 2017年第4期16卷 892-899页

作者： XU Hao-ran HE Lin XIAO Wei SHEN Guang-mao Key Laboratory of Entomology and Pest Control Engineering College of Plant Protection Southwest University Chongqing 400715 P.R.China

Chitin is an important content in the exoskeletons of arthropods, and its hydrolyzation is catalyzed by chitinases during the process of molting, thus, the chitinases are considered as ideal target to interfere the growth of arthropods. This study intends to clarify the characteristic of the chitinases during the development of Tetranychus cinnabarinus, and screen out important genes as potential control targets. The results showed that the total enzyme concentration of chitinases was significantly higher in larva, the first and second nymph than that in egg and adult. Base on the transcriptome data, six unigenes encoding chitinases were identified and their expression patterns in different developmental stages were detected. The expressions of TcCHIT1 and TcCHITIO showed high abundance during the molting process and their expression change during the developmental stages was consistent with the enzyme concentration. The full-length of these two genes were further cloned, and the structural characteristics of their proteins were analyzed by constructing the three-dimensional structure model. The results provide basic information to understand the characteristic of chitinases in T. cinnabarinus and might be considered as target for control.

关键词： Tetranychus cinnabarinus chitinase gene expression protein structure

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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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Functional analysis of UG7207D3 associated with abamectin resistance in Tetranychus cinnabarinus(Boisduval)

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Insect Science 2020年第2期27卷 276-291页

作者： Meng-Yao Wang Xin-Yang Liu Li Shi Jia-Lu Liu Guang-Mao Shen Ping Zhang Wen-Cai Lu Lin He Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina Academy o f Agricultural Sciences Southwest UniversityChongqingChina

Uridine diphosphate(UDP)-glycosyltransferases(UGTS)are widely distributed within living organisms and share roles in biotransformation of various lipophilic endo-and xenobiotics with activated UDP sugars.In this study,it was found that the activity of UGTs in abamectin-resistant(AbR)strain was significantly higher(2.3-fod)than that in susceptible strain(SS)of Tetramychus cinnabarinus.Further analysis showed that 5-nitrouracil,the inhibitor ofUGTs,could enhance the lethal effect of abamectin on mites.From the previous microarray results,we found an UGT gene(UGT201D3)overexpressed in AbR strain.Quantitative PCR analysis showed that UGT20ID3 was highly expressed and more inducible with abamectin exposure in the AbR strain.After silencing the tran-scription of UGT201D3,the activity of UGTs was decreased and the susceptibility to abamectin was increased in AbR strain whereas it was not in sS.Furthermore,UGT201D3 gene was then succefully expressed in Escherichia coli.The recombinant UGT20ID3 exhibited a-naphthol activity(2.810.43 nmol mg protein/min),and the enzyme activity could be inhibited by abamectin(inhibitory concentration at 50%:57.50±3.54 μmol/L).High-performance liquid chromatography analysis demonstrated that the recombinant UGT201D3 could efctively deplete abamectin(15.77%±3.72%)incubating with 150 ug protein for 6 h.These results provided direct evidence that UGT201D3 was involved in abamectin resistance in T cinnabarinus.

关键词： abamectin resistance prokaryotic expression RNA interference Tetrumychus cimnabarins UDP-glycosy ltransferases

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Influence of Host Shift on Genetic Differentiation of the Oriental Fruit Fly,Bactrocera dorsalis

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Journal of Integrative Agriculture 2014年第12期13卷 2701-2708页

作者： WAN Xuan-wu LIU Ying-hong LUO Lin-ming FENG Chuan-hong WANG Sheng MA Li College of Plant Protection Southwest University/Chongqing Key Laboratory of Entomology and Pest Control Engineering Sichuan Plant Protection Station

Invasion of the oriental fruit lfy, Bactrocera dorsalis, into new niches containing different food sources （a process referred to as host shift）, may cause population genetic differentiation and sympatric speciation. To attempt to infer that experimentally, test populations were established by transferring a subset of the original populations, which had been grown on banana for many generations, onto navel orange, and then subculturing the navel orange population and banana population for at least 20 generations. Four pairs of SSR primers with high polymorphism on laboratory strains were used to detect population genetic differentiation. All six tested populations （the 5th, 10th and 15th generations of B. dorsalis fed on banana and navel orange, respectively） were found to have low genetic diversity. Furthermore, the genetic diversity of the navel orange populations was found to decline after being crossed for several generations. Populations initially were deviated from Hardy-Weinberg equilibrium, however, equilibrium was achieved with increasing numbers of generations in both of the host populations. Limited gene lfows were found among the six populations. The Nei’s standard genetic distances between the two host populations of the same generation were initially low, but increased with generation number. Genetic distances between banana and navel orange populations of the same generation were lower than genetic distances between different generations grown on the same host plant. Analysis of molecular distance （AMOVA） results based on generation groups and host groups demonstrated that genetic variation among generations was greater than that between the two host populations. The results indicated that population genetic differentiation occurred after the host shift, albeit at low level. Biogeography and taxonomy of the B. dorsalis complex revealed that speciation of B. dorsalis might be tightly associated with host shift or host specialization of B. dorsalis following dispersal.

关键词： Bactrocera dorsalis oriental fruit lfy host shift population genetic differentiation sympatric speciation

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