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A midgut-specific lytic polysaccharide monooxygenase of Locusta migratoria is indispensable for the deconstruction of the peritrophic matrix

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Insect Science 2022年第5期29卷 1287-1298页

作者： Ming-Bo Qu Xiao-Xi Guo Lin Kong Ling-Jie Hou Qing Yang School of Bioengineering Dalian University of TechnologyDalianLiaoning ProvinceChina State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina Shenzhen Branch Guangdong Laboratory of Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of Agriculture and Rural AffairsAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenGuangdong ProvinceChina

Lytic polysaccharide monooxygenases(LPMOs)are important enzymes that boost the hydrolysis of recalcitrant polysaccharides,such as chitin.They are found extensively in different insect species and are classified as auxiliary activities family 15(AA15)LPMOs(LPMO15).Some of them were identified from the insect midgut and proven to act on chitin.However,knowledge about their physiological roles during insect growth and development remains limited.Here,we found that midgut-specific LPMO15s are widely distributed in different insect orders,such as the orthopteran Locusta migratoria and the lepidopteran Bombyx mori.Using L.migratoria as a model insect,the function of midgut-specific LmLPMO15-3 during development was investigated.Double-stranded RNA-mediated downregulation of LmLPMO15-3 expression at the 4th or 5th instar nymph stage severely decreased the survival rate and resulted in lethal phenotypes.Hematoxylin and eosin staining results indicated that the deficient individuals exhibited incompletely digested peritrophic matrix(PM),which suggested that LmLPMO15-3 is essential for the deconstruction of the PM during molting.This study provides direct evidence of the physiological importance of a midgut-specific LPMO15 during insect development.As L.migratoria is one of the most destructive agricultural pests,LmLPMO15-3 is a potential target for pest management.

关键词： chitin Locusta migratoria lytic polysaccharide monooxygenase midgut peritrophic matrix RNAi

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Lethal giant larvae gene is required for normal nymphal development and midgut morphogenesis in Locusta migratoria

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Insect Science 2022年第4期29卷 1017-1029页

作者： Xuekai Shi Xiaojian Liu Kristopher Silver Kun Yan Zhu Jianzhen Zhang Institute of Applied Biology Shanxi UniversityTaiyuanShanxiChina College of Life Science Shanxi UniversityTaiyuanShanxiChina Department of Entomology Kansas State UniversityManhattanKansasUSA

Disruption of morphogenesis,an essential process in organismal development,can lead to disruption of biological processes,reduction in fitness,or even death of an organism.The roles of lethal giant larvae(Lgl)protein in maintaining tissue organization have been studied extensively in mammals,but little is known about this gene's roles in promoting correct tissue morphogenesis in insects.In this study,we identified an Lgl ortholog in Locusta migratoria.RT-qPCR results revealed that LmLgl was constitutively expressed during third,fourth,and fifth instar nymphs.Furthermore,LmLgl showed highest expression in the ovary followed by wing pads,midgut,hindgut,Malpighian tubules,and foregut of the third-instar nymphs.To examine the role of LmLgl in L.migratoria development,RNA interference was performed during nymphal stages.Silencing of LmLgl increased body size but decreased bodyweight by 9.0%.Histological sections of the midgut revealed abnormal large masses of disordered epithelial cells in dsLmLgl-injected nymphs.In addition,downregulation of LmLgl transcript levels significantly altered the morphological structure in midgut,resulting in the formation of tumor-like structures.Our results indicated that LmLgl may act as a tumor-suppressor gene,which plays an essential role in maintaining a normal morphological structure in the midgut of L.migratoria.Our results also suggest that LmLgl may be explored as a potential target for developing dsRNA-based biological pesticides for managing insect pests.

关键词： development lethal giant larvae Locusita migratoria midgut morphogenesis RNA interference

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Azole selenourea disrupted the midgut and caused malformed development of Plutella xylostella

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Journal of Integrative Agriculture 2023年第4期22卷 1104-1116页

作者： GUO Xue-ying HUANG Zi-hao XIONG Lan-tu DONG Li HUANG Yue-kun WEI Lin-hao TANG Ri-yuan WANG Zhi-lin XU Han-hong Department of Applied Chemistry College of Materials and EnergySouth China Agricultural UniversityGuangzhou 510642P.R.China Key Laboratory of Natural Pesticide&Chemical Biology of Ministry of Education South China Agricultural UniversityGuangzhou 510642P.R.China

Chemical insecticides targeting the digestive system of diamondback moth(DBM),Plutella xylostella,have not been developed.The discovery of an insecticide with novel mode of action is a challenge for the control of DBM.In this study,a class of selenium-and difluoromethyl-modified azoles(fluoroazole selenoureas,FASU)were designed and synthesized for the control of DBM.Of these azoles,compound B4 showed the highest insecticidal activity against DBM.The LC50of third-and second-instar larvae reached 32.3 and 4.6μg mL^(–1),respectively.The midgut tissue of larvae was severely disrupted,and the larval intestinal tissue was dotted with unique red spots after treatment with compound B4.Compound B4 led to disintegration of the peritrophic matrix,swelling of the midgut epithelium,fracture of the microvilli,and extensive leakage of cellular debris in the midgut lumen.Surviving larvae grew very slowly,and the larval duration was significantly prolonged after exposure to compound B4 at sublethal doses(LC10,LC25and LC50).Furthermore,the pupation rate,emergence rate and pupae weight were significantly decreased.Compound B4 also induced abnormal pupae,causing adults to be trapped in the cocoon or failure to fly due to twisted wings.These results demonstrated that FASU could reduce the population of DBM in sublethal doses.FASU is the first synthetic insecticidal lead compound that has been shown to disrupt the midgut tissue of the larvae of DBM,and its mode of action totally differs from that of commercial chemical insecticides.

关键词： Plutella xylostella azole selenium digestive system midgut

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The piRNA response to BmNPV infection in the silkworm fat body and midgut

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Insect Science 2021年第3期28卷 662-679页

作者： Min Feng Anna Kolliopoulou Yao-Hong Zhou Shi-Gang Fei Jun-Ming Xia Luc Swevers Jing-Chen Sun Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding College of Animal ScienceSouth China Agricultural UniversityGuangzhouChina Insect Molecular Genetics and Biotechnology National Centre for Scientific Research DemokritosInstitute of Biosciences and ApplicationsAthensGreece

Bombyx mori nucleopolyhedrovirus(BmNPV)is a DNA virus that causes huge losses to the silkworm industry but the piRNA responses during BmNPV infection in the silkworm remain uninvestigated.Here,silkworm piRNA profiles of uninfected and BmNPV-infected fat body and midgut were determined by high-through sequencing in the early stages of BmNPV infection.A total of 2675 and 3396 genome-derived piRNAs were identified from fat body and midgut,respectively.These genome-derived piRNAs mainly originated from unannotated instead of transposon regions in the silkworm genome.In total,572 piRNAs were associated with 280 putative target genes in fat body and 805 piRNAs with 380 target genes in midgut.Compared to uninfected tissues,322 and 129 piRNAs were significantly upregulated in BmNPV-infected fat body and midgut,respectively.In addition,276 and 117 piRNAs were significantly downregulated.Moreover,differentially expressed(DE)piRNAs during BmNPV infection differed significantly between fat body and midgut.Putative DE piRNA-targeted genes were associated with“response to stimulus”and“environmental information processing”in fat body after infection with BmNPV,which may indicate an active piRNA response to BmNPV infection in fat body.This study may lay the foundation for future research of the potential roles of the piRNA pathway and specific piRNAs in BmNPV pathogenesis.

关键词： BmNPV fat body midgut piRNA silkworm

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Analyses of microRNAs and transcriptomes in the midgut of Spodoptera litura feeding on Brassica juncea

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Insect Science 2021年第2期28卷 533-547页

作者： Xiao-Peng Zou Yi-Guang Lin Yong-Jie Cen Kang Ma Bin-Bin Qiu Qi-Li Feng Si-Chun Zheng School of Life Sciences South China Normal UniversityGuangzhouChina

Spodoptera litura is a destructive agricultural pest in tropical and subtropical areas.Understanding the molecular mechanisms of S.litura adaptation to its preferred host plants may help identify target genes useful for pest control.We used high-throughput sequencing to characterize the expression patterns of messenger RNAs(mRNAs)and microRNAs(miRNAs)in the midgut of S.litura fed on Brassica juncea for 6 h and 48 h.A total of 108 known and 134 novel miRNAs were identified,29 miRNAs and 237 mRNAs were differentially expressed at 6 h of B.juncea feeding,26 miRNAs and 433 mRNAs were differentially expressed at 48 h.For the mRNAs,the up-regulated genes were mostly enriched in detoxification enzymes(cytochrome P450,esterase,glutathione^-transferase,uridine diphosphate-glucuronosyl transferase),while the down-regulated genes were mostly enriched in proteinases and immune-related genes.Furthermore,most detoxification enzymes begin to up-regulate at 6 h,while most digestion and immunerelated genes begin to up-or down-regulate at 48 h.Eighteen and 37 differently expressed transcription factors were identified at 6 h and 48 h,which may regulate the functional genes.We acquired 136 and 41 miRNA versus mRNA pairs at 6 h and 48 h,respectively.Some down-regulated and up-regulated miRNAs were predicted to target detoxification enzymes and proteinases,respectively.Real-time quantitative polymerase chain reaction of nine randomly selected miRNAs and 28 genes confirmed the results of RNA-seq.This analyses of miRNA and mRNA transcriptomes provides useful information about the molecular mechanisms of S.litura response to B.juncea.

关键词： Brassica juncea midgut miRNAs RNA-Seq Spodoptera litura

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Proteomics analysis of Trichoplusia ni midgut epithelial cell brush border membrane vesicles

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Insect Science 2019年第3期26卷 424-440页

作者： Muhammad Afzal Javed Cathy Coutu David A. Theilmann Martin A. Erlandson Dwayne D. Hegedus Saskatoon Research and Development Centre Agriculture and Agri-Food Canada Saskatoon Saskatchewan Canada Summerland Research and Development Centre Agriculture and Agri-Food Canada Summerland British Columbia Canada Department of Food & Bio-Product Sciences University of Saskatchewan Saskatoon Saskatchewan Canada

The insect midgut epithelium is composed of columnar, goblet, and regenerative cells. Columnar epithelial cells are the most abundant and have membrane protrusions that form the brush border membrane (BBM) on their apical side. These increase surface area available for the transport of nutrients, but also provide opportunities for interaction with xenobiotics such as pathogens, toxins and host plant allelochemicals. Recent improvements in proteomic and bioinfbrmatics tools provided an opportunity to determine the proteome of the T. ni BBM in unprecedented detail. This study reports the identification of proteins from BBM vesicles (BBMVs) using single dimension polyacrylamide gel elec? trophoresis coupled with multi-dimensional protein identification technology. More than 3000 proteins were associated with the BBMV of which 697 were predicted to possess either a signal peptide, at least one transmembrane domain or a GPI-anchor signal. Of these, bioinfbrmatics analysis and manual curation predicted that 185 may be associated with the BBMV or epithelial cell plasma membrane. These are discussed with respect to their predicted functions, namely digestion, nutrient uptake, cell signaling, development, cell-cell interactions, and other functions. We believe this to be the most detailed proteomic analysis of the lepidopteran midgut epithelium membrane to date, which will provide information to better understand the biochemical, physiological and pathological processes taking place in the larval midgut.

关键词： brush border membrane vesicles midgut proteomic analysis Trichoplusiani

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Multiple microRNAs control ecdysone signaling in the midgut of Spodoptera litura

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Insect Science 2020年第6期27卷 1208-1223页

作者： Wei Luo Li-Xia Huang Shuang-Kang Qin Xian Zhang Qi-Li Feng Jun Gu Li-Hua Huang Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Guangzhou Key Laboratory of Insect Development Regulation and Application Research Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Hubei Key Laboratory of Application of Plant-Cancer Active Ingredients School of Chemistry and Life SciencesHubei University of EducationWuhanChina

Metamorphosis is one of the most important physiological processes in insects.It is regulated by a serial of ecdysone cascade genes.Recently,lots o f microRNAs(miRNAs)were investigated in insects;however,their function in metamorphosis is largely unknown.In the present study,the dynamics of a small RNA population was investigated by RNA sequencing from the midgut of a lepidopteran pest Spodoptera litura during larval-pupal metamorphosis.A total of 101 miRNAs were identified,and 75 miRNAs were differentially expressed during the metamorphic process.The relationship between these differentially expressed miRNAs and 12 ecdysone cascade genes was analyzed by four classical software programs,and a multiple-to-multiple regulatory network was found to exist between these miRNAs and their targets.Among them,miR-14-3p and its two targets(EcR and E75)were chosen for further validation.MiR-14-3p had higher expression level in the 6 th instar larvae as compared with either the prepupae or pupae,which was opposite to that of both EcR and E75,two ecdysone cascade genes.Luciferase reporter assay confirmed that both EcR and E75 were regulated by miR-14-3p.Interestingly,the 3'untranslated regions are nearly identical to each other among different transcript variants of the ecdysone cascade genes,including EcR,USP,E75,E74,E78,E93,Hr3,Hr4,Hr39,K rhl and F tzfl.Thus,different transcript variants of one ecdysone cascade gene could be regulated by the same mi RNA.The above data suggest that the ecdysone signaling pathway is under the tight control of miRNA.These findings expand our understanding of the mechanism of insect metamorphosis and may also provide a novel possibility for the control of pest insects in the future.

关键词： ecdysone EcR E75 metamorphosis microRNA midgut

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A midgut-specific serine protease, BmSP36, is involved in dietary protein digestion in the silkworm, Bombyx mori

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Insect Science 2017年第5期24卷 753-767页

作者： Hua-Wei Liu You-Shan Li Xin Tang Peng-Chao Guo Dan-Dan Wang Chun-Yan Zhou Qing-You Xia Ping Zhao State Key LaboratoryofSilkworm Genome Biology Southwest University Chongqing China Vitamin D Research Institute Shaanxi Sci-Tech University Hanzhong Shaanxi Province China

Serine proteases play important roles in digestion and immune responses during insect development. In the present study, the serine protease gene BmSP36, which encodes a 292-residue protein, was cloned from the midgut cells ofBombyx mori. BmSP36 contains an intact catalytic triad （H57, D102 and S195） and a conserved substrate-binding site （G189, H216 and G226）, suggesting that it is a serine protease with chymotrypsin- like specificity. The temporal and spatial expression patterns of BmSP36 indicated that its messenger RNA and protein expression mainly occurred in the midgut at the feeding stages. Western blotting, immunofluorescence and liquid chromatography-tandem mass spectrometry analyses revealed secretion of BmSP36 protein from epithelial cells into the midgut lumen. The transcriptional and translational expression of BmSP36 was down- regulated after starvation but up-regulated after refeeding. Moreover, expression of the BmSP36 gene could be up-regulated by a juvenile hormone analogue. These results enable us to better define the potential role of BmSP36 in dietary protein digestion at the feeding stages during larval development.

关键词： Bombyx mori dietary digestion expression profile juvenile hormone midgut serine protease

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Integrated analysis of miRNAs and transcriptomes in Aedes albopictus midgut reveals the differential expression profiles of immune-related genes during dengue virus serotype-2 infection

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Insect Science 2016年第3期23卷 377-385页

作者： Yan-Xia Liu Fen-Xiang Li Zhuan-Zhuan Liu Zhi-Rong Jia Yan-He Zhou Hao Zhang Hui Yan Xian-Qiang Zhou Xiao-Guang Chen Key Laboratory of Prevention and Control of Emerging Infectious Diseases of Guangdong Higher Education Institutes Department ofPathogen Biology School of Public Health and Tropical Medicine Southern Medical University Guangzhou Laboratory of EmergingInfectious Diseases and Division of Laboratory Medicine Zhujiang Hospital Southern Medical University Guangzhou BGI-Shenzhen Shenzhen Guangdong Province China

Mosquito microRNAs （miRNAs） are involved in host-virus interaction, and have been reported to be altered by dengue virus （DENV） infection in Aedes albopictus （Diptera： Culicidae）. However, little is known about the molecular mechanisms of Aedes albopictus midgut--the first organ to interact with DENV--involved in its resistance to DENV. Here we used high-throughput sequencing to characterize miRNA and messenger RNA （mRNA） expression patterns in Aedes albopictus midgut in response to dengue virus serotype 2. A total of three miRNAs and 777 mRNAs were identified to be differentially expressed upon DENV infection. For the mRNAs, we identified 198 immune-related genes and 31 of them were differentially expressed. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses also showed that the differentially expressed immune-related genes were involved in immune response. Then the differential expression patterns of six immune-related genes and three miRNAs were confirmed by real-time reverse transcription polymerase chain reaction. Furthermore, seven known miRNA-mRNA interaction pairs were identified by aligning our two datasets. These analyses of miRNA and mRNA transcriptomes provide valuable information for uncovering the DENV response genes and provide a basis for future study of the resistance mechanisms in Aedes albopictus midgut.

关键词： Aedes albopictus DENV host-pathogen interaction midgut miRNA mRNA

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The Sterile 20-Like Kinase Tao Controls Tissue Homeostasis by Regulating the Hippo Pathway in Drosophila Adult midgut

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Journal of Genetics and Genomics 2014年第8期41卷 429-438页

作者： Xudong Huang Lai Shi Jun Cao Fangfei He Renling Li Yan Zhang Shuang Miao Longjin Jin Jia Qu Zhouhua Li Xinhua Lin School of Optometry and Ophthalmology and Eye Hospital Wenzhou Medical University State Key Laboratory of Biomembrane and Membrane Biotechnology Institute of ZoologyChinese Academy of Sciences School of Laboratory Medicine and Life Science Wenzhou Medical University School of Life Sciences Capital Normal University Division of Developmental Biology Cincinnati Children’s Hospital Medical Center

The proliferation and differentiation of adult stem cells must be tightly controlled in order to maintain resident tissue homeostasis. Dysfunction of stem cells is implicated in many human diseases, including cancer. However, the regulation of stem cell proliferation and differentiation is not fully understood. Here we show that the sterile-like 20 kinase, Tao, controls tissue homeostasis by regulating the Hippo pathway in the Drosophila adult midgut. Depletion of Tao in the progenitors leads to rapid intestinal stem cell （ISC） proliferation and midgut homeostasis loss. Meanwhile, we find that the Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling activity and cytokine production are significantly increased, resulting in stimulated ISC proliferation. Furthermore, expression of the Hippo pathway downstream targets, Diapl and bantam, is dramatically increased in Tao knockdown intestines. Consistently, we show that the Yorkie （Yki） acts downstream of Tao to regulate ISC proliferation. Together, our results provide insights into our understanding of the mechanisms of stem cell proliferation and tissue homeostasis control.

关键词： Stem cell midgut Drosophila Tissue homeostasis Tao Hippo

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