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development and formation of wing cuticle based on transcriptomic analysis in Locusta migratoria during metamorphosis

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Journal of Integrative Agriculture 2024年第4期23卷 1285-1299页

作者： Jing Zhang Zhaochen Wu Shuo Li He Huang Suning Liu Weimin Liu Xiaoming Zhao Jianzhen Zhang Shanxi Key Laboratory of Nucleic Acid Biopesticides Institute of Applied BiologyShanxi UniversityTaiyuan 030006China College of Life Science Shanxi UniversityTaiyuan 030006China Guangmeiyuan R&D Center/Guangdong Provincial Key Laboratory of Insect developmental Biology and Applied Technology South China Normal UniversityMeizhou 514779China

wings are an important flight organ of insects.wing development is a complex process controlled by a series of genes.The flightless wing pad transforms into a mature wing with the function of migratory flight during the nymphto-adult metamorphosis.However,the mechanism of wing morphogenesis in locusts is still unclear.This study analyzed the microstructures of the locust wing pads at pre-eclosion and the wings after eclosion and performed the comparative transcriptome analysis.RNA-seq identified 25,334 unigenesand 3,430 differentially expressed genes(DEGs)(1,907 up-regulated and 1,523 down-regulated).The DEGs mainly included cuticle development(LmACPs),chitin metabolism(Lm Idgf4),lipid metabolism-related genes,cell adhesion(Integrin),zinc finger transcription factors(LmSalm,LmZF593 andLmZF521),and others.Functional analysis based on RNA interference and hematoxylin and eosin(H&E)staining showed that the three genes encoded zinc finger transcription factors are essential for forming wing cuticle and maintaining morphology in Locusta migratoria.Finally,the study found that the LmSalm regulates the expression of LmACPs in the wing pads at pre-eclosion,and LmZF593 and LmZF521 regulate the expression of LmIntegrin/LmIdgf4/LmHMT420 in the wings after eclosion.This study revealed that the molecular regulatory axis controls wing morphology in nymphal and adult stages of locusts,offering a theoretical basis for the study of wing development mechanisms in hemimetabolous insects.

关键词： Locusta migratoria wing development metamorphosis RNA-seq

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Nanocarrier-delivered dsRNA suppresses wing development of green peach aphids

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Insect Science 2022年第3期29卷 669-682页

作者： Yun-Hui Zhang Zhong-Zheng Ma Hang Zhou Zi-Jian Chao Shuo Yan Jie Shen Department of Plant Biosecurity and MOA Key Lab of Pest Monitoring and Green Management College of Plant ProtectionChina Agricultural UniversityBeijingChina

RNA interference(RNAi)has developed rapidly as a potential“green”pest management strategy.At present,most studies have focused on the screening of aphid lethal genes,whereas only a few studies have been conducted on wing development,which is crucial for aphid migration and plant-virus dissemination.Here,the Myzus persicae genes vestigial(vg)and Ultrabithorax(Ubx)related to wing development,were cloned.These two genes were expressed in various tissues of 3rd-instar winged aphids.The mRNA level of vg was high in 3rd-instar nymphs,whereas the expression level of Ubx was high in adults.The nanocarrier-mediated delivery system delivered double-stranded RNAs for aphid RNAi using topical and root applications.The expression levels of vg and Ubx were downregulated by 44.0%and 36.5%,respectively,using the topical application.The simultaneous RNAi of the two target genes caused 63.3%and 32.2%wing aberration rates using topical and root applications,respectively.The current study provided a promising method for controlling aphid migration to alleviate the spread of insect transmitted plant diseases.

关键词： nanocarrier RNA interference Ultrabithorax vestigial wing development

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MicroRNA miR-252-5p regulates the Notch signaling pathway by targeting Rab6 in Drosophila wing development

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Insect Science 2023年第5期30卷 1431-1444页

作者： Do-Hwan Lim Min-Seok Choi Ji Won Jeon Young Sik Lee School of Systems Biomedical Science Soongsil UniversitySeoulRepublic of Korea College of Life Sciences and Biotechnology Korea UniversitySeoulRepublic of Korea Institute of Animal Molecular Biotechnology Korea UniversitySeoulRepublic of Korea

The Notch signaling pathway plays a central role in the development of various organisms.However,dysregulation of microRNAs(miRNAs),which are crucial regulators of gene expression,can disrupt signaling pathways at all stages of development.Although Notch signaling is involved in wing development in Drosophila,the mechanism underlying miRNA-based regulation of the Notch signaling pathway is unclear.Here,we report that loss of Drosophila miR-252 increases the size of adult wings,whereas the overexpression of miR-252 in specific compartments of larval wing discs leads to patterning defects in the adult wings.The miR-252 overexpression-induced wing phenotypes were caused by aberrant Notch signaling with intracellular accumulation of the full-length Notch receptor during development,which could be due to defects in intracellular Notch trafficking associated with its recycling to the plasma membrane and autophagy-mediated degradation.Moreover,we identified Rab6 as a direct target of miR-252-5p;Rab6 encodes a small Ras-like GTPase that regulates endosomal trafficking pathways.Consistent with this finding,RNAi-mediated downregulation of Rab6 led to similar defects in both wing patterning and Notch signaling.Notably,co-overexpression of Rab6 completely rescued the wing phenotype associated with miR-252 overexpression,further supporting that Rab6 is a biologically relevant target of miR-252-5p in the context of wing development.Thus,our data indicate that the miR-252-5p-Rab6 regulatory axis is involved in Drosophila wing development by controlling the Notch signaling pathway.

关键词： Drosophila miR-252-5p Notch signaling Rab6 wing development

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Lmlntegrinβ-PS is required for wing morphogenesis and development in Locusta migratoria

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Insect Science 2021年第3期28卷 705-717页

作者： Xiao-Ming Zhao Niu Niu Jia-Peng Yang Wei-Min Liu Jian-Zhen Zhang Institute of Applied Biology Shanxi UniversityTaiyuanChina College of Life Science Shanxi UniversityTaiyuanChina

wings are an important flight organ of insects and their morphogenesis depends on a series of cell-to-cell and cell-to-extracellular matrix interactions.Integrin as a transmembrane protein receptor mediates cell-to-cell adhesion,cell-to-extracellular matrix interactions and signal transduction.In the present study,we characterized an integrin gene that encodes integrinβ-PS protein in Locusta migratoria.Lmlntegrinβ-PS is highly expressed in the wing pads and the middle stages of 5th instar nymphs.Immunohisto-chemical analysis revealed that the Lmlntegrinβ-PS protein was localized at the cell base of the two layers of wings.After suppression of Lmlntegrinβ-PS by RNA interference,the wing pads or wings were unable to form normally,with a blister wing appearance during nymph to nymph transition and nymph to adult transition.We further found that the dorsal and ventral epidermis of the wings after dsLmlntegrinβ-PS injection were improperly connected and formed huge cavities revealed by hematoxylin and eosin staining.Furthermore,the morphology and structure of the wing cuticle was significantly disturbed which affected the stable arrangement and attachments of the wing epidermis.Moreover,the expression of related cell adhesion genes was significantly decreased in LmIntegrinβ-PS-suppressed L.migratoria,suggesting that Lmlntegrinβ-PS is required for the morphogenesis and development of wings during molting by stabilizing cell adhesion and maintaining the cytoskeleton of these cells.

关键词： cell adhesion integrin Locusta migratoria RNAi wing development

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Bmmp influences wing morphology by regulating anterior-posterior and proximal-distal axes sdevelopment

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Insect Science 2022年第6期29卷 1569-1582页

作者： Yun-Long Zou Xin Ding Li Zhang Li-Feng Xu Shu-Bo Liang Hai Hu Fang-Yin Dai Xiao-Ling Tong State Key Laboratory of Silkworm Genome Biology:Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture and Rural AffairsCollegeof SericultureTextileand Biomass SciencesSouthwest UniversityChongqingChina

Insect wings are subject to strong selective pressure,resulting in the evolution of remarkably diverse wing morphologies that largely determine flight capacity.However,the genetic basis and regulatory mechanisms underlying wing size and shape development are not well understood.The silkworm Bombyx mori micropterous(mp)mutant exhibits shortened wing length and enlarged vein spacings,albeit without changes in total wing area.Thus,the mp mutant comprises a valuable genetic resource for studying wing de-velopment.In this study,we used molecular mapping to identify the gene responsible for the mp phenotype and designated it Bmmp.Phenotype-causing mutations were identified as indels and single nucleotide polymorphisms in noncoding regions.These mutations resulted in decreased Bmmp messenger RNA levels and changes in transcript isoform composition.Bmmp null mutants were generated by clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 and exhibited changed wing shape,similar to mp mutants,and significantly smaller total wing area.By examining the expression of genes critical to wingdevelopment inwildtype and Bmmp null mutants,we found that Bmmp exerts its function by coordinately modulating anterior-posterior and proximal-distal axes development.We also studied a Drosophila mp mutant and found that Bmmp is functionally conserved in Drosophila.The Drosophila mp mutant strain exhibits curly wings of reduced size and a complete loss of flight capacity.Our results increase our understanding of the mechanisms underpinning insect wing development and reveal potential targets for pest control.

关键词： anterior-posterior axis Bmmp CRISPR/Cas9 proximal-distal axis silk-worm wing development

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BmAntp is involved in the regulation of wing development in Bombyx mori

BmAntp is involved in the regulation of wing development in ...

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中国蚕学会第八届青年学术研讨会

作者： Ya-Qun Xin Xiao-Ling Tong Fang-Yin Dai Cheng Lu State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University

Hox genes are well-known master regulators in segmental identity and organ specification during development. The expression and activity of Hox genes are critical to animal body plans in embryonic stage, but their roles in the post-embryonic development remain largely unexplored. Here, we took three wing-defective mutants, Wes, E and Nc, all of which were caused by antp mutation, as material to explore how Antp to be involved in wing development in Bombyx mori. Firstly, the expression patterns of Antp was analyzed during wing disc development in Bombyx mori. We showed that BmAntp was expressed in both fore and hind wings with a similar expression profile. The gene went through a modest expression in fifth instar and then increased gradually after pupation and reached the peak before moth. The Q-PCR indicated that the expression of Antp is higher in the mutants compared with that in the wide-type. EMSA showed that in vitro Antp can bind to ap and vg gene as downstream targets, which indicate that Bm Antp probably be involved in wing formation. Our research has offered a new insight of Hox gene putting in insect wing development, and the Bombyx wing might provide a useful system for understanding how Hox proteins select and regulate their target genes in organ specification.

关键词： Bombyx mori Antp expression wing development

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Characterization and functional analysis of chitinase family genes involved in nymph-adult transition of Sogatella furcifera

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Insect Science 2021年第4期28卷 901-916页

作者： Xibin Yang Cao Zhou Guiyun Long Hong Yang Chen Chen Daochao Jin Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions Institute of EntomologyGuizhou UniversityGuiyangChina College of Tobacco Science of Guizhou University GuiyangChina Scientific Observing and Experimental Station of Crop Pest in Guiyang Ministry of AgricultureGuiyangChina

Chitinase degrades chitin in the old epidermis or peritrophic matrix of insects,which ensures normal development and metamorphosis.In our previous work,we comprehensively studied the function of SfCht7 in Sogatella furcifera.However,the number and function of chitinase genes in S.furcifera remain unknown.Here,we identified 12 full-length chitinase transcripts from S.furcifera,which included nine chitinase(Cht),two imaginal disc growth factor(IDGF),and one endo-β-N-acetylglucosaminidase(ENGase)genes.Expression analysis results revealed that the expression levels of eight genes(SfCht3,SfCht5,SfCht6-1,SfCht6-2,SfCht7,SfCht8,SfCht10,and SfIDGF2)with similar transcript levels peaked prior to molting of each nymph and were highly expressed in the integument.Based on RNA interference(RNAi),description of the functions of each chitinase gene indicated that the silencing of SfCht5,SfCht10,and SfIDGF2 led to molting defects and lethality.RNAi inhibited the expressions of SfCht5,SfCht7,SfCht10,and SfIDGF2,which led to downregulated expressions of chitin synthase 1(SfCHS1,SfCHS1a,and SfCHS1b)and four chitin deacetylase genes(SfCDA1,SfCDA2,SfCDA3,and SfCDA4),and caused a change in the expression level of two trehalase genes(TRE1 and TRE2).Furthermore,silencing of SfCht7 induced a significant decrease in the expression levels of three wing development-related genes(SfWG,SfDpp,and SfHh).In conclusion,SfCht5,SfCht7,SfCht10,and SfIDGF2 play vital roles in nymph–adult transition and are involved in the regulation of chitin metabolism,and SfCht7 is also involved in wing development;therefore,these genes are potential targets for control of S.furcifera.

关键词： chitinase chitin deacetylase chitin synthesis RNA interference Sogatella furcifera wing development

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Cuticular protein gene LmACP8 is involved in wing morphogenesis in the migratory locust,Locusta migratoria

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Journal of Integrative Agriculture 2021年第6期20卷 1596-1606页

作者： ZHAO Xiao-ming YANG Jia-peng GOU Xin LIU Wei-min ZHANG Jian-zhen Institute of Applied Biology Shanxi UniversityTaiyuan 030006P.R.China College of Life Science Shanxi UniversityTaiyuan 030006P.R.China

Cuticular proteins(CPs)are major components of the insect cuticle-associated organs such as integument and wings,although the importance of CPs for wing development and function in hemimetabolous insects remains understudied.In the present study,a wing cuticular protein LmACP8 was identified from Locusta migratoria,which belongs to the RR-2 subfamily of cuticular protein R&R consensus(CPR)chitin-binding proteins.LmACP8 was mainly expressed in the wing pads and showed high expression levels before ecdysis of third-,fourth-,and fifth-instar nymphs,with its encoded protein located in the procuticle of wing pads and adult wings.Depletion of LmACP8 by RNA interference markedly reduced the amount of its protein,which consequently caused abnormal wing morphogenesis in the transition from nymph to adult of L.migratoria.We further demonstrated that the abnormal morphogenesis was caused by severe damage of the endocuticle in the wings.LmACP8 was suppressed by 20-hydroxyecdysone(20 E)in vivo,however,its expression was significantly up-regulated after knocking down the hormone receptor gene LmHR39.Thus,the LmACP8 that is negatively regulated by the LmHR39-mediated 20 E signaling pathway is involved in wing development during the nymph to adult transition.

关键词： cuticular proteins LmACP8,20-hydroxyecdysone wing development

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