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Masculinizer gene controls sexual differentiation in Hyphantria cunea

insect Science 2024年第2期31卷 405-416页

作者： Xiaowei Li Huihui Liu Honglun Bi Yaohui Wang Jun Xu Sufang Zhang Zhen Zhang Ze Zhang Yongping Huang laboratory of evolutionary and Functional Genomics School of Life SciencesChongqing UniversityChongqingChina cas key laboratory of insect developmental and evolutionary biology CAS Centerfor Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyShanghaiChina key laboratory of Forest Protection Research Institute of Forest EcologyEnvironment and ProtectionChinese Academy of ForestryState Forestry AdministrationBeijingChina

The Masculinizer gene,Masc,encodes a lepidopteran-specific novel CCCHtype zinc finger protein,which controls sex determination and dosage compensation in Bombyx mori.Considering the potential application of it in pest control,it is necessary to investigate the function of Masc gene in Hyphantria cunea,a globally invasive forest pest.In the present study,we identified and functionally characterized the Masc gene,HcMasc,in H.cunea.Sequence analysis revealed that HcMASC contained the conserved CCCH-type zinc finger domain,nuclear localization signal,and male determining domain,in which the last was confirmed to be required for its masculinization in BmN cell line.However,expression data showed that unlike male-biased expression in B.mori,Hc-Masc gene expresses in main all developmental stages or tissues in both sexes.Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9-based disruption of the common exons 1 and 3 of the HcMasc gene resulted in imbalanced sex ratio and abnormal external genitalia of both sexes.Our results suggest that the HcMasc gene is required for both male and female sexual differentiation and dosage compensation in H.cunea and provide a foundation for developing better strategies to control this pest.

关键词： CRISPR/cas9 dosage compensation Hyphantria cunea Masc sexual differentiation

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Identification and functional analysis of CG3526 in spermatogenesis of Drosophila melanogaster

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insect Science 2024年第1期31卷 79-90页

作者： Qihao Hu Yanhong Xiao Runnan Wei Ting Tang Liang Wen Yuzhen Lu Xiao-Qiang Yu Guangdong Provincial key laboratory of insect developmental biology and Applied Technology Guangzhou Key Laboratory of Insect Development Regulation and Application ResearchInstitute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhou510631 China

Spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely unknown.In this study,we identified a testis-specific gene CG3526 in Drosophila melanogaster.Bioinformatics analysis showed that CG3526 contains a zinc binding domain and 2 C2H2 type zinc fingers,and it is clustered to the vertebrate really interesting new gene(RING)family E3 ubiquitin-protein ligases.When CG3526 was knocked down by RNA interference(RNAi),the testis became much smaller in size,and the apical tip exhibited a sharp and thin end instead of the blunt and round shape in the control testis.More importantly,compared to the control flies,only a few mature sperm were present in the seminal vesicle of C587-Gal4>CG3526 RNAi flies.Immunofluorescence staining of the testis from CG3526 RNAi flies showed that the homeostasis of testis stem cell niche was disrupted,cell distribution in the apical tip was scattered,and the process of spermatogenesis was not completed.Furthermore,we found that the phenotype of CG3526 RNAi flies’testis was similar to that of testis of Stat92E RNAi flies,the expression level of CG3526 was significantly downregulated in the Stat92EF06346 mutant flies,and the promoter activity of CG3526 was upregulated by STAT92E.Taken together,our results indicated that CG3526 is a downstream effector gene in the JAK-STAT signaling pathway that plays a key role in the spermatogenesis of Drosophila.

关键词： CG3526 JAK-STAT spermatogenesis STAT92E testis

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Hexamerin and allergen are required for female reproduction in the American cockroach,Periplaneta americana

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insect Science 2024年第1期31卷 186-200页

作者： Shiming Zhu Xiaoyi Chen Sishi Xia Qin Li Ziqi Ye Shaoting Zhao Kexin Liu Fangfang Liu Guangdong Provincial key laboratory of insect developmental biology and Applied Technology Institute of Insect Science and TechnologyInstitute of Insect Science and Technology&School of Life SciencesSouth China Normal UniversityGuangzhouChina Guangmeiyuan R&D Center Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied TechnologySouth China Normal UniversityMeizhouGuangdong ProvinceChina Guangdong laboratory for Lingnan Modern Agriculture South China Agricultural UniversityGuangzhouChina

Reproduction is of great importance for the continuation of the species.In insects,the fat body is the major tissue for nutrient storage and involved in vitellogenesis,which is essential for female reproduction.Here,2 proteins,hexamerin and allergen,were separated from the fat bodies of adult female American cockroaches(Periplaneta americana)and identified as storage proteins,encoding for 733 amino acids with molecular weight of 87.88 kDa and 686 amino acids with molecular weight of 82.18 kDa,respectively.The encoding genes of these 2 storage proteins are mainly expressed in the fat body.RNA interference-mediated knockdown of Hexamerin and Allergen in the early stage of the first reproductive cycle in females suppressed vitellogenesis and ovarian maturation,indicating that these storage proteins are involved in controlling reproduction.Importantly,the expression of Hexamerin and Allergen was repressed by knockdown of the juvenile hormone(JH)receptor gene Met and the primary response gene Kr-h1,and was induced by methoprene,a JH analog,in both in vivo and in vitro experiments.Altogether,we have determined that hexamerin and allergen are identified as storage proteins and play an important role in promoting female reproduction in the American cockroach.The expression of their encoding genes is induced by JH signaling.Our data reveal a novel mechanism by which hexamerin and allergen are necessary for JH-stimulated female reproduction.

关键词： fat body juvenile hormone ovary reproduction storage protein vitellogenesis

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Acetyl coenzyme A carboxylase modulates lipogenesis and sugar homeostasis in Blattella germanica

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insect Science 2024年第2期31卷 387-404页

作者： Xiaojin Pei Tiantian Bai Yuan Luo Zhanfeng Zhang Sheng Li Yongliang Fan Tong-Xian Liu Guangdong Provincial key laboratory of insect developmental biology and Applied Technology and Institute of insect Science and Technology School of Life SciencesSouth China Normal UniversityGuangzhouChina State key laboratory of Crop Stress biology for Arid Areas and key laboratory of Integrated Pest Management on Crops in Northwestern Loess Plateau Ministry of AgricultureNorthwest A&F UniversityYanglingShaanxiChina Guangmeiyuan R&D Center Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied TechnologySouth China Normal UniversityMeizhouGuangdong ProvinceChina Institute of Entomology Guizhou UniversityGuiyangChina

Lipid and sugar homeostasis is critical for insect development and survival.In this study,we characterized an acetyl coenzyme A carboxylase gene in Blattella germanica(BgACC)that is involved in both lipogenesis and sugar homeostasis.We found that BgACC was dominantly expressed in the fat body and integument,and was significantly upregulated after molting.Knockdown of BgACC in 5th-instar nymphs did not affect their normal molting to the next nymphal stage,but it caused a lethal phenotype during adult emergence.BgACC-RNA interference(RNAi)significantly downregulated total free fatty acid(FFA)and triacylglycerol(TAG)levels,and also caused a significant decrease of cuticular hydrocarbons(CHCs).Repression of BgACC in adult females affected the development of oocytes and resulted in sterile females,but BgACC-RNAi did not affect the reproductive ability of males.Interestingly,knockdown of BgACC also changed the expression of insulin-like peptide genes(BglLPs),which mimicked a physiological state of high sugar uptake.In addition,BgACC was upregulated when B.germanica were fed on a high sucrose diet,and repression of BgACC upregulated the expression of the glycogen synthase gene(BgGlyS).Moreover,BgACC-RNAi increased the circulating sugar levels and glycogen storage,and a longevity assay suggested that BgACC was important for the survival of B.germanica under conditions of high sucrose uptake.Our results confirm that BgACC is involved in multiple lipid biogenesis and sugar homeostasis processes,which further modulates insect reproduction and sugar tolerance.This study benefits our understanding of the crosstalk between lipid and sugar metabolism.

关键词： acetyl-CoA carboxylase German cockroach lipid biosynthesis sugar tolerance

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Two sex pheromone receptors for sexual communication in the American cockroach

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Science China(Life Sciences) 2024年第7期67卷 1455-1467页

作者： Na Li Renke Dong Huanchao Zeng Yan Zhang Run Huang Wei Liu Fengming Cao Jincong Yu Mingtao Liao Jingyou Chen Wenlei Zhang Zejian Huang Jiahui Wang Li Li Shen Zhu Danyan Huang Zining Li Xiaoshuai Zhang Dongwei Yuan Nan Chen Yongliang Fan Guirong Wang Coby Schal Yufeng Pan Sheng Li Guangdong Provincial key laboratory of insect developmental biology and Applied Technology Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhou 510631China Guangmeiyuan R&D Center Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied TechnologySouth China Normal UniversityMeizhou 514589China Lingnan Guangdong laboratory of Modern Agriculture Genome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518000China State key laboratory of Crop Stress biology for Arid Areas and Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of AgricultureNorthwest A&F UniversityYangling 712100China Department of Entomology and Plant Pathology North Carolina State UniversityRaleigh 27695USA The key laboratory of developmental Genes and Human Disease School of Life Science and TechnologySoutheast UniversityNanjing 210096China

Volatile sex pheromones are vital for sexual communication between males and females.Females of the American cockroach,Periplaneta americana,produce and emit two sex pheromone components,periplanone-A(PA)and periplanone-B(PB).Although PB is the major sex attractant and can attract males,how it interacts with PA in regulating sexual behaviors is still unknown.In this study,we found that in male cockroaches,PA counteracted PB attraction.We identified two odorant receptors(ORs),OR53 and OR100,as PB/PA and PA receptors,respectively.OR53 and OR100 were predominantly expressed in the antennae of sexually mature males,and their expression levels were regulated by the sex differentiation pathway and nutrition-responsive signals.Cellular localization of OR53 and OR100 in male antennae further revealed that two types of sensilla coordinate a complex two-pheromone-two-receptor pathway in regulating cockroach sexual behaviors.These findings indicate distinct functions of the two sex pheromone components,identify their receptors and possible regulatory mechanisms underlying the male-specific and age-dependent sexual behaviors,and can guide novel strategies for pest management.

关键词： sex pheromone odorant receptor sexual attraction sex differentiation cockroach

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The appearance of cytoplasmic cytochrome C precedes apoptosis during Drosophila salivary gland degradation

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insect Science 2024年第1期31卷 157-172页

作者： Shihui Long Wenxin Cao Yongyu Qiu Ruohan Deng Jiali Liu Lidan Zhang Renke Dong Fengxin Liu Sheng Li Haigang Zhao Na Li Kang Li Guangdong Provincial key laboratory of insect developmental biology and Applied Technology Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Guangdong laboratory for Lingnan Modern Agriculture South China Agricultural UniversityGuangzhouChina Guangmeiyuan R&D Center Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied TechnologySouth China Normal UniversityMeizhouGuangdong ProvinceChina School of Food Science and Engineering South China University of TechnologyGuangzhouChina ChemPartner PharmaTech Co. LtdJiangmenGuangdong ProvinceChina

Apoptosis is an important process for organism development that functions to eliminate cell damage,maintain homeostasis,and remove obsolete tissues during morphogenesis.In mammals,apoptosis is accompanied by the release of cytochrome C(Cyt-c)from mitochondria to the cytoplasm.However,whether this process is conserved in the fruit fly,Drosophila melanogaster,remains controversial.In this study,we discovered that during the degradation of Drosophila salivary gland,the transcription of mitochondria apoptosis factors(MAPFs),Cyt-c,and death-associated APAF1-related killer(Dark)encoding genes are all upregulated antecedent to initiator and effector caspases encoding genes.The proteins Cyt-c and the active caspase 3 appear gradually in the cytoplasm during salivary gland degradation.Meanwhile,the Cyt-c protein colocates with mito-GFP,the marker indicating cytoplasmic mitochondria,and the change in mitochondrial membrane potential coincides with the appearance of Cyt-c in the cytoplasm.Moreover,impeding or promoting 20E-induced transcription factor E93 suppresses or enhances the staining of Cyt-c and the active caspase 3 in the cytoplasm of salivary gland,and accordingly decreases or increases the mitochondrial membrane potential,respectively.Our research provides evidence that cytoplasmic Cyt-c appears before apoptosis during Drosophila salivary gland degradation,shedding light on partial conserved mechanism in apoptosis between insects and mammals.

关键词： active caspase 3 apoptosis Cyt-c Drosophila salivary gland

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Pleistocene glaciation advances the cryptic speciation of Stellera chamaejasme L. in a major biodiversity hotspot

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Journal of Integrative Plant biology 2024年第6期66卷 1192-1205页

作者： Santosh Kumar Rana Hum Kala Rana Jacob B.Landis Tianhui Kuang Juntong Chen Hengchang Wang Tao Deng Charles C.Davis Hang Sun cas key laboratory for Plant Diversity and Biogeography of East Asia Kunming Institute of Botany Chinese Academy of Sciences Arkansas Biosciences Institute Arkansas State University School of Integrative Plant Science Section of Plant Biology and the L.H.Bailey Hortorium Cornell University BTI Computational biology Center Boyce Thompson Institute cas key laboratory of Plant Germplasm Enhancement and Specialty Agriculture Chinese Academy of Sciences Department of Organismic and evolutionary biology Herbaria Harvard University

The mountains of Southwest China comprise a significant large mountain range and biodiversity hotspot imperiled by global climate change. The high species diversity in this mountain system has long been attributed to a complex set of factors, and recent large-scale macroevolutionary investigations have placed a broad timeline on plant diversification that stretches from 10 million years ago(Mya) to the present. Despite our increasing understanding of the temporal mode of speciation, finer-scale populationlevel investigations are lacking to better refine these temporal trends and illuminate the abiotic and biotic influences of cryptic speciation. This is largely due to the dearth of organismal sampling among closely related species and populations, spanning the incredible size and topological heterogeneity of this region. Our study dives into these evolutionary dynamics of speciation using genomic and ecomorphological data of Stellera chamaejasme L. We identified four previously unrecognized cryptic species having indistinct morphological traits and large metapopulation of evolving lineages, suggesting a more recent diversification(～2.67–0.90 Mya), largely influenced by Pleistocene glaciation and biotic factors. These factors likely influenced allopatric speciation and advocated cyclical warming–cooling episodes along elevational gradients during the Pleistocene. The study refines the evolutionary timeline to be much younger than previously implicated and raises the concern that projected future warming may influence the alpine species diversity,necessitating increased conservation efforts.

关键词： cryptic speciation glacial-interglacial Hengduan Mountains north-south migration Stellera chamaejasme

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Functional characterization of Bmcap in uric acid metabolism in the silkworm

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insect Science 2024年第1期31卷 147-156页

作者： Linmeng Tang Dehong Yang Zhiwei Liu Yaohui Wang Xu Yang Yujia Liu Dongbin Chen Zheng Tang Yongping Huang key laboratory of insect developmental and evolutionary biology Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghaiChina cas Center for Excellence in Biotic Interactions University of Chinese Academy of SciencesBeijingChina Central Research Institute Shanghai Pharmaceuticals Holding Co.Ltd.ShanghaiChina Departments of Neonatology International Peace Maternity and Child Health Hospital of China Welfare InstitutionSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina

After a millennium of domestication,numerous silkworm mutants have emerged that exhibit transparent epidermis,which is caused by abnormally low levels of uric acid.We identified the Bombyx mori gene Bmcap(BMSK0003832)as the homolog of cappuccino,a subunit of the biogenesis of lysosome-related organelles complex-1(BLOC-1)that has been extensively characterized in human,mouse,and insect species,by analyzing the amino acid sequences of putative purine metabolism genes.Using the clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 system,we disrupted Bmcap,resulting in decreased uric acid levels in the silkworm epidermis and a translucent skin phenotype.In the Bmcap mutant,the purine metabolism,nitrogen metabolism,pyrimidine metabolism,and membrane system were altered compared to the wild type.Biogenesis of lysosome-related organelle complex genes play a role in the pigmentation and biogenesis of lysosome-related organelles(LROs)in platelets,melanocytes,and megakaryocytes.LROs exhibit unique morphologies and functions in various tissues and cells.Investigation of the Bmcap mutant will enhance our understanding of the uric acid metabolic pathway in silkworms,and this mutant offers a valuable silkworm model for LRO studies.

关键词： BLOC-1 subunit Bombyx mori cappuccino translucent mutation uric acid

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Geographical patterns and determinants of insect biodiversity in China

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Science China(Life Sciences) 2024年第6期67卷 1255-1265页

作者： Huizhong Fan Tongyi Liu Youhua Chen Ziyan Liao Jun Chen Yibo Hu Gexia Qiao Fuwen Wei cas key laboratory of Animal Ecology and Conservation biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China cas key laboratory of Mountain Ecological Restoration and Bioresource Utilization&China-Croatia“Belt and Road”Joint laboratory on Biodiversity and Ecosystem Services Chengdu Institute of BiologyChinese Academy of SciencesChengdu 610041China Jiangxi Provincial key laboratory of Conservation biology College of ForestryJiangxi Agricultural UniversityNanchang 330045China University of Chinese Academy of Sciences Beijing 100049China

insects play important roles in the maintenance of ecosystem functioning and the provision of livelihoods for millions of people.However,compared with terrestrial vertebrates and angiosperms,such as the giant panda,crested ibis,and the metasequoia,insect conservation has not attracted enough attention,and a basic understanding of the geographical biodiversity patterns for major components of insects in China is lacking.Herein,we investigated the geographical distribution of insect biodiversity across multiple dimensions(taxonomic,genetic,and phylogenetic diversity)based on the spatial distribution and molecular DNA sequencing data of insects.Our analysis included 18 orders,360 families,5,275 genera,and 14,115 species of insects.The results revealed that Southwestern and Southeastern China harbored higher insect biodiversity and numerous older lineages,representing a museum,whereas regions located in Northwestern China harbored lower insect biodiversity and younger lineages,serving as an evolutionary cradle.We also observed that mean annual temperature and precipitation had significantly positive effects,whereas altitude had significantly negative effects on insect biodiversity in most cases.Moreover,cultivated vegetation harbored the highest insect taxonomic and phylogenetic diversity,and needleleaf and broadleaf mixed forests harbored the highest insect genetic diversity.These results indicated that human activities may positively contribute to insect spatial diversity on a regional scale.Our study fills a knowledge gap in insect spatial diversity in China.These findings could help guide national-level conservation plans and the post-2020 biodiversity conservation framework.

关键词： insect spatial patterns multiple dimensions driving factors vegetation types

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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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