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Low-density lipoprotein receptor-related protein 2(LRP2)is required for lipid export in the midgut of the migratory locust,Locusta migratoria

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Journal of Integrative Agriculture 2024年第5期23卷 1618-1633页

作者： Yiyan Zhao Weimin Liu Xiaoming Zhao Zhitao Yu Hongfang Guo yang yang Hans Merzendorfer kun yan zhu Jianzhen Zhang Research Institute of Applied Biology Shanxi Key Laboratory of Nucleic Acid BiopesticidesShanxi UniversityTaiyuan 030006China College of Life Science Shanxi UniversityTaiyuan 030006China Interfaculty Institute of Cell Biology University of TubingenTubingen 72076Germany Institute of Biology University of SiegenSiegen 57076Germany Department of Entomology Kansas State UniversityManhattanKS 66506USA

Low-density lipoprotein receptor-related protein 2(LRP2)is a multifunctional endocytic receptor expressed in epithelial *** mammals,it acts as an endocytic receptor that mediates the cellular uptake of cholesterol-containing apolipoproteins to maintain lipid ***,little is known about the role of LRP2 in lipid homeostasis in *** the present study,we investigated the function of LRP2 in the migratory locust Locusta migratoria(LmLRP2).The mRNA of LmLRP2 is widely distributed in various tissues,including integument,wing pads,foregut,midgut,hindgut,Malpighian tubules and fat body,and the amounts of LmLRP2 transcripts decreased gradually in the early stages and then increased in the late stages before ecdysis during the nymphal developmental *** immunohistochemistry revealed that the LmLRP2 protein is mainly located in cellular membranes of the midgut and *** RNAi to silence LmLRP2 caused molting defects in nymphs(more than 60%),and the neutral lipid was found to accumulate in the midgut and surface of the integument,but not in the fat body,of dsLmLRP2-treated *** results of a lipidomics analysis showed that the main components of lipids(diglyceride and triglyceride)were significantly increased in the midgut,but decreased in the fat body and ***,the content of total triglyceride was significantly increased in the midgut,but markedly decreased in the fat body and hemolymph in dsLmLRP2-injected *** results indicate that LmLRP2 is located in the cellular membranes of midgut cells,and is required for lipid export from the midgut to the hemolymphand fat body in locusts.

关键词： Locusta migratoria low-density lipoprotein receptor-related protein 2 midgut lipids transport RNAi

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Biochemical characterization of chitin synthase activity and inhibition in the African malaria mosquito, Anopheles gambiae

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Insect Science 2013年第2期20卷 158-166页

作者： Xin Zhang kun yan zhu Department of Entomology Kansas State University Manhattan KS 66506 USA

Chitin synthase （CHS） is an important enzyme catalyzing the formation of chitin polymers in all chitin containing organisms and a potential target site for insect pest control. However, our understanding of biochemical properties of insect CHSs has been very limited. We here report enzymatic and inhibitory properties of CHS prepared from the African malaria mosquito, Anopheles gambiae. Our study, which represents the first time to use a nonradioactive method to assay CHS activity in an insect species, determined the optimal conditions for measuring the enzyme activity, including pH, temperature, and concentrations of the substrate uridine diphosphate N-acetyl-D-glucosamine （UDP- GlcNAc） and Mg＋＋. The optimal pH was about 6.5-7.0, and the highest activity was detected at temperatures between 37℃ and 44℃. Dithithreitol is required to prevent melanization of the enzyme extract. CHS activity was enhanced at low concentration of GIcNAc, but inhibited at high concentrations. Proteolytic activation of the activity is significant both in the 500×g supernatant and the 40 000×g pellet. Our study revealed only slight in vitro inhibition ofA. gambiae CHS activity by diflubenzuron and nikkomycin Z at the highest concentration （2.5μmol/L） examined. There was no in vitro inhibition by polyoxin D at any concentration examined. Furthermore, we did not observe any in vivo inhibition of CHS activity by any of these chemicals at any concentration examined. Our results suggest that the inhibition of chitin synthesis by these chemicals is not due to direct inhibition of CHS in A. gambiae.

关键词： Anophele gambiae chitin synthase diflubenzuron enzyme inhibition

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RNA interference＂ A powerful tool in entomological research and a novel approach for insect pest management

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Insect Science 2013年第1期20卷 1-3页

作者： kun yan zhu Department of Entomology Kansas State University Manhattan KS 66506-4004 USA

RNA interference （RNAi） is a highly conserved posttran- scriptional gene regulatory mechanism that controls gene expression at the mRNA level within living cells （Fire et al., 1998; Bell6s, 2010; zhuang ＆ Hunter, 2012）. RNAi is triggered by small interfering RNA （siRNA）, which is processed by Dicer, an endoribonuclease in the RNase III family, from either exogenous （e.g., infection by a virus or laboratory manipulations） or endogenous （e.g., pre- microRNA） double-stranded RNA （dsRNA）. The siRNAs are short dsRNA fragments of about 20-25 base pairs and carry two base extensions at the 3＇ end of each strand. In the RNAi pathway, siRNAs are incorporated into an RNA-induced silencing complex （RISC） in conjunction with an Argonaute （Ago） multidomain protein. Ago which contains an RNase H-like domain responsible for target degradation cleaves and discards the passenger （sense） strand of the siRNA duplex leading to an active RISC.

关键词： RNA干扰昆虫学双链RNA siRNA Dicer酶工具管理虫害

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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 *** 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 *** this study,we identified an Lgl ortholog in Locusta ***-qPCR results revealed that LmLgl was constitutively expressed during third,fourth,and fifth instar ***,LmLgl showed highest expression in the ovary followed by wing pads,midgut,hindgut,Malpighian tubules,and foregut of the third-instar *** examine the role of LmLgl in *** development,RNA interference was performed during nymphal *** 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 *** addition,downregulation of LmLgl transcript levels significantly altered the morphological structure in midgut,resulting in the formation of tumor-like *** 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 *** 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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A Novel Method for Achieving Gradient Microstructure in a Cu-Al Alloy:Surface Spinning Strengthening (3S)

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Acta Metallurgica Sinica(English Letters) 2017年第3期30卷 212-217页

作者： Chuan-Xi Ren QiangWang Zhen-Jun Zhang yan-kun zhu Zhe-Feng Zhang Shenyang National Laboratory for Materials Science Institute of Metal Research Chinese Academy of SciencesShenyang 110016 China School of Materials Science and Engineering University of Science and Technology of China Hefei 230026 China

A new designed surface strengthening method, surface spinning strengthening （3S）, was applied to achieve gradient microstructure in the surface layer of a Cu-11 at.%A1 alloy. According to the level of grain refinement, the gradient microstructure can be divided into four zones, including nanoscale grain zone, ultra-fine grain zone, fine grain zone and coarse grain zone from the surface to the matrix. Meanwhile, a plenty of grain boundaries and twin boundaries were introduced to inhibit the dislocation motion in the surface layer during the plastic deformation process. Consequently, the hardened layer with a microhardness gradient and high residual compressive stress was produced on the samples, and the yield strength of the Cu-11 at.%A1 alloy was effectively improved after 3S processing due to the strengthening effect caused by the gradient microstructurc.

关键词： Cu-AI alloy Surface spinning strengthening (3S) Strength Microhardness Gradientmicrostructure

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Transcript analysis and expression profiling of three heat shock protein 70 genes in the ectoparasitoid Habrobracon hebetor （Hymenoptera： Braconidae）

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Insect Science 2014年第4期21卷 415-428页

作者： Hao-Liang Chen Hong-Yu Zhang James E. Throne kun yan zhu State Key Laboratory of Agricultural Microbiology Institute of Urban and Horticultural Pests Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory College of Plant Science and Technology Huazhong Agricultural University Wuhan Institute of Plant Protection and Agro-Products Safety Anhui Academy of Agricultural Sciences Hefei China USDA Agricultural Research Service Center for Grain & Animal Health Research Manhattan KS USA Department of Entomology Kansas State University Manhattan KS USA

Heat shock proteins （HSPs） are known as chaperones that help with folding of other proteins when cells are under environmental stresses. The upregulation of HSPs is essential for cold survival during insect diapause. The ectoparasitoidHabrobracon hebetor, a potential biological control agent, can enter reproductive diapause when reared at low temperature and short photoperiod. However, the expression of HSPs during diapause of H. hebetor has not been studied. In this study, we sequenced and characterized the full-length complementary DNAs of three Hsp70 genes （HhHsp70I, HhHsp70II and HhHsp70IIl） from 11. hebetor. Their deduced amino acid sequences showed more than 80% identities to their counterparts from other insect species. However, the multiple se- quence alignment among the three deduced amino acid sequences of HhHsp70s showed only 46% identities. A phylogenetic analysis of the three HhHsp70s and all other known Hsp70 sequences from Hymenoptera clustered all the Hsp70s into four groups, and the three HhHsp70s were distributed into three different groups. Real-time quantitative poly- merase chain reaction analysis showed that the expression of the three HhHspTO genes in H. hebetor reared at different conditions was quite different. HhHspTOI showed higher relative expression when H. hebetor were reared at 27.5℃ than at two lower temperatures （17.5℃ and 20℃） regardless of the photoperiod, whereas HhHspTOII showed higher ex- pression when H. hebetor were reared at 20℃ and 10 ： 14 L ： D than when reared at 17.5℃ and either 16 ： 8 L ： D or 10 ： 14 L ： D. In contrast, HhHSP7OIIIwas expressed at similar levels regardless of the rearing conditions. These results may suggest functional differences among the three HhHspTO genes in H. hebetor.

关键词： ectoparasitoid gene expression Habrobracon hebetor heat shock protein70 phylogenetic analysis quantitative RT-PCR

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Clathrin heavy chain is essential for the development and reproduction of Locusta migratoria

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Insect Science 2022年第6期29卷 1601-1611页

作者： Xuekai Shi Shuai Li Lin yang Xiaojan Liu Hans Merzendorfer kun yan zhu Jianzhen Zhang Institute of Applied Biology Shanxi UniversityTaiyuanChina Collge of Life Science Shanxi UniversiyTaiyuanChina Institute of Biology University of SiegenSiegenGermany Department of Entomology Kansas State UniversityManhattanKansasUSA

Clathrin heavy chain(Chc)is a constituent of clathrin-coated vesicles and serves important functions in endocytosis and intracellular membrane trafficking but ap-pears to have physiological roles also at the organismal *** of what we know about Chc functions originates from studies performed in fungal or vertebrate ***-ever,the physiological functions of Chc in insects remain poorly ***,we identified a Chc ortholog from a Locusta migratoria transcriptome ***-qPCR revealed that LmChc was constitutively expressed in fifth-instar *** this develop-mental stage,LmChc showed the highest expression in the ovary followed by hemolymph,testis,hindgut,midgut,and *** isolated hemocytes,we detected the Chc protein in patches at the plasma *** examine the role of LmChc in *** during development,RNA interference was performed by injecting dsRNA into the early fifth-instar *** of LmChc caused a lethal phenotype with molting defect from nymph to *** addition,silencing of LmChc resulted in abnormal development of the ovaries,the size of which was significantly smaller than that in *** together,our results suggest that LmChc is a vital gene in *** that plays an important role in growth,development,and *** may be used as an efficient RNAi target gene for developing dsRNA-based biological insecticides to manage insect pests.

关键词： clathrin heavy chain development Locusta migratoria ovary RNA inter-ference

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Comparative analysis of cytochrome P450-like genes from Locusta migratoria manilensis： expression profiling and response to insecticide exposure

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Insect Science 2012年第1期19卷 75-85页

作者： yan-Qiong Guo Jian-Zhen Zhang Mei-Ling yang Liang-Zhen yan kun yan zhu Ya-Ping Guo En-Bo Ma Institute of Appfied Biology Shanxi UniversityTaiyuan College of Agriculture Shanxi Agricultural UniversityTaiguShanxi ProvinceChina Department of Entomology Kansas State UniversityManhattanKansasUSA

The cytochrome P450 monooxygenase （cytochrome P450） gene superfamily comprises many genes that may be involved in the biotransformations of pesticides and other xenobiotics. To date, very little is known about cytochrome P450 genes in the oriental migratory locust, Locusta migratoria manilensis. In this study, we carried out a genomewide analysis ofcytochrome P450 genes of the locust to identify putative cytochrome P450 genes and characterize their expression responses to insecticide exposures. We identified 15 cytochrome P450-1ike genes from a locust expressed sequence tag database （LocustDB）. Reverse transcription polymerase chain reaction （RT-PCR） analysis showed that most cytochrome P450-1ike genes displayed different tissue and developmental stage expression patterns. However, most of them were predominantly expressed in the midgnt, gastric caeca, fatbodies, and/or hindgut. Biochemical analysis showed that cytochrome P450 was differentially affected by three different insecticides. Deltamethrin caused significant inductions in 12 h at LD30 （dose to kill 30% of the tested individuals） in the nymphs, whereas malathion and carbaryl did not have significant effect on cytochrome P450 enzyme activity. Further RT-PCR analysis Showed significant increases of transcriptions of several cytochrome P450 genes in deltamethrin-treated locusts. Thus, the increased cytochrome P450 enzyme activity is likely due to increased transcriptions of multiple cytochrome P450 genes in response to deltamethrin exposure. These results are expected to help us better understand the interactions between insecticides and major detoxification enzymes, and possible changes of the susceptibility to other insecticides in deltamethrin-treated insects at various molecular levels.

关键词： cytochrome P450 monooxygenase deltamethrin gene expression profiling insecticide Locusta migratoria manilensis

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The microRNA miR-184 regulates the CYP303A1 transcript level to control molting of Locusta migratoria

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Insect Science 2021年第4期28卷 941-951页

作者： yan-Li Wang Li-Xian Wu Hui-Yong Li Xue-Qin Wen En-Bo Ma kun-yan zhu Jian-Zhen Zhang Research Institute of Applied Biology Shanxi UniversityTaiyuanShanxiChina College of Life Science Shanxi UniversityTaiyuanShanxiChina Department of Entomology Kansas State UniversityManhattanKSUSA

Cytochrome P450 monooxygenases(CYPs)play essential physiological functions in ***303A1 is highly conserved in insect species studied to date,and shows an indispensable role for adult eclosion in both Locusta migratoria and Drosophila ***,how CYP303A1 is regulated to control insect developmental processes remains *** this study,we discovered functional binding sites for miR-184 in the coding sequence of *** luciferase reporter assay showed that miR-184 could target LmCYP303A1 and regulate its expression in ***,overexpression of miR-184 through microinjection of agomir to locusts reduced the transcripts of LmCYP303A1 and led to abnormal molting,which is similar to the phenotype of silencing LmCYP303A1 by direct injection of dsLmCYP303A1 to ***,down-regulation of miR-184 by injection of antagomir increased the LmCYP303A1 transcript and caused molting *** findings suggested that miR-184 could target LmCYP303A1 to regulate the molting process in ***,which might be considered as a novel target for pest control.

关键词： CYP303A1 gene regulation insect molting Locusta migratoria miR-184

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Characteristics and roles of cytochrome b5 in cytochrome P450-mediated oxidative reactions in Locusta migratoria

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Journal of Integrative Agriculture 2020年第6期19卷 1512-1521页

作者： LIU Jiao ZHANG Xue-yao WU Hai-hua MA Wen zhu Wen-ya kun-yan zhu MA En-bo ZHANG Jian-zhen Institute of Applied Biology Shanxi UniversityTaiyuan 030006P.R.China College of Life Science Shanxi UniversityTaiyuan 030006P.R.China Modern Research Center for Traditional Chinese Medicine Shanxi UniversityTaiyuan 030006P.R.China Institute of Plant Protection Shanxi Academy of Agricultural SciencesTaiyuan 030031P.R.China Department of Entomology Kansas State UniversityManhattanKS 66506USA

Cytochrome b5(Cyt-b5)is a small heme protein and known to be involved in a wide range of biochemical transformations,in eluding cytochrome P450 monooxyge nase(CYP)-mediated metabolism of endoge nous and exogenous compo un *** on Cyt-b5 are more con centrated in mammals,but are relatively rare in in *** characteristics and functi on of Cyt-b5 from Locusta migratoria have not been described *** sequeneed the full-length cDNA sequenee of Cyt-b5 from ***(LmCyt-b5)by reverse transcription-PCR(RT-PCR)based on locust transcriptome *** phylogenetic analysis showed that LmCyt-b5 was closely related to the Cyt-b5 from ***-b5 was highly expressed in ovary,Malpighian tubules,midgut,gastric caeca,and fat *** of LmCyt-b5 had no effect on the susceptibility of *** to four different *** of LmCyt-b5 or silencing of both LmCyt-b5 and LmCPR did not significantly change the total CYP activity toward the substrate 7-ethoxycoumarin(7-EC).However,coexpression of LmCYP6FD1 with LmCPR and LmCyt-b5 together in Sf9 cells by using Bac-to-Bac baculovirus expression system significantly increased the catalytic activity of LmCYP6FD1 toward 7-EC as compared with the coexpression of ***6FD1 with cytochrome P450 reductase(LmCPR)or LmCyt-b5 *** results suggest that LmCyt-b5 plays an important role in the catalytic reaction of LmCYP6FD1 toward 7-EC in our in vitro *** study is needed to clarify the role of LmCyt-b5 in CYP-mediated catalytic reactions in ***.

关键词： cytochrome b5 cytochrome P450 cytochrome P450 reductase Locusta migratoria RNA interference

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