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Toxicological and biochemical characterizations of malathion sensitivity in two field populations of Oxya chinensis （Orthoptera： Acridoidea）

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Insect Science 2006年第1期13卷 41-47页

作者： MEI-LING YANG HAI-HUA WU YA-PING GUO en-bo ma College of Life Science and Technology Shanxi University Taiyuan China

We evaluate comparative toxicity of malathion in the two populations of the grasshopper Oxya chinensis, collected from Daixian and Fanshi of Shanxi province, China. General esterases and acetylcholinesterase （ACHE） from the two populations were characterized and compared. LD50 of the Daixian population （7.58 μg/g body weight） was 2.02-fold higher than that of the Fanshi population （3.75μg/g body weight）. General esterase-specific activities in the Daixian population were 1.91,130 and 1.85-fold higher than those in the Fanshi population, when α-NA, α-NB and β-NA were used as a substrate, respectively. Kinetic studies of general esterase showed that Vmax values of general esterases hydrolyzing α-NA,α-NB and β-NA in the Daixian population were 2.15-, 1.12-, and 1.47-fold, respectively, higher than those in the Fanshi population. The AChE activity of the Fanshi population was 1.54-fold higher than that of the Daixian population. Kinetic analysis of AChE showed that significant differences were presented between the two populations in the Km values; and the Vmax value in the Fanshi population was higher than that in the Daixian population. Inhibition studies of AChE indicated that AChE from the Daixian population was 2.56-, 2.80-, and 2.29-fold less sensitive to inhibition by paraoxon, chlorpyrifos-oxon, and demeton-S-methyl, respectively, than that from the Fanshi population. These biochemical characterizations of general esterases and AChE were consistent with malathion bioassay in the two populations. It is inferred that the reduced sensitivity of altered AChE and increased general esterase activities play an important role in the differences of insusceptibility of Oxya chinensis to malathion between the two populations.

关键词： acetylcholinesterase general esterases malathion susceptibility Oxya chinensis

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Identification of LmUAP1 as a 20-hydroxyecdysone response gene in the chitin biosynthesis pathway from the migratory locust, Locusta migratoria

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Insect Science 2018年第2期25卷 211-221页

作者： Xiao-Jian Liu Ya-Wen Sun Da-Qi Li Sheng Li en-bo ma[] Jian-Zhen Zhang Research Institute of Applied Biology College of Life Science Shanxi University Taiyuan China Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

In Locusta migratoria, we found that two chitin biosynthesis genes, UDP N-acetylglucosamine pyrophosphorylase gene LmUAP1 and chitin synthase gene Lm- CHS1, are expressed mainly in the integument and are responsible for cuticle formation. However, whether these genes are regulated by 20-hydroxyecdysone （20E） is still largely unclear. Here, we showed the developmental expression pattern ofLmUAP1, LmCHS1 and the corresponding 20E titer during the last instar nymph stage of locust. RNA interference （RNAi） directed toward a common region of the two isoforms of LmEcR （LmEcRcom） reduced the expression level of LmUAP1, while there was no difference in the expression of LmCHS1. Meantime, injection of 20E in vivo induced the expression of LmUAP1 but not LmCHS1. Further, we found injection-based RNAi of LmEcRcom resulted in 100% mortality. The locusts failed to molt with no apolysis, and maintained in the nymph stage until death. In conclusion, our preliminary results indicated that LmUAP1 in the chitin biosynthesis pathway is a 20E late-response gene and LmEcR plays an essential role in locust growth and development, which could be a good potential target for RNAi-based pest control.

关键词： chitin synthesis Locusta migratoria regulation RNA interference 20-hydroxyecdysone

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Characteristics of Halloween genes and RNA interference-mediated functional analysis of LmCYP307a2 in Locusta migratoria

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Insect Science 2022年第1期29卷 51-64页

作者： Xue-Yao Zhang Qi-Hui He Ting-Ting Zhang Hai-Hua Wu Jian-Zhen Zhang en-bo ma Research Institute of Applied Biology Shanxi UniversityTaiyuanShanxi030006China

Halloween genes are involved in the biosynthesis of the molting hormone, which plays a key role in insect ecdysis, development, metamorphosis, and reproduction. Our previous work identified five Halloween genes from Locusta migratoria, but their functions are currently unknown. In this study, the sequences of these five Halloween genes were analyzed and characterized. LmCYP307a2, LmCYP306a1, LmCYP302a1, and LmCYP315a1 were primarily expressed in the prothoracic glands, while LmCYP314a1 was universally expressed in peripheral tissues, especially in the ovaries and malpighian tubules. All five Halloween genes were mainly expressed from the 5th to the 7th d in 5th-instar nymphs. RNA interference (RNAi) silencing of LmCYP307a2 resulted in severe molting delays and molting failure, which could be rescued by supplementary 20-hydroxyecdysone. A hematoxylin and eosin staining analysis suggested that the RNAi of LmCYP307a2 inhibited the ecdysis process by inhibiting the apolysis and degradation of the old cuticle, and by promoting the synthesis of a new cuticle. Quantitative reverse transcription polymerase chain reaction results showed that the expressions of LmE74, LmCht5, and LmCht10 were dramatically down-regulated, while that of LmChsI was substantially up-regulated, after knockdown of LmCYP307a2. The results suggest that LmCYP307a2 is related to the molt process via regulation of chitin synthesis and degradation.

关键词： Halloween genes Locusta migratoria RNAi silencing Spookier

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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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Heterologous expression and characterization of two chitinase 5 enzymes from the migratory locust Locusta migratoria

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Insect Science 2016年第3期23卷 406-416页

作者： Ying-Long Li Hui-Fang Song Xue-Yao Zhang Da-Qi Li Ting-Ting Zhang en-bo ma Jian-Zhen Zhang Research Institute of Applied Biology College of Life Science Shanxi University Taiyuan China

Insect chitinases are involved in degradation of chitin from the exoskeleton or peritrophic metrix ofmidgut. In Locusta migratoria, two duplicated Cht5s （LmCht5-1 and LmCht5-2） have been shown to have distinct molecular characteristics and biological roles. To explore the protein properties of the two LmCht5s, we heterologously expressed both enzymes using baculovirus expression system in SF9 cells, and characterized kinetic and carbohydrate-binding properties of purified enzymes. LmCht5-1 and LmCht5-2 exhibited similar pH and temperature optimums. LmCht5-1 has lower Km value for the oligomeric substrate （4MU-（GlcNAe）3）, and higher Km value for the longer substrate （CM-Chitin- RBV） compared with LmCht5-2. A comparison of amino acids and homology modeling of catalytic domain presented similar TIM barrel structures and differentiated amino acids between two proteins. LmCht5-1 has a chitin-binding domain （CBD） tightly bound to colloidal chitin, but LmCht5-2 does not have a CBD for binding to colloidal chitin. Our results suggested both LmCht5- I and LmCht5-2, which have the critical glutamate residue in region II of catalytic domain, exhibited chitinolytic activity cleaving both polymeric and oligomeric substrates. LmCht5-1 had relatively higher activity against the oligomeric substrate, 4MU-（GlcNAc）3, whereas LmCht5-2 exhibited higher activity toward the longer substrate, CM-Chitin-RBE These findings are helpful for further research to clarit}g their different roles in insect growth and development.

关键词： chitinase 5 enzyme kinetics heterologous expression Locusta migratoria

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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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RNA interference to reveal roles of β-N-acetylglucosaminidase gene during molting process in Locusta migratoria

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Insect Science 2013年第1期20卷 109-119页

作者： Shuo Rong Da-Qi Li Xue-Yao Zhang Sheng Li Kun Yan Zhu Ya-Ping Guo en-bo ma Jian-Zhen Zhang Research Institute of Appfied Biology Shanxi University Taiyuan Institute of Plant Physiology and Ecology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Shanghai China Department of entomology 123 Waters Hall Kansas State University Manhattan KS 66506 USA

β-N-acetylglucosaminidases are crucial enzymes involved in chitin degrada- tion in insects. We identified a β-N-acetylglucosaminidase gene （LmNAG1） from Locusta migratoria. The full-length complementary DNA （cDNA） of LmNAG1 consists of 2 667 nucleotides, including an open reading frame （ORF） of 1 845 nucleotides encoding 614 amino acid residues, and 233- and 589-nucleotide non-coding regions at the 5＇- and 3＇- ends, respectively. Phylogenetic analysis grouped the cDNA-deduced LmNAG1 protein with the enzymatically characterized β-N-acetylglucosaminidases in group I. Analyses of stage- and tissue-dependent expression patterns of LmNAG1 were carried out by real- time quantitative polymerase chain reaction. Our results showed that LmNAG1 transcript level in the integument was significantly high in the last 2 days of the fourth and fifth instar nymphs. LmNAG1 was highly expressed in foregut and hindgut. RNA interference of LmNAG1 resulted in an effective silence of the gene and a significantly reduced total LmNAG enzyme activity at 48 and 72 h after the injection of LmNAG1 double-stranded RNA （dsRNA）. As compared with the control nymphs injected with GFP dsRNA, 50% of the dsLmNAGl-injected nymphs were not able to molt successfully and eventually died. Our results suggest that LmNAG1 plays an essential role in molting process ofL. migratoria.

关键词： β-N-acetylglucosaminidase chitin degradation Locusta migratoria RNA interference

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A dsRNA-degrading nuclease(dsRNase2)limits RNAi efficiency in the Asian corn borer(Ostrinia furnacalis)

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Insect Science 2021年第6期28卷 1677-1689页

作者： Yun-He Fan Hui-Fang Song Mureed Abbas Yan-Li Wang Tao Li en-bo ma Anastasia M.W.Cooper Kristopher Silver Kun Yan Zhu Jian-Zhen Zhang Institute of Applied Biology Shanxi UniversityTaiyuanChina College of Life Science Shanxi UniversityTaiyuanChina Faculty of Biological Science and Technology Changzhi UniversityChangzhiShanxiChina Department of entomology Kansas State UniversityManhattanKansasUSA

The efficiency of RNA interference(RNAi)varies substantially among different insect *** degradation of double-stranded RNA(dsRNA)by dsRNA-degrading nucleases(dsRNases)has been implicated to cause low RNAi efficiency in several insect *** this study,we identified four dsRNase genes(OfdsRNaseL Ofd-sRNase2,OfdsRNase3 and OfdsRNase4)from the Asian corn borer(Ostrinia furnacalis)transcriptome *** analyses showed that each deduced protein sequence contained endonuclease NS domains and signal *** expression analysis revealed that OfdsRNase2 was exclusively expressed in the midgut of *** efficiency was investigated in 2-d-old fifth-instar larvae(high expression of dsRNase2)and 2-d-old pupae(low expression of dsRNase2)by feeding or injecting dsRNA targeting a marker gene that encodes the lethal giant larvae protein(OfLgl).Our results showed that OfLgl only partially silenced the expression of OfLgl in pupae,but not in larvae,suggesting that OfdsRNase2 could contribute to lower RNAi efficiency in larval *** hypothesis was supported by our RNAi-of-RNAi experiment using a tissue culture technique where the silencing efficiency against the reporter gene,OfHexl,was significantly improved after knockdown of *** double luciferase assays were performed to evaluate the role of the four dsRNases in vitro,only OfdsRNase2 expressed in S2 cells significantly affected RNAi efficiency by degrading *** together,our results suggested that the degradation of dsRNA by OfdsRNase2 in the midgut contributed to low RNAi efficiency in *** larvae.

关键词： dsRNase Ostrinia furnacalis RNA interference RNAi efficiency tissue culture

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PHYLOGenETIC RELATIONSHIP OF ACRIDOIDEA BASED ON THE maLE FOLLICLE MORPHOLOGY （ORTHOPTERA: ACRIDOIDEA）

PHYLOGENETIC RELATIONSHIP OF ACRIDOIDEA BASED ON THE MALE FO...

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第十九届国际动物学大会

作者： en-bo ma Xin Liu and Ya-Ping GuoCollege of Life Science and Technology Shanxi University Taiyuan 030006 China

＜正＞Taxonomy of Acridoidea has been extensively studied, mostly using morphological characteristics-However, the features of the male and female reproductive system may provide useful information to infer the taxono...

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FK506 contributes to peripheral nerve regeneration by inhibiting neuroinflammatory responses and promoting neuron survival

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Neural Regeneration Research 2025年第7期20卷 2108-2115页

作者： Yuhui Kou Zongxue Jin Yusong Yuan bo ma Wenyong Xie Na Han Department of Trauma and Orthopedics Peking University People’s HospitalBeijingChina National Center for Trauma Medicine Peking University People’s HospitalBeijingChina Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina Trauma Medicine Center Peking University People’s HospitalBeijingChina Department of Orthopedics China-Japan Friendship HospitalBeijingChina

FK506(Tacrolimus)is a systemic immunosuppressant approved by the *** and Drug ***506 has been shown to promote peripheral nerve regeneration,however,its precise mechanism of action and its pathways remain *** this study,we established a rat model of sciatic nerve injury and found that FK506 improved the morphology of the injured sciatic nerve,increased the numbers of motor and sensory neurons,reduced inflammatory responses,markedly improved the conduction function of the injured nerve,and promoted motor function *** findings suggest that FK506 promotes peripheral nerve structure recovery and functional regeneration by reducing the intensity of inflammation after neuronal injury and increasing the number of surviving neurons.

关键词： FK506 inflammation motor neurons nerve regeneration neuron peripheral nerve injury sensory neurons

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