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  • Monthly, Founded in 1950
    Supervisor:Chinese Academy of Sciences
    Sponsor:Institute of Zoology,Chinese Academy of Sciences
    The Entomological Society of China
    Domestic postal code: 2-153
    Foreign issuance code: Q61
    ISSN 0454-6296
    CN 11-1832/Q
Table of Content
20 December 2018, Volume 61 Issue 12
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  • RESEARCH PAPERS
    Analysis of circular RNAs and their regulatory networks in the midgut of Apis mellifera ligustica workers
    XIONG Cui-Ling, CHEN Hua-Zhi, CHEN Da-Fu, ZHENG Yan-Zhen, FU Zhong-Min, XU Guo-Jun, DU Yu, WANG Hai-Peng, GENG Si-Hai, ZHOU Ding-Ding, LIU Si-Ya, GUO Rui
    2018, 61(12):  1363-1375.  doi:10.16380/j.kcxb.2018.12.001
    Abstract ( 452 )   PDF (10564KB) ( 153 )      Annex for review
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    Aim Circular RNA (circRNA) plays a key role in alternative splicing, transcription regulation and expression regulation of source genes. This study aims to analyze the quantity, variety, structural characteristics and function of circRNAs in the midgut of Apis mellifera ligustica workers, and to explore the regulatory function of circRNAs via constructing and analyzing regulatory networks. Methods A. m. ligustica workers were reared under laboratory conditions, and the midgut samples from 7- and 10-day-old workers were subjected to deep sequencing using circRNA-seq technology. CircRNAs were predicted from sequencing data after quality control using find_circ software. Source genes of these circRNAs were annotated to GO and KEGG databases to gain function and pathway annotations via BLAST. The target mRNAs of circRNAs and miRNAs were predicted with TargetFinder software, and the regulatory networks between circRNAs and miRNAs and between circRNAs, miRNAs and mRNAs were constructed and visualized using Cytoscape v.3.2.1software. The predicted circRNAs were validated by RT-PCR with the divergent and convergent primers designed. Results Sequencing of midgut samples from A. m. ligustica workers produced a mean of 136 463 071 clean reads, and for each sample over 136 779 122 anchor reads were obtained after removing rRNA. A total of 10 833 circRNAs were predicted, and their length ranged mainly from 15 to 1 000 nt. The types of these circRNAs were abundant, and the largest one was annotated to be exonic circRNA. The number of circRNAs distributed on chromosome 1 of Apis mellifera was the most and that distributed on chromosome 8 was the second. The source genes of circRNAs could be annotated to 45 GO terms including binding, cellular process, and cell, as well as 121 KEGG metabolic pathways including endocytosis, protein processing in endoplasmic reticulum, and ribosome, suggesting that circRNAs may play key roles in such biological processes as growth, development, metabolism, and cellular activity in the midgut of A. m. ligustica workers. Furthermore, the regulatory networks between circRNA and miRNA and between circRNA, miRNA and mRNA were constructed, and the analysis result demonstrated that partial circRNAs could bind microRNAs as competitive endogenous RNAs (ceRNAs). Finally, the true existence of randomly selected three circRNAs was validated by RT-PCR. Conclusion In this study, we predicted, analyzed, and verified circRNAs in the midgut of A. m. ligustica workers. Our findings provide the data of the quantity, variety, structural characteristics, roles, and regulatory network of circRNAs in the midgut, indicating that circRNAs can play roles in the developmental and immune defense processes in the midgut of A. m. ligustica workers through affecting source genes and acting as ceRNAs. This study lays a foundation for further studying the roles of circRNAs in the development and stress response of the midgut of A. m. ligustica.
    Cloning and functional characterization of the NanosO gene promoter in Plutella xylostella (Lepidoptera: Noctuidae)
    WANG Ya-Jun, HUANG Yu-Ping, YU Hui-Hui, XU Xue-Jiao, YANG Guang, AI Qian-Qian, YOU Min-Sheng
    2018, 61(12):  1376-1383.  doi:10.16380/j.kcxb.2018.12.002
    Abstract ( 439 )   PDF (6689KB) ( 159 )     
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    Aim To clone the NanosO gene (PxnosO) promoter from the diamondback moth, Plutella xylostella, and to verify its gonad-specific activity, so as to further study the gene functions and develop transgenic insects for integrated management of P. xylostella and other agricultural pests.MethodsAccording to the sequence information of NanosO gene in the P. xylostella genome, the NanosO gene promoter was cloned by PCR and subjected to sequence analysis. The PxnosO-EGFP expression plasmid was constructed, and then the PxnosO-EGFP and IE1-EGFP expression plasmids were transfected into P. xylostella embryonic cell line (Px-6) and Spodoptera frugiperda ovary cell line (Sf9) by lipofection. The activity of the NanosO promoter in P. xylostella was confirmed by the qualitative and quantitative analysis of EGFP expression by laser confocal fluorescence observation and qRT-PCR, respectively. Results We cloned the PxnosO (Px004767) promoter (with the length of 1 743 bp) of P. xylostella. Sequence analysis showed that this promoter not only contains common core promoter elements like TATA box, and upstream promoter domains TATA box and GC box, but also contains dozens of transcription factor binding sites. Using cell transfection technology, we successfully expressed exogenous gene EGFP driven by this promoter in the Px-6 and Sf9 cell lines. Conclusion We cloned the PxnosO promoter of P. xylostella and verified that it can drive the expression of EGFP gene at the cellular level. This study provides a foundation for analyzing the expression pattern of PxnosO in different developmental stages of P. xylostella and the potential functions of the PxnosO promoter in vivo.
    Characterization of fork head transcription factor 1 gene FOXO1 and its role in sugar and lipid metabolism in the summer prediapause pupae of Delia antiqua (Diptera: Anthomyiidae) (In English)
    XU Yan-Ling, PENG Zhu-Qing, SI Feng-Ling, HAO You-Jin
    2018, 61(12):  1384-1392.  doi:10.16380/j.kcxb.2018.12.003
    Abstract ( 560 )   PDF (594KB) ( 150 )     
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    AimTo clone and identify fork head transcription factor 1 gene in the onion fly, Delia antiqua, so as to explore its roles in sugar and lipid metabolism in the summer prediapause pupae of this insect. Methods The full-length open-reading frame of fork head transcription factor 1 gene was cloned based on the transcriptome data of D. antiqua by 3 RACE method. The sequence characteristics, conserved domains and secondary structure of its encoded protein were analyzed using bioinformatic approaches. A maximum-likelihood phylogenic tree was constructed for the target protein and homologues from other thirteen insect species. After the target gene was silenced in the summer prediapause pupae of D. antiqua by RNA interference, the expression levels of its downstream genes including brummer gene (DaBmm) and phosphoenol-pyruvate carboxykinase gene (DaPepck) were measured by qPCR, and the contents of triacylglycerol (TAG), treaholse and glucose, and the total lipase activity were analyzed. Results The full-length cDNA sequence of the fork head transcription factor 1 gene DaFOXO1 was cloned from D. antiqua, and deposited in the GenBank under the accession number MG813258. It encodes a 619-amino-acid protein with a typical fork head DNA binding domain, nuclear localization signal, two14-3-3binding motifs and a glutamine-rich domain. DaFOXO1 shares 87% sequence identity with FOXO1 of Lucilia cuprina at the amino acid level and they are clustered together. DaFOXO1 knockdown in the summer prediapause pupae of D. antiqua significantly suppressed the expression of DaBmm between 8 h and 20 h after treatment, significantly affected the TAG content and total lipase activity between 12 h and 24 h after treatment, and significantly decreased the expression level of DaPepck between 8 h and 20 h(except 16 h) after treatment. Neither glucose content nor trehalose content in the dsDaFOXO1-treated summer prediapause pupae of D. antiqua significantly changed. Conclusion The results indicate that the accumulation of trehalose and glucose has completed before pupation, and the regulatory role of DaFOXO1 inthe expressions of DaBmm and DaDepck may contribute to the accumulation of lipid in the summer prediapause pupae of D. antiqua. The results also suggest that targeting Bmm-dependent lipolysis and its downstream factors may provide a strategy to disrupt insect diapause.
    Cloning and expression profiling of circadian clock gene HeDbt in Helicoverpa armigera (Lepidoptera: Noctuidae)
    YAN Shuo, LIU Yan-Jun, ZHANG Xin-Fang, ZHU Jia-Lin, LI Zhen, LIU Xiao-Ming, ZHANG Qing-Wen, LIU Xiao-Xia
    2018, 61(12):  1393-1403.  doi:10.16380/j.kcxb.2018.12.004
    Abstract ( 640 )   PDF (1605KB) ( 291 )     
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    Aim The aim of this study is to clone and analyze circadian clock gene Double-time (Dbt) in Helicoverpa armigera, and to examine the diurnal expression pattern of Dbt mRNA levels and its determinants, so as to provide a theoretical basis for studying the mechanisms of action of the circadian clock genes in the compound eyes and understanding the function of circadian clock genes in peripheral tissues in noctuid moths. Methods Dbt was cloned from compound eyes of 2-day-old female adults of H. armigera by RT-PCR and RACE, and its deduced amino acid sequence was analyzed using online sites and software. The expression levels of Dbt among different tissues (head, brain, compound eyes, antennae, thorax, abdomen, leg and wing) of female and male adults were determined and compared by qPCR. The diurnal changes of Dbt mRNA levels in compound eyes and head were measured under a photoperiod of14L10D and constant darkness (DD). The 2-day-old adults were irradiated by sensitive wavelength of UV, blue and green lights, respectively, for 6 h from the beginning of the scotophase, and the changes in the expression levels of Dbt in compound eyes were determined after light exposure. Female and male adults were paired for mating during scotophase, and the changes in the expression levels of Dbt in compound eyes were determined at 0 h and 3 h after copulation, respectively. Results The full-length cDNA of Dbt gene was cloned from the compound eyes of H. armigera and designated as HeDbt (GenBank accession number: KM233159). It has a 1 026 bp open reading frame encoding a predicted protein of 314 amino acids, with a molecular mass of 39.79 kD and a calculated isoelectric point (pI) of 9.55. Its deduced amino acid sequence has no transmembrane topologies, and shares several typical conserved domains of insect DBT. HeDBT shows a high homology with DBT proteins from Spodoptera exigua (99% amino acid sequence identity) and Antheraea pernyi (97% amino acid sequence identity). qPCR results illustrated that HeDbt was expressed in all the tested adult tissues, and had low expression levels in the head, brain and compound eyes and high expression levels in thorax and abdomen. The expression of HeDbt showed no obvious circadian rhythm in both the head and compound eyes under14L10D and DD. The expression of HeDbt was down-regulated in compound eyes after light exposure and copulation, but with similar HeDbt mRNA levels between female and male adults. Conclusion A circadian clock gene HeDbt was successfully cloned from H. armigera. HeDbt had low mRNA levels in the head and compound eyes of H. armigera adults. Its expression in the head and compound eyes showed no obvious circadian rythym, but was influenced by light exposure and copulation of adults. Our study provides a theoretical basis for further studying the function of circadian clock genes in peripheral tissues in noctuid moths.
    Control efficacy of ozone water against Bradysia odoriphaga (Diptera: Sciaridae) larvae and its influence on the growth of Chinese chives
    HU Jing-Rong, SHI Cai-Hua, XU Bao-Yun, WEI Qi-Wen, XIE Wen, LI Chuan-Ren, ZHANG You-Jun
    2018, 61(12):  1404-1413.  doi:10.16380/j.kcxb.2018.12.005
    Abstract ( 585 )   PDF (1167KB) ( 247 )     
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     Aim To determine the optimum irrigation period and application condition for controlling the larvae of the chive maggot, Bradysia odoriphaga, scientifically and rationally with irrigating ozone (O3) water. Methods We investigated the control efficacy of different concentrations of O3 water (5, 10, 20 and 30 mg/L) against B. odoriphaga larvae, the control efficacy of 30 mg/L O3 water against B. odoriphaga larvae in flat land covered with plastic film under different sunlight intensities, in three different irrigation conditions (flat land covered with plastic film, small arch plastic shed, and small arch plastic shed covered with petates), and at various irrigation frequencies of 30 mg/L O3 water under two different irrigation environments (flat land covered with plastic film and small arch plastic shed), respectively. And we also investigated the effects of 30 mg/L O3 water on the yield and root growth of Chinese chives in flat land covered with plastic film. In addition, we analyzed the influence of sunlight intensity on soil temperatures and the control efficacy of irrigating ozone water under different irrigation environments. Results In the flat land covered with plastic film, the optimum concentration of O3 water for the control against B. odoriphaga larvae was 20-30 mg/L. In treatments of irrigating 30 mg/L O3 water to flat land covered with plastic film, the control efficacy of O3 water against B. odoriphaga larvae was positively related to the sunlight intensity on the day of O3 water irrigation, the larval mortality of B. odoriphaga was up to 100% at 1 d after irrigation when the sunlight intensity was over 60 000 lx, while no significant control efficacy was found when the sunlight intensity was below 10 000 lx. When the sunlight intensity was moderate, the prevention and control efficacy of 30 mg/L O3 water against B. odoriphaga larvae was poor in the early stage, and gradually enhanced in the late stage. The effect of soil warming was positively correlated with sunlight intensity and significantly related with irrigation environments. The temperature of5 cmdeep soil under different irrigation conditions was as follows: flat land covered with plastic film> small arch plastic shed> small arch plastic shed covered with petates. The control efficacy of O3 water irrigated into the small arch plastic shed against B. odoriphaga larvae was significantly higher than that irrigated into the flat land covered with plastic film and that irrigated into the small arch plastic shed covered with petates. Repeatedly continuous irrigating of O3 water could improve the control efficacy of O3 water against B. odoriphaga larvae, and the control efficacy in the small arch plastic shed was more remarkable. When O3 water was irrigated, the distance between the inlet and the end of the farmland should not exceed40 m. After irrigating 30 mg/L O3 water in the plat land covered with plastic film, the plant height, weight and the number of fibrous roots of Chinese chives were significantly higher than those of the control (irrigating water), but the number of tillers per cluster of Chinese chive showed no significant difference from the control. Conclusion When the O3 water is irrigated scientifically and reasonably, it not only shows ideal control efficacy against B. odoriphaga larvae, but also can promote the growth of Chinese chives.
    Influence of fatty acids on the necrophoric behavior of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae)
    XU Yan-Qin, CHEN Li, WANG Wen-Kai
    2018, 61(12):  1414-1420.  doi:10.16380/j.kcxb.2018.12.006
    Abstract ( 523 )   PDF (1017KB) ( 147 )     
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    Aim After death, worker corpse is vulnerable to pathogen infection, which is harmful to the health of the entire nest of ants. To avoid horizontal transmission of pathogens, live workers recognize corpses by changes in chemical cues from corpses and removed them to refuse piles. This study aims to test the behavioral response of workers of the red imported fire ant, Solenopsis invicta to six fatty acids including oleic acid, linoleic acid, palmitokic acid, palmitic acid, stearic acid and myristic acid, which may play important roles in its necrophoric behavior. Methods The fatty acid components of body extracts of live and dead ant workers were analyzed with GC-MS. The necrophoric behavioral responses of S. invicta workers to six fatty acids (10 μg/μL), the single component oleic acid or linoleic acid at different concentrations (0.75 and 3 μg/μL) and their mixture at different concentrations (0.01, 0.1, 1 and 10 μg/μL) were tested with filter paper disc method. Results Only linoleic and oleic acids were detected from the corpses of S. invicta worker. The fire ant workers did not significantly respond to four fatty acids, i.e., palmitokic acid, palmitic acid, stearic acid and myristic acid. Both single linoleic acid and oleic acid could trigger significant necrophoric behavior of fire ant workers, which was dose-dependent. The mixture of linoleic acid and oleic acid at the 10 μg/μL concentration could also trigger significant necrophoric behavior of fire ant workers, while showed no significant effect at other lower concentrations.Conclusion The results suggest that oleic acid and linoleic acid, but not other fatty acids, play important roles in triggering necrophoric behavior of S. invicta workers.
    Comparison of female reproductive fitness and offspring life history traits between gamogenesis and parthenogenesis in the camphor sawfly, Mesoneura rufonota (Hymenoptera: Tenthredinidae)
    XU Chuan-Feng, SHI Hao-Ni, YIN Li-Xin, ZHOU Jia-Ying, LIU Xing-Ping
    2018, 61(12):  1421-1429.  doi:10.16380/j.kcxb.2018.12.007
    Abstract ( 724 )   PDF (1121KB) ( 183 )     
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    Aim The camphor sawfly, Mesoneura rufonota, is an important defoliator of the camphor tree Cinnamonum campora. The reproduction strategy of this insect includes gamogenesis and parthenogenesis. This study aims to clarify the biological significance of parthenogenesis in the life history of the camphor sawfly. Methods The differences in female reproductive fitness of M. rufonota including female adult longevity, fecundity and egg hatching rate and offspring life history traits including the duration of various developmental stages, mortality, sex ratio and fecundity of offspring between parthenogenesis and gamogenesis at the constant temperature of25were determined and analyzed. Results The female adult longevity of M. rufonota was longer for parthenogenesis than for gamogenesis, while the fecundity and egg hatching rate were similar between the two reproduction modes. There were no differences in the duration and mortality of various developmental stages and the fecundity of female offspring between parthenogenesis and gamogenesis. However, significant difference was found in the sex ratio of the offspring between parthenogenesis and gamogenesis, being displayed by that parthenogenesis resulted in male-biased sex ratio, while gamogenesis resulted in female-biased sex ratio. Conclusion The parthenogenesis of the camphor sawfly prolongs the longevity of female adult, which has been confirmed as arrhenotoky. These results showed that parthenogenesis of the camphor sawfly not only makes it possible to establish population, but also can provide a large number of males to offset the insufficient male individuals in gamogenesis.
    Effects of the mutualism between ants and lac insects and host plant diversity on various trophic levels of arthropods in lac plantation
    WANG Qing, LU Zhi-Xing, ZHAO Jing-Wen, CHEN You-Qing
    2018, 61(12):  1430-1438.  doi:10.16380/j.kcxb.2018.12.008
    Abstract ( 472 )   PDF (1219KB) ( 140 )   PDF(mobile) (1219KB) ( 11 )     
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    Aim Mutualism between insects and plant diversity play an important role in the ecosystem. The purpose of this study is to investigate the top-down effects of the mutualism between ants and lac insects and the bottom-up effects of plant species diversity on various trophic levels of the arthropod community. Methods The experimental plots were located in a lac plantation in Yayi town, Mojiang county, Puer city, Yunnan province, southwestern China. In August and September, 2016, following a randomized split-plot experiment design, we collected all the arthropods on the host plants under three treatments of ant-lac insect mutualism (presence of mutualism, absence of mutualism, and natural control) and three levels of plant diversity (monoculture, bi-species plantation and tri-species plantation representing plant diversity 1, 2 and 3, respectively) in the experimental plots by using hand-collection, shake-down and sweeping methods, along with classification of all arthropods according to different trophic levels. Results Mutualism between ants and lac insects significantly reduced the consumer abundancepresence of mutualism (14.2±0.95)±1.13)±1.38)(F=56.03, P<0.001), plant diversity increased the consumer abundance significantly plant diversity 1 (13.49±2.54)±2.50)±2.47) (F=4.290, P=0.017). Under the interaction effect of mutualism and plant diversity, consumer abundance in the treatment with the presence of mutualism was significantly lower than that in the treatment with absence of mutualism in all the three levels of plant species diversity (F=6.850, P<0.001). Mutualism had no significant effect on the predator abundance (F=1.277, P=0.284), plant diversity increased the predator abundance significantly plant diversity 1 (1.72±0.49)±0.54)±0.55) (F=10.976, P<0.001), and the interaction between plant diversity and mutualism had no significant effect on the predator abundance either (F=0.428, P=0.788). Mutualism and plant diversity significantly increased the ant abundancenatural control (44.08±8.94)±65.35) (F=48.9, P<0.001), plant diversity 1 (86.31±12.51)±18.80)±21.68) (F=137.85, P<0.001); and the interaction between them had a significant effect on the ant abundance(F=80.31, P<0.001). Conclusion Ant-lac insect mutualism significantly reduces the herbivorous pests on the plant, and has no significant effect on predators, but significantly increases the ant abundance. The plant diversity has a positive effect on various trophic levels of arthropods, and ant-lac insect mutualism and plant diversity have a certain combined effect, causing different effects on each trophic level.
    Diversity and stability of terrestrial insect community in different wetlands in Yinchuan, Ningxia,Northwest China
    HE Yun-Chuan, YANG Gui-Jun, WANG Xin-Pu
    2018, 61(12):  1439-1452.  doi:10.16380/j.kcxb.2018.12.009
    Abstract ( 730 )   PDF (1901KB) ( 236 )     
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    Aim To investigate and analyze the changes in the structural composition, functional diversity and stability of terrestrial insect communities in different wetland types in Yinchuan, Northwest China. Methods Four wetland habitats including Yellow River wetland (HH), Mingcui Lake wetland (MCH), Luhuatai wetland (LHT) and Shuidonggou wetland (SDG) were selected based on the experimental design, and sampled from May to October, 2016. Insects were collected by chessboard sweeping and pitfall trapping approaches. Insect community structure diversity analysis, principal component analysis and cluster analysis were conducted based on the survey data. Results A total of 38 668 insects belonging to 370 species, 97 families and 13 orders were collected from the four wetland habitats in Yinchuan. Among them, Diptera, Homoptera, and Hymenoptera were the dominant groups in all the four habitats. Phytophagous insects accounted for the largest proportion in the family and species level of the insect communities among the four wetlands. The individual number showed a significantly negative correlation between the phytophagous and the neutral insects (Ph-Ne) in the four habitats (P<0.01). In the MCH habitat, the individual number showed a significantly positive correlation between the phytophagous and the predatory insects (Ph-Pr) (P<0.05), while a significantly negative correlation between the neutral and the predatory insects (Ne-Pr) (P<0.01). Additionally, in the LHT habitat, the individual number showed a significantly negative correlation between the neutral and the predatory insects (Ne-Pr) (P<0.05). Finally, the individual number in other combinations among the four functional groups did not show any significant correlation in all the four habitats. The predatory species group was the main component that affected the composition of the insect community based on principal component analysis (PCA). For the diversity analysis of the same functional group in the four habitats, there was a significant difference between the predatory (Pr) and the parasitic (Pa) functional groups based on the Simpson index (P<0.05), while there was no significant difference between the phytophagous (Ph) and the neutral (Ne) functional groups based on the Simpson index (P>0.05). Based on the Shannon-Wiener index, there was no significant difference among the neutral (Ne) functional groups in the four habitats (P>0.05), while there was a significant difference among the neutral (Ne) functional groups in the four habitats based on Pielou evenness index (P<0.05). Based on the Shannon-Wiener indices for the other three functional groups, there were significant differences among the four habitats (P<0.05), while they had no significant differences in the Pielou evenness index (P>0.05). The cluster analysis showed that the four types of wetlands were divided into two categories based on the characteristic index of insect communities, the first was permanent wetlands including HH and MCH, and the second was seasonal wetlands including LHT and SDG. Based on the stability index, the highest stability was found in the MCH habitat. The most stable periods for insect communities in different habitats were in May and September annually. Conclusion Diptera and Hymenoptera are the dominant groups in different wetland habitats in Yinchuan, and can be used as environmental indicators. The insect communities in the permanent lake wetland show the highest stability. The diversities of the insect communities in different habitats change with the seasons, and the most stable periods are in May and September annually. How the climatic factors and environmental factors drive the diversity to change and what factors affect the stability of insect communities in different wetlands remain to be studied.
    Phylogenetic analysis of Apis (Hymenoptera: Apidae) based on COI gene sequences and an evolutionary analysis of its dancing and nesting behavior
    REN Xiao-Xiao, HE Xing-Jiang, GONG Xue-Yang, ZHAO Wen-Zheng, LIU Yi-Qiu, DONG Kun
    2018, 61(12):  1453-1461.  doi:10.16380/j.kcxb.2018.12.010
    Abstract ( 597 )   PDF (3799KB) ( 202 )     
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    Aim To analyze the phylogeny of the genus Apis, and to investigate the ancestral state and the evolutionary process of dance direction, dance sound, nesting environment and comb structure in Apis based on the phylogenetic relationships. Methods The COI gene fragments of A. cerana, A. mellifera, A. dorsata, A. laboriosa, A. florea and A. andreniformis distributed inChinawere amplified by PCR and sequenced. The homologous sequences of COI gene of the above six species in other states or regions and those of A. nulunsis, A. nigrocinta, A. koschevnikovi and A. dorsata binghami were downloaded from NCBI databases. The phylogenetic relationships were established using maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) based on these sequence data. The dance language and nesting behavior of each Apis species mentioned above were coded and treated as trait markers, and added correspondingly into the phylogenetic trees. The ancestral state was traced with the ML model. Results The results of phylogenetic analysis demonstrated that Apis species can be divided into three clusters, i.e., cavity-nesting honeybees (A. cerana, A. mellifera, A. nigrocinta, A. nulunsis, and A. koschevnikovi), giant honeybees (A. dorsata, and A. laboriosa) and dwarf honeybees (A. florea, and A. andreniformis). The dwarf honeybees are more ancestral, and the giant and cavity-nesting species are monophyletic. Our results also suggest that the giant bee A. dorsata binghami is a separate species from A. dorsata and A. laboriosa and closer to A. laboriosa. Prototype inference indicated that ancestral honeybee species nested in the open with a vertical single comb. They disseminate information about food sources and nesting sites using a silent horizontal waggle dance. The behavior of dancing with buzz and nesting with multi-combs appeared subsequently. Conclusion As a molecular marker, COI gene can be used to analyze the ancestral state and the evolution of dancing and nesting of Apis species. The buzzing dance language and multi-comb nesting behavior could be an outcome of adaptive evolution during the later stage.
    Genetic diversity of Apis cerana cerana populations in the YimengMountains,Shandongprovince,East China(In English)
    CHI Xue-Peng, ZHANG Wei-Xing, WEI Wei, CHEN Wen-Feng, LI Zhen-Fang, XIA Zhen-Yu, YU Jing, LIU Jia-Xin, MA Lan-Ting, LIU Zhen-Guo, WANG Hong-Fang, XU Bao-Hua
    2018, 61(12):  1462-1471.  doi:10.16380/j.kcxb.2018.12.011
    Abstract ( 579 )   PDF (982KB) ( 161 )     
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    Aim Apis cerana cerana is an important germplasm resource and pollinator. This study aims to explore the genetic diversity and population structure of A. cerana cerana in theYimengMountains,Shandongprovince,East Chinaso as to provide a theoretical foundation for the utilization and conservation of germplasm resource for A. cerana cerana. Methods In this study, 114 broods of A. cerana cerana from 7 localities in the Yimeng Mountains were selected and 36 morphological indexes were measured according to Ruttners analytical method. And the genetic diversity of A. cerana cerana in theYimengMountainswas evaluated with 11 microsatellite loci. Results The results of morphological analysis showed that the average body length and the average forewing length of A. cerana cerana foraging workers in theYimengMountainswere 12.064-13.351 mmand 8.198-8.694 mm, respectively. Fifty-eight alleles were found in the 11 microsatellite loci. The number of alleles per locus ranged from 2 to 17. The average expected heterozygosity and polymorphism information content (PIC) of all loci were 0.3115 and 0.2872, respectively. The average number of alleles in all the populations ranged from 2.4545 (ST-AQ) to 4.0000 (BHY), and the average expected heterozygosity ranged from 0.1916 to 0.3397. The average genetic differentiation measured as an Fst value was 0.048. The Neis genetic distance ranged from 0.0092 (XL-DLZ to XZ-XLZ) to 0.1000 (XL-DLZ to XL-DJW). Besides, the cluster analysis results showed that all the populations in theYimengMountainscould be grouped into three categories. Conclusion The population structure of A. cerana cerana populations in theYimengMountainsis similar. The results suggest that the gene flow occurs in different populations. Compared with the populations in other positions, A. cerana cerana in theYimengMountainsshows certain specificity in morphology and genetic structure.
    REVIEW ARTICLES
    Matrix metalloproteases in insects
    CHEN Kang-Kang, ZHANG Yi-Qiang, ZHU Qian-Ming, TANG Tai, WANG Ying-Juan, FENG Cong-Jing
    2018, 61(12):  1472-1480.  doi:10.16380/j.kcxb.2018.12.012
    Abstract ( 487 )   PDF (1015KB) ( 131 )     
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    Insects have formed developed metamorphosis and innate immunity system to adapt to the complex habitats during evolution. Some proteins produced in the process of metamorphosis and innate immunity in insects must be degraded in time, so that insects are capable of maintaining the homeostasis of development and immune responses. Matrix metalloproteases (MMPs) are Zn2+-dependent proteases. MMPs can degrade most proteins in the basement membrane and extracellular matrix, and also are involved in the regulations of metamorphosis and innate immunity in insects. They participate in tissue remolding and limitation of excessive immune responses through degradation of key extracellular matrix and peptide hormones. In this article, the functions of MMPs involved in regulation of metamorphosis and innate immunity were summarized, and the directions for further investigations were also discussed.
    Progress and prospects of CRISPR system in the regulation of gene expression in insects
    LIU Su-Ning, LI Sheng, REN Chong-Hua
    2018, 61(12):  1481-1487.  doi:10.16380/j.kcxb.2018.12.013
    Abstract ( 474 )   PDF (1120KB) ( 217 )     
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    The study of gene function in insects is restricted by lacking of relevant tools. However, the emergence of CRISPR/Cas9 system provides great help for the research of gene editing and transcriptional regulation in insects. dCas9 with catalytically dead mutant RuvC and HNH domains of Cas9 has been widely used in the transcriptional regulation of genes in recent years. Meanwhile, CRISPR/dCpf1 and the newly discovered CRISPR/Cas13(a/b) systems provide more options for gene function research. In this article, the mechanism of dCas9, dCpf1 and Cas13(a/b) systems and the progress of their application in the transcriptional regulation in Drosophila were systematically overviewed, aiming to provide a reference for future related studies of insects.
    Progresses in entomic ectosymbiotic microorganisms
    LIN Yong-Wen, HOU You-Ming
    2018, 61(12):  1488-1496.  doi:10.16380/j.kcxb.2018.12.014
    Abstract ( 563 )   PDF (1513KB) ( 322 )     
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    Entomic ectosymbiotic microorganisms refer to the microorganisms that interact with host mutually in vitro. Although ectosymbiotic microorganisms are not as popular as gut microbes in insects, they also play important roles in the growth and development of their hosts. Commonly, entomic ectosymbiotic microorganisms are carried by host via the surface of skin or other organs like mycangi, and move into plant tissues in the specific developmental stage of host. Ectosymbiotic microorganisms emit volatile organic compounds to indicate the location of plants and provide nutrients for their symbiotic host, and even affect in vitro immunity of host insects. The studies on entomic ectosymbiotic microorganisms will advance the knowledge on the interactions between insects and microorganisms, diversify the research of insect symbionts, and uncover the mechanisms of ectosymbiotic microorganisms-mediated immune privilege in insects. In this article, we reviewed the studies of parasitic modes and transmission routes of ectosymbiotic microorganisms and their effects on insect hosts, aiming to provide new insights for integrated pest management.
    CONTENTS
    Contents of Vol. 61 Issue12
    2018, 61(12):  1497-1497. 
    Abstract ( 211 )   PDF (490KB) ( 36 )     
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    General content of Volume 61(1-12)
    2018, 61(12):  1498-1498. 
    Abstract ( 310 )   PDF (1688KB) ( 53 )     
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