【Aim】 To complement the odorant receptor (OR) information of two sibling species of Eucryptorrhynchus, E. scrobiculatus and E. brandti, and to provide theoretical basis for subsequent functional studies by the identification and expression profile analysis of OR genes based on the transcriptome data of these two weevil species at different developmental stages. 【Methods】 The OR gene sequences were screened from the transcriptome databases of different developmental stages of E. scrobiculatus (larva, pupa and adult) and E. brandti (egg, larva, pupa and adult). Interspecific and intraspecific sequence alignments of the screened ORs were performed based on the antennal transcriptome data of the two weevil species. Phylogenetic analysis of the newly identified OR genes was performed using the maximum likelihood method. Expression abundance analysis was performed according to FPKM (fragments per kilobase per million mapped fragments) values of the newly identified OR genes in the transcriptomes of two weevil species at different developmental stages. The expression profiles of the newly identified OR genes at different developmental stages and in different adult tissues of E. scrobiculatus and E. brandti were detected by qPCR. 【Results】 Six and four OR genes with complete open reading frames were identified from the transcriptome databases of different developmental stages of E. scrobiculatus and E. brandti, respectively. According to the sequence alignment results with the antennal transcriptome data, four (EscrOR50-53) and two (EbraOR46-47) OR genes were newly identified from E. scrobiculatus and E. brandti, respectively. We also found a pair of potential homologous genes, EscrOR53 and EbraOR45, between the two weevil species. The phylogenetic tree indicated that the six newly identified ORs belong to the subfamilies 2B and 7 of coleopteran ORs. According to the FPKM data of the transcriptomes of different developmental stages of the two weevil species and the spatiotemporal expression profiles detected by qPCR, three of the newly identified OR genes of E. scrobiculatus and the two newly identified OR genes of E. brandti were all highly expressed in non-olfactory tissues of adults, and also expressed in egg, larval or pupal stages. 【Conclusion】 In this study, the OR genes and their spatiotemporal expression profiles in the transcriptomes of different developmental stages of E. scrobiculatus and E. brandti have been clarified, and it is found that non-olfactory functional ORs possibly exist in the reproductive organs, suggesting that they may play a role in the early stage of development.

【Aim】 Antheraea assama has the typical characteristics of wild silkworms, with its eggs not uniformly hatched, which seriously affects its largescale indoor rearing. This study aims to explore the characteristics of the hatching enzyme gene playing a key role in the egg hatching of A. assamensis and its promoter sequence, and to lay the foundation for further selection of appropriate inhibitors or accelerators to regulate the egg hatching of A. assamensis. 【Methods】 The full-length cDNA sequence of the hatching enzyme gene was cloned from A. assamensis by RACE, and the sequence was analyzed by bioinformatics method. qRT-PCR was used to detect the expression profiles of the hatching enzyme gene in eggs of different days after laying, and in different tissues (silk gland, Malpighian tubules, head, midgut, fat body, epidermis, blood, testis and ovary) of the day-3 and day-4 5th instar larvae of A. assamensis. The promoter sequence of the hatching enzyme gene was cloned from A. assamensis by chromosome walking, and the promoter activity was detected by constructing the insect cell recombinant expression vector and transfecting the BmN cells of Bombyx mori. 【Results】 The full-length cDNA sequence of the hatching enzyme gene AaHE (GenBank accession no.: KT336227.1) was obtained from A. assamensis. It is 993 bp in length, encoding 294 amino acids with the predicted molecular weight of 33.7 kD and the theoretical isoelectric point of 5.17. The amino acid sequence of AaHE contains a signal peptide and a ZnMc domain, and AaHE belongs to a group of zinc-dependent proteolytic enzymes with a HExxH zinc-binding site. The members of this enzyme group are both peptidase and digestive enzyme. AaHE was specifically and highly expressed in the eggs before hatching and the midgut of the 5th instar larva of A. assamensis, being consistent with the properties of peptidase and digestive enzyme. There are multiple transcription factor binding sites in the core region of AaHE promoter, which may be related to the transcription factor’s involvement in regulating the expression of AaHE. Promoter activity analysis showed that the AaHE promoter could initiate the expression of EGFP in the BmN cells of B. mori and had obvious promoter activity. 【Conclusion】 AaHE is a zinc-dependent proteolytic enzyme with multiple transcription factor binding sites in the core region of its promoter. This study provides references for choosing suitable inhibitors or accelerators to regulate the egg hatching of A. assama.

【Aim】 To clarify the potential of a piggyBac (PB) transposon AgoPLE1.1 to be developed as an insect transgenic vector by detecting the transformation activity of AgoPLE1.1 in Drosophila melanogaster. 【Methods】 AgoPLE1.1 transposase helper plasmid (pAgoHsp) and red fluorescent labeled donor plasmid (pXLAgo-PUbDsRed) were constructed. The helper and donor plasmids were mixed at three different ratios (170 ng/μL∶400 ng/μL, 90 ng/μL∶200 ng/μL and 90 ng/μL∶100 ng/μL) and microinjected into fresh W¹¹¹⁸D. melanogaster embryos, respectively. The transgenic offsprings were screened out. Southern blot was used to verify the insertion copy number of AgoPLE1.1 in the transgenic D. melanogaster, and the flanking sequences of AgoPLE1.1 insertion sites were cloned by genome walking to identify the transposition characteristics of AgoPLE1.1. 【Results】 AgoPLE1.1 transposon showed transformation activity in D. melanogaster, and the transgenic frequencies were 1.32%-1.94%. Southern blot analysis showed that there were at least six insertion sites of AgoPLE1.1 transposon in the transgenic flies. The genome walking PCR revealed that four of the insertion sites were located in D. melanogaster 3R, 3L, 2L and X chromosomes, respectively, and the integration of AgoPLE1.1 in D. melanogaster chromosomes contained the skeleton of donor plasmid. 【Conclusion】 The PB transposon AgoPLE1.1 can only perform imprecise excision and transposition in D. melanogaster with a low transformation frequency. Therefore, AgoPLE1.1 transposon has no potential to be developed as a new insect transgenic vector.

【Aim】 Based on the important role of insect gut microorganisms in host health, growth and development, this study aims to preliminarily explore the diversity and differences of gut bacterial communities in different instar larvae and diapause prepupae of Colleles gigas. 【Methods】 The V3-V4 gene fragment of 16S rRNA was amplified by PCR using the bacterial DNA extracted from the gut contents of the 1st-5th instar larvae and diapause prepupae of C. gigas collected from the field, and was sequenced by IlluminaMiseq second-generation high-throughput sequencing technology. Based on the obtained sequence data, the composition, abundance and diversity of gut bacteria of larvae and diapause prepupae of C. gigas were analyzed by bioinformatics methods. 【Results】 Bacteria of 15 phyla, 23 classes, 43 orders, 80 families and 128 genera were detected in the gut bacterial communities of C. gigas larvae, of which the main phylum, order, family and genera were Proteobacteria (accounting for 93.74%), Rickettsiales (accounting for 68.68%), Anaplasmataceae (accounting for 68.64%), and Wolbachia (accounting for 68.64%), respectively. Beta diversity analysis showed that the gut bacterial community changed with the development of larvae, and could be divided into three groups: early instar group (1st-3rd instar larva), late instar group (4th-5th instar larva) and diapause prepupa group. Alpha diversity analysis showed that there was a significant difference in the gut bacterial diversity between the diapause prepupa group and the late instar group and early instar group, while the gut bacterial diversity between the late instar group and the early instar group showed no significant difference. The linear discriminant analysis results showed that there was significantly dominant class in both diapause prepupa group and early instar group, being Alphaproteobacteria and Gammaproteobacteria, respectively, but no dominant class existed in the late instar group. At the order level, Enterobacteriales and Pseudomonadales were dominant in the early instar group, and Rickettsiales was dominant in the diapause prepupa group. At the family level, Enterobacteriaceae and Moraxellaceae were dominant in the early instar group, and Anaplasmataceae was dominant in the diapause prepupa group. At the genus level, Enterobacter and Acinetobacter were dominant in the early instar group, and Wolbachia was dominant in the diapause prepupa group. The results of functional gene annotation also showed the characteristics of the three groups. 【Conclusion】 There are significant differences in the community structure of gut bacteria in different instar larvae and diapause prepupae of C. gigas. The bacterial diversity decreases gradually from the early instar group to the late instar group and then to the diapause prepupa group, and this may be related to the feeding characteristics and adaptation of gut microorganisms to the gut environment. This study lays a foundation for the study of gut microorganisms of soil-nesting wild bees and also provides a new angle and direction for the protection of this kind of wild bees.

【Aim】 To explore the effects of potassium application on the oviposition selection, growth and development, adult longevity and fecundity of Odontothrips loti on alfalfa, and to illustrate the relationship between the nutrient content in alfalfa leaves applied with potassium and the life parameters of O. loti. 【Methods】 The number of eggs, duration and survival rates of various immature developmental stages, and the longevity and fecundity of the second generation adults of O. loti fed with leaves of Medicago sativa cv. Gannong No. 3 applied with different amounts of potassium (40, 60, 80 and 100 mg/kg) (without potassium application as the control) were investigated. Meanwhile the soluble sugar, free amino acid and potassium contents in alfalfa leaves were measured. 【Results】 The number of eggs laid by O. loti eggs in per compound leaf of alfalfa decreased first and then increased with increasing amount of potassium application, and was the lowest at 60 mg/kg potassium treatment, 45.58% lower than the control. The egg hatching rate and the survival rate of the 1st-2nd instar nymphs on alfalfa leaves applied with potassium did not change significantly, but the survival rate of the 3rd-4th instar nymphs and the total immature survival rate of O. loti went down significantly, with the largest decrease at 100 mg/kg and 80 mg/kg potassium treatment, which decreased by 54.36% and 48.48% compared with the control, respectively. There was no significant change in the egg and the 1st-2nd instar nymphal duration of O. loti on alfalfa leaves applied with different amounts of potassium, while the 3rd-4th instar nymphal duration and the total immature developmental duration were prolonged. The fecundity of the second generation adults of O. loti on alfalfa leaves applied with different amounts of potassium was reduced significantly, and the adult longevity was shortened significantly than the control (except 40 mg/kg potassium treatment). After potassium application, both the soluble sugar and potassium contents in alfalfa leaves increased, the free amino acid content decreased, and the sugar to nitrogen ratio increased. The correlation analysis showed that there was no significant correlation between the potassium content in alfalfa leaves and the total immature survival rate and the fecundity of O. loti adults, while the soluble sugar content and the sugar to nitrogen ratio were extremely negatively correlated with the survival rate of the 3rd-4th instar nymphs and the fecundity of adults and significantly negatively correlated with the total immature survival rate of O. loti. 【Conclusion】 Potassium-applied alfalfa has significant antixenosis to the oviposition of O. loti adults. The application of potassium increases the soluble sugar content and the sugar to nitrogen ratio in alfalfa leaves, exerting significant antibiosis to O. loti by affecting the growth and development of nymphs, shortening adult longevity and reducing the fecundity of adults.

 【Aim】As the main vector insect of pine wilt disease by Bursaphelenchus xylophilus in the middle and temperate zone of China, Monochamus saltuarius has caused very serious destruction to pine trees in Liaoning, Northeast China in recent years. In this study the supplementary nutrition of M. saltuarius on Pinus koraiensis was boserved and analyzed so as to provide a theoretical basis to determine the biological characteristics of M. saltuarius, to evaluate its population density and damage degree in the forest, and to determine its transmission law of pine wood nematode. 【Methods】 The newly emerged male and female adults of M. saltuarius were placed in pairs in a transparent glass jar filled with segments and branches of P. koraiensis. Under the condition of temperature 23±3℃, relative humidity 55%±5%, and photoperiod 16L∶8D, the main parts of P. koraiensis fed by M. saltuarius adults, the feeding amount of M. saltuarius adults on different parts of P. koraiensis, the adult longevity, average feeding frequency, average time of each feeding and variation rule of feeding amount with developmental time, and the circadian rhythm of the proportion of feeding frequency of M. saltuarius adults on P. koraiensis branches were continuously observed until their natural death. 【Results】 M. saltuarius adults preferred to feed P. koraiensis needles, followed by branches and buds, with the proportion of the total feeding amount of 7711%, 21.46% and 1.43%, respectively. The average longevity of M. saltuarius adults on P. koraiensis branches was 16.63±6.63 d, being 17.11±5.91 d for females and 16.14±6.77 d for males. The feeding behavior of M. saltuarius adults on P. koraiensis occurred on the 4th day after emergence, and the daily feeding amount increased first and then decreased with the developmental time, and the highest feeding amount was on the 16th day, being 526.48 mm². The feeding behavior of M. saltuarius adults occurred in every hour throughout the day, and the daily feeding frequency of male and female adults showed a state of increasing and decreasing repeatedly. 【Conclusion】 When M. saltuarius adults are reared indoors with P. koraiensis, their feeding starts on the 4th day after emergence, and there is no obvious regularity in the daily feeding amount and feeding rhythm. The feeding characteristics of adult M. saltuarius in the forest need to be further studied.

【Aim】 Over the years, the typical larch pest of the Mongolian Plateau, Erannis jacobsoni, has occurred frequently, and caused severe damage to forest ecosystem. Population density can directly describe the severity of forest pests, and it is extremely important to obtain information on pest population density in a timely and rapid manner. The purpose of this study is to construct a population density estimation method based on the hyperspectral data of larch trees and the population density data of E. jacobsoni in the outbreak areas of E. jacobsoni. 【Methods】 Outbreak areas of E. jacobsoni in four localities in Hangay Province and Khentiy Province in Mongolia were selected as the test areas. Firstly, from the four test areas 110 sample trees of Siberian larch (Larix sibirica) with different degrees of damage were selected, and the population density and the spectral reflectance of the canopy was measured. Secondly, the differential spectral reflectance (DSR) and the modified spectral index (MSI) were calculated through spectral reflectance data. Thirdly, the polynomial curve fitting method was used to analyze the sensitivity of DSR and MSI to population density. Then, the successive projection algorithm (SPA) was used to extract sensitive DSR and MSI. Finally, the sensitive DSR and MSI, polynomial regression (PR) and support vector machine regression (SVMR) algorithms were used to establish the population density estimation model of E. jacobsoni, and the accuracy was evaluated. 【Results】 The sensitive bands of DSR are mainly in the yellow edge and red edge, and the sensitivity at 572 nm was the most significant (R²=0.5821, P<0.001). The most sensitive index of MSI was TVI (R²=0.5386, P<0.001). The TVI (R²_CV=0.6323, RMSE_CV=0.1513) was more accurate than DSR₅₇₂ (R²_CV=0.5581, RMSE_CV=0.1649), while multiple DSRs (R²_CV=0.7309, RMSE_CV=0.1347) had higher estimation potential than multiple MSIs (R²_CV=0.6537, RMSE_CV=0.1453), and the performance of its SVMR model was always better than the PR model, suggesting that SVMR is suitable for population density estimation. 【Conclusion】 MSI and DSR can be used as sensitive indicators for population density estimation, and the polynomial curve fitting method can tap the sensitivity of MSI and DSR to population density. SPA is an effective method for POPD (pest population density) sensitive spectral feature extraction. The extracted DSR sensitivity index and MSI sensitivity index have fully captured the chlorophyll absorption characteristics and water absorption characteristics of larch needles and the reflection characteristics caused by the damage of needle cells. This study not only provides an experimental theoretical basis for the aerial and aerospace remote sensing monitoring of forest pest population density, but also develops a new way for remote sensing and monitoring of forest pests.

 Abstract: 【Aim】 Ficus pumila var. awkeotsang, a special fruit tree, relies on Wiebesia sp. nr. pumilae for pollination. The aim of this study is to investigate the developmental dynamics of galls and W. sp. nr. pumilae, thus providing a foundation for further studies on the developmental biology of fig wasps. 【Methods】 By means of artificially releasing wasps-marking-regularly sampling-microscopic and electron microscopic observations, the synergistic development of W. sp. nr. pumilae with gall, and the source of nutrients and the position change of W. sp. nr. pumilae inside galls were observed, the developmental dynamics of the body length of W. sp. nr. pumilae and the diameter of gall was determined, and the developmental duration of W. sp. nr. pumilae in summer and winter was compared. 【Results】 The development of immature W. sp. nr. pumilae is generally divided into the egg, larval and pupal stages. The larval stage is divided into 5 instars, in which male and female are indistinguishable. The pupal stage is divided into three stages: early stage, middle stage and mature stage. Male and female pupae are differentiated and heteromorphic, and male pupae develop 1.34 d earlier than female pupae. The gall consists of epidermis, protective layer, endodermis and nutritive layer; both the epidermis and endodermis are colloidal and water-rich, providing good humidity guarantee for fig wasps. The protective layer, as the skeleton of the gall shell, protects the larval wasps. The nutritive layer secrets nutritive solution to feed larval wasps. A comparison with the structure of the lean fruit showed that the small wasp-galls changed their protective layer morphology with thickened protective layer, and the nutritive layer is capable to deliver nutritive solution to larval fig wasps. The position of immature fig wasps inside the gall is regularly varying. The body length of larval wasps increases synergistically with the diameter of the gall. Galls grow faster than wasp larvae to provide a bounteous growth space for them. The body length of the fig wasp stops increasing in the pupal stage, while the growth of galls stops after the 4th instar of the larva. In Fujian areas, W. sp. nr. pumilae has two generations a year, its generation time in winter and summer is 253 d and 112 d, respectively, and the duration of various developmental stages in winter is longer than that in summer. Especially, the duration of the 5th instar larva of W. sp. nr. pumilae in winter is 66 d longer than that in summer. W. sp. nr. pumilae overwinters in the form of the 5th instar larva, and a prolonged feeding period is beneficial for its overwintering. 【Conclusion】 The results clarified the nutritive connection between immature W. sp. nr. pumilae and its gall and their synergistic development dynamics, providing a theoretical basis for the study of fig-fig wasp coevolution.

 【Aim】 Monolepta hieroglyphica is a polyphagous pest feeding on a large number of cultivated plant species. The aim of this study is to investigate the genetic diversity, genetic structure, and levels of genetic differentiation and gene flow among geographical populations of M. hieroglyphica distributed in South China, and to clarify the diversity and prevalence of the bacterial endosymbiont Wolbachia in M. hieroglyphica geographical populations in South China. 【Methods】 The mitochondrial COII gene was used as genetic marker. The partial COII gene sequences in a total of 403 individuals from 14 geographical populations of M. hieroglyphica were amplified by PCR and sequenced. The haplotype diversity (Hd), genetic differentiation coefficient (F_st) and gene flow (Nm) between populations were analyzed, and the analysis of molecular variance (AMOVA) and Tajima’s D and Fu’s Fs neutrality tests were performed. Median-joining network and phylogenetic tree were constructed based on haplotype sequences. Wolbachia wsp gene was amplified by PCR to detect population infection rates, and the obtained wsp sequences were used for strain typing and phylogenetic analysis of Wolbachia. 【Results】 For all the 403 test individuals of M. hieroglyphica in this study, 23 COII haplotypes were observed and divided into two clusters in phylogenetic tree. The Hd of total population was 0.748, ranging from 0.394 to 0.782 within each population. The neutrality test results suggested that M. hieroglyphica populations followed the neutral evolution model and there was no evidence of population expansion in recent history. The values of F_st and Nm of total population were 0.2481 and 0.76, respectively. The AMOVA results showed that a high proportion (73.75%) of the total genetic variance attributed to variation within population. There was no significant correlation between genetic distance and geographical distance among populations (R=0.2898, P=0.0640). The Wolbachia infection rates in the 14 geographical populations of M. hieroglyphica ranged from 92.59% to 100%, with an average infection rate of 97.60%. Six Wolbachia strains (named as wMhie1-wMhie6) were identified based on wsp sequences, and these strains all belong to the supergroup A, which is clearly distinguished from other representative stains and forms a unique cluster in the phylogenetic tree. 【Conclusion】 The genetic diversity of M. hieroglyphica populations distributed in South China is comparatively high. There is significant genetic differentiation among most populations and the gene flow is low among populations. No significant correlation exists between genetic differentiation and geographical isolation. High infection rates and diversity of Wolbachia exist in M. hieroglyphica populations in South China.

 In the long evolutionary process, a variety of forms of interaction between microorganisms and insects have been formed. The wide distribution of microorganisms provides background conditions for their contact with insects and influence on insect behavior. To further explore the phenomenon and mechanism of microorganisms influencing insect behaviors, the research progress of the influence of microorganisms on insect behaviors was reviewed in this article. Host location and selection of insects can be influenced by chemical signal substances produced by microorganisms, and the synthesis of semiochemicals in insects and host plants can also be influenced by microorganisms. Microorganisms have also been found to play important roles in intraspecific and interspecific relationships of insects. Microorganisms can even influence the reproductive behavior of insects by altering, for example, insect sex pheromones. Besides, social and aggregation behaviors of insects are also found to be influenced by semiochemicals synthesized by microorganisms. Considering the current research status of the influence of microorganisms on insect behaviors, the following aspects are suggested to be further investigated: (1) How are semiochemicals that influence insect behavior synthesized in the processes of influencing insect behavior by microorganisms? (2) Do microorganisms involve more interspecies interaction in influencing insect behavior? (3) How do host insects acquire and maintain symbiotic microoganisms that influence insect behavior at certain stages?

 The Calyptratae (Diptera: Calyptratae) comprise 20% of the diversity of Diptera, one of the four superradiations of insects. The Calyptratae are distributed widely in the world, exhibit enormously diverse living habits, play vital roles in maintaining the stability of ecosystem, and are not only the hotspot groups in the studies of vectors, forensic medicine, pollinators and natural enemies of insects, but also key groups for exploring the phylogeny of Diptera and its successful adaptive radiation. To trace the evolutionary history of the Calyptratae, numerous dipterologists have conducted studies at various taxonomiclevels. TheCalyptratae arewellsupportedasa monophyletic group, and divided into three
superfamilies, i.e., Hippoboscoidea, Oestroidea, and Muscoidea, with monophyletic Oestroidea nested within paraphyletic Muscoidea, which are a sister group of Hippoboscoidea. At the family level, the Streblidae (Hippoboscoidea), Anthomyiidae (Muscoidea), Calliphoridae (Oestroidea), and Rhinophoridae (Oestroidea) are paraphyletic, and new families have been established, e.g., Polleniidae (Oestroidea) and Ulurumyiidae (Oestroidea). Therefore, the family level relationship of Calyptratae is still insufficiently resolved. Studies have been performed to reconstruct the evolutionary history of Hippoboscidae (Hippoboscoidea), Streblidae, Nycteribiidae (Hippoboscoidea), Muscidae (Muscoidea), Scathophagidae (Muscoidea), Sarcophagidae (Oestroidea), Gasterophilinae (Oestroidea: Oestridae), in terms of the origin and dispersal, host shift, and feeding habit. However, due to the lack of the biology information of some key groups and a well-resolved phylogeny, the evolutionary history of Calyptratae remains open. In this article we reviewed the research progress of classification, phylogeny and evolution of calyptrate flies, being the first review of the progress of the related research subjects of this group in the phylogenomic era.