【Aim】 Long non-coding RNAs (lncRNAs) play important roles in the regulation of development of the silkworm, Bombyx mori. In our previous study, a lncRNA BmlncR2036 was found near the silk fibroin gene P25 of B. mori. This study aims to further explore the molecular mechanism of BmlncR2036 regulating the expression of P25. 【Methods】 qPCR was used to detect the expression profiles of BmlncR2036 in different tissues (cuticle, brain, nerve, testis, ovary, silk gland, Malpighian tubules, hemolymph, fat body and midgut) of the day-3 5th instar larvae of B. mori and in the anterior silk gland, middle silk gland and posterior silk gland during different developmental stages of B. mori. miRNAs that can target both P25 and BmlncR2036 were predicted. The interaction between miRNA and P25 was verified by luciferase assay. Luciferase assay was used to assess the effect of BmlncR2036 on the transcription activity of P25 promoter. Finally, the expression of BmlncR2036 was knocked down in the day-3 5th instar larvae of B. mori by dsRNA injection, and phenotype changes of their silk glands and midgut were observed. 【Results】 The qPCR results showed that BmlncR2036 was highly expressed in the posterior silk glands of the day-3 5th instar larvae and mature 5th instar larvae, showing a expression pattern consistent with that of P25 gene. Meanwhile, BmlncR2036 was also highly expressed in the testis and midgut of the day-3 5th instar larvae of B. mori, while lowly expressed in other tissues, suggesting the diverse roles of BmlncR2036. miR-2739 and miR-279a can not only match with BmlncR2036, but also target the 3′UTR of P25 gene. Luciferase assay results showed that P25, however, was not the real target of miR-2739 and miR-279a, and BmlncR2036 had a negative effect on the promoter activity of P25, extremely significantly decreasing the luciferase activity by 52% after the co-transfection with pcDNA3.1(+)［BmlncR2036］ and pGL3-Enhancer［P25-promoter］ vector. After knocking down the expression of BmlncR2036 in the day-3 5th instar larvae of B. mori, the body color darkened, and a large amount of undigested food was retained in the midgut until its death, indicating that BmlncR2036 might be involved in the regulation of midgut development in B. mori. 【Conclusion】 We found that lncRNA BmlncR2036 can regulate the promoter activity of P25 gene and might also play roles in the regulation of midgut function in B. mori. This study lays the experimental foundation for further exploring the functional roles of lncRNAs in B. mori.

 【Aim】 The objective of this study is to explore the role of Kazal-type serine protease inhibitor KaSPI in the development, digestion, and immune defense processes of the soybean aphid, Aphis glycines. 【Methods】 The cDNA sequence of Kazal-type serine protease inhibitor gene from A. glycines was cloned by PCR based on the transcriptome data of A. glycines. qRT-PCR was used to detect the expression levels of AgKaSPI in the 1st-4thinstar nymphs and adults of A. glycines, and in A. glycines adults at 3, 6, 12, 24, 48 and 72 h after infection by Akanthomyces lecanii. After RNAi of AgKaSPI with siRNA for 3, 6, 12, 24 and 48 h, the RNAi efficiency was detected by qRT-PCR, and the mortality and fecundity of A. glycines adults at 12, 24, 48 and 96 h post RNAi were recorded. The AgKaSPI content and the activities of serine protease, trypsin and chymotrypsin in A. glycines adults were detected by double antibody sandwich method after infection by A. lecanii and RNAi of AgKaSPI. The mortality of A. glycines adults at 12, 24, 48 and 96 h after infection by A. lecanii following RNAi of AgKaSPI for 6 h was observed and recorded. 【Results】 The cDNA sequence of a serine protease inhibitor gene AgKaSPI (GenBank accession no. MK440557) was cloned from A. glycines. AgKaSPI is 1 019 bp in length, with the open reading frame of 324 bp, encoding 107 amino acids. AgKaSPI has the Kazal domain, with the molecular weight of 11.43 kD and isoelectric point of 9.24. AgKaSPI was expressed at different developmental stages of A. glycines. At 24 h after infection by A. lecanii, the expression level of AgKaSPI and the corresponding AgKaSPI content in A. glycines adults were significantly up-regulated, being 4.31-fold and 1.69-fold as high as those of the control group, respectively, while the activities of serine protease, trypsin and chymotrypsin were inhibited. At 6 h after RNAi of AgKaSPI, the expression level of AgKaSPI in A. glycines adults decreased by 71.05%. The KaSPI content decreased by 51.11% and the activities of serine protease and chymotrypsin increased significantly at 12 h after RNAi of AgKaSPI. The fecundity per one hundred individuals decreased by 49 born aphids and the mortality of A. glycines adults increased by 10.12% at 96 h after RNAi of AgKaSPI. 【Conclusion】 AgKaSPI is expressed at different developmental stages of A. glycines, and the expression level of AgKaSPI is significantly up-regulated at 24 h after infection by A. lecanii. AgKaSPI may participate in the immune response of A. glycines to A. lecanii infection by regulating the serine protease activity.

 【Aim】This study aims to clone and identify odorant-binding protein (OBP) genes in the adzuki bean seed weevil, Callosobruchus chinensis and to assay their tissue-specific expression profiles in C. chinensis adults, so as to to lay a foundation for studying the function of OBPs in olfactory perception in C. chinensis. 【Methods】Based on the antenna transcriptome data of C. chinensis, six OBP genes in C. chinensis were cloned by RT-PCR and analyzed by bioinformatics. The expression levels of OBP genes in tissues including head (excluding antennae), antennae, abdomen, legs and wings of female and male adults of C. chinensis were analyzed by qRT-PCR. 【Results】 The open reading frames of six OBP genes from C. chinensis were obtained, and named as CchiOBP1-CchiOBP6 (GenBank accession number: MN832700-MN832703, MN901841-MN901842). CchiOBP5 is a segment of Minus-C OBP with incomplete C-terminus, and the others belong to complete classical OBPs. It was predicted that all the six CchiOBPs contain signal peptides. Phylogenetic analysis showed that CchiOBP1, CchiOBP2 and CchiOBP5 were closely related to OBPs of Chrysomelidae, and CchiOBP3, CchiOBP4 and CchiOBP6 were closely related to OBPs of Cerambycidae. The qRT-PCR results showed that six CchiOBP genes were differentially expressed in the antennae, head (excluding antennae), abdomen, wings and legs of C. chinensis adults. CchiOBP1-4 and CchiOBP6 were highly expressed in antennae of female and male adults of C. chinensis, and their expression levels in antennae were extremely significantly higher than those in the other adult tissues. CchiOBP5 was highly expressed in female antennae and head (excluding antennae), but in males it showed significantly higher expression level in legs than in other adult tissues, with the lowest expression level in antennae. 【Conclusion】 The nucleotide and amino acid sequences of six CchiOBP genes of C. chinensis have been determined, among which five CchiOBP genes are highly expressed in the antennae of female and male adults of C. chinensis, suggesting their important roles in olfactory recognition of host plants.

【Aim】 Gammaaminobutyric acid (GABA) is an important neurotransmitter in animal nervous system. This study aims to identify the GABA receptor (GABAR) familygenesinthewestern flower thrips, Frankliniella occidentalis, and to clarify the role of ionotropic receptor (GABA_AR) in the resistance evolution to spinosad in F. occidentalis. 【Methods】 Based on the genomeand transcriptome data of F. occidentalis, the GABAR genes were identified, cloned and analyzed with bioinformatics tools. The expression patterns of the GABA_AR subunit genes, FoRDL, FoLCCH3, and FoGRD in the spinosad susceptible strain of F. occidentalis at different developmental stages (1st-2nd instar nymphal, pupal, and adult stages), and their expression differences between the spinosad susceptible and resistant strains of F. occidentalis at the adult stage were detected by qPCR. After treatment with 0.250 and 0.400 mg/L spinosad at 24 h post RNAi of FoRDL in the 3-day-old adults of the spinosad susceptible strain of F. occidentalis, the mortality rates of F. occidentalis adults were determined by bioassay. 【Results】 Eight GABAR genes including FoRDL, FoLCCH3, FoGRD, FoGRD-like1, FoGRD-like2, FoB1, FoB2, and FoB-like (GenBank accession numbers: MH148151-MH148158) were annotated and cloned, and their ORF lengths vary from 1 080 to 3 720 bp. Phylogenetic analysis showed that GABAR genes of F. occidentalis were clustered with the corresponding genes of other insect species, indicating high conservativeness. GABA_AR subunits FoRDL, FoLCCH3 and FoGRD all have a typical nitrogen-terminal extracellular region loop structure (loop A-F) and four transmembrane regions (TM 1-4), and exon 3 of FoRDL has mutually exclusive splicing. The expression levels of FoRDL, FoLCCH3 and FoGRD in the spinosad susceptible strain of F. occidentalis increased with the developmental stage of F. occidentalis, and the expression peak occurred at the adult stage. The expression level of FoRDL in the spinosad resistant strain of F. occidentalis at the adult stage was significantly lower than that in the susceptible strain at the adult stage. After treatment with 0.250 and 0.400 mg/L spinosad following RNAi of FoRDL in the susceptible strain of F. occidentalis, the mortality rates of F. occidentalis adults decreased significantly by 55.80% and 43.00%, respectively, as compared to those of the control. 【Conclusion】 Five ionic and three metabotropic GABAR genes have been identified in F. occidentalis. FoRDL may play a role in the resistance evolution to spinosad in F. occidentalis.

of Beauveria bassiana and Orius sauteri for biological control of pests. 【Methods】 We measured the emergence rate of O. sauteri adults after the 5th instar nymphs were treated with two different concentrations of B. bassiana spore suspensions (routine concentration: 1×10⁷ conidia/mL; low concentration: 5×10³ conidia/mL) and spore powder (1×10⁷ conidia/individual) and the predation ability of the 5th instar nymphs of O. sauteri

subjected to the above treatments to female adults of Tetranychus urticae under different prey densities. 【Results】 The emergence rate of O. sauteri adults was significantly reduced after the 5th instar nymphs were treated with B. bassiana spore powder (1×10⁷ conidia/individual), and there was no significant difference in adult emergence rate between the other treatments and control. The predation of the 5th instar nymphs of O.

sauteri in the three treatment groups on female adults of T. urticae fitted with Holling Ⅱ functional response models, and the searching efficiency reduced with the increasing of prey densities. Specifically, the 5th instar nymphs of O. sauteri treated with B. bassiana spore suspension at a low concentration (5×10³ conidia/mL) showed the highest daily maximal predation amount (N_a-max) and the shortest handling time (T_h), and the  gression of searching efficiency showed the lowest decreasing trend with the increasing of prey density. However, the 5th instar nymphs of O. sauteri treated with B. bassiana spore powder had the lowest N_a-max and the longest T_h. 【Conclusion】 Low concentration of B. bassiana spore suspension does not affect the predation ability of the 5th instar nymphs of O. sauteri on female adults of T. urticae, but spore powder treatment causes adverse effect to the predation ability of the 5th instar nymphs of this natural enemy on female adults of T. urticae. This study preliminarily demonstrated the feasibility of the combined application of low concentration of B. bassiana spore suspension and O. sauteri on the control of T. urticae.

 【Aim】 To observe the external morphology and ultrastructure characteristics of Lardoglyphus konoi at different developmental stages. 【Methods】 Based on the individual samples of different developmental stages of L. konoi cultured in the laboratory, the external morphology and ultrastructure of various developmental stages of L. konoi, including egg, larva, nymph and adult, were observed under scanning electron microscope (SEM). 【Results】 The egg of L. konoi is oval. The larva has three pairs of legs, but has no genital organs, genital setae and anal setae. The nymph has four pairs of legs, and the genital area, genital setae and anal setae still look under-developed. The hypopus has a tapering front end and a round and pear-shaped rear end. Sucker plate appears in the end idiosoma, and there are 13 suckers on the sucker plate with 2-2-4-5 distribution. The chelicera of adult mite is slender, scissor-like, and there are several small teeth on its fixed digit and movable digit. The external vertical seta (ve) is about half the length of the internal vertical seta (vi), and situated at the same horizon as the internal vertical seta. Tarsus end of the 3rd leg in the male adult is replaced by two thick, blunt teeth. At the ventral idiosoma, there is a genital region and an anal region with a pair of anal suckers. All legs in the female adult have forked claws. The ventral gonopore is a longitudinal crack. The anus does not reach the posterior margin of idiosoma. 【Conclusion】 The external morphology and ultrastructure of L. konoi at different developmental stages are observed by SEM, and the morphological characteristics of the mite are understood in more details, providing a theoretical basis for further study of its taxonomy and biology.