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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
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Insect symbionts: Research progresses and prospects
LUAN Jun-Bo, WANG Si-Bao
Acta Entomologica Sinica    2023, 66 (10): 1271-1281.   DOI: 10.16380/j.kcxb.2023.10.001
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Insect symbionts are microorganisms that establish enduring and sustained associations with insect hosts. These microorganisms inhabit the body surface, gut, hemocoel, or intracellular cells of insects, participating in the regulation of various physiological functions of their host insects. Research on insect-symbiont interactions involves multidisciplinary collaboration. In-depth exploration of the functions of insect symbionts and their interactions with hosts not only advances our understanding of fundamental mechanisms in the life sciences but also introduces innovative perspectives and methods for pest management, vector-borne disease control, and optimal utilization of beneficial insects. In recent years, Chinese researchers have made noteworthy progress in the insect microbiome and got significant achievements in many research directions. In this article, we provided an overview of the most recent research progress in insect symbionts, introduced the main contents of this special issue, and proposed three noteworthy research directions: (1) the functions of insect intracellular symbionts; (2) the mechanisms by which insects regulate the abundance and transmission of symbionts; and (3) genetic modification and application of insect symbionts.
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Toxicity and risk assessment of the formulations of seven neonicotinoid insecticides to Coccinella septempunctata (Coleoptera: Coccinellidae)
YOU Yong, ZHAO Jian-Wei, TANG Bao-Hua, ZENG Zhao-Hua, LUO Feng-Qiu, CHEN Yi-Xin, YANG Feng-Hua, XIE Miao, LIU Xin-Gang, WEI Hui
Acta Entomologica Sinica    2023, 66 (6): 759-769.   DOI: 10.16380/j.kcxb.2023.06.004
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【Aim】To evaluate the toxicity and ecological risks of the formulations of seven neonicotinoid insecticides to the 7-spot ladybird, Coccinella septempunctata, so as to provide a reference for scientific use of neonicotinoid insecticides and protection of C. septempunctata.【Methods】The acute toxicity of the formulations of seven neonicotinoid insecticides, including 10% imidacloprid WP, 40% acetamiprid SP, 50% dinotefuran WP, 50% clothianidin WG, 40% imidaclothiz WG, 17% flupyradifurone SL and 25% thiamethoxam WG to the 2nd instar larvae of C. septempunctata was determined by using the method of residual film in glass tube, and the ecological risks of these insecticides were assessed. The solutions of the seven insecticides were prepared according to the maximum recommended field application rate, and sprayed on potted plants in the greenhouse to investigate the larval survival rate of C. septempunctata.【Results】The results of the indoor experiments showed that the acute toxicity of the seven neonicotinoid insecticides to the 2nd instar larvae of C. septempunctata was ranked in a descending order: 40% acetamiprid SP>40% imidaclothiz WG>10% imidacloprid WP>50% dinotefuran WP>50% clothianidin WG>25% thiamethoxam WG>17% flupyradifurone SL. The pot experiment in the greenhouse showed that the larval survival rates of C. septempunctata at 7 d after treatment with the seven insecticides at the maximum recommended field application rate were 10.00%-77.50%, and the highest larval survival rate of C. septempunctata was tested in the treatment with 25% thiamethoxam WG, while the lowest survival rate was observed in the treatment with 40% imidaclothiz WG. The results of ecological risk assessment showed that for a single application, the risk of 40% acetamiprid SP was unacceptable with the hazard quotient value in the farm (HQin) of more than 5, and those of the other pesticides were acceptable with the HQin values of less than 5. The hazard quotient values for crop or vegetable field outside the farm (HQoff crop or vegetable) and for fruit tree garden outside the farm (HQoff fruit tree) of these insecticides to C. septempunctata were less than 5, indicating that the risk is acceptable. After the second spray after 7 d, the risks of 40% acetamiprid SP and 50% dinotefuran WP were unacceptable with the HQin values of more than 5, and those of the other insecticides were acceptable with the HQin values of less than 5. The HQoff crop or vegetable and HQoff fruit tree values of the other insecticides except 40% acetamiprid SP were less than 5, indicating that the risks of these insecticides are acceptable.【Conclusion】Acetamiprid and dinotefuran should not be used twice continuously and be used in rotation in order to avoid harm to C. septempunctata.
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Construction of exogenous DNA delivery system based on ovary-delivering peptide and Gal4-UAS binding property in Anopheles sinensis (Diptera: Culicidae)
YANG Xiao-Lin, LING Xia, SUN Quan, CHEN Jie, XIANG Kai, QIU Pin-Pin, HONG Jun-Feng, YAN Zhen-Tian, WANG Rong, CHEN Bin, QIAO Liang
Acta Entomologica Sinica    2023, 66 (6): 723-735.   DOI: 10.16380/j.kcxb.2023.06.001
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【Aim】Based on the features that the P2C can be delivered into ovaries and the Gal4 protein can stably bind to the UAS sequence, to establish an efficient non-embryonic exogenous DNA delivery technical system in Anopheles sinensis.【Methods】The recombinant protein P2C-Gal4-DsRed was injected into the abdomen of female adults of A. sinensis at 20 h after sucking blood. The delivery efficiency of the recombinant protein P2C-Gal4-DsRed in the ovaries was analyzed by frozen section fluorescence observation and Western blot. The recombinant P2C-Gal4 DNA BINDING protein was prepared, transgenic plasmid and helper plasmid containing the 12×UAS repeat motif were constructed, and the in vitro binding between the recombinant protein P2C-Gal4 DNA BINDING and 12×UAS repeat motif was analyzed by electrophoretic mobility shift assay. The complexes P2C-Gal4 DNA BINDING recombinant protein+helper plasmid ITF36-12×UAS and P2C-Gal4 DNA BINDING recombinant protein+transgenic plasmid ITF2-12×UAS afm incubated in vitro were injected into the abdomen of female adults of A. sinensis at 20 h after sucking blood, and the DNA of their ovaries was extracted at 40 h after a blood meal. The delivery of exogenous DNA in vivo was analyzed by PCR amplification with specific primers and sequencing. 【Results】The ovaries of 100% female adults of A. sinensis injected with P2C-Gal4-DsRed showed obvious red fluorescence under the green filter, indicating that the P2C-Gal4-DsRed recombinant protein could be efficiently transferred into the ovaries of female adults of A. sinensis. The recombinant P2C-Gal4 DNA BINDING protein could stably bind to the 12×UAS repeat motif and the plasmid containing this repeat motif fragment. Exogenous DNA fragments were detected in the ovarian tissues of 91% and 93% of female adults of A. sinensis injected with P2C-Gal4 DNA BINDINGP2C-Gal4+ITF36-12×UAS and P2C-Gal4 DNA BINDING+ITF2-12×UAS afm, respectively. 【Conclusion】The exogenous DNA delivery technical system based on the P2C ovary-delivering peptide and the Gal4-12×UAS binding property was successfully established in A. sinensis. Through this technology platform, DNA molecules such as plasmids can be conveniently, rapidly and efficiently delivered into the ovaries of A. sinensis, laying a foundation for further simplifying genetic operations such as transgene, overexpression and gene knock-in.
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Spatiotemporal expression of WntA during the embryonic development in Locusta migratoria manilensis (Orthoptera: Locustidae)
SONG Jia, MA Yu-Jie, PU Xue, JI Rong, KABAK Iliya, YUAN Liang
Acta Entomologica Sinica    2023, 66 (9): 1139-1149.   DOI: 10.16380/j.kcxb.2023.09.001
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 【Aim】 To explore the sequence characteristics of WntA-encoding protein in the Wnt gene family of Locusta migratoria manilensis and its spatiotemporal expression profile during the embryonic development, and lay a foundation for the further functional research of LmmWntA and mining other Wnt gene family members of L. migratoria manilensis. 【Methods】 The Wnt gene family gene LmmWntA of L. migratoria manilensis was cloned by PCR and identified by neighbor-joining (NJ) method. The amino acid sequence characteristics of LmmWntA were analyzed by homologous sequence multiple alignment. Whole-mount in situ hybridization was used to screen the transcriptional signals of LmmWntA in 17 consecutive embryonic developmental stages of L. migratoria manilensis after egg laying (AEL) to 12, 24, 35, 46, 56 and 65 h, and 3, 3.5, 4, 4.5, 5, 5.5, 6.5, 8, 8.5, 9.5 and 11 d. 【Results】 The full-length CDS of LmmWntA (GenBank accession number: MW052768) of L. migratoria manilensis was cloned and is 1 101 bp, encoding 336 amino acids. LmmWntA and WntA proteins of cephalochordates, insects, clawed animals and annelids were clustered into a monophyletic group of WntA subfamily. The middle region and C-terminus of LmmWntA maintain high homology with the WntA protein sequences of the corresponding species, except for the difference in the N-terminal signal peptide region, and LmmWntA was clustered with the AmWntA protein of Apis mellifera in Hymenoptera as a sister group with the amino acid sequence identity of 59.05%. LmmWntA was first expressed in the terminal growth region of the 35 h AEL and continued to be expressed in this region until the 4 d AEL, forming stripe expression in the abdomen of each segment of the neonatal segment, and continued to be expressed in the posterior half of the optic lobe, which develops into compound eyes in the future, from the 46 h AEL to the 8.5 d AEL of L. migratoria manilensis. LmmWntA was continuously expressed in the brain from the 56 h AEL to the 5.5 d AEL. At the 65 h AEL, the stripe-like expression signal of LmmWntA in each segment of the abdomen was gradually transferred to both sides of the midline of the abdomen, and there was a clear expression signal at the base of the antennae. At the 5.5 d AEL, the expression signal of LmmWntA was further transferred to the abdominal nerve. From the 3 d AEL, LmmWntA was expressed at the distal end of the ventral somite, and later transferred to the palate, foot joint and end. As the end of the 4.5 d AEL embryo began to invaginate to form proctodeum, LmmWntA was expressed in the ventral and anterior end of the invaginated proctodeum, and most of them invaginated to the 7th somite of the abdomen. At the 9.5 d AEL, LmmWntA was expressed at the wing germ disc. 【Conclusion】 LmmWntA was dynamically expressed during the embryonic development of L. migratoria manilensis, speculating that LmmWntA is involved in the development and formation of important tissues and organs such as the posterior segment growth of embryo, nervous system (brain and abdominal nerve), compound eye, antennae, posterior end of digestive system (proctodeum), jaw, chest appendages (leg and wing) of L. migratoria manilensis. The results of this study lay the foundation of developmental biology for further research on LmmWntA deficiency.
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Insect odorscape: From odor diffusion to pest control
JIAO Long, TAN Rong-Rong, CHEN Xun, WANG Hong-Juan, HUANG Dan-Juan, MAO Ying-Xin
Acta Entomologica Sinica    2023, 66 (7): 978-991.   DOI: 10.16380/j.kcxb.2023.07.012
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The collective set of odors received by insects is called as insect odorscape. Insects rely on the reception and discrimination of the odorscape to complete life activities, such as the object localization, feeding, mating and oviposition. The behavior of insects can be manipulated by odorscape management for pest control strategy. In this article, we reviewed the research progress in the composition and diffusion of insect odorscape, influences of odorscape on insect behavior, factors affecting odorscape, odorscape reception and discrimination of insects, and the application of odorscape management in pest control. Finally, we analyzed and prospected the development direction and research focus of insect odorscape management in the future. For insects, the odor released by target is dispersed into plumes by air flow and mixed with the background odor carried in the air, which together form the odorscape. Insects search and locate the target along the target odor. The behavior of insects can be affected by the shape, composition and concentration of target odors. The background odor plays a complementary or warning role during the target localization of insects. Different background odors can synergize or interfere with the target localization of insects. The insect odorscape is mainly affected by the temperature, humidity, light, heavy metal elements and plant diseases and insect pests in the environment. Studies showed that the olfactory receptors of insects capture odorscape and transmit those odor signals to olfactory nerve centers such as antennal sensilla lobe along the olfactory nerves. Then, the odorscape is analyzed in the nerve centers by the mode of elemental processing or configural processing. The influences of background odor on insect target localization may be the results of the reciprocal addition, competitive binding or signal crosstalk of different odor molecules during the olfactory sensing and coding. At present, several kinds of green pest control technologies have been developed based on the odorscape management, such as insect behavior regulators, exogenous elicitors, breeding the crop varieties that can release resistant volatiles, “push-pull” technology and plant-mediated support system for natural enemies. In the future, it is necessary to further explore the behavioral, electrophysiological and neurological mechanisms of odorscape discrimination in insects, and optimize and integrate the green control technologies related to odorscape management, so as to build rational and efficient odorscape for insect pest control.
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Structural characteristics and seasonal dynamics of ground-dwelling ant communities in three habitats in Lishui District, Nanjing, East China
GUO Zong-Lin, JIANG Fu-Guo, WANG Ya-Lu, CHEN Xiao-Yi, WANG Guo-Bing
Acta Entomologica Sinica    2023, 66 (6): 816-824.   DOI: 10.16380/j.kcxb.2023.06.010
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【Aim】To investigate the diversity and seasonal dynamics of ground-dwelling ant communities in three habitats in Lishui District, Nanjing, East China.【Methods】A one-year seasonal sampling was conducted from August 2020 to July 2021 in three different sampling habitats (farmland, grassland and plantation forest) in Lishui District, Nanjing, and the individual density, species number, Simpson dominance index, Shannon-Wiener diversity index, Pielou’s evenness index and Jaccard similarity coefficient of grounddwelling ant communities, and the seasonal dynamics of each index in these three habitats were investigated.【Results】During August 2020 to July 2021, the dominant species of ground-dwelling ants in farmland in Lishui District, Nanjing were Tetramorium caespitum (490 individuals, 58.75%), Ochetellus glaber (116 individuals, 13.91%) and Formica japonica (98 individuals, 11.75%), those in grassland were T. caespitum (427 individuals, 58.17%) and Nylanderia bourbonica (110 individuals, 14.99%), and those in plantation forest were T. caespitum (240 individuals, 27.18%), F. japonica (121 individuals, 13.70%), N. sharpii (98 individuals, 11.10%) and Pristomyrmex punctatus (90 individuals, 10.19%). The numbers of species and individuals in ground-dwelling ant communities varied significantly between habitats, with the highest numbers of species and individuals (29 species, 883 individuals) collected from plantation forest and the lowest numbers of species and individuals (19 species, 734 individuals) collected from grassland. The analysis of variance showed that seasonal changes had a significant effect on the number of species and individual density of ground-dwelling ants in this region, while habitat changes had a significant effect on the number of species and no significant effect on the individual density, but there was no interactive effect of both seasonal and habitat changes on the number of species and individual density of ground-dwelling ants in this region. The analysis of community diversity showed that the seasonal trends in the Shannon-Wiener diversity index, Pielou’s evenness index and Simpson dominance index of the ground-dwelling ant communities in the three different habitats were basically the same, with an increasing trend followed by a decreasing trend, while the seasonal trends of the Simpson dominance index and Pielou’s evenness index were basically opposite. The Jaccard similarity coefficients of the ground-dwelling ant communities between farmland and grassland, farmland and plantation forest, and grassland and plantation forest were 0.73, 0.52 and 0.52, respectively, which were moderately similar to each other, while those of the grounddwelling ant communities between different months were low, and most of them were extremely dissimilar. 【Conclusion】The numbers of individuals and species in the ground-dwelling ant communities in Lishui District, Nanjing are relatively rich, and the structure of the ground-dwelling ant communities is significantly influenced by habitat type changes and seasonal changes, so attention should be paid to strengthen the protection of ant species resources in the region in different seasons.
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Molecular characteristics and functional study of heat shock protein gene SmHsp60 in Sitodiplosis mosellana (Diptera: Cecidomyidae)
MA Qian, DONG Jin-Hui, LI Fang-Xiang, ZHU Ke-Yan, CHENG Wei-Ning
Acta Entomologica Sinica    2023, 66 (7): 859-869.   DOI: 10.16380/j.kcxb.2023.07.001
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【Aim】The wheat blossom midge, Sitodiplosis mosellana, an important agricultural pest, survives under temperature extremes during summer and winter by diapause. This study aims to explore the relationship between heat shock protein 60 (Hsp60) gene expression and diapause development and temperature tolerance in the diapause process of S. mosellana.【Methods】 The full-length cDNA sequence of Hsp60 of S. mosellana (SmHsp60) was amplified via RACE and RT-PCR technologies, and analyzed via bioinformatics. qPCR was used to detect the expression levels of SmHsp60 in the S. mosellana larvae at different stages from pre-diapause to post-diapause development including pre-diapause, diapause, post-diapause quiescence and post-diapause development, as well as oversummering larvae under extremely high temperature stress [exposed to water bathes at 34, 40, 45 and 50 ℃ for 1 h, and 35 ℃ for 0 (control), 15, 30, 60 and 120 min] and overwintering larvae under extremely low temperature stress [exposed to 0, -5, -10 and -15 ℃ for 1 h, and -5 ℃ for 0 (control), 15, 30, 60 and 120 min]. The recombinant SmHsp60 protein was obtained by prokaryotic expression and affinity column chromatography, and its ability to suppress the thermal aggregation of mitochondrial malate dehydrogenase (MDH) at 43 ℃ was examined by colorimetry and SDS-PAGE.【Results】 The full-length cDNA sequence of SmHsp60 (GenBank accession no: KR733065) of S. mosellana obtained is 2 270 bp in length, which contains an open reading frame (ORF) of 1 722 bp in length encoding a protein of 573 amino acids with the relative molecular weight of 60.7 kD. Amino acid sequence analysis indicated that SmHsp60 contains the classical signature sequences of mitochondrial Hsp60, and displayed the highest amino acid identity and the closest relationship to Hsp60 from Contarinia nasturtii of Cecidomyiidae. qPCR detection result showed that the expression level of SmHsp60 did not change significantly in the diapause stage, but began to increase gradually in the post-diapause quiescence stage with a peak in early-to-mid phase of post-diapause quiescence (i.e. December and next January) and was significantly higher than those in the other developmental stages. Compared to the untreated control, the expression of SmHsp60 in the oversummering larvae under 35 and 40 ℃for 1 h and 35 ℃ for 30-60 min and that in the overwintering larvae under -5 ℃ for 1 h was significantly induced, while the expression level of SmHsp60 did not change significantly above 45 ℃ or below -10 ℃. The highly purified recombinant SmHsp60 was able to effectively suppress the thermal aggregation of MDH, indicating its significant molecular chaperone function.【Conclusion】 SmHsp60 is involved in diapause regulation of S. mosellana and might play a role in diapause termination, as well as heat tolerance and cold tolerance during diapause.
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Cloning and expression profiling of heat shock protein genes AcHsp83a and AcHsp83b from Arma chinensis (Hemiptera: Pentatomidae) and their response to high or low temperature and UV-B stresses
MENG Jian-Yu, JIN Xin, HE Long-Chun, ZHANG Xue-Xia, YANG Chang-Li, ZHANG Chang-Yu
Acta Entomologica Sinica    2023, 66 (11): 1425-1434.   DOI: 10.16380/j.kcxb.2023.11.001
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【Aim】 To explore the molecular mechanism of the natural enemy insect Arma chinensis in response to high or low temperature and UV-B stresses.【Methods】 The heat shock protein genes AcHsp83a and AcHsp83b of A. chinensis were cloned by RT-PCR, and their sequence characteristics were analyzed by bioinformatics methods. RT-qPCR was used to detect the expression levels of AcHsp83a and AcHsp83b in different developmental stages (egg, 1st-5th instar nymphs, female adult and male adult), different adult tissues (head, thorax, abdomen, wing, antenna, fat body, leg, Malpighian tubules, mouthparts, midgut, ovary and testis), and female and male adults at 0 (CK), 6 and 24 h after exposure to high temperature of 38 ℃, and low temperature of 4 ℃, and at 0 (CK), 6 and 12 h after UV-B stress.【Results】 Two Hsp90 genes were cloned from A. chinensis, and namely AcHsp83a (GenBank accession no.: OP791883) and AcHsp83b (GenBank accession no.: OP791884). Their open reading frames (ORFs) were 2 172 and 2 163 bp in length, encoding 723 and 720 amino acids with the relative molecular weight of 83.12 and 82.90 kD and the isoelectric point (pI) of 4.94 and 4.97, respectively, and the C-terminal sequences both contain the conserved motif EEVD, showing that they belong to the cytoplasmic Hsps. AcHsp83a and AcHsp83b are highly conserved. The expression level of AcHsp83a was the highest in eggs, whereas that of AcHsp83b was the highest in adults. The expression level of AcHsp83a was the highest in the testis of male adults, whereas that of AcHsp83b was the highest in the midgut of female adults. When the female adults were exposed to 38 ℃, 4 ℃ or UV-B, the expression levels of AcHsp83a and AcHsp83b increased firstly and then decreased with treatment time, reaching the peak at 6 h. For the male adults, the expression level of AcHsp83a increased firstly and then decreased with treatment time, and reached the peak at 6 h after exposure to 38 ℃ or UV-B. The expression level of AcHsp83a in male adults decreased firstly and then increased with treatment time, and reached the peak at 24 h after exposure to 4 ℃. Compared with the control group, 38 ℃, 4 ℃ or UV-B exposure significantly decreased the expression levels of AcHsp83b in male adults. 【Conclusion】 The differential expression of AcHsp83a and AcHsp83b in A. chinensis suggests that the two genes play important roles in the growth and development of A. chinensis and contribute to its tolerance to extreme temperatures and UV-B stress.
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Advances in research and application of sex pheromones of the tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae)
LIANG Yong-Xuan, WANG Qi-Jing, GUO Jian-Yang, WANG Yu-Sheng, ZHANG Yi-Bo, YANG Nian-Wan, ZHANG Gui-Fen, ZHOU Qiong, LIU Wan-Xue
Acta Entomologica Sinica    2023, 66 (6): 849-858.   DOI: 10.16380/j.kcxb.2023.06.013
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The tomato leafminer, Tuta absoluta, is native to South America and a quarantine pest in the world nowadays, which can reduce the yield of the main host crop tomato by 80%-100% in severe cases. In over a decade, this insect pest has invaded and spread from its origin to almost the entire continent of Asia, Africa and Europe, becoming a major threat to the world tomato industry. T. absoluta was discovered in Ili, Xinjiang in 2017 and spread to Yunnan, Gansu and other regions in a short time, greatly threatening the healthy development of the tomato industry and other related agricultural industries in China. Internationally, sex pheromone-based monitoring, mass trapping and mating disruption control have become one of the important  measures to control T. absoluta. In order to study the efficient and environmentally friendly population dynamics monitoring and control technology of T. absoluta, we summarized the research and application status of the sex pheromone in mornitoring, early warning and control of T. absoluta in this article. The females of T. absoluta release sex pheromones to attract males to mate mainly in the morning, and adult mating reaches the peak at 7:00 a.m. In 1995 and 1996, the major and minor components of the sex pheromone released from T. absoluta and their synthetic methods were reported successively. The major and minor components of this sex pheromone are (3E, 8Z, 11Z)-tetradecatrienyl acetate (TDTA) and (3E, 8Z)-tetradecadienyl acetate (TDDA), respectively, which constitute the sex pheromone in a ratio of about 90∶10. Bioassay results revealed that TDTA had a strong attraction to males, which could be further enhanced when its mixture with TDDA was used. In recent years, more efficient and convenient synthetic methods of sex pheromone components have been developed, providing good conditions for their large-scale production and application. At present, the sex pheromone of T. absoluta has been widely used in the control of this insect pest in the world, and good results have been achieved. Among the reported sex pheromone traps commonly used for monitoring and mass trapping this insect pest, the water basin trap and triangle trap are more effective. The common doses of sex pheromone contained in commercial lure core are 0.5, 0.8 and 3.0 mg. For field application, the corresponding trap type and sex pheromone dose should be selected according to the actual situation of habitat and growth stage of crops, damage degree, local natural conditions, etc. In addition, the mating disruption by using sex pheromone is also common in the control of T. absoluta, and its successful application requires a highly enclosed environment. Because sex pheromone has the advantages of high efficiency, safety and environmental protection in the monitoring and control of T. absoluta, its related research results can provide important reference for the sustainable control strategies and measures of this insect pest in China.
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Effects of carrying Bursaphelenchus xylophilus on the bacteria in the gut and trachea of adult Monochamus alternatus (Coleoptera: Cerambycidae)
GU Yu-Tong, ZHU Hao-Cheng, CHANG Yue, ZHOU Jiao, FAN Jian-Ting, ZHAO Li-Lin
Acta Entomologica Sinica    2023, 66 (10): 1319-1328.   DOI: 10.16380/j.kcxb.2023.10.006
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 【Aim】 To investigate the effects of Bursaphelenchus xylophilus in Monochamus alternatus on the bacteria in the gut and trachea of M. alternatus.【Methods】The whole gut and trachea were isolated from the collected M. alternatus adults carrying B. xylophilus in the field to extract DNA for sequencing and splicing of the 16S rDNA gene. The composition, structure, abundance and diversity of bacteria in the gut and trachea of M. alternatus adults carrying B. xylophilus were analyzed by bioinformatics methods. 【Results】A total of 15 phyla, 26 classes, 66 orders, 110 families, 201 genera, 296 species and 444 operational taxonomic units (OTUs) of bacterial flora in the gut and trachea of M. alternatus adults carrying B. xylophilus were detected. There was no significant change in the dominant bacterial flora in the gut between M. alternatus adults carrying B. xylophilus and M. alternatus adults without carrying B. xylophilus, and their dominant bacterial flora were Proteobacteria and Enterobacterales. Proteobacteria and Enterobacterales were the dominant bacterial flora in the trachea of M. alternatus adults carrying B. xylophilus, while Firmicutes and Lactobacillales were the dominant bacterial flora in the trachea of M. alternatus adults without carrying B. xylophilus. The diversity and abundance of bacteria in the trachea of M. alternatus adults carrying B. xylophilus were higher than those of M. alternatus adults without carrying B. xylophilus, and the bacterial community structure in the trachea of M. alternatus adults carrying B. xylophilus was significantly different from that of M. alternatus adults without carrying B. xylophilus. Serratia and Enterobacter were enriched in the gut and trachea of M. alternatus adults carrying B. xylophilus. 【Conclusion】The introduction of B. xylophilus has significant effects on the composition, structure, abundance and diversity of bacterial flora in the trachea of M. alternatus adults. Serratia and Enterobacter in the trachea of M. alternatus adults may be closely related to the B. xylophilus- M. alternatus  complex. The increase of Serratia in the trachea of M. alternatus adults carrying B. xylophilus may be associated with the suppressed tracheal immunity of M. alternatus. This study provides a theoretical basis for the study of bacterial diversity in the gut and trachea of M. alternatus, and a new direction for the prevention and control of B. xylophilus.
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Nuclear receptor gene SfUSP regulates the molting development of the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)
ZHOU Cao, GONG Ming-Fu, YANG Xi-Bin, YANG Hong, LONG Gui-Yun, JIA Ze-Yan, ZENG Qing-Hui, JIN Dao-Chao
Acta Entomologica Sinica    2023, 66 (5): 609-618.   DOI: 10.16380/j.kcxb.2023.05.001
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【Aim】To explore the function of nuclear receptor ultraspiracle protein (USP) in the molting development of Sogatella furcifera nymph and its regulatory relationship with chitin synthesis and degradation. 【Methods】Based on the genome data of S. furcifera, the full-length cDNA sequence of SfUSP was amplified by RT-PCR. The expression levels of SfUSP in different developmental stages (1st-5th instar nymphs, 5th instar nymph prior to ecdysis, 5th instar nymph during ecdysis and female adult), tissues (head, integument, fat body, gut and leg) of the 5th instar nymph, female adult tissues (integument, wing, fat body, leg and ovary) and the day-1 5th instar nymph of S. furcifera after treatment with 100 ng/individual of 20E were detected by RT-qPCR. After the targeted silencing of SfUSP by microinjection of dsRNA into the day-1 5th instar nymph, the nymphal survival rate was calculated, the lethal phenotype of nymphs was observed and the expression levels of the key genes in chitin synthesis and degradation pathways were determined by RT-qPCR. 【Results】The full-length cDNA sequence of SfUSP (GenBank accession number: ON209396) of S. furcifera was obtained by cloning, with an open reading frame of 1 263 bp in length, encoding 420 amino acids. The predicted molecular weight of SfUSP is 47.27 kD with the theoretical isoelectric point of 7.18. Sequence analysis result showed that SfUSP contains five conserved domains of nuclear receptor family, and DNA-binding domain (DBD) and ligand-binding domain (LBD) are highly conserved. The developmental stage expression profiles showed that SfUSP was highly expressed in the 1st instar nymph, and the 5th instar nymphs prior to ecdysis and female adult. The tissue expression profiles showed that SfUSP was expressed highly in the head, integument, fat body and gut of the 5th instar nymphs, and in the wing, leg and integument of the female adult. In addition, at 12 h after microinjection of 20E (100 ng/individual), the expression level of SfUSP in the 5th instar nymph was significantly increased as compared with that in the control group. After targeted silencing of SfUSP expression, the nymphal survival rate of S. furcifera was significantly reduced compared with the control group (injected with dsGFP) and only 18.01% of that of the control group at 6 d after microinjection of dsRNA, with 51.46% individuals unable to molt successfully. In addition, after targeted silencing of SfUSP expression, the expression levels of the key genes SfCHS1, SfCHS1a, SfUAP and SfGFAT in chitin synthesis pathways and SfCht7, SfNAG1, SfNAG2, SfCDA1, SfCDA2 and SfCDA4 in chitin degradation pathways were significantly inhibited, but those of SfG6PI and SfCht10 were significantly increased as compared with those in the control group microinjected with dsGFP. 【Conclusion】SfUSP is a key gene in the growth and development of S. furcifera, which can affect the synthesis and degradation of chitin, and then regulates the ecdysis development of S. furcifera.
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Research advances in the structure and function of insect fat bodies and their application in human disease models
MA Zhen-Gang, HE Yu-Bo, MA Qiang, WANG Jing-Lin, ZHOU Ze-Yang
Acta Entomologica Sinica    2023, 66 (9): 1246-1257.   DOI: 10.16380/j.kcxb.2023.09.011
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Insect fat bodies are widely distributed in the body of insects, and they are a dynamically loose tissue that includes peripheral fat bodies and visceral fat bodies. The structure and function of insect fat bodies are closely related, and their structure changes with the changes of different life stages, and their functions also change. However, the most basic function of insect fat bodies is to act as the core tissue of insect life metabolism to synthesize and store substances. In addition, fat bodies, which are the target organs of various hormones, have many important functions and are related to the innate immunity, longevity, growth and development of insects. Therefore, the study on insect fat body is very important for the analysis of the important life phenomena of insects. In recent years, a large number of studies have taken insect fat bodies as research models for human diseases and drug development, and applied them in many fields such as human immunology, pathogenesis of human diseases, and new drug research and development. Here, in order to systematically deepen the understanding of the structure and function of insect fat bodies, the morphological structure, formation and metamorphosis, biological function, and their application and prospect in the study of human disease models were reviewed. Insect fat bodies would have great application potential in the construction of major human disease models and the exploration of disease pathogenesis.
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Regulation of adipokinetic hormone on the growth and development of the Bactrocera dorsalis (Diptera: Tephritidae) larva
HOU Qiu-Li, ZHANG Han-Qiao, JIANG Hong-Bo, WANG Jin-Jun
Acta Entomologica Sinica    2023, 66 (5): 628-635.   DOI: 10.16380/j.kcxb.2023.05.003
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【Aim】Adipokinetic hormone (AKH), a type of neuropeptide, plays central roles in regulating development and behavioral events in diverse insects. Investigating the functions of AKH on the growth and development of Bactrocera dorsalis larva will contribute to a better understanding of the conservation and functional diversity of neuropeptides in insects, and may shed light on evaluating the potential of AKH signal system as the target of novel insect control agents. 【Methods】The full-length cDNA sequence of BdAKH of B. dorsalis was cloned, and characterized with bioinformatics methods. The expression levels of BdAKH in different developmental stages (egg, 1st-3rd instar larvae, pupa, virgin female adult and male adult), and different tissues [central nervous system, corpora cardiaca/corpus allatum (CC/CA), fat body, Malpighian tubules, midgut, hindgut, cuticle, ovary, testis and hemocytes] of the late 3th instar larva of B. dorsalis were detected by qPCR. Immunohistochemistry technique was used to localize the neurons expressing BdAKH in the 3rd instar larva of B. dorsalis. RNAi combined with biological observation was used to analyze the regulation of BdAKH on the growth and development of B. dorsalis larva.【Results】The full-length cDNA sequence of BdAKH (GenBank accession no. KY073319) of B. dorsalis was cloned. This cDNA of 572 bp contains an open reading frame (ORF) of 252 bp, which encodes 83 amino acid residues. BdAKH of B. dorsalis has conserved structure that consists of one mature peptide (QLTFSPDWamide). The expression levels of BdAKH were higher in adult. BdAKH was mainly expressed in the CC/CA of the late 3rd instar larva. Compared with the control group (dsGFP), BdAKH knockdown in the 3rd instar larva of B. dorsalis caused a delay in pupation and decreased pupation rate, while the pupal weight and phenotype had no remarkable difference. 【Conclusion】AKH plays important roles in regulating the larval growth and development of B. dorsalis, which could be viewed as new targets of novel insect control agents.
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Analysis of the functional mechanism of ATP synthase subunit α in the metamorphosis of Helicoverpa armigera (Lepidoptera: Noctuidae) larvae
GENG Zi-Chen, CHANG Yan-Peng, ZHANG Bo, AN Shi-Heng, ZHAO Wen-Li
Acta Entomologica Sinica    2023, 66 (5): 619-627.   DOI: 10.16380/j.kcxb.2023.05.002
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 【Aim】This study aims to analyze the functional mechanism of ATP synthase subunit α in the metamorphosis and development of Helicoverpa armigera.【Methods】The open reading frame (ORF) of ATPS-α gene of H. armigera was amplified by PCR and analyzed by bioinformatics. qRT-PCR was employed to detect the expression levels of HaATPs-α in the cuticle, midgut and fat body of larva at the 5th instar molting stage and day-1-5 of the 6th instar, and the cuticle and fat body of the 6th instar larva after the induction of exogenous 20-hydroxyecdysone (20E)(0.1 mg/mL). The 6th instar larvae of H. armigera were injected with dsHaATPs-α, and the effects of knocking down the expression level of HaATPs-α by RNAi on the development and metamorphosis of larvae, and the ATP content, trehalose content, glucose content and trehalase activity in the larva were analyzed. 【Results】The ORF length of HaATPs-α of H. armigera is 1 677 bp. The amino acid sequence identity of ATPs-α in H. armigera, Spodoptera frugiperda, S. litura and Trichoplusia ni of Noctuidae of Lepidoptera was 96.56%, and their evolution relationship was close. The expression pattern analysis result showed that the expression levels of HaATPs-α in the cuticle and midgut of the day-3 6th instar larvae were the highest and also peaked in the fat body of the 5th instar larvae at the molting stage. Treatment with 20E(0.1 mg/mL) significantly upregulated the expression level of HaATPs-α in the 6th instar larvae compared with the control. Compared with the control group (injected with dsGFP), knocking down the expression level of HaATPs-α by RNAi delayed the larval development, significantly reduced larval weight, significantly increased larval mortality, significantly decreased pupation rate and adult emergence rate, significantly decreased ATP content and trehalase activity, significantly increased trehalose content, and significantly decreased glucose content. 【Conclusion】HaATPs-α not only controls ATP production, but also affects the contents of trehalose and glucose in H. armigera. Therefore, HaATPs-α plays an important role in larval metamorphosis. Furthermore, our results also provide experimental evidence and theoretical basis for using ATPs-α as a new target for pest control in the future.
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Detection and recognition of grooming behaviors of Bactrocera dorsalis (Diptera: Trypetidae) based on YOLO V7 and TSM model
LIU Hu, ZHAN Wei, HE Zhang-Zhang, TANG Jian-Tao, JIANG Zhen-Yu, SUN Yong
Acta Entomologica Sinica    2023, 66 (6): 787-796.   DOI: 10.16380/j.kcxb.2023.06.007
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【Aim】 Statistical analysis and study of insect grooming behaviors are important for pest control and human health. In view that the traditional method of manually recording grooming behaviors is time-consuming and error-prone, we proposed a computer vision and deep learning-based method for the detection and recognition of grooming behaviors of the Oriental fruit fly, Bactrocera dorsalis. 【Methods】 First, we processed the B. dorsalis video data to obtain frame images, and screened 3 000 images as the training dataset. We built the YOLO V7 target detection algorithm to detect B. dorsalis target in video data, and framed the target and cropped it by video processing algorithm. Finally, we migrated the pre-training weights to the training model by transfer learning method, and recognized seven grooming behaviors (foreleg grooming, head grooming, fore-midleg grooming, mid-hindleg grooming, hindleg grooming, wing grooming and stationary) of B. dorsalis using the temporal shift module (TSM) deep learning model based on non-local attention improvement. 【Results】 The accuracy and recall rate of the original video of the B. dorsalis dataset trained by YOLO V7 target detection algorithm were 99.2% and 99.1%, respectively. Applying this research algorithm to process the video dataset, and then recognizing and counting grooming behaviors through an improved TSM model based on the non-local attention module, we got the final average accuracy of over 97% with a standard deviation of less than 3%. Compared with the other four deep learning models (I3D, R2+1D, SlowFast and Timesformer), this research method had a ~9.76% improvement in accuracy, ensuring the accuracy of B. dorsalis grooming behavior detection and recognition. 【Conclusion】 The method proposed in this study greatly reduces the time of manual observation, and ensures the accuracy of grooming behavior recognition of B. dorsalis, providing new ideas and methods for researching insect behavior and contributing to the modern development of intelligent agriculture.
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Effects of light intensity on the oviposition preference of Grapholita molesta (Lepidoptera: Tortricidae)
YANG Xiao-Fan, JIAO Hui-Tan, LI Xu-Zhao, LU Zi-Yun, RAN Hong-Fan, MA Ai-Hong, WEI Guo-Shu, LI Jian-Cheng
Acta Entomologica Sinica    2023, 66 (7): 918-924.   DOI: 10.16380/j.kcxb.2023.07.006
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【Aim】Grapholita molesta is an important and frequent pest of fruit trees. This study aims to investigate the roles of light intensity on the oviposition preference of G. molesta, and to further reveal the dim-light vision ability in G. molesta. 【Methods】The numbers of eggs of G. molesta laid on leaves on the sunny side and shaded side in the peach tree canopy were investigated in spring, summer and autumn, and the difference in light intensity between the sunny side and shaded side at sunset was measured. Light-dark two-choice experiments were carried out indoor to test the oviposition preference behavior response of G. molesta adults to light intensity. 【Results】 In spring, summer and autumn, G. molesta adults preferred to lay eggs on leaves on the sunny side in the peach tree capony with significantly higher light intensity, the numbers of eggs laid on the sunny side were 4.24, 9.30 and 5.82 times as high as those on the shaded side, respectively. In the light-dark two-choice experiments, G. molesta adults preferred to lay eggs to light than to dark, with the oviposition selection rates of 87.48%, 83.68%, 82.92%, 80.08% and 84.84%, respectively, under 10 000, 100, 1 and 0.01 lx and natural light conditions. The oviposition preferences between to strong light and to weak light revealed that when the light contrast (strong light/weak light) was 10 and 2, respectively, G. molesta adults showed a preference to strong light under 10 000, 100, 1 and 0.01 lx and natural light conditions. Moreover, this preference to strong light did not decrease significantly with the decrease of light intensity, and the oviposition selection rates were higher than 75%.【Conclusion】 Light intensity can affect the oviposition preference of G. molesta adults. Female adults show obvious oviposition preference to strong light, and can still discriminate well the difference in light intensity even under dim light, exhibiting a well dim-light vision ability. The results provide a basis for new physical trapping techniques based on visual behavior regulation.
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Resistance analysis of the antimicrobial peptide SAAP-148 against the pathogenic bacteria of bacterial intestinal disease in Bombyx mori
WANG Jue, SHEN Dong-Xu, PENG Yun-Wu, QIAN He-Ying, ZHAO Qiao-Ling, CHEN An-Li
Acta Entomologica Sinica    2023, 66 (6): 736-746.   DOI: 10.16380/j.kcxb.2023.06.002
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【Aim】Bacterial intestinal disease is one of the main diseases of Bombyx mori. The purpose of this study is to explore the inhibitory effects of the broad-spectrum antimicrobial peptide SAAP-148 on the pathogenic bacteria of bacterial intestinal disease in B. mori, and to provide references for further use of antimicrobial peptides instead of antibiotics to prevent bacterial diseases in B. mori.【Methods】The expression levels of antimicrobial peptide genes in the midgut of healthy and bacterial intestinal disease-infected individuals of the day-4 5th instar larvae of B. mori were detected by qRT-PCR. The bacterial flora in the midgut of the day-4 5th instar larvae of B. mori infected with bacterial intestinal disease was isolated, and the phylogenetic tree was constructed by maximum likelihood method. The pathogenicity of the eight isolated bacteria and four common pathogenic bacteria of bacterial disease including Enterococcus faecalis, Bacillus bombyseptieus, B. thuringiensis and Serratia marcescens, to the healthy 5th instar larvae of B. mori was determined by feeding method (1×1010 and 1×1014 CFU/mL bacterial suspensions) and puncture inoculation method (2×108 and 2×109 CFU/mL bacterial suspensions). The inhibitory effects of the antimicrobial peptide SAAP-148 on the isolated pathogenic bacteria of intestinal disease and other common pathogenic bacteria of bacterial disease such as B. bombyseptieus were analyzed by agar plate diffusion method.【Results】The expression levels of the detected antibacterial peptide genes of B. mori in the midgut of the day-4 5th instar larvae of B. mori infected with bacterial intestinal disease were significantly up-regulated as compared with those of the healthy individuals, indicating that the antibacterial peptides in B. mori play a certain role in resisting bacterial intestinal disease. A total of eight pathogenic bacterial species (Mammaliicoccus sciuri, Aeromonas hydrophila, Enterobacter cancerogenus, E. bugandensis, E. hormaechei, Citrobacter freundii, Klebsiella pneumoniae and K. variicola) were isolated from the midgut of the day-4 5th instar larvae of B. mori infected with bacterial intestinal disease. The puncture results showed that the eight bacterial species were pathogenic. Among the eight bacterial species, M. sciuri, A. hydrophila, E. cancerogenus and E. bugandensis had strong pathogenicity, and E. hormaechei, C. freundii, K. pneumoniae and K. variicola were less pathogenic. The antibacterial peptide SAAP-148 had in vitro inhibitory effects on the eight isolated pathogenic bacteria and other four common pathogenic bacteria of B. mori bacterial disease, and the inhibitory effect increased with the increase of SAAP-148 concentration.【Conclusion】The antibacterial peptide SAAP-148 not only has obvious inhibitory effects on the pathogenic bacteria that may cause bacterial intestinal disease of B. mori, but also has inhibitory effects on the pathogenic bacteria that cause black chest sepsis, prodigiotic sepsis, bacterial poisoning of B. mori, and E. hormaechei which has produced resistance to ampicillin. SAAP-148 can be used as an ideal antibacterial agent to prevent the occurrence of bacterial disease of B. mori.
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Morphological characteristics and nesting behavior of Trypoxylon melanocorne (Hymenoptera: Sphecidae)
HU Jun-Jie, FANG Quan-Bo, WANG Xiang, SUN Kai-Li, LIU Xiao-Yu, HE Chun-Ling
Acta Entomologica Sinica    2023, 66 (6): 805-815.   DOI: 10.16380/j.kcxb.2023.06.009
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 【Aim】 As carnivorous insects preying on spiders, different species of Trypoxylon possess obviously different nesting strategies. This study aims to clarify the morphological characteristics and nesting behavior of T. melanocorne, so as to reveal the nesting and survival strategies of Trypoxylon. 【Methods】Man-made nest-trap boxes were set up by choosing different habitats in the Yugong Forest Farm and Nanshan Forest Farm in Jiyuan City, Henan Province, Central China from 2019 to 2020, and then the trap-nests that were nested by T. melanocorne adults were regularly collected. The internal structure data of the trap-nests were measured by dissecting trap-nests, and the development of offspring was observed and the parasitic enemies were collected in the laboratory. 【Results】 A total of 111 trap-nests nested by T. melanocorne adults were collected in the Yugong Forest Farm and Nanshan Forest Farm in Jiyuan City, Henan Province during 2019-2020, and 511 brood cells were dissected. The average number of eggs laid by females in a single trap-nest was 4.60±2.39 grains, and the majority of offspring was female (female to male ratio=2∶1). The average length of nests and length of brood cells were (123.38±46.45) and (16.01±5.57) mm, respectively. The numbers of intercalary cells and nests with vestibular cells accounted for 5.04% and 6.09%, respectively, of the total number of cells. The number of brood cells was significantly affected by the length of nests, and the length of cells of females was significantly longer than that of males. Two parasitic enemies of T. melanocorne, Trichrysis pellucida (Chrysidoidae) and Amobia quatei (Sarcophagidae) were found, with the parasitism rates of 196% and 1.57%, respectively. 【Conclusion】 The results of this study suggest that the population size of T. melanocorne distributed in the two forest farms in the southern Taihang Mountains, Henan Province is small, female progeny are more numerous than males, and one of the reasons for the small population size probably is low adult eclosion rate and sex ratio imbalance.
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Gene cloning, expression profiling and ligand binding characteristics of GmolNPC2 from Grapholita molesta (Lepidoptera: Tortricidae)
LI Chun-Qin, ZHANG Yu-Xi, XU Shi-Cai, LI Bo-Liao, CHEN Xiu-Lin, LUO Kun, LI Guang-Wei
Acta Entomologica Sinica    2023, 66 (7): 870-884.   DOI: 10.16380/j.kcxb.2023.07.002
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【Aim】The purpose of this study is to provide a theoretical reference for the further study on the function of the Niemann-Pick type C2 protein (NPC2) of Grapholita molesta GmolNPC2 in olfactory sensing.【Methods】Based on the antennal transcriptome data of G. molesta, the full-length cDNA sequence of GmolNPC2 was amplified by RT-PCR, and the phylogenetic analysis and 3D structure model prediction of GmoNPC2 were performed. The relative expression levels of GmolNPC2 in different developmental stages (egg, 1st-5th instar larvae, pupa, and female and male adults) and different tissues (antenna, head without antenna, thorax, abdomen, leg, and wing) of the 3-day-old male and female adults were detected by RT-qPCR. The inhibitory constant (Ki) values of the recombinant GmolNPC2 to 44 odorant ligands including six sex pheromones and 38 host plant volatile compounds were determined by fluorescence competitive binding assays to analyze the binding ability of GmolNPC2 to odorant ligands. By using molecular docking simulations, the binding energy of GmolNPC2 with different odorant ligands was calculated and the key amino acid residues of protein-ligand interactions were predicted.【Results】The full-length cDNA sequence of GmolNPC2 (GenBank accession no.: OQ054801) of G. molesta was obtained and the open reading frame (ORF) is 438 bp in length, encoding 145 amino acids. Multiple sequence alignment showed that GmolNPC2 has the typical structural characteristics of NPC2 of insects. Phylogenetic analysis showed that the reported NPC2s of lepidopteran insects were clustered into two branches, and the GmolNPC2 sequence was highly identic to the NPC2 sequences of Leguminivora glycinivorella, Pectinophora gossypiella, and Manduca sexta, which were clustered into one branch. RT-qPCR detection result showed that the expression levels of GmolNPC2 in the male adult and egg were significantly higher than those in other developmental stages. The expression level of GmolNPC2 was the highest in the wing of the 3-day-old male and female adults followed by that in the antenna, and the lowest in the thorax, abdomen, and leg. The binding spectrum of the recombinant GmolNPC2 was narrow with only 17 out of 44 tested odorant ligands, among which, the recombinant GmolNPC2 exhibited strong binding abilities to (Z)-8-dodecenyl acetate, benzyl alcohol, and cis-3-hexenyl acetate, with the Ki values of (4.117±0.046), (4.845±0.079) and (3.979±0.167) μmol/L, respectively. The molecular docking simulation result showed that the binding energy between GmolNPC2 and each odorant ligand was different, which was consistent with the results of the above fluorescence binding assays. Hydrophobic amino acid residues and hydrogen bonds play an important role in GmolNPC2 binding with different odorant ligands.【Conclusion】 GmolNPC2 has the conserved structure of the insect NPC2 family and is highly expressed in the egg and adult, and the wing and antenna of adult, speculating that GmolNPC2 participates in different physiological processes in G. molesta. The recombinant GmolNPC2 selectively binds sex pheromones and host plant volatile compounds, indicating the involvement of GmolNPC2 in the detection and transportation of volatile semiochemicals.
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Analysis of the adult antennal transcriptome and olfaction-related genes of Pieris rapae (Lepidoptera: Pieridae)
XUE Zeng-Sheng, YAN Xi-Zhong, ZHAO Jing-Yu, SONG Cheng-Fei, LIU Dan, WANG Han, HAO Chi, MA Li
Acta Entomologica Sinica    2023, 66 (5): 642-652.   DOI: 10.16380/j.kcxb.2023.05.005
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 【Aim】To establish the antennal transcriptome database of the Pieris rapae adult, so as to deeply mine the gene data of P. rapae by using bioinformatics and molecular biology methods.【Methods】High-throughput sequencing platform Illumina NovaSeq 6000 was used to perform antennal transcriptome sequencing, sequence assembly, functional annotation and differentially expressed gene analysis of P. rapae adults. qRT-PCR was used to verify nine differentially expressed olfaction-related genes including PrapOR1, PrapOR2, PrapOR5, PrapOBP1, PrapOBP4, PrapOBP5, PrapSNMP1, PrapSNMP2 and PrapSNMP3.【Results】By sequencing of the antennal transcriptome of adults of P. rapae, a total of 17.65 GB sequencing data (NCBI registration number: PRJNA869896) were obtained. A total of 116 317 transcripts were obtained through filtering and sequence splicing. Then, 43 390 unigenes were obtained by Corset hierarchical clustering. BUSCO evaluation showed that the stitching quality had good integrity and high accuracy. The databases with the largest number of unigene annotations in a descending order were NT, NR, Pfam, GO, Swiss-Prot, KEGG and KOG/COG. Furthermore, 176 olfaction-related genes were screened by gene functional annotation analysis, among them 19 genes were differentially expressed, including 15 genes highly expressed in female adult antennae and four genes highly expressed in male adult antennae. qRT-PCR verification results showed that PrapOR1 and PrapOR2 were highly expressed in male adult antennae, PrapOR5, PrapOBP1, PrapOBP4, PrapOBP5, PrapSNMP1, PrapSNMP2 and PrapSNMP3 were highly expressed in female adult antennae, indicating the consistency with the transcriptome sequencing results.【Conclusion】In this study, a transcriptomic database of adult antennae of P. rapae was established, the olfaction-related genes were screened and the differential expression of olfaction-related genes was analyzed. The results provide a theoretical basis for further studies on the gene function and olfactory molecular mechanism of P. rapae.
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Research progress in insect gut microbes and the methods for studying their functions
MA Ling, CAO Jing-Yu, BAI Jian-Yang, XU Zhe, LI Lu, ZHANG Yue, MIN Meng-Ru
Acta Entomologica Sinica    2023, 66 (10): 1415-1424.   DOI: 10.16380/j.kcxb.2023.10.014
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 Insects are the most abundant and widespread group of animals in the world that harbor complex and diverse gut microbes. Different insects differ in gut structure, gut environment, edibility, age and external environment, and the composition and abundance of gut microbes also show difference. Insect gut microbes are mainly transferred vertically and horizontally between populations and individuals, and play a vital role in nutrient metabolism, physiological behaviour, defence, detoxification and many other functions in insect hosts. Insect gut microbes can be isolated from culture media by in vitro culture methods and rapidly identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and sequencing technologies such as 16S rRNA gene sequencing. The combination of metagenomics, proteomics, metabolomics and other omics technologies makes the identification and functional prediction of gut microbes more efficient. Microbial functions can be more accurately verified by in vitro experiments, microbial supplementation, microbiota transplantation and silencing of genes associated with microbial members. Sterile insects can be obtained by high temperature treatment, lysozyme treatment, sterile feeding and antibiotic treatment for functional verification experiments. However, the most widely used antibiotic method still has limitations in application. By exploiting the characteristics of gut microbes, genetic engineering of symbiotic bacteria can be used to control pests and insect-borne infectious diseases. At present, insect gut microbes play an important role in the fields of ecology, economy, energy and environmental protection. With the development and integration of new technologies, more insect-microbe interaction mechanisms will be revealed, and pest control methods through insect gut microbes will become more diverse, environmentally friendly and efficient.
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Cloning  of Cu/Zn-SODl, Cu/Zn-SOD2 and Fe/Mn-SOD in Liposce1is bostrychophilα  (Psocoptera:  Liposce1ididae)   and their responseto  high and1ow temperature stresses

WANG Xiao, XU De-Jun, ZHU Bin-Jian, XU Jun-Ting, AO Guo-Hong, ZHANG Chang-Yu, HAN Kai-Yu
Acta Entomologica Sinica    2024, 67 (1): 1-8.   DOI: 10.16380/j.kcxb.2024.01.001
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【Aim】 To reveal the role of the superoxide dismutase genes in Liposcelis bostrychophila in response to high and low temperature stresses. 【Methods】The cDNAs of three superoxide dismutase genes Cu/Zn-SOD1Cu/Zn-SOD2 and Fe/Mn-SOD of L. bostrychophila were cloned by RT-PCR, and their sequence characteristics were analyzed by bioinformatics methods. RT-qPCR was used to detect the relative expression levels of Cu/Zn-SOD1Cu/Zn-SOD2 and Fe/Mn-SOD in adults at 0, 1 and 2 h under high temperature (42 ℃) and low temperature (4 ℃) stresses. 【Results】 LbCu/Zn-SOD1LbCu/Zn-SOD2 and LbFe/Mn-SOD (GenBank accession numbers are OQ938782, OQ938783 and OQ938784, respectively) of L. bostrychophila were cloned, with the open reading frames (ORFs) of 465, 630 and 636 bp in length, encoding 154, 209 and 211 amino acids with the relative molecular weights of 15.85, 22.33 and 23.72 kD, and the isoelectric points of 6.17, 7.68 and 6.79, respectively. LbCu/Zn-SOD1 and LbCu/Zn-SOD2 had one and two Cu/Zn superoxide dismutase signatures, respectively. LbFe/Mn-SOD had one Fe/Mn superoxide dismutase signature. The results of phylogenetic analysis showed that Cu/Zn-SOD1, Cu/Zn-SOD2 and Fe/Mn-SOD were highly conserved in insects. The expression of LbCu/Zn-SOD1LbCu/Zn-SOD2 and LbFe/Mn-SOD in adults of L. bostrychophila was induced by high temperature42 ℃ stress, and the expression levels of LbCu/Zn-SOD1 and LbCu/Zn-SOD2 inadults of L. bostrychophila were significantly higher than that of the control at 1 and 2 h. The expression level of LbFe/Mn-SOD in adults of L. bostrychophila was significantly lower than that of the control at 1 h and significantly higher than that of the control at 2 h under42 ℃stress. There was no significant difference in the expression levels of LbCu/Zn-SOD1 and LbCu/Zn-SOD2 inadults of L. bostrychophila at 1 h, while that of LbFe/Mn-SOD decreased significantly at 1 h and those of LbCu/Zn-SOD1LbCu/Zn-SOD2 and LbFe/Mn-SOD increased significantly at 2 h under 4 ℃stress as compared with that of the control. 【Conclusion】 The superoxide dismutase genes LbCu/Zn-SOD1LbCu/Zn-SOD2 and LbFe/Mn-SOD are involved in the tolerance of L. bostrychophila to extreme temperature stress.


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Research progress of behavioral manipulation techniques of Tuta absoluta (Lepidoptera: Gelechiidae)
LI Xiao-Wei, YI Song-Wang, CHEN Li-Min, CHENG Jiang-Hui, ZHOU Shu-Xing, LV Yao-Bin
Acta Entomologica Sinica    2023, 66 (6): 835-848.   DOI: 10.16380/j.kcxb.2023.06.012
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The South American tomato pinworm, Tuta absoluta, is a worldwide devastating pest on tomatoes. This insect pest invaded China in 2017 and showed a trend of spread, posing a serious threat to tomato production in China. Unreasonable use of insecticides for control of this insect pest has led to the increasing level of resistance and the decrease of control efficacy. Consequently, there is an urgent need to develop novel green control technologies. Physical control trapping technique and olfactory behavior manipulation technique are important components of the integrated pest management program. In this article, the research progress of physical control trapping (light traps and color traps) and olfactory behavior manipulation (sex pheromone and plant volatiles) in T. absoluta were summarized. Color significantly affects the trapping efficiency of pheromone traps and sticky color traps. White, black and blue traps attract more T. absoluta adults in the field. The black light traps and LED traps with the wavelength of 470 nm have the best trapping efficiency on T. absoluta, and also have synergistic effects on sex pheromone. The sex pheromone of T. absoluta includes the major component (3E, 8Z, 11Z)-tetradecatrienyl acetate and the minor component (3E, 8Z)-tetradecadienyl acetate, in a ratio of ~9∶1. The optimal trapping blend for mass trapping is (3E, 8Z, 11Z)-tetradecatrienyl acetate and (3E, 8Z)-tetradecadienyl acetate in a ratio of 95∶5, and delta traps with sex pheromone lure show the highest trapping efficiency in comparison with other trap types. The sex pheromone of T. absoluta has been widely used for monitoring, mass trapping and mating disruption. Plant volatiles and traditional food attractant volatiles affect the host selection and oviposition of T. absoluta. Octyl acetate, phenylacetaldehyde+acetic acid blend, acetic acid+3-methyl-1-butanol blend, and a blend consisting of limonene, β-ocimene, α-terpinene, δ-elemene and (E)-β-caryophyllene show attractive activities to T. absoluta. p-Quinone, 2-carene, d-curcumene and 1,2-diethylbenzene in tomato can serve as oviposition stimulants to T. absoluta. By contrast, 1-fluorododecane, caryophyllene, curcumene, elemene, humulene, zingiberene, tridecan-2-one, undecan-2-one, (Z)-3-hexenyl propanoate and methyl salicylate have adverse effects on host selection, oviposition or growth and development of T. absoluta. Finally, current problems of behavioral manipulation techniques of T. absoluta, such as unclear mechanisms, unstable effects, and lack of combined application technology with other green control strategies, were discussed, and potential research directions were prospected, which will provide a reference for the development of green control technologies based on insect behavioral manipulation in this pest.
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Screening and expression profiling of highly expressed genes in the hypopharyngeal glands of Apis mellifera (Hymenoptera: Apidae) foragers
ZHANG Juan, GAO Yan, WANG Ruo-Hong, LI Qiu-Fang, YANG Shang-Ning, ZHOU Shi-Wen, SHI Dan-Dan, SU Song-Kun, NIE Hong-Yi
Acta Entomologica Sinica    2023, 66 (7): 896-908.   DOI: 10.16380/j.kcxb.2023.07.004
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【Aim】 This study aims to screen genes highly expressed in the hypopharyngeal glands (HGs) of Apis mellefera foragers, and to provide a basis for further studying the genes related to honey bee foraging behavior.【Methods】 Differentially expressed genes (DEGs) in the HGs were screened based on the previously sequenced HGs transcriptome data of the 3-day-old workers (3 d_W), 10-day-old nurses (10 d_N), 10-day-old foragers (10 d_F), 21-day-old nurses (21 d_N), 21day-old foragers (21 d_F) and the DEGs highly expressed in the HGs of the 10-day-old foragers and 21-day-old foragers of A. mellefera were subjected to GO and KEGG enrichment analysis. Meanwhile, the expression levels of four highly expressed DEGs (LOC409889, LOC406114, LOC408453 and LOC100578735) in the HGs of foragers in different developmental stages (egg, larva, pupa, newly emerged workers, 10-day-old nurses and 21-day-old foragers) and tissues (abdomen, thorax, antenna, mandibular gland, hypoharyngeal gland, sting, leg, gut, wing and brain) of the 21-day-old foragers of A. mellefera were examined using qPCR.【Results】Comparative transcriptome result showed that the expression levels of 164 up-regulated DEGs in the HGs of the 21-day-old foragers were obviously increased as compared with those of the 3-day-old workers, 10-day-old nurses and 21-day-old nurses, and those of the 10-day-old foragers were also higher than those of the 10-day-old nurses. While the expression trend of the 105 down-regulated DEGs was opposite with that of the above 164 DEGs. GO analysis result showed that the 164 highly expressed DEGs in the HGs of the 10-/21-day-old foragers were enriched significantly in sulfate transmembrane transporter activity, sulfur compound transmembrane transporter activity, iron ion binding and oxidoreductase activity, and the 105 down-regulated DEGs were enriched significantly in ribosome, intracellular ribonucleoprotein complex and ribonucleoprotein complex. KEGG pathway analysis result revealed that the 105 down-regulated DEGs were significantly enriched in two pathways ribosome, and starch and sucrose metabolism. qPCR detection result indicated that the expression levels of LOC409889, LOC406114, LOC408453 and LOC100578735 were the highest in the 21-day-old foragers, and those of LOC406114 and LOC100578735 were the highest in the HGs of the 21-day-old foragers and showed low expression level or no expression in other tissues.【Conclusion】 The highly expressed DEGs identified in the HGs of A. mellifera forager were screened in this study via comparative transcriptome, which provides a valuable source of data for the function study of forager HGs, and also provides a new perspective for further studying the foraging behavior of A. mellefera and the molecular breeding selection of bee species with strong foraging ability.
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Analysis on the stability and influencing factors of protogyny of Spodoptera frugiperda (Lepidoptera: Noctuidae)
BI Si-Jia, HU Ben-Jin, HU Fei, XU Ting-Ting, XU Li-Na
Acta Entomologica Sinica    2023, 66 (9): 1201-1209.   DOI: 10.16380/j.kcxb.2023.09.007
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【Aim】 Protandry was more common in studies on developmental duration of sex dimorphic insects, and protogyny was less reported. The fall armyworm, Spodoptera frugiperda is a severe migratory pest on maize, and there is a phenomenon of protogyny in its developmental duration. The purpose of this study is to clarify the stability of this phenomenon and explore its influencing factors, so as to provide a theoretical basis for the accurate monitoring and early warning of S. frugiperda.【Methods】 The growth and development of S. frugiperda under three photoperiods (16L∶8D, 14L∶10D and 8L∶16D) and three food conditions (maize leaves, peanut leaves and artificial diet) were analyzed, and the ovaries of S. frugiperda reared under different photoperiods and food conditions were dissected. Moreover, the growth and development of Mythimna separata, S. litura and S. frugiperda in Noctuidae were compared to evaluate whether the protogyny was the commonness of Noctuidae insects with migratory habits.【Results】 In the comparison of three Noctuidae migratory insects, only the female adults of S. frugiperda eclosed significantly earlier (1.28 d ahead) than the male adults, and the eclosion phenomenon was stable under different photoperiods and food conditions, and only showed a weakening trend when S. frugiperda fed on peanut leaves. It was found that the ovaries of the newly eclosed adults of S. frugiperda were always immature under different photoperiods and food conditions, and did not change with external conditions. The development of female pupa of S. frugiperda was significantly faster than that of male pupa, which was the main reason for the difference in eclosion time between the two sexes of S. frugiperda, and the difference was positively correlated with the pupal weight of the two sexes: Compared with the female pupal weight, the male pupal weight of S. frugiperda increased significantly by 9.88%.【Conclusion】 The phenomenon of protogyny of S. frugiperda is relatively stable. Photoperiod and food will not change this phenomenon, but will affect the difference of eclosion time between the two sexes, and protogyny is not the commonness of Noctuidae insects with migratory habits.
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Isolation and identification of actinomycete strain BYC-18 and its antimicrobial metabolites from the gut of Odontotermes formosanus (Isoptera: Termitidae)
YIN Cai-Ping, BAI Xue-Yan, Naeem ABBAS, SUN Lu-Lu, YIN Xin-Ran, ZHANG Ying-Lao
Acta Entomologica Sinica    2023, 66 (10): 1282-1288.   DOI: 10.16380/j.kcxb.2023.10.002
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【Aim】To determine the antimicrobial activities of the fermentation products of actinomycetes from the gut of Odontotermes formosanus and analyze their antimicrobial components, so as to find novel antimicrobial lead compounds.【Methods】 The gut actinomycetes of O. formosanus were isolated by spread plate method. The antimicrobial activities of the fermentation broth extracts against four pathogens (Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Candida albicans) were tested by the Oxford cup method, and the active strain BYC-18 was screened. The taxonomic status of BYC-18 was determined according to its morphological features and 16S rRNA sequence analysis. The antimicrobial activities of different polar solvent extracts of BYC-18 fermentation solution were determined by filter paper method. Active compounds were isolated and purified from ethyl acetate crude extract by various chromatographic methods, and the chemical structure of the isolated compound was identified by mass spectrometry and nuclear magnetic resonance spectroscopy. The antimicrobial activities of the isolated compound were determined by filter paper method and minimum inhibitory concentration method. 【Results】 BYC-18 was identified as a strain of Streptomyces sp. The fermentation broth of BYC-18 had antimicrobial activities against all the four pathogens, and its ethyl acetate extract had an obvious inhibitory effect on S. aureus, with an inhibition zone diameter of 11.1 mm. A monomer compound BYC-18-1 was isolated from the ethyl acetate extract and identified as β-rubromycin. The antimicrobial activity test showed that β-rubromycin had a strong inhibitory effect on S. aureus at the test concentration of 30 μg/6 mm filter paper, and its inhibition zone diameter was 13.2 mm, which was close to that of the positive gentamicin sulfate (inhibition zone diameter: 16.6 mm). 【Conclusion】The Streptomyces strain BYC-18 has the potential to be developed as a new bactericide.
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Relationship between the symbiotic microbial community and insecticide resistance in wild Anopheles sinensis (Diptera: Culicidae) 
QIU Xin, MENG Chen, WANG Ming-Bin, CHEN Bin, ZHANG Yu-Juan
Acta Entomologica Sinica    2023, 66 (9): 1183-1191.   DOI: 10.16380/j.kcxb.2023.09.005
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【Aim】 To investigate the relationship between the symbiotic microbial communities in Anopheles sinensis and the insecticide resistance of An. sinensis in different regions of the wild environment. 【Methods】 Wild female adults of An. sinensis were collected from Chongqing, Yunnan, and Anhui in China. WHO standard in vitro bioassay and 0.05% deltamethrin impregnated paper were used to conduct insecticide susceptibility tests on the female adults of An. sinensis, classifying them as insecticide-sensitive (FS) or insecticide-resistant (FR) to perform high-throughput sequencing of the whole genomes through Illumina Hiseq 2000 platform. The sequences of 16S rRNA and 18S rRNA genes were aligned to identify symbiotic microbiota in the FS and FR An. sinensis. Alpha diversity analysis, beta diversity analysis, clustering analysis, principal coordinates analysis (PCoA) and analysis of symbiotic microbial community differences were conducted. 【Results】 A total of 3 284 species of 52 genera of 14 phyla of symbiotic microbiota were identified from the female adults of the FS and FR wild An. sinensis from Chongqing, Yunnan and Anhui. The diversity difference and community richness of the FR wild An. sinensis female adults from Anhui were the highest, and those of the FS wild An. sinensis female adults from Chongqing were the lowest. The symbiotic microbial communities in the female adults of wild An. sinensis were first clustered by region, and the diversity of the symbiotic microbial communities in the female adults of An. sinensis from Chongqing and Yunnan were more similar. A total of 10 strains including two strains of Cyanobacteria, three strains of Halobacteria, one strain of Haliangium, three strains of Ruminococcaceae and one strain of Streptococcus of bacteria in the symbiotic microbiota in the FR wild An. sinensis female adults were more abundant.【Conclusion】 The symbiotic microbial communities of the wild An. sinensis exhibit regional and resistance differences. The identified symbiotic microbiota play a positive role in the insecticide resistance of An. sinensis. The results of this study provide insights into understanding the insecticide resistance of An. sinensis.
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Identification and pathogenicity determination of Beauveria bassiana strain JCF from Holotrichia oblita (Coleoptera: Scarabaeidae)
CAO Wei-Ping, CHENG Jia-Xu, FENG Shuo, ZHAO Ju-Ying, CHEN Dan, SONG Jian
Acta Entomologica Sinica    2023, 66 (8): 999-1008.   DOI: 10.16380/j.kcxb.2023.08.001
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【Aim】 Holotrichia oblita is an important insect pest of various crops such as peanuts, fruit forests and nurseries, and it has been mainly controlled by chemical control for a long time. In this study, the pathogenicity of Beauveria bassiana strain JCF to the larvae and adults of H. oblita was determined to provide fungal resources for the biological control of H. oblita. 【Methods】 The parasitic fungi on the dead body of the diseased H. oblita were isolated and rejoined, and the pathogenic strain JCF was isolated and purified. The morphological characteristics of the strain were observed by light microscope. The rDNA-ITS sequence of JCF was amplified by PCR and then sequenced, and the phylogenetic tree of JCF was constructed for molecular identification. The virulence of the JCF strain to the adults and larvae of H. oblita was determined by immersion method and field simulation test. 【Results】 The colony of the strain JCF with colorless back is white fluffy at the initial stage, then becomes yellow or light yellow flocculent at the late stage. The conidiogenous cells are clustered on the conidiophore in a bottle shape, and the conidiophore is spike-shaped and attached to the vegetative hypha. Conidia are solitary, transparent, subglobose, and (2.0-3.5) μm×(1.5-2.0) μm in diameter. The molecular identification of rDNA-ITS sequence showed that the strain JCF was B. bassiana. The medium lethal concentration (LC50) value of B. bassiana strain JCF against H. oblita adults was 2.124×107 conidia/mL, and the medium lethal time (LT50) values of this strain at the concentrations of 2×108, 1×108 and 5×107 conidia/mL against H. oblita adults were 6.267, 6.852 and 7.455 d, respectively. The LC50 values of B. bassiana strain JCF against the 1st, 2nd and 3rd instar larvae of H. oblita were 5.47×106, 1.59×108 and 2.50×108 conidia/mL, respectively. The LT50 values of B. bassiana strain JCF at the concentration of 1×108 conidia/mL against the 1st and 2nd instar larvae of H. oblita were 8.6 and 18.6 d, respectively, and those of B. bassiana strain JCF at the concentration of 5×108 conidia/mL against the 2nd and 3rd instar larvae were 10.3 and 13.2 d, respectively. In the field simulation test, the spore suspension of B. bassiana strain JCF (2×108 conidia/mL) exhibited good pathogenic effects on both adults and larvae of H. oblita.【Conclusion】 The B. bassiana strain JCF has high pathogenicity to the adults and larvae of H. oblita, providing a new strain resource for the development of B. bassiana formulations to control Melolonthidae pests. And it has potential application value for biological control of Melolonthidae pests.
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Morphology and development of the internal reproductive system of Monochamus alternatus (Coleoptera: Cerambycidae) adults
CHEN Jin, LI Hui, HU Tian-Yi, YANG Hua-Lei, WEI Jia-Hang, JIN Lin, HAO De-Jun
Acta Entomologica Sinica    2023, 66 (9): 1210-1220.   DOI: 10.16380/j.kcxb.2023.09.008
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【Aim】To clarify the structure and developmental process of the internal reproductive system of Monochamus alternatus adults and provide a theoretical basis for its reproductive mechanism research and population dynamics monitoring.【Methods】The internal reproductive system of M. alternatus adults at different ages (day-old) was anatomically observed under optical microscope to clarify its structural characteristics and developmental classification criteria, and the developmental process of the internal reproductive system of female and male adults was determined by divison of developmental levels. 【Results】The female internal reproductive system of M. alternatus adults is composed of one pair of ovaries, one pair of lateral oviducts, common oviduct, copulatory pouch, spermathecal duct, spermatheca, spermathecal gland , ovipositor and genital cavity. According to the morphology and color of ovarioles and spermathecal glands, the sexual maturation process of female adults can be divided into the following stages: Transparent stage (stage Ⅰ), primary vitellogenesis stage (stage Ⅱ), secondary vitellogenesis stage (stage Ⅲ) and egg maturation stage (stage Ⅳ). Ovarioles grow continuously during the ovarian development, and the color changes from transparent milky white to yellow. The morphology and color of the spermathecal gland changes from flat and translucent to full and yellow. The male internal reproductive system of M. alternatus adults is composed of one pair of testes, two pairs of accessory glands, one pair of vas deferens, one pair of vesicula seminalis, one pair of ejaculatory ducts, circulator and aedeagus. According to the morphology and color of the testes, vesicula seminalis and accessory glands, the sexual maturation process of male adults can be divided into four stages including transparent stage (stage Ⅰ), initial stage of spermiogenesis (stage Ⅱ), peak period of spermiogenesis (stage Ⅲ) and maturing stage (stage Ⅳ). The testicular long diameter changes little. The color of the testis, vesicula seminalis and circulator changes from milky white and translucent to white. The morphology and color of the accessory gland changes from small and translucent to swollen and transparent. The development of the internal reproductive system of both female and male adults is mainly concentrated at the stages Ⅱ-Ⅲ. In terms of the developmental process of the internal reproductive system, male adults develop more rapidly and their internal reproductive system is already mature at the 12-day-old, while the internal reproductive system of female adults is mature at the 15-day-old. 【Conclusion】 This study has clarified the basic structure and developmental processes of the internal reproductive system of female and male adults of M. alternatus, not only providing a theoretical basis for further research on the reproductive mechanism of M. alternatus, but also providing a reference for predicting the peak ovioposition period of M. alternatus in production practice.
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Research progress of the population genetic differentiation and environmental adaptation mechanisms in Apis cerana cerana (Hymenoptera: Apidae)
CHEN Bing, LUO Jia-Yu
Acta Entomologica Sinica    2023, 66 (9): 1258-1270.   DOI: 10.16380/j.kcxb.2023.09.012
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 Apis cerana cerana is a critical and important pollinator in China. This bee species has a wide distribution in various habitats, and exhibits several advantages as pollinator, e.g., agility in flight, long nectar gathering period and strong adaptability. However, in recent years, A. c. cerana has been facing an unprecedented decline in population diversity. In order to protect the genetic resources of these specific populations, researchers studied the physiology and mechanism of genetic differentiation and environmental adaptation in A. c. cerana based on geometric morphology, molecular biology and genomics technologies. Meanwhile, the diverse populations of A. c. cerana in China provided rich materials for analyzing their adaptive evolution. In this article, we summarized the research progress from the four aspects: The correlation between population genetic differentiation and environmental changes, the morphological variation and environmental adaptation, the enviroment-adaptive physiological and behavioral changes, and the genetic mechanisms behind these phenotypic changes of A. c. cerana populations. Previous studies showed that changes in physical barriers and ecological environment, especially those related to altitude and latitude, were the main reasons for the differentiation of A. c. cerana populations. Among the climatic factors, temperature, oxygen, radiation and humidity had important effects on the morphological development and eco-physiological traits of A. c. cerana. The morphological changes were mainly explained by variation in body size and color. Changes in metabolic physiology and behaviors have been evolved as a crucial adaption strategy. Population genetics and genomics based on modern genomics and molecular biology techniques showed that genes and pathways related to social division of labor, learning and perceiving behavior, information perception, growth and development, thermal adaptation and metabolism are subject to natural selection. These findings provide molecular evidence for the ability of A. c. cerana to adapt to different habitats and the evolution of bee species. However, the specific molecular evolution mechanisms for the environmental adaption of bees await further investigation. Our review on the mechanisms of genetic differentiation and environmental adaption of A. c. cerana will deepen our understanding of the evolutionary history and genetic diversity of ancient bee species, and lay a foundation for further studies on the adaptive mechanisms of social insects to different environments and the development of effective conservation strategies.
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Effects of chronic exposure to LC20 of dinotefuran on the growth and development of Sitobion avenae (Hemiptera: Aphididae)
SHI Hai-Chun, JI Qin-Long, MA Jian-Xiong, LIANG Yu-Ting, LI Guang-Kuo, ZHANG Hai-Ying, LIU Yong-Gang
Acta Entomologica Sinica    2023, 66 (5): 685-693.   DOI: 10.16380/j.kcxb.2023.05.009
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【Aim】 The effects of chronic exposure to the sublethal concentration (LC20) of dinotefuran on the growth and development of Sitobion avenae were studied in order to provide a theoretical basis for the scientific use of dinotefuran to control S. avenae and delaying the development of the resistance to the pesticide and extending the useful life of pesticide. 【Methods】 The LC20 of dinotefuran to the 3rd instar nymphs of S. avenae was determined by using the leaf-dip method andusedforchronic exposure on S.
avenae for 15 generations. The resistance ratio of S. avenae to dinotefuran was determined, the developmental duration, reproduction and survival rates of S. avenae of two successive generations which were raised with wheat seedlings in test tube were recorded, and the body length, body width and body weight of adults of F0 generation were measured. The differences in the body length, body width, body weight, survival rate, adult longevity and number of offspring produced between lowly resistant strain of various generations and the susceptible strain were analyzed by population specific life table and DPS software. 【Results】 After chronic exposure to LC20 of dinotefuran for 15 generations, the resistance of S. avenae reached low level (6.54-fold). The developmental duration of the 2nd, 3rd and 4th instar nymph of F0 generation of the lowly resistant strain was significantly longer than that of the susceptible strain, and the body length, body width and body weight of F0 generation of adults of the lowly resistant strain were significantly increased, but the number of offspring produced per adult and adult longevity of F0 generation of the lowly resistant strain were significantly reduced or shortened as compared to those of the susceptible strain. The adult survival rate, number of offspring produced per adult and adult longevity of F1 generation of the lowly resistant strain were significantly reduced, the 2nd instar nymphal duration and nymphal duration of F1 generation of the lowly resistant strain were significantly shortened as compared to those of the susceptible strain. The net reproductive rate (R0), intrinsic rate of increase (Rm) and finite rate of increase (λ) of F0 and F1 generations of the lowly resistant strain were significantly lower than those of the susceptible strain, while the population doubling time (DT) of F0 and F1 generations and the mean generation time (T) of F1 generation of the lowly resistant strain were significantly prolonged as compared to those of the susceptible strain. 【Conclusion】 Chronic exposure to LC20 of dinotefuran results in the increase in the resistance of S. avenae to dinotefuran and individual size of adults of F0 generation, while inhibits the longevity and fecundity of F0 and F1 generations of S. avenae.
Key words: 
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Mitochondrial genomes of Caerulea coeligena (Lepidoptera: Lycaenidae) and Dodona maculosa (Lepidoptera: Riodinidae) and phylogenetic relationships in Papilionoidea based on mitochondrial genes
YAN Zhen-Tian, LUO Si-Te, FAN Zhen-Huai, CHEN Bin
Acta Entomologica Sinica    2023, 66 (7): 946-958.   DOI: 10.16380/j.kcxb.2023.07.009
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【Aim】 To sequence and analyze the complete mitochondrial genomes (mtgenomes) of Caerulea coeligena and Dodona maculosa, and to explore the phylogenetic relationships in Papilionoidea based on the known mitochondrial genome data. 【Methods】 The complete mtgenomes of C. coeligena and D. maculosa were sequenced using Illumina NovaSeq 6000 sequencing platform, assembled and annotated, and the secondary structure of tRNA genes was predicted. Based on the nucleotide sequences of 13 protein-coding genes (PCGs) from the mtgenomes of 48 reported butterfly species belonging to six families in Papilionoidea obtained from the NCBI database, the selective pressures on these 13 PCGs in 50 species were analyzed. The phylogenetic relationships among these 50 species, as well as their respective families and subfamilies, were analyzed using maximum likelihood (ML) and Bayesian inference (BI) methods. 【Results】 The mtgenomes of C. coeligena and D. maculosa (GenBank accession numbers are MZ489120 and MZ 489121, respectively) are 15 164 and 15 486 bp in length, respectively. Their genome size, structure and composition, gene arrangement, nucleotide composition, codon usage, and predicted tRNA structure of mitogenomes of C. coeligena and D. maculosa were similar to those of the known closely related butterfly species. The mtgenomes in Papilionoidea exhibited a high level of purifying selection, with the ratios of nonsynonymous substitution rate to synonymous substitution rate (Ka/Ks) for the 13 PCGs obviously lower than 1. The phylogenetic relationships in Papilionoidea were (Papilionidae+Hesperiidae+Pieridae+(Nymphalidae+ (Lycaenidae+Riodinidae))).【Conclusion】 The reconstructed phylogenetic tree based on the mtgenomes of 48 reported butterfly species in Papilionoidea and two newly sequenced species in this study suggests that Riodinidae and Lycaenidae be monophyletic, and sister each other. The sister group would be closely related to Nymphalidae in phylogenetics. Moreover, Papilionidae may represent one of the earliest lineages in Papilionoidea. This study provides new molecular data to support further research on the systematic evolution and refinement of the classification system for butterflies.
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Effects of introduction of Bacillus spp. on the microbiota and growth and development of the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)
WANG Wei-Xia, ZHU Ting-Heng, LAI Feng-Xiang, WEI Qi, WAN Pin-Jun, HE Jia-Chun, FU Qiang
Acta Entomologica Sinica    2023, 66 (10): 1289-1301.   DOI: 10.16380/j.kcxb.2023.10.003
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【Aim】To identify the species of culturable symbiotic bacteria in the brown planthopper, Nilaparvata lugens, explore the effects of the symbiotic Bacillus spp. on the microbiota and growth and development of N. lugens. 【Methods】 The culturable symbiotic bacteria were isolated from two different virulent populations of N. lugens (the susceptible population TN1 and the virulent population IR56) by in vitro culture. The obtained cuttural symbiotic bacteria were identified by 16S rDNA sequencing technology. On this basis, the distribution of symbiotic bacteria in N. lugens was studied by in situ hybridization. The effects of reducing symbionts and supplementation of Bacillus spp. on the growth and development of N. lugens, as well as the abundance of symbionts, were studied using artificial diet added with antibiotics or symbiotic Bacillus spp. The effects of introducing symbiotic Bacillus spp. through feeding and microinjection methods were compared, and the correlation between Bacillus spp. colonization and the virulence of TN1 population was investigated. 【Results】 In total 15 strains of different symbiotic bacteria were obtained from N. lugens by in vitro culture, including 2 strains of the symbiotic Bacillus spp. (BPH-S36 and BPH-S33) from the highly virulent population IR56. The result of in situ hybridization showed that symbiotic bacteria were distributed in the salivary glands, gut, fat body and female internal genitalia of N. lugens, but rarely in the male internal genitalia. Symbionts are crucial for the growth and development of N. lugens. The reduction of symbiotic bacteria led to a significant decrease in the survival rate of N. lugens on the 3rd and 6th day, while supplementing the symbiotic Bacillus BPH-S33 or BPH-S36 resulted in a significant increase in the survival rate of N. lugens on the 6th day. The relative abundance of Bacillus spp.in N. lugens increased by 23.2-24.5-fold by feeding on the artificial diet supplemented with the symbiotic Bacillus spp., but not increased through microinjection into nymphs. With the increase of the symbiotic Bacillus spp., the relative abundance of yeast-like symbiont (YLS) also significantly increased by 5-12-fold. Introduction of Bacillus strain BPH-S36 significantly increased the survival rate of N. lugens TN1 population on the resistant rice IR56 from 52.1%±1.5% to 64.2%±3.0%. 【Conclusion】Symbiotic bacteria are widely distributed in the salivary glands, gut and ovaries of N. lugens. Antibiotic treatment can significantly reduce the abundance of symbiotic bacteria in their bodies and result in a decline in survival rate. The exogenous symbiotic bacteria colonize successfully in N. lugens through artificial feeding. Bacillus strain BPH-S36 derived from the IR56 population can improve the population virulence to the resistant rice variety IR56.
Key words: Nilaparvata lugens; symbiotic bacteria; yeast-like symbiont; virulence; Bacillus spp.; adundance
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Functional analysis of Bmttv in the wing development of the silkworm, Bombyx mori
LIANG Yan, LI Jing-Wen, CHEN Yin-Lin, JIA Wen-Yi, XIA Qing-You, HUA Xiao-Ting
Acta Entomologica Sinica    2023, 66 (11): 1435-1442.   DOI: 10.16380/j.kcxb.2023.11.002
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 【Aim】 To analyze the function of glycosyltransferase gene Bmttv of Bombyx mori in the wing development.【Methods】 The full-length sequence of the Bmttv CDS of B. mori was cloned by RACE and analyzed by bioinformatics. The expression levels of Bmttv in different developmental stages (5th instar larva, larva at the wandering stage, prepupa, pupa and adult) and different tissues (cuticle, wing disc, testis, Malpighian tubules, trachea, silk gland, head, hemolymph, fat body and midgut) of the 5th instar larva of B. mori were detected by qRT-PCR. The overexpression vector was constructed and transiently transfected into BmE cells. Subcellular localization of BmTTV was performed by immunofluorescence. qRT-PCR was used to detect the expression levels of BmHippo, Bmwg, BmDpp, Bmsotv and BmHh, which are the key genes in the wing development of B. mori. 【Results】The CDS sequence of Bmttvof B. moriwas successfully cloned and is 2 228 bp in full-length. BmTTV has two special structural domains, both of which are related to the synthesis of heparan sulfate proteoglycans (HSPGs), with about 45% amino acid sequence identity to TTVs of Drosophila melanogaster and Homo  sapiens, and closer affinity to TTVs of lepidopteran insects. Bmttv was expressed at high levels from the pupal stage and in the midgut, testis and wing disc of the 5th instar larva. BmTTV is mainly localized in the cytoplasm of BmE cells, and BmHippo, which is associated with the wing development, was significantly down-regulated in the expression after the overexpression of Bmttv. 【Conclusion】BmTTV may play an important role in the wing development of B. mori by participating in Hippo signaling pathway, and the results of this study lay a foundation for further research.
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Vitellogenin determines the abundance of the symbiotic bacterium  Rickettsia  in  Bemisia tabaci  (Hemiptera: Aleyrodidae) by regulating autophagy
SUN Xiang, LIU Bing-Qi, CHEN Zhan-Bo, LI Huan, LI Chu-Qiao, HONG Ji-Sheng, LUAN Jun-Bo
Acta Entomologica Sinica    2023, 66 (10): 1302-1310.   DOI: 10.16380/j.kcxb.2023.10.004
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【Aim】 To explore the molecular mechanism by which vitellogenin (Vg) regulates the abundance of the symbiotic bacterium Rickettsia in Bemisia tabaci. 【Methods】 Female adult of B. tabaci MEAM1 was microinjected with dsRNA to conduct RNAi of BtVg, the expression levels of BtVg and the autophagy gene BtAtg8 in the female adult were detected by qRT-PCR, the mortality rate of female adults was recorded, and the abundance of Rickettsia in the female adult was quantified by qPCR. The localization and expression of BtVg and BtAtg8 in ovarioles and the abundance of Rickettsia in the ovarioles and midguts of the female adult were examined by immunofluorescence microscopy. After the autophagy induction by feeding the female adult of B. tabaci MEAM1 with the artificial diet containing rapamycin (10 μmol/L), the expression and localization of BtAtg8 in ovarioles were investigated by immunofluorescence microscopy and the abundance of Rickettsia in ovaries was detected by qPCR. 【Results】 The expression levels of BtVg and BtAtg8 in the female adult of B. tabaci MEAM1 were significantly decreased and significantly increased, respectively, the mortality rate of female adults of B. tabaci MEAM1 was significantly increased, the abundance of Rickettsia in the female adult of B. tabaci MEAM1 was significantly decreased, the expression levels of BtVg and BtAtg8 in ovarioles were obviously decreased and obviously increased, respectively, and the abundance of Rickettsia in the ovarioles and midgut obviously decreased at 3 d after microinjection of dsBtVg compared to the control (microinjected with dsGFP). The expression level of BtAtg8 in the ovarioles of the female adult of B. tabaci MEAM1 was increased significantly at 5 d after feeding with the artificial diet containing rapamycin (10 μmol/L), and the abundance of Rickettsia in the ovarioles of the female adult of B. tabaci MEAM1 was significantly decreased at 3 and 5 d after feeding with the artificial diet containing rapamycin (10 μmol/L) compared with the control fed with the artificial diet containing DMSO. 【Conclusion】 These results suggest that BtVg can protect Rickettsia from degradation by autophagy, and demonstrate the role of BtVg in regulating the abundance of Rickettsia in B. tabaci.
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Research progress in protein-carbohydrate regulation in insects based on geometric framework model
SUN Shao-Lei, JING Xiang-Feng
Acta Entomologica Sinica    2023, 66 (5): 713-722.   DOI: 10.16380/j.kcxb.2023.05.012
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 Protein and carbohydrate are two essential nutrients for insects. Protein is the source of amino acids for insect growth, while carbohydrate is the source of energy for activity. These two nutrients play important roles in insect growth, development and reproduction. In the nature, all insects experience heterogeneous nutritional landscapes, and the contents of proteins and carbohydrates in their food including plants or preys generally vary upon species, tissues or developmental stages. Thus, insects may have some strategies to find the food that contains the proper nutritional composition in their living environment so as to make their growth and development reach the optimal status. Geometric framework model has been demonstrated to be a powerful tool to investigate nutritional physiological regulation mechanisms in insects. This model has been extensively used in how insects coordinate different nutrients in food to achieve the balance of life processes, such as growth, development and reproduction. In herbivores, it is shown that the protein/carbohydrate ratio in food combining with environmental factors can affect the growth and development, as well as the immune response of insects. In predatory and social insects, it is shown that they can strictly regulate the intake of protein and carbohydrate nutrients to meet their needs. In model insects, it is shown that protein/carbohydrate ratio in food may have a balancing effect on lifespan and reproduction in insects. In this review, we introduced the working principle of geometric framework model and systematically summarized its application in insects, aiming to provide new ideas for the study of insect nutrition physiology and ecology.
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Difference in the temperature adaptability between the sympatric Ostrinia furnacalis and O. nubilalis (Lepidoptera: Crambidae) in Xinjiang, northwest China
DING Xin-Hua, JIA Zun-Zun, WANG Xiao-Wu, FU Kai-Yun, WANG Yang-Zhou, AERZIGULI·Rouzi, TUERXUN·Ahemati, LIU Xiao-Xia, GUO Wen-Chao
Acta Entomologica Sinica    2023, 66 (5): 694-703.   DOI: 10.16380/j.kcxb.2023.05.010
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【Aim】By clarifying the difference in the temperature adaptability of the main biological characters between two closely related species of corn borers, the Asian corn borer (Ostrinia furnacalis) and European corn borer (O. nubilalis), living in the only one sympatric region in the world and accompanied by fierce interspecific competition, this study aims to reveal the biological driving mechanism of the rapid replacement of O. nubilalis population by O. furnacalis in the sympatric region in Xinjiang, Northwest China.【Methods】 At 15, 20, 25, 30 and 35 ℃, the egg duration, egg mass hatching rates, egg hatching rates per egg mass and mortalities of egg masses of the O. furnacalis and O. nubilalis populations isolated from the same region in Xinjiang, and at 20, 25, 30 and 35 ℃, their adult longevity, pre-oviposition period, oviposition period, numbers of eggs laid per female, larval and pupal duration, larval and pupal weight etc. were measured and compared. 【Results】 The results showed that the mortalities of egg masses of O. nubilalis were significantly higher than those of O. furnacalis at 15, 20, 25 and 35 ℃, and the average egg hatching rates per egg mass of O. nubilalis were significantly lower than those of O. furnacalis at 20, 25, 30 and 35 ℃. Furthermore, the hatching ability of egg masses of O. nubilalis decreased significantly with the increase of temperature, until the temperature increased to 35 ℃, the mortality of egg masses was the highest (73.42%±2.28%) and the average egg hatching rate per egg mass was the lowest (48.46%±2.64%), while those of O. furnacalis were 41.88%±4.33% and 67.99%±2.52%, respectively. At the same temperature, there was no significant difference in the pre-oviposition period, female adult longevity, male adult longevity and oviposition period between sympatric O. furnacalis and O. nubilalis. However, the main fecundity indexes of O. nubilalis, such as the number of egg masses laid per female, average number of eggs per egg mass laid by a single female and average number of eggs laid per female, were significantly lower than those of O. furnacalis. Compared with O. furnacalis, O. nubilalis had significantly heavier 1st-4th instar larval weight at the low temperature of 20 ℃. However, at higher temperatures (30 and 35 ℃), the 2nd and 3rd instar larval weight of O. nubilalis was significantly lighter than that of O. furnacalis and the duration of various instar larvale and pupal duration of O. nubilalis showed no significant difference from those of O. furnacalis. 【Conclusion】Compared with O. nubilalis, O. furnacalis has better temperature adaptability. Especially at high temperatures (30 and 35 ℃), O. furnacalis has higher survival rate of egg masses, better hatching ability, higher fecundity and heavier body weight, showing stronger competitiveness.
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Evaluation of reference genes for real-time quantitative PCR in the pea aphid, Acyrthosiphon pisum (Hemiptera: Aphididae)
GAO Yu-Qing, MA Zi-Qi, LI Zhen-Xiang, CHEN Zhen-Zhen, LIU Fang-Hua, KANG Zhi-Wei, XU Yong-Yu
Acta Entomologica Sinica    2023, 66 (5): 663-675.   DOI: 10.16380/j.kcxb.2023.05.007
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【Aim】The aim of this study is to lay a foundation for the gene expression analysis of Acyrthosiphon pisum by identifying the expression stablility of reference genes in A. pisum under different experimental conditions. 【Methods】The expression levels of 14 candidate reference genes commonly used in insects (EF1α, Tubulin, NADH, RPL12, SDHB, 18S rRNA, 28S rRNA, 16S rRNA, ATPase, Actin, TATA, RPL32, GAPDH and RPL7)in different developmental stages (1st-4th instar nymphs and adult), winged and wingless parthenogenetic adults, different tissues (head, thorax and abdomen) of parthenogenetic wingless adults, parthenogenetic wingless adults of different geographical populations (American population, Gansu population, Yunan population and Delingha population), parthenogenetic wingless adults fed on different host plants (alfalfa, clover and broad bean), parthenogenetic wingless adults reared under different photoperiods (24L∶0D, 0L∶24D and 16L∶8D), parthenogenetic wingless adults reared under different temperatures (4, 18 and 35 ℃) and parthenogenetic wingless adults treated with 200 g/L imidacloprid were detected by qPCR. The expression stability of the above 14 reference genes was evaluated using RefFinder, ΔCt method, GeNorm, NormFinder, and BestKeeper. Using CYP6CY3 as the target gene to explore the influence of different reference genes on its expression level analysis in parthenogenetic wingless adults treated with 200 g/L imidacloprid. 【Results】 18S rRNA and GAPDH were recommended as the most stably expressed reference genes, whereas 16S rRNA and Actin showed the least expression stability under biotic conditions (developmental stage, wing morph, tissue, geographical population and host plant) through comprehensive analysis for the results of ΔCt method, GeNorm, NormFinder and BestKeeper by RefFinder, based on the qPCR results. Meanwhile, 18S rRNA and EF1α were recommended as the most stably expressed reference genes, while Tubulin and TATA showed the least expression stablility under abiotic conditions (photoperiod, temperature and insecticide). According to the GeNorm analysis data of the optimal number of reference genes and analysis of the influence of different reference genes on the expression of target gene CYP6CY3, 18S rRNA and EF1α were recommended as the most stably expressed reference genes for further studies in A. pisum. 【Conclusion】It is recommended to use the combination of 18S rRNA and EF1α in qPCR analyses in A. pisum.
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Identification and localization analysis of intestinal mucin AcMucin5AC-1 in Apis cerana cerana (Hymenoptera: Apidae)
LI Xiao-Qing, GUO Yue, ZHANG Jie, ZHOU Ze-Yang, DANG Xiao-Qun
Acta Entomologica Sinica    2023, 66 (9): 1150-1160.   DOI: 10.16380/j.kcxb.2023.09.002
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【Aim】 The purpose of this study is to identify the structure, distribution and effect of intestinal mucin AcMucin5AC-1 on the midgut of Apis cerana cerana, so as to provide a theoretical basis for analyzing the physiological function of the honeybee midgut. 【Methods】 Bioinformatics was used to compare and analyze the sequence characteristics of Mucin5AC protein in A. cerana cerana. RT-qPCR was used to detect the expression levels of AcMucin5AC-1 in the midgut and cuticle of the 4-day-old larva of A. cerana cerana and in the midgut of the 2-day-old larva of A. cerana cerana at 0, 12, 24, 48 and 72 h after infection with Chinese sacbrood virus (CSBV). The prokaryotic expression system was used to express AcMucin5AC-1. The recombinant AcMucin5AC-1 was purified by Ni2+ affinity column and the polyclonal antibody to AcMucin5AC-1 was prepared. Western blot was used to detect the expression of AcMucin5AC-1 in the 3-6-day-old larvae, and the midgut, cuticle and peritrophic membrane of the 4-day-old larva of A. cerana cerana. Using indirect immunofluorescence assay, the localization of AcMucin5AC-1 in the 4-day-old larva of A. cerana cerana was analyzed. The interference efficiency of RNAi was analyzed at 12, 24, 48 and 72 h after silencing AcMucin5AC-1 in the 2-day-old A. cerana cerana larva by RNAi through feeding method. Hematoxylin-eosin (HE) staining method was used to explore the effects at 24, 48 and 72 h after silencing AcMucin5AC-1 in the 2-day-old larva by RNAi on the overall histomorphology of A. cerana cerana larva.【Results】 There are eight AcMucin5AC genes in the genome of A. cerana cerana, and their amino acid sequences all contain mucin domains. The results of RT-qPCR showed that the expression level of AcMucin5AC-1 was higher in the midgut of the 4-day-old larva of A. cerana cerana than in the cuticle, and was significantly down-regulated in the midgut of the 2-day-old larva at 12 and 72 h after infection with CSBV as compared to that in the control group. The recombinant protein AcMucin5AC-1 about 50 kD and polyclonal antibody were successfully obtained. Western blot result showed that AcMucin5AC-1 could be detected in the total protein of the 3-6-day-old larvae, and the midgut, cuticle and peritrophic membrane of the 4-day-old larva of A. cerana cerana. Indirect immunofluorescence experiment result showed that AcMucin5AC-1 was mainly localized in the midgut and peritrophic membrane of the 4-day-old larva. The detection result of RNAi efficiency showed that the expression level of AcMucin5AC-1 in A. cerana cerana was down-regulated by 92% at 24 h by 2.0 μg/individual dsAcMucin5AC-1 compared with the dsEGFP control group. HE staining detection result showed that the compactness between cells of the whole intestinal lumen of A. cerana cerana larva was weakened, and the morphological structure was disordered at 72 h after RNAi. 【Conclusion】 AcMucin5AC-1 is characterized as a mucin located in the midgut and peritrophic membrane of A.cerana cerana larvae. Down-regulation of AcMucin5AC-1 affects the morphology of the midgut of A. cerana cerana larva, suggesting that this gene may play an important role in the development of the A. cerana cerana larval midgut.
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Observation of the sensilla on the adult antennae of Cotesia ruficrus (Hymenoptera: Braconidae), a parasitoid of Spodoptera frugiperda (Lepidoptera: Noctuidae), under scanning electron microscope
WANG Wen-Wen, HE Peng-Yang, LIU Tong-Xian, JING Xiang-Feng, ZHANG Shi-Ze
Acta Entomologica Sinica    2023, 66 (8): 1105-1116.   DOI: 10.16380/j.kcxb.2023.08.011
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【Aim】 To clarify the types, morphology, quantity and distribution of sensilla on the female and male adult antennae of Cotesia ruficrus, an indigenous parasitic natural enemy against Spodoptera frugiperda, a major invasive agricultural pest, and provide theoretical support for exploring the function and olfactory mechanism of various types of sensilla of C. ruficrus.【Methods】 The sensilla on female and male adult antennae of C. ruficrus were observed with scanning electron microscope. 【Results】 The subsegments of antennal flagellum of the male adult of C. ruficrus are long moniliform, and the distal 5 segments of the antennae of female adults are short moniliform. The total length of the male antennae is significantly longer than that of the female. Eleven types of sensilla including Böhm’s bristles, sensilla trichodea, sensilla placodea, sensilla basiconica, sensilla chaetica, sensilla coeloconica, sensilla campaniformia, sensilla squamous, sensilla gemmiformia, sensilla cavity and sensilla cylindric, and smell pores were found on female and male adult antenna. Among them, sensilla trichodea, sensilla placodea and sensilla basiconica have two subtypes. Sensilla trichodea and sensilla placodea are the main sensilla on the antennae of C. ruficrus, with a wide distribution and a large number. Sensilla cavity are only distributed in pairs on the pedicel, and sensilla cylindric were only found on the flagellum of male adult. Sensilla gemmiformia were found for the first time in Braconidae.【Conclusion】We identified eleven types of antennal sensilla in female and male adults of C. ruficrus, more than those in other known braconid species, and sensilla gemmiformia were found for the first time in braconid species, speculating that they may play an important role in host positioning and searching for spouses.
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