Acta Entomologica Sinica

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Research and applications of genetic pest management techniques

SUN Hao, GAO Cong-Fen, WU Shun-Fan

Acta Entomologica Sinica 2024, 67 (3): 404-421. DOI: 10.16380/j.kcxb.2024.03.011

Abstract （502）

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Insect vectors cause significant economic losses and human casualties worldwide each year by transmitting parasites and viruses such as malaria, Zika virus, and dengue. Agricultural pests cause huge losses of crop yield every year and seriously threaten global food security. However, the current control methods based on chemical agents are insufficient to completely control the occurrence and damage of pests. At the same time, the use of chemical pesticides will induce resistance and result in environmental pollution and pesticide residues, etc. Therefore, there is an urgent need to develop new pest control strategies in production. In recent years, with the development of genome sequencing and gene editing techniques, the genetic control technology for the target pest population and their specific target genes has been rapidly developed. Compared with traditional pest control methods such as chemical control, genetic control strategies for pests have the advantages of species specificity, environmental friendliness, and efficient control. In this article, we reviewed several widely studied genetic control techniques for pests, including sterile insect technique (SIT), release of insects carrying a dominant lethal (RIDL), and gene drive (GD) technology. Finally, we presented several prospects for the research of genetic control technology for pests and its application in agricultural pest control: (1) to establish stable and efficient genetic manipulation systems; (2) to identify efficient promoters in germ cells or other tissues to improve the efficiency of gene editing or gene transformation; and (3) to elucidate the sex determination pathway of pests and excavate the key genes involved in the reproductive development of pests.

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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

Abstract （480）

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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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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

Abstract （267）

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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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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

Abstract （261）

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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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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

Abstract （252）

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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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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

Abstract （239）

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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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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

Abstract （238）

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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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Effects of the gustatory receptor gene EscrGR8 on the fecundity of female adults of Eucryptorrhynchus scrobiculatus (Coleptera：Curculioniclae)

GUO Xiao-Li, WEN Chao, WEN Jun-Bao, WEN Xiao-Jian

Acta Entomologica Sinica 2024, 67 (3): 307-317. DOI: 10.16380/j.kcxb.2024.03.001

Abstract （227）

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【Aim】 To analyze and elucidate the molecular characteristics and function of the gustatory receptor gene EscrGR8 in Eucryptorrhynchus scrobiculatus, and to reveal its role in the regulation of the fecundity of female adult. 【Methods】 Based on the antennal transcriptome database of E. scrobiculatus, the cDNA full-length sequence of EscrGR8 was cloned using RACE. The expression levels of EscrGR8 in different developmental stages (egg, 1st－6th instar larvae, pupa, female adult, and male adult), and female and male adult tissues (head without antennae and proboscis, antennae, mouthparts, midgut, forefoot, testicles, ovaries, male copulatory organ and female ovipositor) of E. scrobiculatus were detected by qRT-PCR. The expression levels of EscrGR8 at 0, 6, 12, 24, 36, 48 and 72 h after RNAi through microinjection of dsRNA into female adults were detected by qRT-PCR. The oviposition selection rates, total numbers of eggs laid and hatching rates of eggs laid by female adults under different soil moisture conditions (0-10%, 11%-20%, 21%-40%, 41%-60%, 61%-80%, and 81%-100%) at 1-5 d and 6-11 d after dsEscrGR8 microinjection were determined, and the effects of inhibition of the EscrGR8 expression on the oviposition preference and fecundity of female adults were studied. 【Results】 The full-length cDNA sequence of EscrGR8 (GenBank accession no.: OR836580) of E. scrobiculatus was successfully cloned with the open reading frame (ORF) of 1 251 bp in length, encoding 416 amino acids. EscrGR8 has six transmembrane domains. Multiple sequence alignment and phylogenetic analysis results showed that EscrGR8 has a low homology to the GRs of other insects, and the amino acid sequence identity with AgraGR64f of Anthonomus grandis is 30.96%. qRT-PCR result showed that there were significant differences in the expression levels of EscrGR8 in different developmental stages and adult tissues of E. scrobiculatus. EscrGR8 had the highest expression level in female adults, and the lowest expression level in eggs. EscrGR8 was highly expressed specifically in the female ovaries. Microinjection of dsEscrGR8 not only significantly reduced the expression level of EscrGR8 within a certain period of time, but also had a significant impact on the oviposition preference of female adults. At 1-5 d after microinjection of dsEscrGR8, the oviposition selection rate and hatching rate of eggs laid by female adults under soil condition with the moisture content of 21%-40% were significantly reduced by 2466% and 1583%, respectively, compared with those of the dsGFP-microinjected control group. The oviposition selection rate increased significantly by 28.39% under soil condition with the moisture content of 81%-100%, and almost all of the eggs laid by female adults failed to hatch. 【Conclusion】 This study has confirmed the effect of gustatory receptor gene EscrGR8 on the oviposition preference and fecundity of female adults of E. scrobiculatus, which is helpful for understanding the diversity and functional specificity of gustatory receptor genes in insects.

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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

Abstract （226）

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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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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

Abstract （218）

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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-SOD1, Cu/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-SOD1, Cu/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-SOD1, LbCu/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-SOD1, LbCu/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-SOD1, LbCu/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-SOD1, LbCu/Zn-SOD2 and LbFe/Mn-SOD are involved in the tolerance of L. bostrychophila to extreme temperature stress.

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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

Abstract （210）

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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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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

Abstract （204）

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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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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

Abstract （194）

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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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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

Abstract （181）

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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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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

Abstract （180）

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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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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

Abstract （175）

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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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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

Abstract （173）

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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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Age-stage, two-sex life table of Tuta absoluta (Lepidoptera: Gelechiidae) at different humidities

ZHANG Bo-Chen, XU Shuang-Ye, WU Zi-Qing, ZHANG Rui, ZHENG Wei-Feng, LI Ya, HAO Chi, YU Qin

Acta Entomologica Sinica 2024, 67 (1): 78-89. DOI: 10.16380/j.kcxb.2024.01.009

Abstract （173）

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【Aim】To clarify the impact of humidity change on the growth, development and fecundity of Tuta absoluta.【Methods】At the relative humidities of 15%, 35%, 55%, 75% and 95% indoor, the developmental parameters including developmental duration, pre-oviposition period, generation time, larval survival rate, adult survival rate, and number of eggs laid per female of T. absoluta were measured. The age-stage, two-sex life table s of T. absoluta were constructed, and population parameters including survival rate, fecundity, life expectancy, and reproduction value were calculated using the age-stage, two-sex life table.【Results】At the relative humidities of 15%－95%, the egg duration at the relative humidity of 15% and 95% were significantly longer than those at the relative humidities of 35%, 55%, and 75%. The 4th instar larval duration first shortened and then lengthened with the relative humidity increased. The male adult longevity increased with the rising relative humidity, reaching the maximum at the relative humidity of 95%. The female adult longevity was the longest at the relative humidity of 75%. The generation time was significantly longer at relative humidity of 95% than those at other relative humidities. The number of eggs laid per female increased with the increasing relative humidity. The initial oviposition time of female adults was affected by the relative humidity, the initial oviposition time were 31, 27, 27, 27 and 31 d at the relative humidities of 15%, 35%, 55%, 75% and 95%, respectively. The population age specific net maternity and age-stage-specific reproductive value increased with the rising relative humidity, reaching the maximum at the relative humidity of 95%. The intrinsic rate of increase and finite rate of increase initially increased and then decreased with the rising relative humidity, the net reproductive rate increased with the rising relative humidity. The mean generation time initially decreased and then increased with the rising relative humidity. According to the parameters of the population life table, the prediction of population dynamics within next 60 d indicated that the relative humidity of 95% was conducive to the increase of population size, relative humidities of 15% and 95% were unfavorable to the increase of the population generation number. The impacts of different relative humidities on the survival rates of eggs and larvae were significant through linear regression fitting. 【Conclusion】Within the test setting relative humidity range, the optimal relative humidities for the growth and development of T. absoluta are 55% and 75%, and the optimum relative humidity for reproduction are 95%.

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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

Abstract （172）

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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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Functional analysis of the G-protein β-subunit gene in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

XU Hang-Peng, REN Peng-Peng, ZHANG Chuan-Xi, LU Jia-Bao

Acta Entomologica Sinica 2024, 67 (4): 456-467. DOI: 10.16380/j.kcxb.2024.04.002

Abstract （169）

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【Aim】This study aims to identify the G-protein β-subunit gene (NlGβ) in the brown planthopper, Nilaparvata lugens, and analyze its function, so as to supply a potential novel target gene for RNAi-based control of N. lugens. 【Methods】The coding sequence (CDS) of NlGβ of N. lugens was cloned and verified using PCR, and subjected to bioinformatics analysis. The spatiotemporal expression patterns of NlGβ were analyzed based on its transcriptome expression profiles in different developmental stages (egg, 1st-5th instar nymphs, and female and male adults) and adult tissues (head, leg, gut, cuticle, fat body, female reproductive system, and male reproductive system) of N. lugens. Microinjection of dsRNA against NlGβ was performed on the 2nd and 5th instar nymphs to silence NlGβ. Individual and female reproductive system phenotypes were observed, and the survival rate, number of eggs laid per female, and egg hatching rate were counted. Additionally, transmission electron microscopy was employed to observe the lateral oviduct intumescent region. 【Results】 The CDS of NlGβ (GenBank accession no.: XP_022200908.1) of N. lugens was 948 bp in length. The NlGβ protein comprised seven WD40 domains and four WD_REPEATS_1 motifs, indicating its conservation. Apart from the homologous proteins found in orthopteran insects, homologous NlGβs from other insect orders clustered well within the same evolutionary branch. The expression level of NlGβ exhibited periodic change during the 1st-3rd instar nymphal stages, and those in the 5th instar female nymphs and female adults were higher than those in the 5th instar male nymphs and male adults. NlGβ was expressed in various tissues of adults, with the highest expression level in the fat body, and the expression level of NlGβ in female reproductive system was higher than that in male reproductive system. Silencing NlGβ in the 2nd instar nymphs resulted in molting difficulty, leading to a significant decrease in the survival rate compared to the dsGFP control group. Silencing NlGβ in the 5th instar nymphs resulted in abnormal abdominal swelling in adult females, malformed ovarian development, a significant decrease in the number of eggs laid per female compared to the dsGFP control group, failure of egg hatching, an increase in secretion within the lateral oviduct intumescent region, and degradation of epithelial cells. 【Conclusion】 NlGβ is closely associated with the growth and development of N. lugens and the reproduction of its female individuals.

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Analysis of differences in the midgut bacterial composition and function between different Bombyx mori varieties at the 5th instar larval and pupal stages

LI Qing-Rong, YANG Qiong, XING Dong-Xu, ZOU Yu-Xiao, ZHANG Wei-Long, XIAO Yang

Acta Entomologica Sinica 2023, 66 (8): 1020-1030. DOI: 10.16380/j.kcxb.2023.08.003

Abstract （166）

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【Aim】 To analyze the differences in the midgut bacterial composition among different varieties of Bombyx mori with different vitalities and cocoon shell ratios, and their effects on the related traits such as vitality and cocoon shell ratio. 【Methods】 Based on the results of the long-term feeding survey on B. mori resource, the variety 932G with high vitality and the variety 2041J with high silk yield were selected as experimental materials, the midguts of the 5th instar larva and pupa were collected, and the 16S rDNA sequences of the midgut bacteria were sequenced and analyzed by the high-throughput sequencing platform to compare the differences in the composition and function of the midgut bacteria between different varieties and different developmental stages of B. mori. 【Results】 A total of 399 and 453 operational taxonomic units (OTUs) of the midgut bacteria were obtained from 932G and 2041J at the 5th instar larval stage, and 138 and 162 OTUs of the midgut bacteria were obtained from 932G and 2041J at the pupal stage, respectively. The dominant phyla of the midgut bacteria of B. mori at the 5th instar larval stage were Proteobacteria, Firmicutes and Actinobacteriota, and the dominant genus was Methylobacterium with the highest abundance, followed by Staphylococcus. However, there were significant differences in the relative abundance of some genera between the two varieties. The relative abundance of some genera including Devosia, Ralstonia, Nitrospira, Brachybacterium, Rothia, Lawsonella etc. in the midgut of the 5th instar larva of 932G was significantly higher than that in the midgut of the 5th instar larva of 2041J. The relative abundance of Pseudomonas, Methylobacterium, Acinetobscter, Cloacibacterium, Leuconostoc, Propionibacteriaceae, Psychrobacter, Sphingobium, Bacteroides etc. in the midgut of the 5th instar larva of 2041J was significantly higher than that in the midgut of the 5th instar larva of 932G. There were 77% and 78% functional genes in the midgut bacteria of the 5th instar larvae of 932G and 2041J enriched on the metabolic pathways of KEGG, respectively, followed by those of the function genes enriched on the environmental information processing and genetic information processing. The proportions of the midgut bacterial communities involved in nitrate reduction, nitrogen respiration, nitrate respiration, nitrite respiration and nitrogen fixation in the 5th instar larva of 932G were higher than those in 2041J. The proportions of the midgut bacterial communities involved in chemical heterotrophic, urea decomposition and methanol oxidation in the 5th instar larva of 2041J were higher than those in the 5th instar larva of 932G. The difference in the midgut bacterial composition between the 5th instar larva and pupa was significant. Erwinia of Proteobacteria in the midgut of the pupa was the dominant genus, and there was no significant difference in the relative abundance of Erwinia between the two varieties at the pupal stage. The relative abundance of functional genes involved in amino acid transport and metabolism, extracellular secretion and transport, carbohydrate transport and metabolism, and inorganic salt transport and metabolism of the midgut bacteria in the pupa was significantly higher than that in the 5th instar larva of 932G.【Conclusion】 There were significant differences in the midgut bacterial composition and predicted function between the varieties 932G with high vitality and 2041J with high silk yield of B. mori at the 5th instar larval stage. The composition and function of the midgut bacteria of B. mori at the pupal stage were significantly different from those of B. mori at the 5th instar larval stage, and there was no significant difference in the relative abundance of midgut bacteria between the two varieties at the pupal stage. These research results can provide a reference for further exploring the role of gut microorganisms in stress resistance, drug resistance, disease resistance, protein synthesis and transformation of B. mori and variety breeding.

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Rice pests: Research progresses and prospects

LI You-Zhi, FANG Ji-Chao

Acta Entomologica Sinica 2024, 67 (4): 443-455. DOI: 10.16380/j.kcxb.2024.04.001

Abstract （164）

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Rice, as a major staple crop in China, is directly threatened by disease and pest infestations, which jeopardize its safe production. Rice pests are diverse, and their outbreak mechanisms are complex, making the development of green control technologies challenging. Indepth research into rice pest monitoring and early warning technologies, developmental and reproductive regulation mechanisms, chemical communication mechanisms, and novel control technologies not only helps elucidate the outbreak mechanisms of rice pests but also provides new effective control targets and techniques for integrated pest management. In recent years, Chinese scholars have made significant progress in the field of rice pest research, gaining important achievements in multiple research directions, and some recent research findings were presented in this special issue. In this article, we outline the latest progresses in rice pest research at home and abroad, introduce the main research contents of this special issue, and propose the following three research directions worthy of further attention: (1) intelligent monitoring and early warning of rice pests; (2) mechanisms and evolution patterns of rice pest outbreaks; and (3) exploration of green control targets for rice pests and the construction and application of sustainable control systems.

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Effects of oviposition times and host larval instars on the biological characters of Cotesia ruficrus (Hymenoptera: Braconidae), an indigenous parasitoid of Spodoptera frugiperda (Lepidoptera: Noctuidae)

HE Peng-Yang, LI Xian, LIU Tong-Xian, ZHANG Shi-Ze

Acta Entomologica Sinica 2023, 66 (8): 1095-1104. DOI: 10.16380/j.kcxb.2023.08.010

Abstract （163）

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【Aim】 To evaluate the parasitism potential of Cotesia ruficrus, an indigenous parasitic natural enemy, against the larvae of Spodoptera frugiperda, a major invasive agricultural pest. 【Methods】The parasitism preference of the mated 24 h-old female adults of C. ruficrus to the 1st-3rd instar larvae of S. frugiperda was performed by selection test. The developmental duration, numbers of cocoons, eclosion rates, sex ratios (proportions of females), adult longevity and adult size of the offspring of C. ruficrus were determined when its mated 24 h-old female adults parasitized the 1st, 2nd or 3rd instar larvae of S. frugiperda, respectively, by non-selection test. The oviposition times of the mated 24 h-old female adults of C. ruficrus on the 2nd instar larvae of S. frugiperda were determined by the direct observation method, and then the effects of different oviposition times on the developmental duration, number of cocoons, eclosion rate, sex ratio, adult longevity and adult size of the offspring of C. ruficrus were measured. The parasitic functional response of the mated 24 h-old female adults of C. ruficrus to the 2nd instar larvae of S. frugiperda was determined with small-leaf dish method. 【Results】 When the mated 24 h-old female adults of C. ruficrus parasitized the 1st-3rd instar larvae of S. frugiperda, the parasitism rates, egg-larval duration, pupal duration, numbers of cocoons per host, eclosion rates, and proportions of females of C. ruficrus were 34%-54%, 8.7-9.9 d, 3.9-4.1 d, 19.7-41.2 cocoons, 84.4%-92.0% and 31.5%-38.8%, respectively, and the female adult size of the offspring was larger than the males. The female adult longevity of the offspring of C. ruficrus was significantly longer than that of the males when the 1st or 3rd instar larvae of S. frugiperda were parasitized. The oviposition times of the mated 24 h-old female adults of C. ruficrus on the 2nd instar larvae of S. frugiperda had a significant effect on the biological characters of the offspring of C. ruficrus. With the increase of the oviposition times, the male adult longevity of the offspring of C. ruficrus was shortened, and the male adult size of the offspring became smaller. Compared with parasitization once, after the 2nd instar larvae of S. frugiperda were parasitized by the mated 24 h-old female adults of C. ruficrus for three times, the egg-larval duration of the offspring of C. ruficrus was significantly prolonged by 0.7 d, the pupal duration was shortened by 0.4 d, the number of cocoons per host significantly increased by 25 and the proportion of females significantly decreased by 58.8%. The parasitic functional response of the mated 24 h-old female adults of C. ruficrus to the 2nd instar larvae of S. frugiperda was Holling Ⅱ type, the daily maximum parasitic number was 17.5, the instantaneous attack rate was 0.043 and the average handling time was 1.368. 【Conclusion】 This study evaluated for the first time the biological control potential of C. ruficrus against S. frugiperda larvae, and indicated that the parasitoid was an effective native natural enemy of S. frugiperda, providing technical reserves and theoretical basis for further utilizing local natural enemies to control invasive pests.

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Community composition and functional gene annotation of the symbiotic microbes in Ips nitidus (Coleoptera: Scolytinae)

LIU Cai-Xia, LI Gang, LIANG Ling-Yu, HAN Fu-Zhong, WANG Zheng, WANG Hui-Min, LÜ Quan

Acta Entomologica Sinica 2023, 66 (10): 1329-1342. DOI: 10.16380/j.kcxb.2023.10.007

Abstract （162）

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【Aim】To reveal the community composition and functional characteristics of the symbiotic microbes in Ips nitidus. 【Methods】Metagenomics and correlation network analysis were conducted on the symbiotic fungi and bacteria in the gut and other tissues of I. nitidus from Maixiu Nature Reserve in Qinghai Province, Northwest China to compare the community composition of the symbiotic fungi and bacteria in the gut and other tissues of I. nitidus, to analyze the interrelationships of the microbial communities and to annotate the functional genes of the symbiotic microbes by non-redundant protein sequence alignment method. 【Results】A total of 3 520 species of symbiotic microbes were found in the gut and other tissues of I. nitidus. Fungi and bacteria possessed the highest numbers of species (accounting for 23.21% and 69.01% of all species, respectively), with the average relative abundance of 9.69% and 13.79%, respectively. For fungal symbionts, Erysiphales showed higher relative abundance and species diversity in other tissues than in the gut, and Saccharomycetales showed lower relative abundance and species diversity in other tissues than in the gut. Ophiostomatoid fungi in other tissues showed higher species diversity than those in the gut (16 species of 6 genera, and 13 species of 5 genera, respectively). The average relative abundance of ophiostomatoid fungi in other tissues and gut occupied 0.016% and 0.013%, respectively. The relative abundance and species diversity of Enterobacterales in other tissues were higher than those in the gut. Enterobacter, Erysiphe, Rhizophagus and Wolbachia were the core genera in the gut and other tissues. Some communities of fungi and bacteria showed opposite interrelationships in the gut and other tissues, suggesting that there may be complex interactions between the symbionts in I. nitidus. A total of 7 010 and 6 483 functional genes were identified from symbiotic fungi and bacteria, respectively. The number of functional genes related to xenobiotics biodegradation and metabolism in bacteria was higher than that in fungi. However, 11 pathways such as mRNA surveillance pathway, propanoate metabolism, ubiquitin-mediated proteolysis and so on had more significant differential genes in the gut than in other tissues. The annotation results for functional genes related to limonene degradation and sterol synthesis showed that bacteria may degrade limonene significantly compared with fungi, but fungi may play a more crucial role in promoting sterol biosynthesis than bacteria. A total of 74 functional genes of ophiostomatoid fungi are mainly involved in oxidative phosphorylation, metabolic pathway and amino acid metabolism. Moreover, ophiostomatoid fungi were of higher diversity in other tissues and mainly distributed in Ophiostoma, etc. 【Conclusion】There are some differences in the community composition, diversity and function of the symbiotic microbes in the gut and other tissues of I. nitidus. Enterobacter, Erysiphe, Rhizophagus and Wolbachia are the most important core flora, and they might play an important role in the community assembly.

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Regulatory mechanism of intestinal microbial homeostasis in insects

SONG Yang, FAN Lin-Lin, SHENTU Xu-Ping, YU Xiao-Ping

Acta Entomologica Sinica 2023, 66 (10): 1404-1414. DOI: 10.16380/j.kcxb.2023.10.013

Abstract （159）

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A stable intestinal microbial internal environment is the result of the interaction between intestinal microbes and intestinal immune response. During the continuous feeding, the species and number of insect intestinal microbes constantly change, so a complex and dynamic homeostatic mechanism is formed between intestinal microbes and intestinal epithelial cells. Insect intestinal epithelial cells can sense beneficial and harmful conditions and use immunoregulatory pathways for dynamic regulation of microbial population homeostasis, such as the dual oxidase-reactive oxygen species (Duox-ROS) system and immunodeficiency (Imd) signaling pathways, which can sense the changes in the number of intestinal microbies and participate in the regulation of intestinal microbial homeostasis. In addition, the intestinal microbes can also indirectly play a role in homeostatic regulation by releasing corresponding effectors to regulate the behavior of the flora through quorum sensing (QS). Thus, in this article, we reviewed the roles of physical defense, immune signaling pathways and QS in the maintenance of insect intestinal microbial homeostasis, which will deepen the understanding of the interactions between intestinal tissues and intestinal microbes. In the future, we will continue to reveal the regulatory mechanisms of microbial homeostasis in more species of insects and the relationship between the regulatory mechanisms. In addition, new pesticides will be designed and developed to alter the homeostasis of intestinal microbes based on the regulatory mechanisms, which will provide new targets and ideas for the pest control.

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Screening and simulated field efficacy evaluation of Beauveria bassiana with high virulence to Holotrichia parallela (Coleoptera: Scarabaeidae) larvae

ZHU Chang-Jian, MA Yu-Meng, LI Bin, LIANG Jia-Hui, CAO Fu, YANG Yang, LIU Su, LI Mao-Ye

Acta Entomologica Sinica 2023, 66 (8): 1009-1019. DOI: 10.16380/j.kcxb.2023.08.002

Abstract （157）

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【Aim】 To screen Beauveria bassiana strains with high virulence against Holotrichia parallela larvae and to evaluate the simulated field efficacy of the strain with the highest virulence in combination with azadirachtin and phoxim, respectively.【Methods】 The virulence of 10 strains of B. bassiana to the 3rd instar larvae of H. parallela was evaluated based on the corrected mortality and medium lethal time (LT₅₀) value using the insect dipping method. Furthermore, the medium lethal concentration (LC₅₀) values of the screened Bb14 strain against the 1st and 3rd instar larvae of H. parallela were determined. In the pot experiments, the conidial suspension of Bb14 (1×10⁸ conidia/mL) was applied in combination with azadirachtin (3 mg/L) and phoxim (200 mg/L), respectively, and the control efficacy against the 3rd instar larvae of H. parallela on tobacco seedlings was evaluated in greenhouse.【Results】 Among the 10 strains of B. bassiana at the concentration of 1.0×10⁸ conidia/mL, Bb14 had the highest virulence against the 3rd instar larvae of H. parallela, causing the cumulative corrected mortality of 76.67% in 17 d and with the LT₅₀ value of 11.14 d. The LC₅₀values of Bb14 infecting the 1st and 3rd instar larvae of H. parallela were 0.32×10⁷ and 0.68×10⁷ conidia/mL, respectively. In the pot experiments, the control efficacy of Bb14, azadirachtin and phoxim against the 3rd instar larvae of H. parallela was 42.86%, 64.29% and 82.14%, respectively, at 25 d after application. At 25 d after applying Bb14 in combination with azadirachtin and phoxim, respectively, the control efficacy against the 3rd instar larvae of H. parallela reached 78.57% and 92.86%, respectively. 【Conclusion】 Among the tested 10 strains of B. bassiana, Bb14 has the highest virulence to H. parallela larvae. The application of Bb14 in combination with azadirachtin and phoxim leads to improved control efficacy against H. parallela larvae.

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Effects of the symbiont Serratia symbiotica on the development and reproduction of the pea aphid, Acyrthosiphon pisum (Hemiptera: Aphididae)

LI Yue-Ming, ZHANG Yong-Dong, GONG Hou-Yan, LÜ Zhi-Qiang

Acta Entomologica Sinica 2023, 66 (10): 1311-1318. DOI: 10.16380/j.kcxb.2023.10.005

Abstract （156）

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【Aim】 To clarify the effects of antibiotics on the obligate symbiont in the pea aphid, Acyrthosiphon pisum and investigate the impacts of the symbiont Serratia symbiotica on the growth, development and reproduction of A. pisum. 【Methods】 The type of facultative symbionts in adults of the green morph of A. pisum used in this study was detected by diagnostic PCR. The facultative symbionts were removed by injecting and feeding antibiotics to establish A. pisum line without facultative symbionts. The copy numbers of specific gene fragment of the obligate symbiont Buchnera aphidicola in A. pisum adults with and without S. symbiotica were determined by quantitative PCR. The body weight, rates of ecdysis, mortality rates and fecundity of A. pisum with and without S. symbiotica were recorded. 【Results】The A. pisum adults used in this study harbored only one facultative symbiont, S. symbiotica. The facultative S. symbiotica in A. pisum adults could be removed by injecting antibiotics, while the injection of antibiotics had no effect on the content of the obligate symbiont B. aphidicola. Compared with S. symbiotica-harboring A. pisum adults, S. symbiotica-removed A. pisum adults gained weight very slowly, and had less than one-third of the weight of S. symbiotica-harboring A. pisum adults at 9 d after birth. S. symbiotica-removed A. pisum adults had longer 1st instar nymphal duration, higher mortality rate and fewer offspring produced by per adult. 【Conclusion】The laboratory green A. pisum line carries the symbiont S. symbiotica. The removal of the symbiont S. symbiotica from A. pisum does not affect the content of the obligate symbiont B. aphidicola but has a negative effect on the growth, development and reproduction of A. pisum.

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Cloning and identification of two GST genes in Pardosa astrigera (Araneae: Lycosidae) and their expression in response to deltamethrin stress

JIAO Li-Ya-Lin, ZHAO Meng-Meng, WANG Mei, NIU Yue, WANG Xi, LI Rui

Acta Entomologica Sinica 2023, 66 (9): 1161-1170. DOI: 10.16380/j.kcxb.2023.09.003

Abstract （155）

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【Aim】To clarify the spatiotemporal expression patterns of two genes of glutathione S-transferases (GSTs) in Pardosa astrigera and to investigate whether the two genes can respond to deltamethrin stress.【Methods】 Based on the transcriptome database of P. astrigera, the full-length cDNA sequences of the GST genes PaGSTd1 and PaGSTd2 of P. astrigera were cloned by PCR and bioinformatically analyzed. RT-qPCR was used to detect the expression levels of PaGSTd1 and PaGSTd2 in different developmental stages (2nd-6th instar nymphs and adult), female and male adult tissues (cephalothorax, abdomen and leg)， and in male adult of P. astrigera stressed by deltamethrin at different concentrations ［LC₁₀(5.151 mg/L), LC₃₀(8.619 mg/L) and LC₅₀ (12.311 mg/L)］ for different time (6, 12, 18, 24 and 48 h)through residual film method.【Results】 The full-length cDNA sequence of PaGSTd1 (GenBank accession number: OR096398) of P. astrigera is 708 bp in length with an open reading frame of 645 bp in length, encoding 214 amino acids. The full-length cDNA sequence of PaGSTd2 (GenBank accession number: OR096399) of P. astrigerais 793 bp in length with an open reading frame of 645 bp in length, encoding 214 amino acids. The detection results of RT-qPCR showed that PaGSTd1 and PaGSTd2 were expressed in different developmental stages and adult tissues of P. astrigera, and the expression levels of PaGSTd1 and PaGSTd2 were the lowest in the 5th instar nymph. The expression level of PaGSTd1 was the highest in the 4th instar nymph of P. astrigera,and that of PaGSTd2 was the highest in adult of P. astrigera. The expression levels of PaGSTd1 and PaGSTd2 were the highest in the leg, and the expression levels in other tissues except for that in the abdomen were significantly higher in male adults than in female adults of P. astrigera. PaGSTd1 and PaGSTd2 in male adult of P. astrigera were induced at 24 h, and at 6 and 18 h, respectively, after stress by LC₁₀ of deltamethrin. PaGSTd1 and PaGSTd2 in male adult of P. astrigera were induced at 18, 24 and 48 h, and at 18 h, respectively, after stress by LC₃₀ of deltamethrin. Both PaGSTd1 and PaGSTd2 were induced in male adult of P. astrigera at 24 h after stress by LC₅₀ of deltamethrin.【Conclusion】 In this study, two GST genes PaGSTd1 and PaGSTd2 were cloned from P. astrigera. Both PaGSTd1 and PaGSTd2 were most highly expressed in the leg of P. astrigera, suggesting that these two GST genes in the leg of P. astrigera play an important role in the detoxification of exogenous substances. These two GST genes could be induced to express by deltamethrin, suggesting that these two GST genes PaGSTd1 and PaGSTd2 may be involved in the stress response of P. astrigera to deltamethrin, providing clues for subsequent functional studies of GSTs.

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Research progress on senescence and longevity regulation of honeybees

JIN Meng-Jie, ZENG Zhi-Jiang

Acta Entomologica Sinica 2023, 66 (11): 1527-1534. DOI: 10.16380/j.kcxb.2023.11.012

Abstract （154）

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Honeybee queens share the same genetic background with the workers, and they both are developed from fertilized eggs. Nutritional and spatial variances during the development lead to significant morphological, physiological and behavioral dimorphism between the two castes. The insulin signaling pathway (IIS) regulates the behavior of worker bees, thus influencing their longevity. Longer longevity of queen bees is associated with increased oxidative stress and enhanced stress defense. Vitellogenin (Vg) interacts with juvenile hormone (JH), and the relatively high level of Vg and low level of JH usually lead to longer longevity. Telomerase activity and telomere length are influenced by the development and caste of honeybees. Queens inherit longer telomere length and maintain higher telomerase activity than workers. Overwintering worker bees live longer and show higher telomerase activity than the summer worker bees. Mitochondrial damage is a sign of senescence, while the mitochondrial function of the aged queen remains vigorous. Senescence is closely related to DNA methylation, and DNA methylation and histone modification play important roles in the regulation of plasticity in social insects. With the increase in population aging and the high prevalence of senescence-related diseases, “healthy aging” has triggered a series of concerns in life sciences and social sciences. Studies on honeybee senescence and longevity regulation will provide an important reference for the biology of senescence.

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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

Abstract （153）

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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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Cloning, identification and functional analysis of the mucin-like protein EfMLP genes in Empoasca flavescens (Hemiptera: Cicadellidae)

HE Xue-Yi, LEI Yu-Huan, SONG Shi-Yue, XIA Lu-Xia, WANG Shi-Yu, MA Cheng-Wen, WEI Ke-Xin, WANG Meng-Xin, PAN Cheng, HAN Bao-Yu

Acta Entomologica Sinica 2024, 67 (2): 151-162. DOI: 10.16380/j.kcxb.2024.02.001

Abstract （150）

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【Aim】This study aims to investigate the molecular characteristics, expression patterns, and biological functions of the mucin-like protein EfMLP genes of Empoasca flavescens. 【Methods】Based on the transcriptome data of E. flavescens, the full-length cDNA sequences of four EfMLP genes were cloned by PCR and analyzed by bioinformatics. qRT-PCR was used to detect the expression levels of EfMLP genes across different developmental stages (egg, 1st-5th instar nymphs, and newly emerged female and male adults), and in different tissues (integument, fat body, salivary gland, gut, ovary, and testis) of the newly emerged adults. EfMLP2 and EfMLP4 in the 5th instar nymph were silenced by RNAi through feeding method, and the survival rates of E. flavescens after silencing the EfMLP genes by RNAi were determined by bioassay. 【Results】 The full-length cDNA sequences of four EfMLP genes of E. flavescens were obtained, and named EfMLP1, EfMLP2, EfMLP3 and EfMLP4 with the GenBank accession numbers of OR504428, OR504429, OR504430 and OR504431, respectively. The obtained four EfMLPs all contain highly repetitive tandem repeat sequences, which are rich in O-linked glycosylation sites, forming the mucin domain (MD). Among them, both EfMLP3 and EfMLP4 contain a conserved type-2 chitin binding domain (CBD). Phylogenetic analysis result revealed that EfMLPs were divided into two different branches belonging to two different MLP types, which showed no correlation with insect taxonomy, but might be considered to be related to their functions. EfMLP1 and EfMLP2 exhibited specifically high expression in the newly emerged female and male adults and the salivary glands of the newly emerged adults. In contrast, the expression of EfMLP3 and EfMLP4 was identified in various developmental stages, including egg, nymphal and adult stages, as well as in diverse tissues such as the fat body of the newly emerged adult. Inhibition of the expression of EfMLP2 and EfMLP4 in E. flavescens by feeding dsEfMLP2 and dsEfMLP4 significantly reduced the survival rate of E. flavescens compared with the control group fed with dsGFP. 【Conclusion】 EfMLPs play an important role in the feeding of E. flavescens and can be used as a potential target in control of this pest insect based on RNAi strategies.

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Life cycle and overwintering rules of Lissorhoptrus orvzophitus (Coleoptera: Curculionidae) in Guangdong province, South China

YUAN Long-Yu, LEI Hao-Lin, LI Yan-Fang, XIAO Han-Xiang, WEI Hong-Yi, ZHANG Zhen-Fei

Acta Entomologica Sinica 2023, 66 (9): 1192-1200. DOI: 10.16380/j.kcxb.2023.09.006

Abstract （149）

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【Aim】 To clarify the life cycle, overwintering habitats, occurrence regularity, host plant species and natural enemy species of the rice water weevil (Lissorhoptrus orvzophitus) in Guangdong province, South China, so as to provide scientific guidance for the prevention and control of this insect pest.【Methods】 We investigated the life cycle and overwintering habitats of L. orvzophitus in Guangdong through field surveys, cage observation experiments and ovary dissection from March, 2019 to April, 2021, identified its overwintering transitional host plants through molecular detection of the midgut contents in adults, and detected its main natural enemies using sequence characterized amplified region (SCAR) marker technology.【Results】 L. orvzophitus had two generations a year in the double cropping rice area of Guangdong province, and had overwintering and oversummering habits during March, 2019 and April, 2021. A complete generation of L. orvzophitus needed (140.86±1.05) d, and the larval duration was about 20 d in Guangdong. L. orvzophitus overwintered as adult in Guangdong province. The overwintering habitat pattern was paddy field, transitional zone (grassland), and swamp (abundant water grass). It was found that the ovary of L. orvzophitus adults during the overwintering period underwent diapause, and the ovarian grade was level Ⅰ. Through molecular detection of the midgut contents in L. orvzophitus adults, it was found that during the overwintering period, the main transitional host plants of L. orvzophitus were regenerated rice (Oryza sativa) and Leersia hexandra, and the main natural enemy was Pardosa laura, a nomadic spider, as revealed by the result of SCAR-based PCR.【Conclusion】 L. orvzophitus has two generations a year in Guangdong, and mainly overwinters in the swamp. The main transitional hosts during the overwintering period are Leersia hexandra and regenerated rice, and the main natural enemy in overwintering habits is P. laura. These findings have theoretical guiding significance for the accurate prevention and control of L. orvzophitus epidemic in Guangdong.

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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

Abstract （148）

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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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Molecular properties, spatio-temporal expression profiles and antibody preparation of AmAGO1 protein of Apis mellifera

YE Ya-Ping, LIU Zhi-Tan, LI Qi-Ming, ZANG He, FENG Pei-Lin, WANG Ning, WANG Jie, HUANG Zhi-Jian, CHEN Da-Fu, GUO Rui

Acta Entomologica Sinica 2024, 67 (1): 18-28. DOI: 10.16380/j.kcxb.2024.01.003

Abstract （148）

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【Aim】 As a highly conserved protein family in evolution, Argonaute (AGO) family mainly engages in the formation of RNA-induced silencing complex (RISC) in eukaryotes to silence gene expression and further participate in numerous biological processes. Currently, studies on AGO proteins of honey bees are lacking. This study aims to provide the reference and basis for further performing study on function and mechanism of AGO1 of Apis mellifera AmAGO1 by predicting the physicochemical and molecular properties of AmAGO1, analyzing the spatio-temporal expression profiles of AmAGO1, and preparing the polyclonal antibodies against AmAGO1.【Methods】 The coding sequence (CDS) of AmAGO1 of A. mellifera was amplified using PCR amplification followed by predicting the the physicochemical and molecular properties of AmAGO1 protein using bioinformatics. The expression levels of AmAGO1 inthe egg, 3-day-old larva, 7-day-old prepupa, 8-day-old prepupa, 12-day-old pupa, 1-, 2-, 6-, 12-, 15- and 18-day-old adult A. mellifera workers, and in the antenna, hypopharyngeal gland, brain, cuticle, midgut, fat body and venom gland of adult workers were detected by RT-qPCR. After constructing prokaryotic expression plasmid, AmAGO1 fusion protein was induced and its expression form was identified. The polyclonal antibody against AmAGO1 was prepared and the titer, sensitivity and specificity of the antibody were further detected by ELISA, Western blot and immunoprecipitation (IP), respectively. 【Results】The CDS of AmAGO1 was successfully cloned from A. mellifera. AmAGO1 contains 928 amino acids, with the molecular formula C₄₆₂₄H₇₃₃₂N₁₃₁₆O₁₃₂₅S₅₁, the molecular weight of about 104.2 kD, isoelectric point of 9.31, average hydrophilic coefficient of –0.2965, 86 phosphorylation sites, and the typical domains PAZ and PIWI, without typical signal peptides. There was a high amino acide sequence identity of AGO1 proteins of Homo sapiens, Danio rerio, Drosophila melanogaster, Bombyx mori, A. mellifera, A. cerana, and Bombus terrestris. AGO1s from A. mellifera and A. cerana were clustered into one branch, with the highest homology. AmAGO1 was differentially expressed in egg, larva, prepupa, pupa, and adult of A. mellifera. The expression levels of AmAGO1 inthe 3-day-old larva and 7-day-old prepupae were significantly lower than that in the egg, while those in the 8-day-old prepupae and 12-day-old pupae were significantly higher than that in the egg of A. mellifera workers. AmAGO1 was differentially expressed in the 1-, 2-, 6-, 12-, 15-, and 18-day-old adults. The expression levels of AmAGO1 inthe 2-, 6-, 12-, 15- and 18-day-old adults were significantly lower than that in the 1-day-old adults. AmAGO1 was differentially expressed in the antenna, venom gland, brain, midgut, hypopharyngeal gland, fat body and cuticle of worker adults. The expression level of AmAGO1 inthe antenna was significantly higher than those in venom gland, brain, midgut, fat body and cuticle but as the same as that in the hypopharyngeal gland. The expression form of AmAGO1 fusion protein was inclusion body. The prepared AmAGO1 polyclonal antibody had high titer, sensitivity and specificity. 【Conclusion】 AmAGO1 may be a hydrophilic intracellular protein which contains typical PAZ and PIWI domains. AmAGO1 plays a potentially important role in different tissues and developmental stages of A. mellifera workers. AGO1 polyclonal antibody with high titer, high sensitivity, and strong specificity was successfully prepared.

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Analysis of expression patterns of CCE family genes in Anopheles sinensis (Diptera: Culicidae)

GU Xin-Yao, SI Feng-Ling, CHEN Bin

Acta Entomologica Sinica 2024, 67 (3): 318-326. DOI: 10.16380/j.kcxb.2024.03.002

Abstract （146）

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【Aim】 Carboxylesterases (CCEs) are an important hydrolase superfamily and have a role in the metabolism of various exogenous substances in insects. This study aims to lay a foundation for further research on the potential functions of CCEs in different physiological processes by analyzing the expression patterns of CCE genes from transcriptome datasets of Anopheles sinensis at different developmental stages, in different tissues, and before and after blood meal. 【Methods】 Based on 50 CCE genes collected from the obtained transcriptome data of the An. sinensis laboratory strain, the expression patterns of CCE genes in An. sinensis at different developmental stages (egg, 1st-4th instar larvae, male pupa, female pupa, male adult and female adult), in different adult tissues (antennae, salivary gland, midgut, Malpighian tubules, testis, ovary, cuticle and fat body), and female adults before and after blood meal (at 1, 3, 6, 12, 24 and 48 h) were analyzed by bioinformatics. 【Results】 CCE genes were mainly highly or specifically expressed in the larval or adult stages. AsAe12, AsAe4 and AsBe4 were highly expressed during all developmental stages. The expression patterns of CCE genes were also tissue specific and mainly expressed in antennae, cuticle and testis of adults. Moreover, five CCE genes (AsAe13, AsAe12, AsAe6, AsAe4 and AsBe4) were highly expressed in the midgut, Malpighian tubules and fat body of adults, suggesting their potential involvement in xenobiotic metabolism within these detoxification organs. After feeding female adults with blood meal, the expression levels of most CCE genes changed, and their expression patterns were different, suggesting that the blood digestion is a complex process. 【Conclusion】 The results of this study have enriched the knowledge of CCE genes in An. sinensis, providing a valuable reference for further research on the potential function of CCE family genes in the growth and development of An. sinensis.

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Circadian rhythms of movement, calling and mating behavior of the tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae)

SHI Qian-Rui, DONG Yi-Xia, NIE Zhen, GU Xing-Feng, LIU Zhen-Dong, GUO Qian-Shuang, DU Yong-Jun,

Acta Entomologica Sinica 2024, 67 (3): 393-403. DOI: 10.16380/j.kcxb.2024.03.010

Abstract （146）

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【Aim】To make better use of sex pheromones in the monitoring and control of the tomato leafminer, Tuta absoluta, and to further study the circadian rhythms of its movement, calling and mating behavior. 【Methods】For T. absoluta adults, video recording and two-dimensional trajectories were used to analyze their movements. Solid-phase microextraction (SPME) adsorption extraction and gas chromatography-mass spectrometry (GC-MS) were used to analyze the sex pheromone titer in female adults. In tomato greenhouses in Xichang City, Sichuan Province from August to September 2022, field trapping experiments were carried out to analyze the relationship between the sex pheromone titers of T. absoluta adults and their day-old age and circadian rhythm based on the trapping results. The mating behavior was observed in indoor insect cages, and the reproductive system of the male adult was observed by dissection, and the relationship between development and day-old age was analyzed. 【Results】The results showed that in the indoor natural environment, the movement of female and male adults occurred from the 5th hour of the scotophase to the 2nd hour of the photophase, with the peak at the 5th to 7th hour of the scotophase, and there was no significant difference in the time and distance of movement between females and males, and between the 1-7-day-old assayed. The sex pheromones of T. absoluta can be detected in adults at the 1-11-day-old, but there was no significant difference in the sex pheromone titer between day-old ages. The sex pheromones released by female adults were detected during the entire 24 h period, and the difference in the sex pheromone titer between different periods was not significant. Mating did not significantly affect the sex pheromone titer in female adults. However, the male adults in the field were trapped from the 7th hour of the scotophase to the 2nd hour of the photophase. T. absoluta adults were able to mate on the same day of emergence, and the mating rate was the highest when the male adults were at the 3-day-old, and then decreased, but there was no significant difference in the mating duration between day-old ages. The peak time of mating occurred at the 7th and 10th hour of the scotophase, and there was no new mating after the photophase. The testicular volume of male adults decreased as the day-old age increased, and based on the testicular volume of male adults caught by sex pheromone trapping, it was calculated that the age of those trapped males was 3-7-day-old. 【Conclusion】The calling and mating time of T. absoluta adults occurred at the end period of the scotophase. The sex pheromone titer of female adults remained at a consistently high level at different day-old ages and circadian rhythms, and mating did not significantly affect sex pheromone titer, resulting in older male adults being caught by pheromone trapping, and their testes were the same size as those of the 7-day-old male adults, and most of them may have already mated. The data here give a more clear description of the calling and mating behavior of T. absoluta adults, thus, they can provide technical bases and parameters for the development and application of mass trapping and mating disruption by sex pheromones.

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Analysis of expression patterns of genes involved in pigment pathways in the tergum of the red- and black-backed Apis cerana cerana adult workers

WANG Ruo-Hong, YANG Zhen-Hui, ZHOU Shi-Wen, WU Yu-Jia, LI Qiu-Fang, LIANG Li-Qiang, SHI Dan-Dan, YANG Shang-Ning, MIAO Liu-Chang, SU Song-Kun, NIE Hong-Yi

Acta Entomologica Sinica 2024, 67 (1): 9-17. DOI: 10.16380/j.kcxb.2024.01.002

Abstract （145）

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【Aim】To analyze the differential expression of genes involved in the pigment pathway of red-backed Apis cerana cerana and reveal the molecular mechanism of pigment formation of red-backed A. cerana cerana. 【Methods】 The differences in the adult worker body color between the newly emerged red- and black-backed (normal individuals) A. cerana cerana were observed using stereomicroscope. Homologous genes of 8 genes related to melanin metabolism pathway (PAH, TH, DDC, ebony, tan, aaNAT, yellow-y and laccase 2), 4 genes related to pterin pathway (GTPCH I, SPR, PTPS and GC-1), 2 genes related to ommochrome pigment pathway (vermilion and cinnabar), and 4 genes related to urate transport protein (BLOS2, HPS5, OK and Varp) were identified in adult A. cerana cerana workers via BLAST. The relative expression levels of the above genes involved in pigment pathways in the thoracic tergum and abdominal integument of the red- and black-backed A. cerana cerana adult workers were detected by fluorescence quantitative PCR. 【Results】 The color difference in the adult worker body color between the red- and black-backed A. cerana cerana was found on thoracic tergum. The thoracic tergum of the red-backed A. cerana cerana is in brownish red, while that of the black-backed A. cerana cerana is in black. The fluorescence quantitative PCR results showed that the expression levels of tan, laccase 2, SPR, vermilion, cinnabar, BLOS2 and OK and that of OK in the thoracic tergum and abdominal integument had significant difference, respectively, between the red-backed adult A. cerana cerana workers and black-backed adult A. cerana cerana workers. 【Conclusion】 Red-backed A. cerana cerana and black-backed A. cerana cerana have obvious body color difference on the thoracic tergum. This phenomenon of body color differentiation is influenced by the combined effects of genes related to melanin, pterin and ommochrome pigment pathways, and urate transport in honeybee.

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Sequencing and analysis of the complete mitochondrial genome of Eurytoma acutibialis (Hymenoptera: Eurytomidae), an important natural enemy insect of Bradybatus sp. (Coleoptera: Curculionidae) in China

LIU Hui-Hui, LI En-Jie, CAO Liang-Ming, BAO Qing-Chun, WANG Xiao-Lin, XIN Xue-Bing, YANG Zhong-Qi

Acta Entomologica Sinica 2023, 66 (9): 1233-1245. DOI: 10.16380/j.kcxb.2023.09.010

Abstract （141）

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【Aim】 Eurytoma acutibialis Yang, Liu et Cao is a parasitic wasp of Bradybatus sp., a serious seed-eating pest of Acer truncatum in China, and is a new species found during the investigation of Bradybatus sp. and can play an important role in controlling Bradybatus sp. To further understand the new parasitoid, the complete mitochondrial genome of E. acutibialis was sequenced and analyzed so as to provide molecular biological evidence for the systematic taxonomy and phylogenetic relationship of hymenopterous wasps (Hymenoptera: Chalcidoidea: Eurytomidae).【Methods】 The complete mitochondrial genome of E. acutibialis was sequenced on Illumina HiSeq ^TM 4000 sequencing platform and then assembled, annotated and analyzed. Based on the nucleotide sequences of 13 protein-coding genes (PCGs) from 40 species of hymenopteran insects in the Chalcidoidea, the phylogenetic tree was constructed by Bayesian inference (BI) and maximum likelihood (ML) methods, and the phylogenetic relationship between E. acutibialis and other insects in the Chalcidoidea was analyzed.【Results】The mitochondrial genome of E. acutibialis is 15 849 bp in full-length, with the A+T content of 82.00%, and consists of 37 genes including 13 PCGs, 2 rRNA genes, 22 tRNA genes, and a control region (CR). Among the 13 PCGs, the amino acid usage frequencies from high to low are Leu，Ile, Met， Phe and Asn. All the 13 PCGs initiate with the standard start codons of ATN (ATT, ATG and ATA) and terminate with the typical stop codons TAA/TAG. Among the 22 tRNA genes, trnI lacks receptor arm, trnF lacks TΨC-arm, trnS1 lacks DHU arm and DHU arm of trnS2 forms a simple loop. The remaining tRNA genes have a typical cloverleaf structure. There is a significant gene rearrangement, with an inversion occurring in the nad2-nad1-cytb-nad6-nad4L-nad4-nad5 cluster and rearrangements of 16 tRNA genes. The phylogenetic analysis highly supported the monophyly of the subfamily Eurytomidae, and E. acutibialis was the closest to Eurytoma sp. ZJUH 2016013.【Conclusion】The complete mitochondrial genome sequence of E. acutibialis was firstly obtained, and the phylogenetic analysis based on the mitochondrial genome supported its taxonomic position in the genus Eurytoma in the family Eurytomidae in the order Hymenoptera and proved to be consistent with the morphological taxonomy.

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Analysis of the function and upstream regulatory sequence activity of the male determining factor gene MoY in Zeugodacus cucurbitae (Diptera: Tephritidae)

FU Jun-Kai, WEN Jian, CAO Feng-Qin, YAN Ri-Hui, LIN Xian-Wu

Acta Entomologica Sinica 2024, 67 (3): 339-345. DOI: 10.16380/j.kcxb.2024.03.004

Abstract （138）

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【Aim】The aim of this study is to identify the upstream regulatory sequence of the male determining factor gene MoY of the melon fly, Zeugodacus cucurbitae， and to explore the impact of this sequence on the sex determination of Z. cucurbitae by directing the expression of MoY in the early embryonic stages, so as to provide a reference basis and available elements for the construction of the subsequent Z. cucurbitae transgenic strains.【Methods】We amplified and sequenced the upstream sequence of MoY linked to the Y chromosome of Z. cucurbitae using the genome-walking, and connected the obtained upstream sequence to the CDS region of MoY to construct a plasmid that drives the expression of MoY. We injected the plasmid expressing MoY into the fresh embryos of Z. cucurbitae, and after hatching, extracted the genomic DNA based on the observed adult phenotype and sex ratio to amplify MoY so as to determine whether the upstream regulatory sequence of MoY had the activity of directing MoY expression and analyze the impact of MoY expression in embryos on the gender determination. 【Results】The upstream sequence of 1 660 bp of MoY of Z. cucurbitae was cloned and obtained, and the plasmid p1660 that drives the MoY expression was constructed. Eighteen male and 13 female adults developed from the injected embryos of Z. cucurbitae, and three adults were found to have abnormal genitalia, with negative amplification results for MoY, confirming them as intersex individuals. 【Conclusion】In this study, we found that the upstream regulatory sequence of MoY of Z. cucurbitae obtained by genome-walking amplification has the activity of directing the expression of MoY. When MoY is expressed in the early stages of Z. cucurbitae embryos, the gender reversal in the previously developed female individuals can be happened.

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Research progress of the insect secretion-mediated interaction between insects and plants

CAI Xiang-Yun, WANG Ya-Ru, YAO Yang, WANG Jin-Da, HOU You-Ming

Acta Entomologica Sinica 2024, 67 (2): 284-306. DOI: 10.16380/j.kcxb.2024.02.013

Abstract （137）

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Insects and plants are important components of biological community in nature and have evolved complex interactions during the long-term evolution process. For example, plants have evolved complex defense strategies to resist insect damage, meanwhile, insects have also evolved more adaptive ways to get more resources for survival. Therefore, studies that focus on the mutual adaptation between insects and host plants are important for effective pest control and cultivation of insect-resistant plants. In the study of the interactions between insects and host plants, the effective components of the oral secretions from herbivorous insects such as saliva or regurgitant liquid, egg secretions, intestinal compounds and microorganisms as the intermediate media to connect insects and plants not only play important roles in inducing or enhancing plant defense, but also have some functions of inhibiting or weakening plant defense. Therefore, according to the role played by the effective components, the oral secretions from herbivorous insects can be divided into elicitors and effectors. In this review article, we introduced the main types of insect effectors including calciumbinding proteins, enzymes and venom proteins, and elicitors including fatty acid amino acid conjugates, polypeptides and enzymes, and the main ways of mediating plant defense from the perspective of insect secretion, clarified the different strategies of herbivorous insects adapting to plant defense, which not only contribute to the in-depth analysis of the interaction mechanism between insects and plants, but also provide new ideas for green and effective pest control and breeding of plant resistant varieties.

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