【Aim】 In this study, we aim to identify the function of phenoloxidase in the immune response in Blattella germanica induced by Escherichia coli. 【Methods】 A full-length cDNA of B. germanica phenoleoxidase gene (BgPO) was cloned using homology-based cloning and rapid amplification of cDNA ends (RACE). The phylogenetic tree of BgPO and PO proteins from other insects was constructed by the software MEGA 5.1. The tissue expression patterns of BgPO and its transcription levels in hemocytes at different time points post E. coli infection were detected by RT-PCR. The antibacterial activity of hemolymph was measured by Hultmark’s method and the BgPO enzyme activity was determined by catechol method. 【Results】The full-length cDNA sequence of BgPO is 2 252 bp (GenBank accession number: KJ789157), which contains an ORF of 2 085 bp encoding 695 amino acid- residues. The deduced protein has a calculated molecular weight of 79.7 kDa and the theoretical pI of 6.19. The sequence alignment analysis showed that BgPO has high sequence identities with the homologies of other insect species, and shares an 80% amino acid sequence identity with the PO from Coptotermes formosanus. Phylogenetic tree analysis indicated that BgPO has the closest genetic relationship with the PO of C. formosanus. Gene expression profiling showed that BgPO was expressed mainly in the hemocytes and epidermis. The transcription level of BgPO mRNA increased from 24 h post E. coli infection, and reached a peak at 36 h p.i. The antibacterial activity and phenoloxidase activity increased after bacterial induction during 6-36 h p.i. The antibacterial activity showed significant differences 〖JP4〗between the bacteria-infected and the PBS-induced samples during 6-48 h p.i. (P<0.05), and the phenoloxidase activity also showed significant differences between the bacteria-induced and the PBS-induced samples during 24-36 h p.i. (P<0.05). 【Conclusion】Our findings revealed that BgPO is mainly expressed in the hemocytes and epidermis. BgPO may participate in the immune response against E. coli invasion. The results provide a good foundation for the further research on immune response of BgPO in B. germanica.

【Aim】 The fitness of Helicoverpa armigera is dependent on different pepper (Capsicum annuum L.) varieties. This study aims to explore the effect of different pepper varieties on the expression of salivary gland proteins and the corresponding genes in the 5th instar larvae of H. armigera.【Methods】 We used twodimensional polyacrylamide gel electrophoresis (2-DE) and real-time quantitative PCR (qPCR) to examine the changes in the expression profiles of salivary gland proteins and the corresponding genes in the 5th instar larvae of H. armigera feeding on artificial diets containing fruits of commercial pepper varieties with different levels of capsaicinoids, respectively.【Results】 The results indicated that the expression of proteins in the salivary glands of the 5th instar larvae of H. armigera was strongly influenced by the different pepper varieties, significant difference in the expression levels of 28 proteins was found among different treatments through quantitative analysis (P<0.05) and 15 differentially expressed proteins were successfully determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF MS). Real-time PCR analysis manifested that the higher level of capsaicin promoted the expression of β-fructofuranosidase, and vice versa. Continuous exposure to four pepper treatments inhibited the expression of hsp70 all the time and inhibited the expression of eIF5A to a certain extent. 【Conclusion】 The functional studies of β-fructofuranosidase, hsp70 and  eIF5A have been enriched and widened. The results revealed the adaptive mechanism of H. armigera in different pepper varieties, providing a sound basis for selection of pepper varieties resistant to  insects.

【Aim】 To select the suitable reference genes in Phenacoccus solenopsis after exposure to different temperature stress by using quantitative real-time PCR (RT-qPCR). 【Methods】 In this study, we investigated the mRNA expression stability of six candidate reference genes (α-tub, GAPDH, rpl32, β-tub, SDHA and TBP) in different developmental stages (2nd instar nymph, 3rd instar nymph and female adult) of P. solenopsis under seven different temperature regimes (constant temperature 18℃ and 32℃, and temporary heat shock (1 h) at 37℃, 39℃, 41℃, 43℃ and 45℃ and then recovery at 26℃ for 1 h) by RT-qPCR method. Three software-based approaches (geNorm, Bestkeeper and Normfinder) and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes, and evaluate the expression stability of these genes. 【Results】 In the 2nd instar nymphs, the M values of six reference genes stability in the ascending order were α-tub (0.579)<GAPDH (0.654)<TBP (0.663)<β-tub (0.668)<rpl32 (0.675)<SDHA (0.755), and the SD values were TBP (0.31)<α-tub (0.37)< rpl32 (0.38)<SDHA (0.52)<β-tub (0.53)<GAPDH (0.57). In the 3rd instar nymphs, the M values in the ascending order were α-tub (0.542)<β-tub (0.596)<TBP (0.618)< GAPDH (0.655)<SDHA (0.668)<rpl32(0.749), while the SD values were α-tub (0.29)<β-tub (0.40)<rpl32 (0.45)<TBP (0.49)<GAPDH (0.50)<SDHA (0.55). The M values in female adults in the ascending order were rpl32 (0.554)<α-tub (0.635)<GAPDH (0.662)<β-tub (0.666)<SDHA (0.704)<TBP (0.837), and the SD values were rpl32 (0.31)<α-tub (0.35)<GAPDH (0.44)<SDHA (0.44)<β-tub (0.48)<TBP (0.71). According to the comprehensive rank of RefFinder, α-tub was the most stable for the 2nd and 3rd instar nymph, while rpl32 for female adult under seven different temperature regimes. 【Conclusion】α-tub can be used as a reference gene for the 2nd and 3rd instar nymph, and rpl32 could be used as the reference gene for female adult. This study provides an accurate reference for future study of the heat shock protein gene expression in P. solenopsis under heat stress.

【Aim】Recent warmer climate caused persistent outbreaks of Siberian grasshopper, Gomphocerus sibiricus (L.), in Xinjiang. This study aims to explore the potential physiological and biochemical mechanisms of G. sibiricus exposed to high temperature. 【Methods】The contents of trehalose, glycerine, oleic acid, linoleic acid, and linolenic acid accumulated in adult grasshopper bodies were measured after these locusts had been placed in climate cabinet under consistent temperature regimes of 24-42℃ at 3℃ intervals for 4 h. 【Results】The contents of trehalose, glycerine, oleic acid, linoleic acid, and linolenic acid in G. sibiricus increased with temperature ranging from 24 to 30℃, and reached the peak at 30℃, with the contents of 18.691 μg/g, 261.432 μg/g, 79.063 mg/g, 78.664 and 227.593 mg/g, respectively. The contents of trehalose, glycerine, oleic acid, linoleic acid, and linolenic acid in G. sibiricus began to decrease as the temperature rose further, and reached the lowest at 42℃, with the contents of 18.218 μg/g, 104.588 μg/g, 4.343 mg/g, 3.093 mg/g, and 11.067 mg/g, respectively. The accumulation rates of these stress resistant substances were different with temperature increasing. The content of linolenic acid increased dramatically (832.189%) while that of trehalose accumulated increased slightly (0.893%). The content of linoleic acid had a significant decrease (63.988%) while that of trehalose reduced slightly (0.224%). 【Conclusion】The higher the temperature above 30℃, the higher the locust mortality. This study demonstrated that with the temperature increasing in a certain range, G. sibiricus can adapt to higher temperature conditions by regulating the contents of stress resistant substances, especially the unsaturated fatty acids, and thus remains a serious pest to the steppe in Xinjiang under the trend of climate warming.

【Aim】The distinctive arrow-shaped morphology and large size of the spermatozoa of Gampsocleis gratiosa makes it a useful model to explore spermiogenesis in Tettigoniidae. In order to better understand the mechanism of spermiogenesis, especially the mechanism of nucleus morphogenesis and arrow-shaped acrosome formation in katydids, the nuclei of spermatid and spermatozoa of G. gratiosa were observed in the present study. 【Methods】 The testes of G. gratiosa in sexually mature males were used as the materials. We observed the nuclei of spermatid and spermatozoa by transmission electron microscopy, ordinary optical microscopy, fluorescence microscopy and with H&E and DAPI (DNA specific probe) staining method. 【Results】 According to the morphology and structure, the nuclei during spermiogenesis of G. gratiosa were divided into four phases: rounded, leaf, prismatic and mature phases. During the spermiogenesis of G. gratiosa, there are two globular structures at the rounded phase, one is the nucleus and the other is pro-acrosome. The cytoplasm droplet, which would be derelict along the tail of spermatozoa at the mature phase, contains DNA. 【Conclusion】 The observation revealed that spermatid nucleus of G. gratiosa undergoes dramatic morphological changes during its spermiogenesis. The nucleus shaping is driven by cytoskeletal microtubules, and the chromatin reorganization is together with nucleus shaping. This study will be the foundation to clarify the molecular mechanism of spermiogenesis in Orthoptera.

【Aim】 The western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an important invasive insect pest. It is also the most effective vector for tomato spotted wilt virus (TSWV) which has a certain influence on the growth and development of F. occidentalis. Spinosad is one of the most effective insecticides in F. occidentalis control. However, it has been reported that F. occidentalis has developed resistance to spinosad in the field. Whether TSWV has an impact on the resistant strain and how high the impact is are not clear. This study would provide a basis for further understanding of the impact of TSWV on F. occidentalis.【Methods】 Age-stage and two-sex life tables were used to study the biological characteristics of the spinosad-resistant and susceptible strains of F. occidentalis with TSWV treated or nontreated. The susceptibilities of the spinosad-resistant and susceptible strains of F. occidentalis to three insecticides (spinosad, emamectin benzoate and chlorfenapyr) were investigated by using leaf-tube residue method. 【Results】 For the resistant strain of F. occidentalis treated by TSWV, the developmental period shortened, the female longevity and fecundity increased, but they showed no significant difference from those of the non-treated thrips (P>0.05). The intrinsic rate of increase (r, 0.0433 d^-1) and the net reproduction rate (R₀, 2.210) of the resistant strain treated by TSWV were significantly higher than those in the control group (0.0356 d^-1 and 1.972, respectively) (P≤0.001). As for the susceptible strain treated by TSWV, the developmental period shortened, the adult longevity increased significantly (P≤0.001), and the fecundity increased as well. The R₀ of TSWV-treated susceptible strain (4.125 ) was significantly higher than that in the control group (3.979 ) (P≤ 0.001). The susceptibilities of the spinosad-resistant and susceptible strains of F. occidentalis treated by TSWV to spinosad had no obvious change compared with those of the corresponding nontreated thrips. However, their susceptibilities to emamectin benzoate and chlorfenapyr decreased significantly. 【Conclusion】 TSWV has directly positive effect on the biological characteristics of both the spinosadresistant and susceptible strains of F. occidentalis. The TSWV treated thrips have shortened developmental period, enhanced fecundity and longer adult longevity, and decreased susceptibility to insecticides.

【Aim】Although polyandry is common among Coccinellidae, the data attained in various studies are insufficient to explain the general adaptive significance (or fitness consequences) of female remating and polyandry. Temperature was used as a stressor in the study. This study aims to evaluate whether certain benefits of polyandry in terms of increased fitness are also passed on to the progeny. 【Methods】The present study examined the fitness consequences of three mating treatments in Coelophora saucia (Mulsant) (Coleoptera: Coccinellidae): monandry (five matings with same male; one mating/day), sequential polyandry (five matings with five different males with one mating/day with a new male), and simultaneous polyandry (five matings with five males introduced together allowing female to select male to mate; one mating/day). Changes in fecundity and egg fertility, and offspring development and survival of the different mating treatments under different temperatures (25, 27 and 30℃) were observed. 【Results】The results revealed that females subjected to simultaneous polyandry and thus allowed mate choice or competition amongst males had maximum reproductive performance and offspring best suited to developing and surviving at a wider temperature range. However, sequential polyandrous females had similar reproductive performance as the monandrous female. 【Conclusion】This indicates that in the absence of mate choice or male competition conditions, benefits of polyandry are not evident. This could be either due to sperm competition amongst subsequent male ejaculates or female cryptic choice. The lack of benefits of polyandry in absence of mate choice as observed in this study has not been previously observed in insects to the best of our knowledge.

【Aim】 The tobacco aphid Myzus persicae (Sulzer) is an important pest found on cultivated crops across the world. This study aims to evaluate the effects of brief exposure to high temperatures on the development, reproduction, and feeding behavior of M. persicae.【Methods】Nymphs and adults of M. persicae were exposed to high temperatures of 30, 35 and 40℃ for 1, 2 and 4 h every day, and the developmental duration and fecundity were measured. The electronic penetration graph (EPG) technique was used to record the feeding behavior after the adults were exposed to high temperatures of 30℃ and 35℃ for 1 and 2 h as well as at constant temperatures of 25, 28, 30 and 35℃. 【Results】 Brief exposure to high temperatures of 30-35℃ had little effect on the nymphal duration. However, the developmental duration was significantly prolonged after the nymphs were exposed to a high temperature of 40℃ for 2 h (P<0.01). The decrease in the survival rate of the nymphs, adult longevity and duration of reproduction of adults was proportional to both temperature and exposure duration. The reproduction rate was little influenced at 25-35℃ with the increase of temperature and duration of exposure, while significantly decreased at the high temperature of 40℃ for 2 and 4 h (P<0.01). Brief exposure to high temperatures had a significant effect on feeding behavior. After brief exposure to high temperatures, the number (P<0.05) and total duration (P<0.01) of nonprobing waveform (np) significantly decreased while the total duration of phloem waveform (E) significantly increased (P<0.05) with an increase in temperature and exposure duration. However, the number and total duration of np decreased initially and then increased, while that of E initially increased and then decreased with an increase in temperature under constant temperature conditions【Conclusion】Brief exposure to high temperatures has no significant effect on the developmental rate of M. persicae nymphs. However, the survival rate of nymphs, adult longevity and duration of reproduction of adults decreased after brief exposure to high temperatures. In addition, brief exposure to high temperature plays a role in inhibiting the reproduction of M. persicae adults, and promoting their feeding behavior. These results will aid in understanding the mechanisms underlying changes in M. persicae populations in summer between different years.

【Aim】This study aims to understand the influence of alternating temperature on the growth and development of experimental populations of Athetis lepigone. 【Methods】We measured and calculated the developmental threshold temperature, effective accumulated temperature and life table parameters to study the influence of different combinations of alternating temperatures (20/24℃, 20/28℃, 20/32℃, 24/28℃, 24/32℃ and 28/32℃) on the mean developmental duration, survival rate and fecundity of A. lepigone.【Results】The results showed that the duration of different developmental stages of A. lepigone shortened as the average temperature in each alternating temperature combination increased. The generation survival rate of A. lepigone decreased in the order of 24/28℃>20/32℃>28/32℃>20/28℃>24/32℃>20/24℃. The fecundity was the highest at 24/28℃, with 390.17 eggs laid per female. The developmental threshold temperature and effective accumulated temperature of the whole generation was 10.37℃ and 663.17 degree-days, respectively. The intrinsic rate of natural increase （r_m） initially increased and then decreased as the average temperature of each alternating temperature combination increased. At 24/28℃, the intrinsic rate of natural increase (r_m), net reproductive rate (R₀), the finite rate of increase (λ) were the highest, while the doubling time (t) and generation time (T) were the minimum.【Conclusion】The results suggest that the temperature combination 24/28℃ is the optimal combination of alternating temperatures for the development of experimental populations of A. lepigone. These results provide a scientific basis for the prediction and integrated manangement of A. lepigone.

【Aim】 In order to study what factors impact the diapauses of Microplitis tuberculifer, the effect of temperature and photoperiod on diapause induction and storage condition of diapause cocoons of M. tuberculifer were studied in the field during the late summer and in the laboratory. 【Methods】 The diapause rates of M. tuberculifer were investigated in the field cages when the parasitized host larvae were released into the cages every 5 days from August 31 to September 25 and in the laboratory under the conditions of five temperatures （16, 18, 20, 22 and 24℃）and seven photoperiods (6L:18D, 8L:16D, 10L:14D, 12L:12D, 14L:10D, 16L:8D and 18L:6D). 【Results】 The field tests in the central of Hebei Province showed that when the average daily temperature was 24.4℃ and the average daylength was 12 hours and 51 minutes, only a few of M. tuberculifer pupae (5.08%) entered diapause; and when the average daily temperature was below 17.2℃ and the average daylength was shorter than 11 hours and 36 minutes, almost all of M. tuberculifer pupae (99.70%) entered diapause. No diapause was observed when the temperature was > 22℃ regardless of the photoperiod. Only a small part of population (6.0%-25.9%) entered diapause at 20℃ under short daylengths of 6L:18D to 12L:12D. In contrast, when the larvae were exposed to 16, 18 and 20℃ combined with a photoperiod of 10L:14D, the percentages of parasitoids that entered pupal diapause were 100%, 89.75% and 29.58%, respectively. The emergence rate and the parasitic ability showed no significant difference between adults emerged from the diapause cocoons stored at 0℃ for 240 d and 5℃ for 180 d and those from the nondiapause cocoons without cold storage (P>0.05). After cold storage at 0℃ for 300 d, the emergence rate of the diapause cocoons could still reach to 79%. 【Conclusion】 Temperature and photoperiod, or their interaction, are the most common diapauseinducing stimuli of the parasitoid. The results of this study would be valuable for mass rearing and storage of diapause cocoons of M. tuberculifer.

【Aim】Superparasitism often occurs in parasitoids. In this study, we aimed to investigate the effects of superparasitism on the offspring growth and development of Cotesia ruficrus and to determine the factors that contribute to superparasitism. 【Methods】 The effects of oviposition times of C. ruficrus on the survival rate of the host （the 3rd instar larvae of Cnaphalocrocis medinalis） and the growth and development of the parasitoid offsprings, and the effects of exposure time and parasitoid density on superparasitism were detected in this study. 【Results】Laboratory experiments demonstrated that the superparasitism occurred in C. ruficrus when it parasitized C. medinalis larvae. Female wasps, irrespective of the experience of oviposition, attacked hosts that had been parasitized by themselves or other individuals. The number of parasitoid cocoons increased with oviposition times, while the number of the dead parasitoid larvae increased when the host was parasitized 3 to 5 times. The mortality of the host before emergence of parasitoid adults increased with the oviposition times, and reached to 50% when the host was parasitized 5 times. Superparasitism prolonged the developmental duration, lowered the emergence rate and the female/male ratio of the parasitoid offsprings. The female body size of the parasitoid offsprings significantly decreased as the oviposition times increased, and the superparasitism rate increased with the parasitoid density and exposure time. 【Conclusion】Ovipositing twice on host larvae shows to be the most beneficial to the growth and development of C. ruficrus offsprings, and ovipositing thrice causes superparasitism. Although superparasitism increases the number of parasitoid offsprings, it is detrimental to the progeny development. The superparasitism, therefore, should be avoided in mass rearing of C.ruficrus in laboratory, by means of controlling exposure time and density of the parasitoids.

【Aim】This study aims to illustrate the effects of using frozen-preserved housefly (Musca domestica) pupae as hosts on the reproduction of Spalangia endius.【Methods】The survival number, offspring number and sexes of S. endius on fresh housefly pupae and the housefly pupae frozen killed and preserved at -20℃, respectively, were recorded daily, and the life tables were built. The effects of host types on the parasitism rate, offspring number and sex ratio, and parameters of life table were compared and analyzed. 【Results】The results showed that the parasitism rate, offspring number, and male proportion of S.endius on the frozen-killed housefly pupae were lower than those of wasps on the fresh housefly pupae (P<0.01). The longevity and oviposition duration of adults were similar between the two groups (P>0.05). The parasitism rate and offspring number in both groups decreased with the increasing of female age (P<0.01). The net reproductive rate (R₀) of S. endius on the fresh housefly pupae and frozen-killed pupae was 34.91 and 20.16, and their intrinsic natural increase (r_m) was 0.17 and 0.11, respectively. R₀ and r_m were both larger on the fresh housefly pupae than on frozen-killed housefly pupae (P<0.01). The mean generation time (T) and doubling time (DT) of S. endius on the frozen-killed housefly pupae were prolonged compared with those of wasps on the fresh pupae (P<0.01). 【Conclusion】Our results indicate that S. endius can use frozen-killed housefly pupae as hosts to finish life history. We believe that the method of frozen-killed hosts would be useful for massively culturing of S. endius.

【Aim】In order to provide the important scientific basis for building effective prevention and control system of aquatic leaf beetle Donacia provostii Fairmaire, the diurnal rhythm of adult behaviors were studied. 【Methods】The adults were observed for 5 consecutive days and recorded in a 30 min interval by single rearing separately and rearing in pairs outside, and data was collected and analyzed. All behaviors were divided into six types: resting without contacting water (ROCW), resting with contacting water (RWCW), foraging (FG), moving (MG), mating (MT) and spawning (SG). 【Results】These results showed the adult behaviors of D. provostii were significantly different at various time points or in various time periods (P<0.001). The proportions of four behaviors were significantly different in different spatial positions (P<0.001), and the behaviors on leaf of Nymphoides peltatum (Gmel.) Kuntze or water surface were also significantly different at different time points (P<0.001). 【Conclusion】Both temporal and spatial variation could significantly influence the behaviors of the waterleaf beetle, and the effect of spatial variation is more significant than that of the temporal variation.

【Aim】The aim of the study is to test the validity of the VspI-, StyI- or StuI-based cleaved amplified polymorphic sequence (CAPS) markers of mitochondrial cytochrome oxidase I gene (mtCOI) for identification of Bemisia tabaci cryptic species Q and B. 【Methods】In this study, the mtCOI fragments from B. tabaci from China and abroad were sequenced and used to identify cryptic species. Based on the analyses of mtCOI sequences of 464 and 98 individuals from cryptic species Q and B, respectively, the restriction endonuclease enzyme VspI-, StyI-, or StuI-based CAPS markers was used to differentiate cryptic species Q and B for assessing its utility. The mtCOI sequences of B. tabaci cryptic species Q and B in GenBank were downloaded and the distribution of the recognition sites of VspI-,StyI-, and StuI- was analyzed. 【Results】The results showed that the individuals from China and abroad could be differentiated by using VspI-based CAPS markers effectively, but not by StyI- or StuI-based CAPS markers. The mtCOI sequence analysis showed that VspI-, StyI- or StuI-based CAPS markers could not differentiate cryptic species Q and B. 【Conclusion】VspI-, StyI- or StuI-based CAPS markers has limitations in identifying cryptic species Q from B.

【Aim】In order to explain the different pathogenicity of Lecanicillium lecanii to adults, pupae and larvae of Boettcherisca peregrina, the adhesion capacity of L. lecanii conidia on body surface of B. peregrina at the three developmental stages and its relationship with the surface structures of insect body were analyzed. 【Methods】The number of conidia adhered to the surface of adults, pupae and larvae of B. peregrina were measured by hemocytometer, and the distribution of outgrowths and conidia on the body surface were observed by scanning electron microscope (SEM) and other microscopies.【Results】 There was significant difference in the adhesion number of L. lecanii conidia on the surface of three developmental stages of B. peregrina (P=0.0001). The adhesion number of L. lecanii conidia was (1.35±0.12)×10⁶, (0.57±0.09)×10⁶ and (0.45±0.06)×10⁶ on each adult, pupa and larva, respectively. Many L. lecanii conidia adhered to microchaetae of adults. Only few L. lecanii conidia were found to adhere to "rope" folds on the bottom of the thorns in pupae. No adhered L. lecanii conidia on larvae were found. 【Conclusion】 The adhesion capacity of L. lecanii conidia is closely associated with the body surface structures of adults, pupae and larvae of B. peregrina, resulting in the different pathogenicity of L. lecanii  to these different developmental stages of B. peregrina.