【Aim】 This study aims to explore the cold tolerance strategies and overwintering stage of Athetis dissimilis (Hampson). 【Methods】 The supercooling point (SCP), freezing point (FP), and the contents of water, fat, protein, glycerin, sorbitol and trehalose in the 3rd, 4th, 5th, 6th instar and mature larvae, and pupae of A. dissimilis were measured in the laboratory. 【Results】 The SCP and FP had obvious differences among different instar larvae of A. dissimilis, and the SCP and FP of young larvae were higher than those of old larvae. The SCP and FP of mature larvae dropped to the lowest (-18.20℃ and -11.72℃, respectively). There were significant differences in the contents of biochemical substances between different instar larvae. The water and glycerin contents declined with the increase of developmental stage. The lowest water and glycerin contents were found in pupae (65.33% and 154.90 μg/mg, respectively). There was a trend of increase of fat and protein contents with the increase of developmental stage. The fat and protein contents in pupae (25.36% and 2 298.37 μg/mg, respectively) were higher than those in larval stages. The trehalose content in mature larvae was relatively high (57.28 mg/g), while the sorbitol content was relatively low (15.49 mg/g). 【Conclusion】 The strongest two cold-tolerant stages of A. dissimilis were mature larva and pupa, which may be more suitable for overwintering than other developmental stages. Fat, protein and trehalose may be the important cold-resistant substances in A. dissimilis.

【Aim】 To clarify the physiological mechanisms of the red and green color morphs of the pea aphid, Acyrthosiphon pisum (Harris), in response to elevated CO₂ concentration. 【Methods】 Two elevated levels of CO₂ concentration (moderate: 550±27 μL/L; doubling: 750±37 μL/L) were manipulated in incubators with ambient CO₂ (375±18 μL/L) as the control. Vicia faba seedlings were planted in the above incubators, nymphs of the pea aphid within 6 h were introduced later, and molting and the survival in various developmental stages were recorded until the adult. Nutrient contents and digestive enzyme activities of two color (red and green) morphs of the pea aphid in three successive generations and nutrient contents in V. faba leaves were assayed. 【Results】 Elevated CO₂ concentration significantly enhanced the contents of soluble protein, soluble carbohydrates, free amino acids and starch in V. faba leaves. Under the moderate CO₂ concentration (550±27 μL/L) condition, the nutrient contents in F₀ generation of the two color morphs of the pea aphid had no significant difference as compared with the control, while the contents of protein, total amino acids, soluble carbohydrates and glycogen in F₂ generation all increased and the contents of total lipid and free fatty acids decreased significantly, and moreover the content of soluble carbohydrates in F₂ generation of green pea aphid was significantly higher than that in red pea aphid. Under the doubling CO₂ concentration (750±37 μL/L) condition, the contents of protein, total amino acids, soluble carbohydrates and glycogen in three successive generations of the two color morphs all increased significantly and the contents of total lipid and free fatty acids decreased significantly as compared with the control, and moreover the contents of protein, total amino acids, and glycogen in the F₁ and F₂ generations of green pea aphid were significantly higher than those in red pea aphid. Under the elevated CO₂ concentration (550±27 and 750±37 μL/L) conditions, the digestive enzyme activities in F₀ generation of two color morphs of the pea aphid had no significant difference as compared with the control, while the pepsin and trypsin activities in F₂ generation decreased significantly and those of lipase and amylase increased significantly, and moreover the digestive enzyme activities had no significant difference between the two color morphs.【Conclusion】 Under the elevated CO₂ concentration, the change in nutrient composition in broad bean leaf causes the change in nutrient composition and digestive enzyme activities in pea aphids feeding the disposed V. faba leaves. Additionally, the changes in contents of protein, total amino acids, soluble carbohydrates and glycogen in the green morph of the pea aphid are more significant than those in the red morph. These results provide some references for controlling the pea aphid, the main pest in leguminous plants, under the elevated atmospheric CO₂ concentration condition in the future.

【Aim】 To explore the mechanisms of the NaF injury to the gonad of the lepidopteran model insect-the silkworm, Bombyx mori, we studied the effects of NaF on the testis and ovarian tissue structure, and the antioxidant enzyme activities and their gene expression in the silkworm. 【Methods】 B. mori larvae were fed with mulberry leaves soaked in different concentrations of NaF solution (0, 25, 50, 100 and 200 mg/L). The malondialdehyde (MDA) content, and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) and their corresponding mRNA levels in the gonad of the 5th instar larvae of both sexes were evaluated, and the change in the anatomical structure of gonad was observed by paraffin section and Hematoxylin-Eosin (H&E) staining technique. 【Results】 The MDA contents in male and female larvae fed with mulberry leaves soaked in different concentrations of NaF solution significantly changed in a concentration-dependent manner, increasing with the NaF concentration, as compared with those in the control group. The activities of both SOD and CAT increased in B. mori larvae fed with mulberry leaves soaked in low dose of NaF, but successively decreased after high dose of NaF was added; the activities in the treatment groups, however, were still higher than those in the control group. According to the analysis of Pearson correlation, the SOD activity was positively correlated with the CAT activity (male: R=0.865, P=0.000; female: R=0.766, P=0.001), and the CAT activity had an obvious correlation with its mRNA level. The GSH-Px activities in gonads of male and female larvae changed in a concentration-dependent manner, increasing with the NaF exposure concentration, and the changes in GSH-Px activity showed a positive correlation with its mRNA level. The changes in the antioxidant enzyme activities in male larvae were more responsive than those in female larvae when they were fed with mulberry leaves soaked in NaF solution. Changes in the structure of gonads of male and female larvae were observed by using H&E staining. Such phenomena as abnormal morphology, enlarged cell vacuoles and relative less reproductive cells were observed. With the increase of NaF concentration, gonads were more seriously damaged. 【Conclusion】 NaF in mulberry leaves fed by B. mori larvae not only causes changes in gonadal structure of its male and female larvae, but also results in changes in the antioxidant enzyme activities and their mRNA transcriptional level in larval gonads.

【Aim】 The process of the growth and differentiation of wing discs of the silkworm, Bombyx mori is accompanied with the disappearance of such tissues as hematopoietic organs and wing bursa. The study aims to analyze whether apoptosis and autophagy occur during the growth and differentiation of wing discs in the silkworm. 【Methods】 Apoptosis in wing discs of B. mori at the pupal stage was analyzed by TUNEL. The total RNA of wing discs at day-3 and day-6 5th instar larval stage, day-1 wandering stage, pre-pupal stage, and day-0 and day-3 pupal stage were extracted and then reversely transcribed into cDNA. The expression patterns of apoptosis and autophagy related genes were detected by qRT-PCR. The activities of Caspase 3 and acid phosphatase were analyzed. 【Results】 It was observed that apoptosis occurs in the partial wing discs cells at day-0 pupal stage. The apoptosis related genes Caspase 3 and Nc had higher expression levels during the 5th instar larval stage, while IAP had higher expression level at the late larval stage than at the pupal stage. The activity of Caspase 3 reached the peak at day-1 wandering stage. The autophagy related genes Atg5, Atg6, Atg8 and Atg12 had higher expression levels during the late larval stage. The acid phosphatase activity was higher during the late larval stage. 【Conclusion】 Apoptosis might occur during the larval stage to inhibit the cell proliferation of wing discs and to prevent wing discs from growth and differentiation. Apoptosis and autophagy might occur at the late larval stage to promote the degeneration of wing bursa, to inhibit the cell proliferation of wing discs, and to push forward the development of pupal wing from wing discs.

【Aim】 Juvenile hormone response region (JHRR) is a 120 bp nucleotide sequence identified from the promoter regulatory region of Krüppel homolog1 (Kr-h1) in Drosophila. This study aims to detect the effects of three E-box like motifs (two B boxes and one C box) on the binding capacity of JHRR to nuclear proteins, and to illustrate the regulatory effects of JH, JH receptor-Methoprene-tolerant (Met) and heat shock protein Hsp83 on the binding capacity of JHRR to nuclear proteins. 【Methods】 The electrophoretic mobility shift assay (EMSA) was used to analyze the binding capacity of JHRR with two B boxes or one C box mutants to nuclear proteins in Kc cells. Nuclear proteins of fat body tissues, which were isolated from JH deficient flies (Aug21-GAL4>UAS-Grim), Met/Gce double mutant flies (Met²⁷gce^2.5k),Met overexpressed flies (Lsp2-GAL4>UAS-Met-V5) and Hsp83 homozygous mutant flies (Hsp^8308445) at the early wandering stage when the JH titer was high, were extracted, and the binding capacity of JHRR to nuclear proteins was detected by EMSA. 【Results】 EMSA results showed that both B box and C box mutation decreased the JH-induced binding capacity of JHRR to nuclear proteins in Kc cells, and the inhibitory effect of C box mutation was even more efficient. The binding complex of JHRR and nuclear proteins which were extracted from fat body tissues of JH deficient or Met/Gce double mutant larvae at the wandering stage was almost undetectable, and decreased significantly when the nuclear proteins were extracted from Hsp83 homozygous mutant larvae. In contrast, the formation of JHRR-nuclear protein complex was enhanced significantly when the nuclear proteins were extracted from fat body tissues of Met overexpressed larvae. 【Conclusion】 B box and C box are essential for JHRR to bind to nuclear proteins, and the binding activity of JHRR to nuclear proteins is dependent on JH and Met and regulated by Hsp83.

【Aim】 The study aims to clone the SRP9 gene from the Chinese honeybee, Apis cerana cerana, to analyze its sequence, and to gain the purified recombinant protein, so as to lay the foundation for the study of its protein function. 【Methods】 The cDNA sequence of the SRP9 gene was cloned from A. c. cerana by RT-PCR. Then the phylogenetic tree was constructed by MEGA5.1, and the protein structure was predicted and analyzed by software PrediceProtein and SWISS-MODEL. The SRP9 gene of A. c. cerana was expressed in Escherichia coli. The recombinant protein was purified by urea elution. 【Results】 Sequence analysis revealed an open reading frame (ORF) of 237 bp, which was named AccSRP9 and encodes 78 amino acids. The deduced relative molecular weight of the encoded protein is 9.36 kD and the isoelectric point is 9.17. Phylogenetic analysis showed that the SRP9 proteins of A. c. cerana, A. mellifera and A. florea cluster together. Protein secondary structure prediction showed that AccSRP9 contains two α-helix and three β-fold regions. By using homology modeling, the predicted three-dimensional structure of AccSRP9 was obtained. The AccSRP9 ORF was ligated into the expression vector pGEX-6P-1 and then transformed to E. coli BL21(DE3) for prokaryotic expression and the recombinant protein was purified. The results showed that the protein was expressed in inclusion bodies. Using the method of washing inclusion bodies, the N-terminal GST tagged recombinant protein pGEX-6P-1-SRP9 was obtained. 【Conclusion】 This study successfully cloned the SRP9 gene AccSRP9 from A. c. cerana, analyzed its sequences and obtained the purified recombinant protein, providing referencs and materials for further study of the functions of the gene.

【Aim】 To clarify the types, morphology, quantity and distribution of sensilla on the female adult antenna and larval head of Agrotis ypsilon. 【Methods】 The sensilla on female adult antenna and the head of the 5th instar larva of A. ypsilon were observed with scanning electron microscope. 【Results】 Five types of chemoreceptors, including sensillum trichodeum, sensillum chaeticum, sensillum coeloconicum, sensillum auricillicum and sensillum basiconicum, were found on female adult antenna of A. ypsilon. For the 5th instar larva of A. ypsilon, the head is oval and hypognathous with chewing mouthparts; the larval head comprises of antenna, ocelli, ecdysial line, setae, clypeus, frons, labrum, mandibles, maxillae, maxillary palpi, labium, labial palpi and fusula; one sensillum trichodeum and four sensilla basiconica were found on the antenna; one sensillum trichodeum and one sensillum chaeticum were found on the mandible, and three sensilla trichodea, three sensilla chaetica, four sensilla basiconica and one sensillum styloconicum were found on the maxilla and maxillary palpus; one sensillum styloconicum and one sensillum basiconicum were found on labial palpus, and one sensillum chaeticum was found on the front of each side of fusula. 【Conclusion】 The quantity of sensilla on female adult antenna of A. ypsilon is more than that on its larval head. It is supposed that female adults use these sensilla to choose host plants and oviposition environment. The sensilla on larval mouthparts are associated with gustation and olfaction, and larvae use them to judge the varieties and suitability of foods.

【Aim】 This study aims to evaluate the effects of syringic acid on the lethal and sublethal effects of Lymantria dispar nucleopolyhedrovirus (LdNPV) against larvae of the gypsy moth, L. dispar, so as to provide a scientific basis for the prevention and control of this insect pest. 【Methods】 Diet plus was used to assess the lethal and sublethal effects of LdNPV with and without syringic acid on the 2nd instar larvae of L. dispar. 【Results】 The median lethal concentration (LC₅₀) values of LdNPV to the 2nd instar larvae of L. dispar fed on the artificial diets with and without syringic acid were 91.1 OBs/μL and 194.8 OBs/μL, respectively. The median lethal time (LT₅₀) values of LdNPV at the concentration of 2 400 OBs/μL to the 2nd instar larvae fed on the artificial diets with and without syringic acid were 9.3 and 10.1 d, respectively. The median lethal time (LT₅₀) values of LdNPV at the concentration  of 24 000 OBs/μL to the 2nd instar larvae fed on the artificial diets with and without syringic acid were 7.8 and 8.1 d, respectively. The higher the concentration of LdNPV, the lower the LT₅₀ value. Syringic acid added in the artificial diet enhanced the lethal effects of LdNPV against L. dispar larvae, and caused the lower LC₅₀ and LT₅₀ values of LdNPV against L. dispar larvae as compared with the diet without syringic acid. When the larvae were fed the artificial diet, the sublethal effects of LdNPV on larval duration, pupal duration and female adult longevity were not significant; when the LdNPV concentration was higher than 240 OBs/μL, however, the male adult longevity was shortened and the number of eggs laid per female was less than those of the control. The sublethal effects of LdNPV on L. dispar larvae fed on the artificial diet with syringic acid and high concentration of LdNPV were significant: the female and male larval duration was prolonged, the male adult longevity was shortened, and the male adult emergence rate and the number of eggs laid per female reduced. 【Conclusion】 The results suggest that syringic acid can enhance the virulence of LdNPV against L. dispar larvae and its sublethal effects on L. dispar.

【Aim】 To determine a scientific and reasonable method of ozone (O₃) water to control the chive gnat, Bradysia odoriphaga, and to make maximum use of it. 【Methods】 A series of concentrations of O₃ water (10, 15, 20 and 30 mg/L) were irrigated into the Chinese chive (Allium tuberosum) outdoor fields. Three concentrations of O₃ water (5, 10 and 15 mg/L) were irrigated into the Chinese chive fields covered with plastic film on the ground and with arch plastic shed. Different developmental stages of B. odoriphaga (adult, larva and pupa) and plants of Chinese chives were treated with ozone water (10-15 mg/L), and Chinese chive seeds were soaked with O₃ water (2 and 10 mg/L) in the laboratory. Then, the control efficacy of ozone water against B. odoriphaga was assayed, and the influences of ozone water on the growth and seed germination of Chinese chives were evaluated. 【Results】 O₃ water irrigated at the concentration of 10-15 mg/L in Chinese chive fields covered with plastic film had a significant control efficacy against B. odoriphaga, even as high as 100%. However, O₃ water irrigated in open fields had no obvious control efficacy on B. odoriphaga. O₃ water irrigated in Chinese chive fields of various cultivation types did not aggravate the damage of Chinese chives by B. odoriphaga. In the laboratory conditions, ozone water led to malformation or death of adults, larvae and pupae, even the body surface was corroded obviously. O₃ water promoted the early growth of Chinese chives, increased the plant height, but was not conducive to regerminate new roots or grow the clipped fibrous roots. When the seeds of Chinese chives were soaked with O₃ water and then washed instantly with clean water, they normally germinated. However, when the seeds were soaked with O₃ water but not washed with clean water, the seed germination was seriously inhibited. 【Conclusion】 Control of B. odoriphaga with ozone water is worthy to be recommended, and the highest efficiency can be achieved under closed conditions.

【Aim】 The meadow moth, Loxostege sticticalis L., exhibits intermittent population dynamics. This study aims to reveal the causes for the termination of the 2nd and 3rd outbreak periods of L. sticticalis in China. 【Methods】 We analyzed the migration dynamics of L. sticticalis in 1983 and 2009 using trajectory analysis, explored the climatic background of population fluctuation by calculating anomalies of temperature and precipitation and checking the atmospheric circulation background responsible for abnormal climatic condition. 【Results】 In June of 1983, extremely low temperature and excessive precipitation occurred in Northeast China and East Siberia of Russia, and severe drought occurred in North China, resulting in a large-scale population collapse event of L. sticticalis with the overwintering area being limited within the high-altitude area in northern Shanxi province in autumn of 1983. The parasitism rate of natural enemies in northern Shanxi province reached a rather high level in spring of 1984, further suppressing the population of L. sticticalis, and leading to the termination of the 2nd outbreak period. In June of 2009, Northeast Asia experienced extremely bad climate similar to that in June of 1983, and then a largescale population collapse event of L. sticticalis occurred again in Northeast China, North China, and East Siberia and Far East of Russia, resulting in the termination of the 3rd outbreak period. Abnormal atmospheric circulation pattern was the direct cause of these two extreme climate events. 【Conclusion】 Extreme climate event has become an important factor causing the termination of outbreak periods of L. sticticalis in China. Due to large uncertainty of extreme climate event, long-term prediction for the termination of outbreak periods of the meadow moth in China is difficult to be made in the current stage.

【Aim】 The aim of the present study is to develop an optical remote sensing technology to automatically monitor insect pests in agricultural fields day and night. 【Methods】 A small reflective telescope was arranged facing a distant dark box, providing a low-lightlevel base line. When insects flied into the telescope field of view, they scattered light from the sun, and the optical information could be collected by the recording system attached to the telescope. At night we instead employed a strong lamp for insect illumination. 【Results】 Insect counting, wing-beat frequency analysis and reflectance spectra recordings were all successfully achieved. This illustrates the possibility to monitor agricultural insect pests with a dark-field back-scattering detection system. Each released known insect species showed a unique reflectance spectrum character. Several insect species, in particular Conogethes punctiferalis (Guenée) and Xenocatantops brachycerus (Willemse) were successfully distinguished from each other by employing the method we introduced. 【Conclusion】 The dark-field back-scattering detection system is practical for monitoring agricultural insect pests in the field through day and night. The wingbeat frequency as well as the spectral contents could be used for pest insect recognition as anticipated.

【Aim】 Chongqing of southwestern China features hills and low mountains in which floral resource plants are abundant. As a consequence, there exists a rich resource of the Chinese honeybee, Apis cerana cerana in different geographical regions of Chongqing. Our study comprehensively surveyed the morphological and haplotype diversity of A. cerana cerana populations collected from different habitats in different regions of Chongqing, which will contribute to the resource conservation of A. cerana cerana. 【Methods】 Samples of one hundred and thirty-nine A. cerana cerana populations from different mountainous regions of Chongqing and surrounding areas were collected, and six morphological characteristics of these samples were measured. The fragments of mtDNA tRNA^leu-COⅡ non-coding region were amplified by PCR and then sequenced to evaluate their genetic diversity. 【Results】 Morphological cluster analysis showed that honeybee populations from Wuling and Ta-lou mountains with subtropical monsoon climate could be distinguished from the honeybee populations from Ta-pa mountains with subtropical mountain climate, and the northern and western regions of Chongqing. The analysis of genetic structure of mtDNA non-coding region showed that gene flow occurred among the honeybee populations from different mountainous habitats. Three new haplotypes of A. cerana cerana were identified, i.e., CqH4 from Ta-pa Mountains, CqH7 from Ta-lou Mountains and CqH13 from western hills. 【Conclusion】 The morphological characteristics of different geographical populations of A. cerana cerana in Chongqing show a degree of diversification and are consistent with the geographical distribution. The common haplotypes are usually shared between the A. cerana cerana populations from different mountains and regions of Chongqing, suggesting that gene flow occurs and thereby increases the genetic complexity of the Chinese honeybee.

【Aim】 To assess the silence efficiency of Knickkopf (Knk) gene at the mRNA and protein levels in the migratory locust, Locusta migratoria, by doublestranded RNA (dsRNA), so as to provide basic information for the gene function analysis in the migratory locust by using RNAi technology.【Methods】 dsRNA was synthesized and injected into the body cavity of the day-1 2nd instar nymphs, and the integuments of abdomen were dissected after dsRNA injection into the locusts at 48, 72 and 96 h, respectively. Half of abdomen integuments were collected and used for the mRNA expression detection by RT-qPCR, while the other half were prepared and used for the protein expression detection by Western blotting. 【Results】 Compared with the control, the relative expression level of Knk mRNA in the treatment group decreased by 78.3%, 96.2% and 95.1%, respectively, while the relative expression level of Knk protein decreased by 61.2%, 33.3% and 52.9%, respectively, at 48, 72 and 96 h after dsRNA injection into the locusts. 【Conclusion】 The Knk expression can be significantly suppressed at both mRNA and protein levels in the migratory locust after dsKnk injection. However, the silence efficiency at the mRNA level is much stronger than that at protein level.