Antennal binding proteins (ABPs) represent a sub-class of odorant binding proteins (OBPs), and thus are assumed to play a role in insect olfaction. In order to explore the role of ABPs in olfaction, the full-length cDNA of an antennal binding protein 2 gene from Spodoptera exigua (SexigABP2) (GenBank accession no. HQ234486) was identified by transcriptome analysis and RACE technology. The sequence analysis showed that SexigABP2 contains a 444 bp open reading frame that encodes 148 amino acids including the six conserved cysteine residues of typical OBPs. SexigABP2 shares the highest amino acid identity (up to 72%) with an ABP2 from Heliothis virescens (HvirABP2). The results of real-time quantitative PCR showed that SexigABP2 was highly expressed in male and female antennae, but weakly expressed in proboscis, legs, and wings of both sexes. The expression levels in female antennae and legs were significantly higher than those in male antennae and legs, respectively. SexigABP2 was further expressed in a prokaryotic expression system, and the protein was purified. By fluorescence competitive binding assay, the affinities of SexigABP2 with 35 odorant compounds were tested. Among the tested ligands, (Z)-9-tetradecenol (a sex pheromone component of S. exigua) and farnesol (a plant volatile compound) showed the highest affinity, with the Ki values of 8.24 μmol/L and 8.14 μmol/L, respectively. Affinity comparison indicated that long carbon-chain compounds with unsaturated bond(s) exhibited the higher affinities than short ones without unsaturated bond; among the unsaturated long carbon-chain compounds, however, alcohols displayed higher affinities than acetates. The results suggest that SexigABP2 might be involved in perception of plant volatile compounds with a long carbon-chain and unsaturated bonds.

E75D is one of the important early transcription factors in the molting process of insects. The cDNA of E75D was cloned from Spodoptera litura by RT-PCR and RACE technology for the first time in this experiment and named Sli-E75D (GenBank accession no. JQ266225). Sli-E75D consists of a 1 836 bp open reading frame encoding 611 amino acids, with a 341 bp 5′ untranslated regions (UTR) and a 358 bp 3′UTR. E75, especially its 3′UTR, is highly conserved among the Lepidoptera insects; however, it shows significant difference in 5′UTR among four isoforms because of the difference of their promoter. The deduced amino acid sequence of E75D in S. litura share 98.4%, 79.3% and 76.5% identity with the homologues in Spodoptera littoralis, Manduca sexta and Bombyx mori, respectively. The miRNAs miR-14, miR-33 and miR-87, which are the most possible regulator of E75, were predicted by PITA and RNAhybird programs based on the highly conserved 3′UTR of E75. The recombinant vector pET28a-Sli-E75D was constructed and transformed into Escherichia coli BL21(DE3) and Transetta(DE3), respectively, to study the effects of rare codons on E75D expression. SDS-PAGE analysis of prokaryotic protein showed that the Transetta(DE3) which transformed pET28a-Sli-E75D recombinant vector expressed much more prokaryotic recombinant E75D protein (consisting of 67.19 kD Sli-E75D, and 7.6 kD T7·Tag and His·Tag) than BL21(DE3). The result of SDS-PAGE analysis demonstrated that Transetta(DE3) is better for the prokaryotic expression of E75D because it supplies tRNAs corresponding to six rare codons in E. coli. The expression levels of Sli-E75 in developmental stages from the last instar larva to adult were detected by qRT-PCR. The results of qRT-PCR revealed that Sli-E75 was expressed at a low level in 6th instar larva, and its expression increased rapidly from prepupa and reached the peak in the middle pupal stage. Then, the expression level of Sli-E75 decreased quickly in the last stage of pupa and rebounded again in the adult. These results can contribute to the in-depth study of E75 in molting process in insects.

Glutathione S-transferases are a ubiquitous superfamily of multifunctional enzymes, in which Omega-class GST is responsible for important physiological function in insects. In this study, the full-length cDNA of a GST Omega gene from Locusta migratoria was cloned by RT-PCR technique, and named as LmGSTo1 (GenBank accession no. JQ750592). The open reading frame (ORF) of LmGSTo1 is 738 bp in length coding 245 amino acids. LmGSTO1 consists of an N-terminal domain and a C-terminal domain. The N-terminal domain includes five β-sheets and three α-turns, while the C-terminal domain is composed of eight α-turns. Temporal expression pattern revealed that LmGSTo1 was constitutively expressed in all developmental stages of the locust. Tissue expression profiling further indicated that LmGSTo1 was slightly expressed in midgut and gastric caecum and had higher expression levels in foregut, Malpighian tubules, fat body and muscle. The expression level of LmGSTo1 was significantly decreased after deltamethrin treatment. The results provide a basis for further studying the function of LmGSTo1.

Infective juveniles (IJs) of Steinernema ceratophorum strain D43 are all ensheathed in a transparent sheath. In order to investigate the effect of sheaths on infectivity of S. ceratophorum D43 and determine the roles of sheath proteins (SPs) in immune suppression of Galleria mellonella larvae, we first obtained the desheathed IJs with chemical exsheathment method. And then, the infection rate of desheathed and ensheathed IJs to G. mellonella larvae was compared according to the host mortality and number of penetration sites on the host. Moreover, the SPs from IJs were extracted with ethanol and characterized by two dimensional electrophoresis (2-DE) and MALDI-TOF-MS. Finally, the influence of SPs on hemocyte number and phenoloxidase (PO) activity of G. mellonella larvae was evaluated. The results indicated that over 95% ensheathed nematodes lost their sheaths after being exposed to 0.5% sodium hypochlorite at room temperature for 20 min and kept alive. Desheathed nematodes caused lower mortality of G. mellonella larvae, longer lethal time and less penetration sites compared to ensheathed nematodes. The SPs extracted in cold 35% ethanol showed six protein spots on 2-DE, and only one was successfully identified as a serine protease by peptide mass fingerprinting. The hemocyte number and PO activity in G. mellonella larvae injected with the extracted SPs were significantly reduced and suppressed, respectively. These results suggest that sheaths have an important role in pathogenicity of S. ceratophorum D43, and the SPs are implicated in the suppression of host immune responses.

The objective of this research was to study the composition of chemical elements and screen out the kinds of chemical elements with geographical characteristics in the body of adults of the brown planthopper (BPH), Nilaparvata lugens (Stal), in different geographic populations. Semi-quantitative analysis of inductively coupled plasma mass spectrometry (ICP-MS) was employed to determine the content of chemical elements existing in the body of adult BPH, and the differences in content of chemical elements among geographical populations were tested by means of analysis of variance (ANOVA) with nested design. Furthermore, full-quantitative analysis of ICP-MS and ANOVA with nested design was utilized to screen and verify the chemical elements which had significant differences among geographical populations but had no significant difference between years. The results showed that there were 57 chemical elements in body of adult BPH, and 27 kinds of chemical elements were filtered out by nested design ANOVA. Twelve elements (Ce, Nd, Pr, Sm, Gd, Th, V, Tl, Mo, Cs, Fe and Mn), which can be used to discriminate geographical origins of BPH, were screened out by means of ICP-MS full-quantitative detection analysis and nested design ANOVA. There are significant differences in content of some chemical elements among different geographical populations of BPH, which would contribute to recognizing the origin of long-distance immigration of BPH, and providing probably a new way to trace back geographic origins of BPH.

【Aim】 To understand the effects of defense responses in tomato plants induced by Frankliniella occidentalis (Pergande) on behavioral responses of Bemisia tabaci (Gennadius) B-biotype, and to explore the interspecific competition mechanisms between F. occidentalis and B. tabaci B-biotype. 【Methods】 Using Y-tube olfactometer and choice test methods, we studied the differences of host preference and behavioral responses of B. tabaci on different plants under different treatment combinations, including the undamaged plants (CK) vs the plants damaged by B. tabaci (B), CK vs the plants damaged by F. occidentalis (F) and the plants damaged by B. tabaci plants (B) vs the plants damaged by B. tabaci and F. occidentalis (B+F), and identified the volatiles of tomato plants in different treatments by GC-MS. 【Results】 The olfactory responses of female B. tabaci adults were not significantly different between CK vs B and CK vs F treatment combinations, whereas strong selection reaction of female adults of B. tabaci to tomato plants damaged only by B. tabaci was found in B vs B+F treatment combination (P<0.01). The results of choice test showed that B. tabaci adults were both strongly attracted by the undamaged plants and oviposited on them in CK vs B and CK vs F treatment combinations (P<0.01), whereas B. tabaci adults prefered the plants damaged only by B. tabaci and oviposited on them in B vs B+F treatment combination (P<0.01). The results showed that 9 kinds of plant volatiles were collected, 5 of which were alkenes, accounting for 90% of the total volatiles by GC-MS. β-Myrcene, which shows strong attractiveness to B. tabaci, remarkably reduced and β-phellandrene, which shows rejection to B. tabaci, remarkably increased in the plants damaged by both herbivores, compared to the plants damaged only by B. tabaci. 【Conclusion】 The defense responses in tomato plants induced by F. occidentalis have a great effect upon host preference and behavioral responses of B. tabaci.

The peach fruit borer, Carposina sasakii, is an important pest of deciduous fruit trees in northern China. In order to explore the influence of host plants such as apricot (Armeniaca vulgaris), plum (Prunus salicina), peach (Amygdalus persica), jujube (Ziziphus jujuba), apple (Malus pumila), and pear (Pyrus sorotina) on the development and reproduction of this pest, experiments designed to examine the developmental duration and reproduction of the pest feeding on various host plants were carried out under laboratory conditions of 23±1℃, RH 80%±7% and a photoperiod of 15L∶9D. The results showed that the peach fruit borer exhibited significant differences in the developmental duration and reproduction among various host plants. Larval duration was the longest on pear (19.15 d) and the shortest on plum (12.48 d), while the preimaginal survival rate was the lowest on pear (17.91%) and the highest on plum (50.54%). Adult females derived from larvae reared on jujube and peach laid significantly more eggs (214.50 and 197.94 eggs laid per female, respectively) compared with those reared on the other host plants. Lifetable parameters were calculated for each host plant and compared by jackknife procedures. Analysis indicated that the net reproduction rate (R₀) was the greatest on jujube (117.49), the mean generation time (T) were the longest on pear (41.31 d) and apple (41.51 d), while the intrinsic rate of increase (r_m) was the greatest on plum (0.1294), followed by jujube and apricot (0.1201 and 0.1128, respectively). The results will help to understand the population dynamics of this pest feeding on various host plants.

To illustrate the influences of temperature on development and reproduction of the yellow peach moth, Conogethes punctiferalis (Guenée), the development and fecundity of C. punctiferalis were investigated at five temperatures (15, 19, 23, 27 and 31℃), with chestnut served as food. The results showed that temperature had significant effects on the developmental duration, survival rate, pupal weight and reproduction of C. punctiferalis. The developmental duration of every stage of C. punctiferalis reduced with increasing temperatures from 15 to 27℃, and there was a positive relationship between developmental rate and temperature. Further increase of temperature to 31℃, however, is unfavorable to its larval development. The larval developmental duration at 31℃ was 1.11 d longer than that at 27℃, while the developmental durations of egg, pupa and preoviposition stages kept shorter at high temperature (31℃) than those at low temperatures. In addition, the development of the 5th instar larva ceased at 15℃, indicating that a higher threshold temperature was needed for the development of mature larva than lower instar larva. The generation survival rate of the yellow peach moth at different temperatures decreased in the order of 23℃>27℃>19℃>31℃, of which the survival rate was high (54.44%-63.56%) at 23-27℃ and low (4.30%) at 31℃, suggesting that excessively high or low temperature was unfavorable to its survival. The oviposition amount of C. punctiferalis was the largest (averagely 55.00 eggs laid by per female) at 23℃, followed by those at 19℃ and 27℃ (43.30 and 39.70 eggs laid by per female, respectively), and the least was 20.90 eggs per female at 31℃. Based on the direct optimal method, the developmental threshold temperatures for egg stage, larval stage, pupal stage, preoviposition stage and the whole generation were 10.37, 10.06, 14.27, 7.47 and 11.85℃, respectively, and the corresponding effective accumulated temperatures were 70.84, 287.71, 118.42, 58.33 and 509.06 degree-days, respectively. The results provide the basis for forecasting the occurrence of the yellow peach moth and are applicable in agricultural practices.

Lymantria dispar is an international quarantine forest insect pest. Ooencyrtus kuwanae (Howard) is an important egg parasitoid of L. dispar (L.) and effective against the host population in some areas. In order to establish the mass rearing methods in practice, the relationship between development of O. kuwanae and environmental temperature was studied through rearing at 5 different environmental temperatures. Since eggs of Antheraea pernyi Guerin-Meneville are cheap and easily available, it is used as the substitute host in mass rearing of the O. kuwanae in the laboratory. The quality of offsprings emerged from O. kuwanae was evaluated by comparison with offsprings emerged from the natural host L. dispar. The results showed that the developmental threshold temperature and effective accumulated temperature of O. kuwanae are 10.50±1.41℃ and 260.74±25.09 day degrees, respectively. The regressive equation between temperature and developmental rate is T=10.50+260.74V. When O. kuwanae adults were fed with 30% honey, the mean life-span of males and females emerged from eggs of L. dispar (15.95 and 20.94 d, respectively) was longer than that of males and females emerged from eggs of A. pernyi （10.38 and 15.01 d, respectively）. The body size of females emerged from eggs of A. pernyi was significantly larger than that of females from L. dispar eggs. The female to male ratio of O. kuwanae emerged from eggs of L. dispar and A. pernyi was similar, 2.42∶1 and 2.57∶1, respectively. The results indicate that O. kuwanae offsprings mass-reared with the substitute host A. pernyi could be used as one of effective biological control agents for controlling L. dispar in fields.

The oak longhorn beetle, Massicus raddei (Blessig) (Coleoptera: Cerambycidae) is an important wood borer pest in natural oak forests (Quercus spp.) in northeastern China. This beetle takes 3 years to complete one generation and is difficult to be controlled due to a highly concealed life history in its larval and pupal stages. Determination of larval instar number and duration is the important foundation for the insect pest forecasting and the scientific management. The number of larval instars was determined using statistical method of frequency analysis through sampling periodically by dissecting infestation trees at Kuandian County, Liaoning Province during 2008-2011, during which the oak longhorn beetle larvae at different developmental stages in forests were collected and five morphological variables of the longhorn beetle larvae, including length of mandible, distance between main ocelli, width of the prothoracic plate, length of the mesothoracic spiracle, and body length, were measured under binocular microscope with an eyepiece micrometer. The results showed that the larvae of oak longhorn beetle have 6 instars without difference between male and female. The three variables including mandible length, distance between main ocelli and width of the prothoracic plate can be used for the separation of larval instars as the sclerotized structures, while length of the mesothoracic spiracle and body length were not reliable due to high variation. The average duration of 1-6 instars measured by the method of median population stadia in fields were 9.25, 266.85, 48.09, 51.29, 260.33 and 385.71 d, respectively. The total larval stage duration was over 1 021.52 d in fields. In northeastern China, the occurrence of oak longhorn beetle was very uniform and the development of population was highly synchronous in natural conditions. The larvae of the oak longhorn beetle need to undergo 3 winters to complete one generation. In the first year larvae overwinter as 2nd-3rd instar stage, the next year the 4th-5th instar larvae overwinter and the third winter all larvae enter the fullgrown larval stage. These findings further reveal the larval biology of the oak longhorn beetle and provide valuable information for the rational control of the pest in practice.

Epicephala lativalvaris Li, Wang & Zhang has been found obligately pollinating Breynia fruticosa and B. rostrata. Based on the field observation of the flower-visiting behavior of E. lativalvaris, the indoor rearing of the E. lativalvaris larvae and the dissection of the B. fruticosa and B. rostrata fruits, we made a detailed study for the first time on the morphological characters, the habit of each developmental stage and the life history of E. lativalvaris. The results show that the life history of E. lativalvaris varies under different climatic conditions. It has 4-5 generations and 4 emergence peaks annually in Xiamen, Fujian Province, and the emergence peaks occur during the following periods: early to middle May, middle to late June, August and late September to early October, and a small number of adults emerge during middle to late November and overwinter in eggs or pupae during early December to middle April of the following year; while it has 6 generations annually and does not overwinter in Yinggeling, Hainan Province. Adults of E. lativalvaris actively pollinated female flowers of B. fruticosa or B. rostrata and laid eggs into their ovaries; larvae fed on seeds of the host plants and developed to maturity by consuming all six seeds, whereas 20.37%-77.63% intact fruits were left to keep the stabilization of mutualisms in each population. The ripening rate of B. fruticosa was similar to that of B. rostrata, but the consumption frequency of E. lativalvaris on B. fruticosa fruits was 2.05 times as high as that on B. rostrata. The results indicate that in the E. lativalvaris-Breynia fruticosa and B. rostrata obligate pollination mutualism, the two host plants obtain about equal benefits from E. lativalvaris, whereas E. lativalvaris benefits more from B. rostrata than from B. fruticosa. This study provides reference for the biological study of other Epicephala species and lays a basis for understanding the mechanism to sustain the stabilization of the obligate pollination mutualism in E. lativalvaris-Breynia fruticosa and B. rostrata.

To attest the feasibility of math-morphological characters (MMCs) of moth wings for classification and identification of Noctuidae, we used digital technology to acquire and process insect images. Thirteen MMCs, such as rectangularity, elongation, lobation, eccentricity, sphericity, roundness and movement invariants including Hu1 and Hu2, which are invariants to the image size and direction, were selected. The characters from right forewings of moths from six noctuid species were extracted in this study. The methods including variance analysis, stepwise discriminant analysis and cluster analysis were used to study the feasibility, reliability and importance of each MMC as classification attribute of insect classification. The kinship among the six noctuids was analyzed from the perspective of mathematical morphology. The analysis results showed that rectangularity and elongation had no significant contribution to classification and identification of the six noctuids. Therefore, 11 morphological characters were selected as the classification variables. The contribution of these variables could be ranked as follows: (eccentricity, Hu5, Hu7)＞Hu2＞roundness＞sphericity＞Hu3＞(lobation, Hu1, Hu6)＞Hu4. Classification and discrimination of the six noctuids were realized by using the character parameters of moth wings. The phylogenetic relationship of the six noctuids based on the character parameters was consistent with that based on the traditional morphological observation. The results indicate that MMCs of moth wings can be used to identify moths rapidly and will be useful in gradually realizing the automatic recognition of moths in the future.

Effective management of the Chinese citrus fruit fly, Bactrocera (Tetradacus) minax (Enderlein) (Diptera: Tephritidae), is hindered by a lack of knowledge about its movements. By measuring the wing loading and extra loading capacity of adults of the Chinese citrus fruit fly, we determined the effect of weight of extra loading electronic tags on the ratio of upward to downward flights of flies, in order to provide technical parameters for making the feasibility of electronic tag for tracking natural movement of B. minax using harmonic radar technology. The wing loading of the Chinese citrus fruit fly was found not to decrease with increasing size over a wide range of individual sizes and independent of sex. The results indicate that an average wing loading of adults of Chinese citrus fruit fly is about 11 mg after normal food uptake alternated with food deprivation (only water is supplied). It is estimated from our results in a large screened cage that 7.3 mg (representing about 23% of adult’s weight) extra loading for the technique has no or minimal impact on the number of upward flights of adults of Chinese citrus fruit fly. The results further suggest that when we determine the feasibility of tags for the tracking of natural movement of B. minax using harmonic radar technology, the weight of electronic tags should be no more than 7.3 mg.

In order to investigate the possible impacts of different tillage practices on occurrence of the orange wheat blossom midge, Sitodiplosis mosellana (Géhin), surveys of wheat midges including removing the larvae from soils, net-trapping and stripping glumes of ears were arranged in wheat field transects with different tillage practices at Luancheng Agro-ecosystem Station, Chinese Academy of Sciences, in Hebei Province in 2010 and 2011. The results showed that the average amount of larvae in transects with different tillage practices was in the following sequence: straw mulch and no-tillage (SMNT) > straw standing and notillage (SSNT) > straw mulch and rotary tillage (SRRT) in 2010 and 2011; the average amount of larvae per 100 tillers by stripping glumes, however, was in the reverse sequence: SRRT > SSNT > SMNT. In 2011, catches of net-trap in SRRT was the highest among three transects. Analysis showed that fields with notillage practices would be beneficial to larval over-wintering, but wheat paddy with this tillage practice would get less damage n wheat ears. Fields with rotary tillage practice would be harmful to larval over-wintering, but wheat paddy with this tillage practice would get much more damages in wheat ears. For the fields with rotary tillage practice, flood irrigation could cause more serious occurrence of wheat midges. The results may help realize the influences of tillage practices on occurrence of the orange wheat blossom midge with the transition of tillage practices in large scale.

Aimed to provide a reference to the chemical and biological control of Nilaparvata lugens (Stal), the effects of the chlorantraniliprole on the functional response of Cyrtorhinus lividipennis (Reuter) preying on N. lugens were studied with rice stem dipping method. The results revealed that the functional response models of C. lividipennis preying on N. lugens all conformed to Holling’s type Ⅱ curve after C. lividipennis were pretreated with the labeled dose of chlorantraniliprole (LDC). However, the parameters in the models had some changes. The instantaneous attacking rate (a′) of C. lividipennis pretreated with the insecticide declined. The handling time (Th) on eggs, 1st instar nymphs and 2nd instar nymphs of N. lugens by C. lividipennis nymphs pretreated with LDC was extended by 163.03%, 104.21% and 95.11%, and 81.97%, 102.45% and 99.52% by C. lividipennis adults pretreated with LDC, respectively. The maximal predatory capacities (Na*) of C. lividipennis pretreated with the insecticide reduced. The searching efficiency (S) of C. lividipennis pretreated with LDC reduced as compared to that of untreated predators. These results suggest that chlorantraniliprole at labeled dose has harmful influence on the predation of C. lividipennis.