Midgut is the main target for Bacillus thuringiensis (Bt) action, and a number of insect midgut proteins have been proposed as putative Bt toxin receptors. In order to study the resistance mechanism of the cotton bollworm, Helicoverpa armigera to Bt, we constructed a larval midgut cDNA library of the cotton bollworm using the Switching Mechanism at 5′ end of the RNA Transcript (SMART）technique. The total RNA of 5th instar larval midgut was extracted and the double-stranded cDNA synthesized. After the normalization treatment, cDNAs were digested and ligated into vector, and then the recombinants were transformed into competent cells. The titer was tested and the cDNA library was amplified and sequenced. The quality evaluation showed that the library had a complexity of 2×10⁶ pfu/mL, and the recombination rate was 100%. The average length of inserted cDNA fragments was over 1 000 bp, and 50% fragments were in the full-length form. A total of 1 098 expressed sequence tags (ESTs) were generated successfully after editing and trimming the vector and ambiguous sequences, and 789 unigene sequences were identified, including 132 contigs and 657 singlets. The assembled 789 ESTs were analyzed with Blast in NT, NR and SWISSPORT database of NCBI. The Blast analysis showed that 218 ESTs (27.62%) had no comparable sequences in databases, 119 ESTs (15.08%) had no definite annotations, and the rest 452 ESTs (57.29%) had high homologies with the available sequences, which had definite annotation with over 300 protein products. Through this study, a high-quality cDNA library of the larval midgut of H. armigera has been constructed, which will be a useful tool for studing gene functions in H. armigera midgut.

RNA and export factor binding proteins (Aly/REF) play important roles in RNA stability, processing and nuclear export. Referring to the sequence of Bombyx mori aly gene (GenBank accession no.DQ497195.1), the aly/ref gene (Bmaly/ref） from the silkworm, B. mori, was cloned via RT-PCR. The sequencing result showed that the open reading frame(ORF) of Bmaly/ref is 765 bp in length, encoding 254 amino acid residues, and the amino acid sequence of BmAly/REF showed 49.7% and 52.7% identities with the homologs of Drosophila melanogaster and Mus musculus, respectively. The structure prediction displayed that BmAly/REF contains RNA binding domain (RRM), REF-N and REF-C motif of REF subfamily. Phylogenetic analysis showed that the insect Aly/REF proteins clustered to a group and the BmAly/REF was close to Aly/REF proteins of Apis mellifera and Tribolium castaneum. The BmAly gene was inserted into expression vector pGS21a (+), and then the recombinant protein was expressed in Escherichia coli and used to immunize mice to prepare the antibody against BmAly/REF. Immuno-fluorescence examination showed that BmAly/REF was distributed in both cytoplasm and nucleus of the cell, but mainly located in the nucleus. Microarray data analysis showed that Bmaly/ref gene was highly expressed in tissues of the day- 3 5th instar larvae of the silkworm. The results suggest that BmAly/REF is likely to play important roles in RNA nuclear export, which lays a good foundation to further investigate the function of BmAly/REF.

In order to better understand the role of arginine kinas (AK) gene and seek new molecular targets for insect pest control, we cloned AK cDNA sequence from fat body of Helicoverpa assulta by RT-PCR and RACE methods. The AK cDNA was named HassAK (GenBank accession no. HQ336337). The expression pattern of HassAK at different developmental stages (from 1st day of 4th instar larva to 1-d-old pupa), in different tissues (head, midgut, fat body, cuticle and abdominal legs) and after treatment with different temperatures was further determined by RT-PCR. The results of sequencing and sequence analysis showed that the fulllength open reading frame of HassAK is 1 068 bp, encoding 355 amino acid residues with the predicted molecular weight and isoelectric point of 40.0 kD and 5.76, respectively. Amino acid sequence analysis showed that the HassAK sequence has the typical characteristics of arginine kinase, which contains the active sequence, the active site and a pair of highly conserved amino acids that form an ion pair of AK. Sequence comparison results showed that HassAK has more than 70% amino acid sequence identity with AKs from other insects. The results of fluorescent quantitative analysis revealed that HassAK was expressed in the head, midgut, fat body, cuticle and abdominal legs of larva. Among them, the expression level of HassAK was higher in abdominal legs and the midgut. Temporal analysis indicated that the expression level of HassAK reached the peak in the pre-pupation stage. In addition, high and low temperatures led to increased expression of HassAK, suggesting that HassAK gene may be involved in insect resistance to adverse external environments.

Rieske iron-sulfur protein (RISP) is one of key subunits in mitochondria complex Ⅲ that plays an important role in respiratory electron-transport chain. The full-length Spodoptera litura RISP cDNA (SlitRISP) of 4 413 bp was cloned and characterized by RT-PCR and RACE technique, which contains three introns. The open reading frame (ORF) of SlitRISP is 816 bp encoding 271 amino acid residues. The sequence analysis indicated that SlitRISP shares overall 83% and 79% amino acid sequence identity to the RISP sequences from Bombyx mori and Plutella xylostella， respectively. The qRT-PCR result revealed that the expression level of SlitRISP was significantly different in different stages of S. litura, which in larval and adult stages was significantly higher than that in egg and pupal stages, while the expression levels in 4th and 5th instars were clearly higher than that in the early instars and 6th instar. This study provides a foundation for understanding the SlitRISP function and its application as a new target for respiratory electron transport blocking agent in designing new and efficient insecticides.

 The sand field cricket, Gryllus firmus, is a wing-polymorphic insect, a model insect for studying fitness of reproduction and dispersal within species. In this study, we qualitatively and quantitatively analyzed titers of juvenile hormone (JH) and fatty acids in flight muscles between the long wing ［LW(f), flight-capable］ and the short wing (SW, flightless) females by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). The results showed that JH titers at 4 h before flight on day 5 and 7 of LW(f) morph exhibited a daily rhythm with about 3.4- and 2.7-fold increases, respectively (P<0.05) compared with those in SW morph, that is, from 386.52±68.40 ng/g to 1 327.53±277.98 ng/g on day 5 and from 630.36±37.26 ng/g to 1 685.77±143.95 ng/g on day 7, respectively. By contrast, there was no significant daily variation of JH titers on day 5 and 7 of SW morph, and day 1 of both morphs. Further, the 14-methyl-pentadecanoic acid, a C16 fatty acid, was also found to exhibit a daily rhythm on the day 5 and 7 of the LW(f) morph with temporal consistency with the JH peak, which did not occur in the flightless morph. The experiments also prove that the daily rhythm of JH titers is not due to the variation in flight muscle mass or the ratio of flight muscle mass. These results help us to understand the relationship between JH titer and flight behavior.

To determine the distribution and dynamic excretion of metal elements, the test insects, adults of the Sogatella furcifera (Horváth) were subjected to feeding on deionized water for 0 h, 24 h, 48 h, and 96 h, respectively. The body weight changes were measured and the contents of twelve metal elements (Cu, Zn, Cd, Mn, Ca, Fe, Na, As, U, Mg, K and Pb) in S. furcifera were detected using ICP-MS technique. At the same time, we denote the entire body weight, the organic tissue weight, the weight of the intestinal contents and their excretion rate of S. furcifera by W, O, E and r, respectively. The body weight of S. furcifera at different time can be expressed by the modified exponent equation W_t=O+E(1-r)^t . Considering the content of a certain element in S. furcifera (WC_t )  should be the sum of the content of that element in the organic tissues (OC) and in the intestinal contents (EC), the statistical model is WC_t=OC·OP_t+EC·EP_t, where OP_t is the percentage of O to body weight at time t and EP_t is the percentage of E. The results of simulation modeling showed that the contents of Cu, Zn, Cd, Mn and Ca are significantly higher in the organic tissues than those in the intestinal contents (P<0.05), suggesting that these elements can be absorbed or accumulated by organic tissues. The contents of seven elements (Fe, Na, As, U, Mg, K and Pb) have no significant difference between in the organic tissues and in the intestinal contents (P>0.05), suggesting that the contents of these elements in the organic tissues are balanced with those in the intestinal contents and their contents in insect body are not significantly changed. The simulation modeling method can provide reference to other studies involving metal distribution, accumulation and excretion in other small or micro-insects.

 To understand the physiological mechanism of insecticide-induced enhancement of flight capacity of the brown planthopper, Nilaparvata lugens (Stål), we examined the changes of trehalose content and the trehalase activity in the 3rd and 5th instar nymphs and the adults of N. lugens under the stress of sublethal dose of three insecticides (imidacloprid, triazophos and deltamethrin).The results showed that the trehalose content and the trehalase activity in the 3rd instar nymphs treated with three insecticides were not significantly different from those of the control (P>0.05). The trehalose content in the 5th instar nymphs treated with 40 mg/L triazophos was significantly lower than that in the control (P<0.05), decreased by 24%, while the trehalase activity in the 5th instar nymphs treated with 20 and 40 mg/L triazophos was significantly higher than that in the control (P<0.05), increased by 100% and 129%, respectively. Compared to the controls, the trehalose content in brachypterous female and male adults was significantly reduced in treatments of 10 mg/L imidacloprid, 20 and 40 mg/L triazophos, and 3 and 6 mg/L deltamethrin (P<0.05), decreased by 36%, 53%, 67%, 58% and 69% in female adults, and 59%, 71%, 65%, 70% and 77% in male adults, respectively. However, the trehalase activity in brachypterous female and male adults was significantly increased in treatments of 40 mg/L triazophos and 3 and 6 mg/L deltamethrin, compared to the controls (P<0.05), increased by 124%, 100% and 88% in female adults, and 146%, 132% and 118% in male adults, respectively. The trehalose content in macropterous female and male adults treated with 10 mg/L imidacloprid, 40 mg/L triazophos and 3 mg/L deltamethrin was significantly lower than that in the control (P<0.05), decreased by 44%, 34% and 37% in female adults, and 48%, 54% and 43% in male adults, respectively. However, the trehalase activity in macropterous female and male adults treated with 5 and 10 mg/L imidacloprid, was significantly higher than that in the control (P<0.05), increased by 170% and 97% in female adults, and 317% and 300% in male adults, respectively. The results suggest that the trehalase activity of N. lugens is increased by treatments of sublethal dose of the three insecticides, while the trehalose content is reduced. The present work provides some valuable clues for understanding the mechanism of insecticide-induced enhancement of flight capacity of the N. lugens.

The aromatic nature of pine and cinnamon oils has established them as good adult repellents but their larvicidal efficacy against mosquitoes has not been explored much. Keeping this in view, laboratory studies were conducted to uncover the larvicidal potential of commercially available pine (Pinus longifolia) oil and cinnamon (Cinnamomum zeylanicum) oil against the early 4th instar larvae of Aedes aegypti, originating from Delhi, India. The larvicidal effects were investigated in terms of the larval mortality, behavioural and morphological changes, if any. Our studies revealed the larvicidal efficiency of both the oils against 4th instar larvae of Ae. aegypti, the pine oil proving to be more effective than the cinnamon oil. The LC₅₀ and LC₇₀ values exhibited by pine oil were 0.33093 mg/L and 0.54476 mg/L, respectively, while the values obtained with cinnamon oil were 0.63159 mg/L and 0.77736 mg/L, respectively. It was further observed that at LC₉₀ the larvicidal potential of cinnamon oil surpassed that of the pine oil, exhibiting a value of 1.11879 mg/L as in comparison to 1.04915 mg/L shown by pine oil. The behavioural changes as excitation, restlessness, tremors, and convulsions followed by paralysis observed in the treated larvae suggest a probable effect of the oils on their neuromuscular system. Microscopic study of morphological alterations in the treated larvae revealed that most of their organs had a normal structural appearance as that of controls except the little internal shrinkage in anal gills leading to the structural deformity. This indicates the anal gills as the probable action sites of the oil extracts and dysfunction of the gills leading to larval mortality. The potential of oils as new types of larvicides for the control of mosquitoes are explored.

 Tetraphleps galchanoides Ghauri is an important predator of the hemlock woolly adelgid (HWA), Adelges tsugae Annand, in China. Its biology, habitats and prey habit were studied through breeding observation in the laboratory and field investigation. It occurs 2 generations per year without generation overlapping in Tianshengqiao Forest Farm, Lanping County, Yunnan Province. The 1st generation occurs in late May to late October. Egg period lasts 13.8±1.6 d, nymph period lasts 97.6±7.4 d, and adult period lasts 55.2±4.7 d. The 2nd generation occurs in mid November to late April of next year. Egg period lasts 11.3±1.1 d, nymph period lasts 105.7±8.5 d, and adult period lasts 60.4±5.3 d. The nymphs have 5 instars. The 2nd-generation adults emerge from overwintered 5th instar nymphs under bark of hemlock branch or ground litter. The sex ratio of female to male is 8.5∶1. Adults fly slowly usually at the lower canopy, and their flight was affected by mating, oviposition, and feeding. The optimum habitat for T. galchanoides is at the elevation range from 2 851 to 2 980 m, with forest coverage of 0.61-0.70, and with hemlock density 13.3-15.5/100 m. Food choice test was conducted among HWA and other 3 adelgids, i.e., Aphrastasia pectinatae (Cholodkovsky), Pineus armandicola Zhang, and Adelges laricis Vall. The results showed that there was a significant difference in its preference between HWA and the other adelgids. HWA was the most preferred prey. T. galchanoides can be used as one of the major natural enemies for the biological control of HWA.

The sand sagebrush carpenterworm, Holcocerus artemisiae Chou et Hua, is a severe pest of Artemisia ordosica and A. sphaerocephala, with the larval stages boring into the roots of the host plants. Morphological and biological characteristics of H. artemisiae in Ningxia were studied through field survey and laboratory breeding and observation. The results showed that a generation takes two years in fields of Ningxia. Larvae of all instars were found to overwinter in the host plant roots. Mature larvae begin to pupate in mid May in the soil around the base of plant stems, with the average pupal stage lasting 19.5±3.5 d. Moths emerge from the beginning of June to the end of August, with three peaks of eclosion observed in the early June, early July and early August, respectively. Adult emergence occurs mainly in the afternoon (between 14:00-17:00), and takes 30-45 min. Copulation peaks in the late evening (around 20:00-21:00). The longevity of adults is 2-3 d for males and 1-3 d for females, with a sex ratio of 1∶2.16 (female ∶male) in Ningxia fields. Female moths start laying eggs near mid June, and the eggs hatch in late June. Larvae consume a single root alone in irregular tunnels before transferring to another. They were found to cause 11%-44% damage (average 24%) on A. ordosica from May to September, with a population density of 0.13 individuals per root. There was a significant difference in the selectivity of H. artemisiae larvae to tree age (1-4 years) and ground diameter (16-34 mm). The systematic research of biological characteristics of H. artemisiae provides scientific evidence for its control.

The growth and survival of a geometrid defoliator, Hyposidra infixaria Walker were studied on four popular clonal varieties (TV1, TV9, TV25 and Teen ali 17) of tea Camellia sinensis in the laboratory with an aim to find out the most suitable clonal variety of tea for the insect. The results showed that the larval period of H. infixaria on TV25 clone was significantly shorter (15.78 d) than that on TV1 (18.14 d), TV9 (18.00 d) and Teen ali 17 (17.00 d) clones. There were two variations in the larval instar numbers, those being the five- or six-instar type. The incidence of the six-instar type fed on TV25 was higher than those fed on other three clones. Larval and pupal weights were also significantly higher on TV25 than other three clones. Similarly, the survival of immature stages of H. infixaria fed on TV25 was significantly higher than other three clones. Based on growth parameters and survival, TV25 was found to be the most preferable clonal variety of tea for H. infixaria.

 Cacopsylla chinensis (Yang et Li) is one of the main pests of pear. In order to estimate the genetic variability within populations and genetic differentiation among populations of C. chinensis at the molecular level, genetic diversities of 16 geographic populations of C. chinensis in China were analyzed with 7 microsatellite loci. The results showed that the number of effective alleles and the polymorphism information content (PIC) per locus were 2.2927-10.0610 and 0.5073-0.8735, respectively. The average expected heterozygosity of all C. chinensis populations was 0.7876. The genetic distances between different populations were 0.0951-1.0139, the Nei’s expected heterozygosity 0.4771-0.7892, and the Shannon information indexes 0.8396-1.9989. The average value of inbreeding indexes (F_ST) and gene flow were 11.61% and 2.2236, respectively. The results indicated that the seven microsatellite loci are of high polymorphism, C. chinensis in China keeps a low level of population genetic differentiation and high degree of gene flow, and genetic variability mainly exists between individuals within populations. The results provides some basic information on molecular biology for the development of effective control strategies of the psyllid.

Monolepta hieroglyphica (Motschulsky) (Coleoptera: Chrysomelidae) has become more serious on maize in some maize planting areas of North China since 2001. To investigate the level of genetic diversity, genetic differentiation and gene flow among different populations and within populations of M. hieroglyphica in North China, partial sequences (484 bp) of mtDNA COⅡ (mitochondrial cytochrome oxidase subunit Ⅱ) gene in 26 geographic populations from North China were sequenced and analyzed. In the total 515 individuals, 28 variable sites and 15 haplotypes were observed. Phylogenetic tree of 15 COⅡ haplotypes exhibited two main clusters. The mean haplotype diversity of all populations was 0.257, varied from 0.100 to 0.515 within populations. The overall Fst value was 0.585, Gst was 0.417, and the average gene flow (Nm) among the 26 populations was 0.35. Molecular variance (AMOVA) analysis indicated that a high proportion of the total genetic variance was attributable to variations among populations (58.58%). The Tajima’s D neutrality test results suggest that for M. hieroglyphica in North China there might be no population expansion. These results indicate that the level of gene flow among different populations is very low,  and there is obvious differentiation among M. hieroglyphica populations in North China.

The genus Cotysoides was erected in 2000, recognizable through the following morphological characters: size small; anterior margin of vertex protruding beyond eyes; antennae inserted in the lower one third level of anterior margin of eyes; disc of pronotum densely covered with small tubercles; median keel undulated, with a series projections; hind process not reaching the top of hind femur; lateral lobe of pronotum turning over outwards, posterior angles truncate; hind wing not reaching the top of hind process; the first segment of hind tarsus longer than the third. This paper reports six species of the genus Cotysoides Zheng et Jiang, including a new species, Cotysoides interruptus sp. nov. The new species is allied to Cotysoides guangxiensis Zheng et Jiang, 2000, but differs in: 1) width of vertex 1.5 times of an eye; 2) top of anterior angle of lateral lobe of pronotum obtuse angular; 3) hind wing reaching the knee of hind femur; 4) width of midfemur narrower than the width of tegmina; 5) hind margin of subgenital plate of female with a triangular projection in the middle. A key to species of Cotysoides is provided. Type specimens are kept in Institute of Zoology, Shaanxi Normal University and the Museum of Southwest Forestry University.

This paper revises the taxonomy of the genus Gibbotettix Zheng from China. Nine species are reported, in which two are new to science: Gibbotettix lativertex sp. nov. and Gibbotettix guizhouensis sp. nov. A key to the known species of Gibbotettix Zheng is given. G. lativertex sp. nov. is allied to G. hongheensis Zheng, 1992 and G. hupingshanensis Fu et Zheng, 2003. G. guizhouensis sp. nov. is allied to G. lativertex. The type localities of the two new species are  Sangzhi of Hunan and Daozhen of Guizhou, respectively. The type specimens are deposited in the Institute of Zoology, Shaanxi Normal University. The generic characters are revised based on the specific characters of new species.

To determine the localization of cross-reactivity allergens in the body of Tarsonemus granarius, paraffin sections of T. granarius stained by hatmatoxylin-eosin (HE) were observed by optical microscope. Serum specific IgE antibodies from patients who were sensitive to the house dust mite served as probe, and the cross-reactivity allergens in the body of T. granarius were localized by the method of immunohistochemistry. The digestive system was found to occupy large parts of body cavity of T. granarius by HE staining, which consists of oropharynx, midgut, hindgut, anus, salivary gland and so on. The cross-reactivity allergens of T. granarius mostly occurred in the midgut tissue, gut contents, cuticle and reproductive system in the immunostained sections. The body of T. granarius has allergens which can trigger IgE antibodies of the human body and the distribution of allergens in its body is consistent with that in the house dust mite, Dermatophagoides pteronyssinus.