piggyBac is a DNA transposon widespread in many organisms. IFP2 is the first piggyBac element found and has been developed as a gene vector most widely used in transgenesis of insects and mammals. Therefore, the study of piggyBac transposons in various organisms has attracted extensive interests and attention. CsuPLE, a novel member of piggyBac transposons, was cloned from Chilo suppressalis in this study with the GenBank accession no. JX392388. The sequence is 2 537 bp in length and contains a single open reading frame (ORF) of 1 914 bp encoding a transposase of 637 amino acids. Within the transposase, there is a conserved DDD-domain. The full-length sequence of CsuPLE contains a pair of perfectly symmetrical 13 bp inverted terminal repeats (ITRs) and a pair of non-completely symmetric 21 bp internal repeats (IRs). CsuPLE sequence is integrated in the “TTAA” target site duplication (TSD) of the genome. CsuPLE with similar structure was found in different geographic populations of C. suppressalis from China. This study not only provides insight into the relationship between structure and function of the piggyBac transposon but also sets up a theoretical basis for utilization of the CsuPLE transposon in transgenic research in C. suppressalis.

To understand the bacterial diversity and the dominant types of bacteria in the spiraling whitefly, Aleurodicus dispersus Russell, bacterial communities present in both sexes of A. dispersus collected from Psidium guajava in Hainan were characterized using 16S rDNA-polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and 16S rRNA gene clone libraries. The partial bacterial 16S rRNA gene fragment was amplified with PCR, and the clone libraries were constructed. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed by digestion of the 16S rRNA gene, and each unique restriction fragment polymorphism pattern was designated as an operational taxonomic unit (OTU). A total of 10 OTUs were identified from samples of both sexes of A. dispersus. Phylogenetic trees of bacterial 16S rRNA nucleotide sequences were constructed. Phylogenetic analysis revealed that Zymobacter, Arsenophonus, Pantoea, and Pseudomonas are the most dominant groups in both male and female adults of A. dispersus. Candidatus Portiera aleyrodidarum and Arsenophonus sp., possibly the endosymbionts of A. dispersus, were detected in all samples. These bacteria may play synergetic roles in development, reproduction and sex-ratio control of the whitefly.

Wolbachia and Cardinium are both maternally inherited intracellular bacteria that infect a wide range of arthropods and are associated with various reproductive abnormalities in their hosts. Infection with bothWolbachia and Cardinium is reasonably common, yet it is relatively difficult to find the individuals infected with single symbiont in doubly infected hosts naturally. In this study, Wolbachia and Cardinium were detected in two populations of the whitebacked planthopper, Sogatella furcifera (Horváth), collected from Wenshan of Yunnan (YN) and Sanya of Hainan (HN). Using the Hainan population of doubly infected S. furcifera as the material, we developed a new technique of microinjecting different concentrations of penicillin G into nymphs of the planthopper. The results showed that the rate of individuals infected with only Wolbachia was very low in natural populations of S. furcifera, and no singly Wolbachia-infected individual was found in the Hainan population. We got singly-infected individuals by using microinjection of penicillin G in the laboratory. The singly infected lines can be produced most efficiently by injecting 0.2% (w/v) penicillin G into the 5th instar nymphs. The result of PCR detection of F5 progenies showed that the singly infected lines obtained by microinjection can be inherited stably. These results here will provide a speedy method to get singly infected lines, and this method can be used to establish lines with different infection type for other arthropods doubly infected with Wolbachia and Cardinium.

To explore why the fungus Ascosphaera apis only infects the honeybee stretched larvae some hours before and after sealing and the related infection mechanism, the inoculation experiments were carried out with different dosages of A. apis spores (0, 1.0×10², 1.0×10³, 1.0×10⁴, 1.0×10⁵ and 1.0×10⁶ spores/mL), inoculation time (3rd, 4th, 5th and 6th instar larva) and low-temperature treatment of 28℃. Furthermore, larvae in different infection stages were sectioned to investigate the infection process of A. apis. The results showed that A. apis pathogenicity is dose-dependent, A. mellifera ligustica larvae could resist the infection of A. apis at the dosages lower than 1.0×10³ spores/mL and the corresponding infection rates at these dosages had no significant difference from the control (P>0.05). The differences of morbidity in different instars of larvae inoculated are due to the difference in spore dosage ingested by different instar larvae. Chilling treatment at the transformation stage significantly increased the infection (P<0.05). In addition, pathologic section showed that the spores in the midgut could not grow due to lack of oxygen, unless they could arrive the hindgut accompanying with faeces. The mycelium in hindgut broke through the gut wall, massively proliferated in body cavity and finally outgrew from the cuticle. Larvae in transformation with the maximal nutrient substances stored and significantly decreased immunity provide A. apis the best opportunity to infect by increasing the possibility of success and decreasing the cost. Altogether, the present work provides a foundation to elucidate the mechanism of A. apis infection and the mechanisms of insect-pathogen interaction

【Aim】 To study the combining effects of two or more insect-resistance genes in plants, as well as to screen and confirm double Bt transgenic 741 poplar lines with high resistance against coleopteran and lepidopteran insects. 【Methods】 Eight lines with three transgenes (Cry3Aa+Cry1Ac+API), one line with double transgenes (Cry1Ac+API) and three lines with single transgene (Cry3Aa) of transgenic 741 poplar were used as experimental materials. Comparative studies were conducted from three aspects: PCR detection of exogenous gene, the expressions of insecticidal proteins and the assessment of insect-resistance ability of plant. 【Results】 The expected electrophoretic bands from the transgenic lines appeared in PCR amplification. ELISA detection showed that the expression of insecticidal proteins was consistent with the exogenous genes in each line. Toxicity tests were performed in the laboratory with Plagiodera versicolora and Hyphantria cunea on fresh detached leaves. Transgenic poplar lines carrying different insect-resistance genes demonstrated selective resistance to target insects, but showed no toxic effects towards non-target insects. Transgenic 741 poplar lines with double Bt transgenes had double insect-resistance ability, and individual lines showed resistance ranging from high, medium to low. Five lines (pCCA1, pCCA2, pCCA5, pCCA6 and pCCA9) selected with high resistance against P. versicolora showed higher toxicity than three single Cry3Aa gene lines with high resistance (pCC11, pCC53, and pCC84). As regard to resistance toward H. cunea, seven lines (pCCA2-pCCA7 and pCCA9) exhibited similar effectiveness as the single Bt line (pB29) and only one line (pCCA1) showed an extremely low level of resistance. 【Conclusion】 The combination of multiple insectresistance genes in 741 poplars exerts gene stacking effects which not only expand its insect-resistance spectrum but also improve its insect-resistance ability largely.

More and more attention was paid to the induced resistance in plant pest control. In order to explore a new control method for wolfberry pests, the induced defense response in wolfberry and the defense mechanism were studied. The changes of activities of protease inhibitors (PIs), superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL), and the contents of polysaccharide and xylogen in wolfberry (Lycium barbarum) leaves subjected to exogenous JA treatment and damage by gall mite Aceria pallida were tested. The results showed that the activities of PIs, SOD and PAL and the xylogen content increased significantly in wolfberry leaves subjected to treatments with JA at the concentrations of 0.001 and 0.1 mmol/L and the damage by A. pallida compared with the control not subjected to JA treatment and the damage by A. pallida， but the polysaccharide content was dramatically reduced. The JA treatment or the damage by A. pallida caused significant increase in activities of chymotrypsin inhibitors (CI) and trypsin inhibitors (TI) and the change extent differed among the three treatments (two JA treatments and one damage treatment by A. pallida) (P<0.05). Activities of CI and TI increased 96% and 128%, respectively, in wolfberry leaves treated with 0.1 mmol/L JA, and 94% and 122%, respectively, in wolfberry leaves damaged by A. pallida. The polysaccharide content in wolfberry leaves treated with JA or gall mite reduced significantly (P<0.05), and the reduction was related with the JA concentrations, the higher the JA concentration, the less polysaccharide content. The xylogen content increased significantly in wolfberry leaves treated by 0.1 mmol/L JA, gall mite and 0.001 mmol/L JA (P<0.05), and reached the peak at 5 d after treatment, which increased by 70%, 41% and 36%, respectively. The activities of SOD and PAL increased significantly in wolfberry leaves treated by JA or gall mite (P<0.05), and also reached the peak at 5 d after treatment. The SOD activity in wolfberry leaves treated with 0.1 mmol/L JA increased from 70.77 to 128.98 U, and PAL activity from 10.91 to 20.59 U at 5 d after 0.01 mmol/L JA treatment. The activities of both enzymes decreased at 10 d after 0.01 mmol/L JA treatment but still higher than that of the control. These results suggest that both exogenous JA treatment and gall mite damage could induce defense responses in wolfberry, which are concentration- and time-dependent.

In order to clarify the effects of cold acclimation on development and fitness of Harmonia axyridis (Pallas) offsprings, based on age-stage, two-sex life tables, we investigated the developmental duration, pre-oviposition period, fecundity, adult life span and population parameters of the offsprings from parental populations with cold acclimation in the laboratory condition. The results showed that for offsprings, the egg developmental period increased while the larval (1-4 instar) and pupal durations were shortened. Variation of body length and body weight gain decreased with the duration of cold acclimation prolonged. Cold acclimation of parents could significantly delayed pre-oviposition period of the offsprings, lowered fecundity, and shortened adult life span. For population parameters of offsprings from the acclimated parental generation, the intrinsic rate of increase (r), the finite rate of increase (λ), the net reproduction rate (R₀) and the age-specific survival rate (l_x) significantly decreased whereas the proportion of females increased. In addition, the type of cold acclimation had a dramatic impact on growth and development of offsprings, with longer longevity and higher l_x for offsprings from the acclimated parental generation at the fluctuating temperature than at the constant temperature 5℃, but lower fecundity and population parameters (r, λ and R₀). The results suggest that plastic responses to chilling are not only realized within the parental generation but also extend across generations, which is very important for H. axyridis to survive and reproduce at low temperature.

The non-diapausing (ND), winter-diapausing (WD) and summer-diapausing (SD) pupae have distinguishable physiological and developmental process. In the present study, the developmental duration and morphological changes of ND, WD and SD pupae were comparatively investigated with the onion maggot Delia antiqua as the model species through anatomic and photographed observation and measurement, with emphasis on the head evagination- and diapause-related characteristics. The study aims to understand the differences of developmental and morphological characteristics of ND, WD and SD pupae, and lays a morphological base for the distinguishment of diapause developmental stage and study of diapause molecular mechanism. The developmental durations of pre-diapause, diapause and post-diapause of WD pupae were 4, 85 and 27 d, respectively, while those for SD pupae were 2, 8 and 22 d, respectively. The pre-diapause begins at the pupation stage, and ends when the free fat body appears in the central area of eyes, which happens 10 h after the completion of head evagination. The completion of head evagination is the premise of diapause occurrence, the head evagination of ND, SD and WD pupae happened at 48, 36 and 83 h after pupation, and there was no morphological difference observed in the developmental process of head evagination between these three kinds of pupae. During the process of head evagination, firstly, the head capsule and thoracic appendages turned out from thorax capsule, and the head got to the location and shape of matured pupae; then, the abdominal muscles continued contraction to push the haemolymph and fat body into the head capsule and thoracic appendages. The pupae entered SD and WD when approximate 15% total effective accumulated temperature was obtained at the pupal stage. In the duration of diapause, the pupal morphology remained in the form of the appearance of free fat body in the central area of eyes, and no difference was observed between these two kinds of diapausing pupae in morphology in the diapause stage. The WD pupae had the largest body length, width and weight, followed by SD pupae and then ND pupae. At the post-diapause stage, malpighian tubules of ND pupae were green and clearly visible during the existence of the yellow body, whereas they were almost invisible in the diapausing pupae. This study provides basic data for understanding diapause physiology and inferring diapause development process of insects through developmental duration and morphology.

Acorns of oaks are usually attacked by weevils. Although the interaction between acorns and weevils has been well studied, how acorns affect the fitness of weevil larvae remains unclear. In this study, we attempted to investigate the evolutionary adaptation of weevils to the development of acorns through carefully investigating the seed fall processes and the characteristics of larval emergence from the trapped acorns of the Oriental white oak, Quercus aliena. Our results indicated that seed crop of Q. aliena was 51.92 ± 29.26 nuts/m² with an infestation rate estimated of 42.4% in 2009. The infested acorns were much larger than the sound ones. About 65% of the infested acorns were superparasitized by weevil larvae. Weevil larvae emerged earlier were much larger than those exited later from infested acorns after seed fall. Early emerged weevil larvae were much larger than those coming out later from the same individual superparasitized acorn. Weevil larvae parasitized in the early abscised acorns were smaller than those in later abscised ones. Our results demonstrated that dry weight per weevil larva was decreased significantly with the increase in the number of weevil larvae in single acorn. The number of weevil larvae was closely and positively correlated with length and maximum width of acorns, respectively. Although the fitness of weevil larvae was reduced in the superparasitized acorns, a preference of female weevil for larger acorns to oviposition may counter this disadvantage. Our results may suggest that smaller acorns may benefit from superparasitization because large acorns attract female weevils to oviposit more eggs.

To investigate the thrips species on potted ficus and to understand the damage difference by dominant thrips to Ficus and non-Ficus potted  pl ants mixing planted in the same greenhouse, we collected the leaves of potted Ficus plants from the Ficus Planting-Demonstration Base of Zhangzhou, Fuzhou and Quanzhou, surveyed the thrips species and determined the dominant species, and the damage degree by Gynaikothrips uzeli Zimmerman on different host plants including Araceae Philodenron, Bignoniaceae Radermachera, Bombacaceae Pachira and Moraceae Ficus potted plants was also investigated. The results showed that there were 9 thrips species belonging to eight genera of two families in two suborders on potted ficus, of which G. uzeli was the dominant species, Androthrips ramachandrai Karny and Mesothrips jordani Zimmermann were the common species, and the other six thrips species were the occasional species. A narrow host range of G. uzeli was found. G. uzeli had the most serious damage to Moraceae Ficus, but had the weaker damage to Philodendron, Radermachera, and Pachira plants. Of the six Ficus species, the damage degrees of G. uzeli reached higher than level three to Ficus benjamina Linn., F. microcarpa Linn. f. and F. benjamina cv. Golden Princess, of which G. uzeli caused the highest damage to the F. benjamina Linn., the second highest damage to F. microcarpa Linn. f. and the milder damage to F. benjamina cv. Golden Princess. The damages of the pest were only level one to F. microcarpa cv. Golden Leaves, F. elastica cv. Deocora Burgundy and F. elastica var. variegata. G. uzeli had higher fecundity on F. microcarpa Linn. f. F. benjamina Linn. and F. benjamina cv. Golden Princess， and could complete one generation on them, but had the best developmental performance on F. benjamina Linn. G. uzeli had lower fecundity on F. microcarpa cv. Golden Leaves, F. elastica cv. Deocora Burgundy and F. elastica var. variegata, and could not complete one generation on them. These results offer the basic information for the control of thrips on potted ficus using host preference.

Plant virus diseases are the “cancer” of the crops, and up to now there is still no effective method to control them. Currently 80% of the known plant virus diseases depend on insect vectors, and the transmission of plant viruses by insects is an interaction of insects, viruses and host plants. The procedure of plant virus transmission has several processes, such as acquisition, retention and inoculation, and a series of virus receptors or proteins are involved in the process. The ways of plant virus transmission consist of stylet-borne, foregut-borne and circulative types, which are nonpersistent, semipersistent and persistent, respectively. The acquisition access period, retention site and inoculation access period are different depending on different insect vectors. The procedure can be affected by many factors including sex and age of insect vectors, host plants, environmental conditions, and symbionts of insects. The main related proteins are as follows: coat protein, minor coat protein, GroEL, helper component and underside-jaw protein. In recent years, the research of plant virus genome has made a great progress, and the mechanism of the virus transmission has attracted an extensive attention. This article reviews the recent studies and developments of this field, including the ways of plant virus transmission vectored by insects, the factors influencing the efficiency of virus transmission, and the mechanisms of virus transmission, especially the possible receptors of the insects related to the virus transmission.

Entomological radar enables us to visualize individual insects from great distances and to scan huge volumes very rapidly, which contribute much to our understanding of insect migration. Vertical-looking insect monitoring radar (VLR) was developed as a new tool for monitoring high-altitude insect migration in the 1970s. In contrast to the early entomological scanning radar, VLR can provide information about the displacement speed and direction, heading orientation, size and shape of targets. Therefore, VLR has more precise capacity in target identification. Furthermore, VLR systems allow continuous and autonomous monitoring of pest migrations under the control of microcomputer, and this make it practicable to carry out routine monitoring with VLRs. In this article, the history of VLR technologies, their configurations and procedures for signal analysis are reviewed, the limitations and related suggestions are discussed, and finally the perspective of using VLRs in future insect monitoring network is presented.

3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is a key enzyme of the mevalonate pathway. Obtaining HMGR gene information is the basis of studying the relationship between cantharidin biosynthesis and the mevalonate pathway. In this study, a fulllength cDNA of the HMGR gene was cloned from the blister beetle Epicauta mannerheimi (Maklin) by RACE technology, which was named EmHMGR (GeneBank accession no. JQ690539). The full-length of EmHMGR is 3 118 bp with an ORF of 2 526 bp, containing a 5′-untranslated region (5′-UTR) of 178 bp and a 3′UTR of 414 bp. It encodes a deduced protein of 842-amino acid residues, with a predicted molecular mass of 92.8 kDa and isoeletric point (pI) of 6.0. Its predicted molecular formula is C₄₁₃₅H₆₆₀₄N₁₀₉₈O₁₂₁₆S₅₀. Bioinformatical analysis showed that its instability index is 43.37 and the GRAVY 0.091, suggesting that EmHMGR is an unstable hydrophobic protein. The deduced protein EmHMGR has the conserved functional domain of HMGR_ClassⅠ and the sterol-sensing domain, and shares more than 50% amino acid identity with HMGRs from other insects. Phylogenetic analysis showed that EmHMGR has the closest relationship with HMGRs of chrysomelids. This study provides the basis for further research on the biosynthetic pathway of cantharidin in the blister beetles.

The pathological changes in midgut tissues of larvae of the cotton bollworm, Helicoverpa armigera (Hübner), fed on artificial diet containing Vip3Aa protein were observed using transmission electron microscope, and the changes induced by Cry1Ac were also investigated in order to ascertain the action mechanism of Vip3Aa. The results showed that the goblet cells changed gradually after H. armigera larvae fed on artificial diet containing Vip3Aa protein, and the major symptoms included: microvilli swelled and dropped off, the boundaries of karyotheca in cell nucleus blurred and the chromatins unevenly distributed, the numbers of mitochondria and endoplasmic reticulum reduced, the inner cristae of mitochondria blurred, and the endoplasmic reticulum mixed disorderly. The pathological changes of goblet cells in the midgut of H. armigera larvae induced by Vip3Aa were slower than that by Cry1Ac. The obvious pathological changes were detected at 12 h after feeding Vip3Aa, and the pathological changes became more and more evident with extension of feeding time. The obvious pathological changes of H. armigera induced by Cry1Ac, however, appeared at 2 h after Cry1Ac treatment. This study may enrich the theoretical basis for better using Vip3Aa as new toxin strategy in IPM of the cotton bollworm.

The ultramorphology of sensilla on the proboscis in three pest noctuid moths of agricultural significance, the cotton bollworm Helicoverpa armigera (Hübner), the Oriental tobacco budworm moth H. assulta (Guenée), and the silver looper moth Argyrogramma agnata (Staudinger), were investigated using scanning electron microscopy and compared to reveal the ultramorphological differences of the proboscis sensilla among these noctuid moths. The results showed that sexual dimorphism is not found in types of the proboscis sensilla in these species. The proboscises in H. armigera and H. assulta are similar in sensillum types and morphology, sharing similar sensilla trichodea, sensilla basiconica, and sensilla styloconica. Sensilla styloconica bear prominent ridges and are densely borne on the distal part of the proboscis. The proboscis in A. agnata, however, bears sensilla coeloconica in addition to the three types of sensilla mentioned above; sensilla styloconica are slender, lack prominent ridges, and are sparsely distributed on the distal part of the proboscis. The results suggest that the proboscis sensilla can be used in the classification and phylogenetic analysis of the Noctuidae.