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Densoviruses (DNVs) infecting arthropods are members of the family Parvoviridae . Here we report the complete genome sequence of a novel DNV with a monosense genome that infects cotton bollworms ( Helicoverpa armigera ), named HaDNV-1. Alignment and phylogenetic analysis revealed that HaDNV-1 showed high identity with the genus Iteravirus .

Two transgenic lines of the soybean Glycine max, MON87701 expressing the Cry1Ac protein and MON87701RR2Y expressing Cry1Ac + EPSPS proteins, were evaluated for their resistance to four lepidopteran pests in the laboratory using detached-leaf bioassays throughout the soybean growth seasons (before anthesis, during anthesis and after anthesis) in China. Enzyme-linked immunosorbent assays (ELISAs) were used to monitor the Cry1Ac expression in soybean leaves.The bioassay results revealed that both transgenic soybean lines exhibited significantly high resistance against Helicoverpa armigera (Hübner) throughout the soybean growing seasons. The survival rates of H. armigera larvae ranged from 5.4 to 24.4% when feeding on the transgenic soybean leaves, significantly lower than the survival rates when feeding on control leaves (71.1-94.9%). Limited resistance was found for both transgenic soybean lines against Spodoptera litura (Fabricius), although the survival rates and weight of S. litura larvae as well as female fecundity were significantly decreased when feeding on Bt soybean leaves compared with feeding on control leaves. In contrast, both transgenic soybean lines provided almost no resistance to Spodoptera exigua (Hübner) and Agrotis ypsilon (Rottemberg). Cry1Ac expression in the leaves of both transgenic soybean lines was relatively stable throughout the soybean growing season, with a peak occurring at V6 -8 and V11 -12 before anthesis. The ELISA results were positively correlated with the results from the insect bioassays.The results show that, while Cry1Ac-expressing Bt soybeans may provide good protection against H. armigera, alternative control measures are required to manage S. exigua, S. litura and A. ypsilon.

Insect chemosensory proteins (CSPs) have been proposed to capture and transport hydrophobic chemicals from air to olfactory receptors in the lymph of antennal chemosensilla. They may represent a new class of soluble carrier protein involved in insect chemoreception. However, their specific functional roles in insect chemoreception have not been fully elucidated. In this study, we report for the first time three novel CSP genes (AlinCSP1-3) of the alfalfa plant bug Adelphocoris lineolatus (Goeze) by screening the antennal cDNA library. The qRT-PCR examinations of the transcript levels revealed that all three genes (AlinCSP1-3) are mainly expressed in the antennae. Interestingly, these CSP genes AlinCSP1-3 are also highly expressed in the 5(th) instar nymphs, suggesting a proposed function of these CSP proteins (AlinCSP1-3) in the olfactory reception and in maintaining particular life activities into the adult stage. Using bacterial expression system, the three CSP proteins were expressed and purified. For the first time we characterized the types of sensilla in the antennae of the plant bug using scanning electron microscopy (SEM). Immunocytochemistry analysis indicated that the CSP proteins were expressed in the pheromone-sensitive sensilla trichodea and general odorant-sensitive sensilla basiconica, providing further evidence of their involvement in chemoreception. The antennal activity of 55 host-related semiochemicals and sex pheromone compounds in the host location and mate selection behavior of A. lineolatus was investigated using electroantennogram (EAG), and the binding affinities of these chemicals to the three CSPs (AlinCSP1-3) were measured using fluorescent binding assays. The results showed several host-related semiochemicals, (Z)-3-hexen-1-ol, (E)-2-hexen-1-al and valeraldehyde, have a high binding affinity with AlinCSP1-3 and can elicit significant high EAG responses of A. lineolatus antennae. Our studies indicate the three antennae-biased CSPs may mediate host recognition in the alfalfa plant bug A. lineolatus.

Evolution of pest resistance reduces the efficacy of insecticidal proteins from Bacillus thuringiensis (Bt) used in sprays or in transgenic crops. Although several pests have evolved resistance to Bt crops in the field, information about the genetic basis of field-evolved resistance to Bt crops has been limited. In particular, laboratory-selected resistance to Bt toxin Cry1Ac based on recessive mutations in a gene encoding a toxin-binding cadherin protein has been identified in three major cotton pests, but previous work has not determined if such mutations are associated with field-selected resistance to Bt cotton. Here we show that the most common resistance alleles in field populations of cotton bollworm, Helicoverpa armigera , selected with Bt cotton in northern China, had recessive cadherin mutations, including the deletion mutation identified via laboratory selection. However, unlike all previously studied cadherin resistance alleles, one field-selected cadherin resistance allele conferred nonrecessive resistance. We also detected nonrecessive resistance that was not genetically linked with the cadherin locus. In field-selected populations, recessive cadherin alleles accounted for 75–84% of resistance alleles detected. However, most resistance alleles occurred in heterozygotes and 59–94% of resistant individuals carried at least one nonrecessive resistance allele. The results suggest that resistance management strategies must account for diverse resistance alleles in field-selected populations, including nonrecessive alleles.

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a key insect pest of cotton in Xinjiang cotton-planting region of northwest China. In this region, cotton is grown on average ≈ 1.65 million ha (1.53 ≈ 1.80 million ha) annually in largely monoculture agricultural landscapes, similarly to cropping systems in the United States or Australia. Under such cropping regimes, naturally occurring refuges (with non-Bt crops) may be insufficient to prevent H. armigera resistance development to Bt toxins. Therefore, we assessed frequency of alleles conferring resistance to Cry1Ac toxin of F(1) and F(2) offspring of H. armigera isofemale lines from two distinct localities in the region during 2005-2009. More specifically, a total of 224 isofemale lines was collected from Korla County (≈ 70% Bt cotton adoption) and 402 lines from Shache County (≈ 5% Bt cotton planting). Subsequent offspring was screened on Cry1Ac artificial diet. From 2005 to 2009, resistance gene frequency in Korla fluctuated between 0.0000 and 0.0040, while being 0.0000-0.0008 in individuals collected from Shache, and there were no significant increases in both counties from 2005 to 2009. Relative average development rates (RADRs) of larvae in F(1) tests showed significant increases from Korla, but not in Shache. RADR of F(1) larvae is significantly correlated with RADR of F(2) offspring, indicating genetic variation in response to toxin in field H. armigera population. Although the occurrence of Cry1Ac resistance alleles was low in Xinjiang cotton-planting region of China, particular attention should be given to H. armigera resistance development in Korla County.

International audience; Over the past 16 years, vast plantings of transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have helped to control several major insect pests(1-5) and reduce the need for insecticide sprays(1,5,6). Because broad-spectrum insecticides kill arthropod natural enemies that provide biological control of pests, the decrease in use of insecticide sprays associated with Bt crops could enhance biocontrol services(7-12). However, this hypothesis has not been tested in terms of long-term landscape-level impacts(10). On the basis of data from 1990 to 2010 at 36 sites in six provinces of northern China, we show here a marked increase in abundance of three types of generalist arthropod predators (ladybirds, lacewings and spiders) and a decreased abundance of aphid pests associated with widespread adoption of Bt cotton and reduced insecticide sprays in this crop. We also found evidence that the predators might provide additional biocontrol services spilling over from Bt cotton fields onto neighbouring crops (maize, peanut and soybean). Our work extends results from general studies evaluating ecological effects of Bt crops(1-4,6,12,13) by demonstrating that such crops can promote biocontrol services in agricultural landscapes.

The Dachshund homolog 1 (Dach1) gene encodes a conserved co-repressor protein that, through its interaction with other nuclear proteins in the context of the local chromatin, participates in the regulation of genes essential for precursor cell proliferation and survival. DACH1 has been implicated in breast, prostate, ovarian and endometrial tumorigenesis. In breast cancer, reduced expression of DACH1 correlates with poor prognosis. Analysis of the mechanisms involved discovered that DACH1 binds and represses a subset of genes with important functions in tumorigenesis and tumor progression. By inhibiting chemokine expression, DACH1 reduces cellular migration, invasion, and metastatic capability of breast cancer cells. DACH1 also represses several AP-1-responsive genes, including the cell cycle regulator Cyclin D1. Recent studies demonstrated that endogenous DACH1 targets key regulators of stem cell identity in breast cancer cells. The present chapter summarizes the evidence that DACH1 regulates breast tumor progression and cancer stem cell expansion. We discuss the mechanisms involved.

The pink bollworm Pectinophora gossypiella is an invasive pest insect that has successfully established populations in many cotton growing regions around the world. In this study, the genetic diversity and population structure of Chinese populations of P. gossypiella were evaluated using mitochondrial DNA sequence data (COII and Nad4). For comparison, individuals of Pakistan and America were also sequenced at the same two mtDNA regions. Extremely low genetic variation was observed in the two mitochondrial regions among all populations examined. Most of the populations harbored only one to two haplotypes. Although the Nad4 region showed relatively high haplotype diversity and nucleotide variation, ranging from 0.363 to 0.591 and from 0.00078 to 0.00140, respectively, there were only three haplotypes observed in this region. COII and Nad4 haplotype networks shaved one or two common haplotype(s) forming the center of a star-shaped phylogeny. Pairwise tests showed that most of the populations were not significantly differentiated from each other. The Chinese populations were differentiated from the Pakistani and American populations in the Nad4 region. The low level of population genetic variation of P. gossypiella is attributed to invasion bottlenecks, which may have been subsequently strengthened by its nonmigratory biology and the mosaic pattern of agricultural activities.

The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most important pests in tropical and subtropical agriculture and is a key pest in greenhouse production worldwide. Current management of B. tabaci relies upon frequent applications of insecticides. Insecticide use not only directly affects pest populations through acute toxicity but also has indirect (sublethal) effects on pest physiology or behavior. In this study, we described sublethal effects of imidacloprid on adult feeding, immature development, adult fecundity, and F1 development of B. tabaci. Honeydew excretion of adults feeding on leaves treated with LC20 and LC40 concentration was significantly lower than that on untreated leaf discs. Egg production of B. tabaci adults subject to LC20 and LC40 concentrations also was less than untreated individuals. Upon transfer to untreated leaves, honeydew excretion and egg production recovered well within 24 and 48 h, respectively. Exposure to LC20 and LC40 concentrations significantly affected developmental time of B. tabaci eggs and nymphs, whereas it did not affect adult molting rate. We did not find sublethal effects on longevity and fecundity of B. tabaci adults when exposed to LC20 and LC40 concentrations for 24 h, and on egg hatching rate, nymphal mortality, and molting rate of the subsequent F1 generation. Exposure to imidacloprid at LC40 concentration significantly decreased the number of females in the F1 generation. Imidacloprid negatively affects development and reproduction of exposed individuals, and sex ratio of subsequent (F1) generation of B. tabaci, which probably disrupts B. tabaci population dynamics, slows population increase, and reduces infestation levels. Therefore, it is necessary to consider potential impact from imidacloprid for integrated management of the pest.

The lacewing Chrysoperla sinica (Tjeder) (Neuroptera: Chrysopidae) is an important predator of several insect pests in China and has considerable potential as a biological control agent. An inoculative approach would be the releasing adults early in the season to ensure that populations are present before pest densities increase. However, an understanding of adult flight activity under different conditions is necessary to develop appropriate release strategies. Therefore, we used a 32-channel, computer-monitored flight mill system to determine the effect of age on the flight activity of unmated female and male adults. Both sexes had high total flight activity levels as well as the longest individual flight bouts 2 and 3 d after emergence. The effects of temperature (between 13 and 33 degrees C at 75% RH) and relative humidity (between 30 and 90% RH at 23 degrees C) on the flight activity of 3-d-old unmated adults also were determined. Flight activity declined at the lowest (13 degrees C) and highest (33 degrees C) temperatures tested, as well as at the lowest relative humidity (30% RH). These findings are discussed within the context of selecting the appropriate environmental conditions for releasing C. sinica.

Use of transgenic maize expressing phytase in seeds as feedstuff can greatly increase phosphate availability to livestock and poultry. Because phosphorus is an essential mineral for all living organisms, growing of phytase transgenic maize may affect the performance of the arthropod community in maize fields. We conducted a preliminary study to assess the potential effects of phytase transgenic maize (BVLA430101) on two herbivore species, Ostrinia furnacalis (Guenee) and Helicoverpa armigera (Hubner), both of which are directly exposed to high concentrations of phytase caused by ingestion of transgenic maize kernels. Our results showed that for both species, survival and duration of the first and second instars and fresh weight of the third instar were not affected when fed transgenic phytase maize kernels compared with those fed nontransformed near isoline kernels. Similarly, there was no statistical difference detected for the same life table parameters when the herbivores were fed artificial diet containing either transgenic phytase maize meal or nontransformed maize meal. In addition, the nutrition utilization of the two species was evaluated with the same diet treatments by comparing the following indices: relative food consumption rate (RCR), relative metabolic rate (RMR), efficiency of approximate digestibility (EAD), efficiency of conversation of ingested food (ECI), and efficiency of conversation of digested food (ECD). No statistical difference was detected for any index of either species between transgenic maize and nontransformed maize treatments. These results provide useful baseline information for further studies to assess the potential effects of phytase transgenic maize on other arthropods in maize fields.

The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here (CsCad) showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region (CR7), which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin Cry1Ab. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.

Insect odorant binding proteins (OBPs) are required for insect olfaction perception and play a key role in transporting hydrophobic semiochemicals across the sensillum lymph to the olfactory receptors (Ors). We constructed two high-quality cDNA libraries from the male and female antennae of the lucerne plant bug, Adelphocoris lineolatus (Goeze) (Hemiptera: Miridae), a hemipteran species in a large and economically important phylogenetic group of phytophagous insects. A total of 1538 male ESTs and 1576 female ESTs were sequenced and analyzed, which produced 2915 high-quality ESTs for further analysis. The 2915 ESTs were assembled to 1423 unigenes. Of the 1423 unigenes, 895 (63%) showed no significant similarity with any known GenBank entry. The most prevalent transcripts in the cDNA libraries are OBPs, chemosensory proteins (CSPs), protein takeout precursors, antennae-specific proteins. We identified 14 genes encoding 12 “classical OBPs” with only six conserved cysteines and 2 “Plus-C OBPs” with two additional conserved cysteines and a conserved proline immediately after the sixth cysteine. AlinOBP4 has a very high amino acid identity of 89% to LAP, a well studied OBP of the tarnished plant bug Lygus lineolaris. The expression profiles of the 14 OBPs in different tissues (antennae, heads, thoraxes, abdomens, legs and wings) were measured by real-time qPCR. The results revealed some OBP genes are highly and differentially expressed in male and female antennae, and four OBP genes have an extremely high transcript level in the legs. Of two Plus-C OBP genes, one (AlinOBP14) is highly expressed only in the heads and another (AlinOBP7) is expressed in all olfactory tissues with much lower transcript levels comparing to other OBPs. The possible physiological functions of these OBPs are discussed.

The dachshund (dac) gene was initially described as a mutant phenotype in flies featuring extremely short legs relative to their body length. Functioning as a dominant suppressor of the ellipse mutation, a hypermorphic allele of the Epidermal Growth Factor Receptor (EGFR), the dac gene plays a key role in metazoan development, regulating ocular, limb, brain, and gonadal development. In the Drosophila eye, dac is a key component of the Retinal Determination Gene Network (RDGN) governing the normal initiation of the morphogenetic furrow and thereby eye development. Recent studies have demonstrated an important role for human Dachshund homologue (DACH1) in tumorigenesis, in particular, breast, prostate and ovarian cancer. The molecular mechanisms by which DACH1 regulates differentiation and tumorigenesis are discussed herein.

Aminopeptidase N (APN) and cadherin-like proteins have been previously identified as Cry1Ac-binding proteins in Helicoverpa armigera (Hubner). In this study, a proteomic approach was used to identify novel Cry1Ac-binding proteins in H. armigera. Brush border membrane vesicles (BBMV) of H. armigera were extracted and separated by two-dimensional gel electrophoresis (2-DE). Cry1Ac-binding proteins were detected using antisera against Cry1Ac. Peptide mass fingerprinting (PMF) was used to identify Cry1Ac-binding proteins. In total, four proteins were identified as candidate Cry1Ac-binding proteins in H. armigera: vacuolar ATP synthase (V-ATPase) subunit B, actin, heat shock cognate protein (HSCP), and a novel protein. © 2009 Wiley Periodicals, Inc.

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is an important lepidopteran pest of cotton, Gossypium hirsutum L., in Asia. Transgenic cotton expressing the cry1Ac gene from Bacillus thuringiensis (Bt) has been widely planted for control of this pest. For managing the potential risk from resistance evolution in this pest, a new transgenic Bt cotton containing cry1Ac and cry2Ab genes (gene pyramided strategy) was designed as a successor of cry1Ac cotton. This article reports on levels of Cry2Ab tolerance in H. armigera populations from CrylAc cotton planting region in China based on bioassays of F1 and F2 offspring of isofemale lines. In total, 572 isofemale families of H. armigera from Xiajin County of Shandong Province (an intensive Bt cotton-planting area) and 124 families from Anci County of Hebei Province [a multiple-crop system, including corn (Zea mays L.), soybean (Gycine max (L.) Merr., peanut (Arachis spp.), and Bt cotton] were screened with both Cry1Ac- and Cry2Ab-containing diets in 2008. The bioassays results indicated that relative average development rates (RADR) of F1 full-sib families from field-collected female moths on Cry1Ac- and Cry2Ab-containing diet were positively correlated. The same correlation was found in the F2 generation, indicating cross-tolerance between Cry1Ac and Cry2Ab in field populations of H. armigera in Yellow River cotton-farming region of China. This cross-tolerance must be considered in evaluating the utility of pyramiding Bt genes in cotton for delaying evolution of resistance.

The Dachshund (dac) gene, initially cloned as a dominant inhibitor of the Drosophila hyperactive EGFR mutant ellipse, encodes a key component of the cell fate determination pathway involved in Drosophila eye development. Analysis of more than 2,200 breast cancer samples showed improved survival by some 40 months in patients whose tumors expressed DACH1. Herein, DACH1 and estrogen receptor-α (ERα) expressions were inversely correlated in human breast cancer. DACH1 bound and inhibited ERα function. Nuclear DACH1 expression inhibited estradiol (E2)-induced DNA synthesis and cellular proliferation. DACH1 bound ERα in immunoprecipitation-Western blotting, associated with ERα in chromatin immunoprecipitation, and inhibited ERα transcriptional activity, requiring a conserved DS domain. Proteomic analysis identified proline, glutamic acid, and leucine rich protein 1 (PELP1) as a DACH1-binding protein. The DACH1 COOH terminus was required for binding to PELP1. DACH1 inhibited induction of ERα signaling. E2 recruited ERα and disengaged corepressors from DACH1 at an endogenous ER response element, allowing PELP1 to serve as an ERα coactivator. DACH1 expression, which is lost in poor prognosis human breast cancer, functions as an endogenous inhibitor of ERα function. [Cancer Res 2009;69(14):5752–60]

Transgenic cotton expressing the Bacillus thuringiensis (Bt) Cry1Ac toxin has been commercially cultivated in China since 1997, and by 2000 Bt cotton had almost completely replaced non-transgenic cotton cultivars. To evaluate the impact of Bt cotton planting on the seasonal population patterns of cotton bollworm, Helicoverpa armigera, the dynamics of H. armigera moths were monitored with light traps from four locations (Xiajin, Linqing and Dingtao of Shandong Province; Guantao of Hebei Province) in high Bt density region and five locations (Anci and Xinji of Hebei Province; Dancheng and Fengqiu of Henan Province; Gaomi of Shandong Province) in low Bt density region from 1996 to 2008. A negative correlation was found between moth densities of H. armigera and the planting years of Bt cotton in both high and low Bt density areas. These data indicate that the moth population density of H. armigera was reduced with the introduction of Bt cotton in northern China. Three generations of moths occurred between early June and late September in the cotton regions. Interestingly, second-generation moths decreased and seemed to vanish in recent years in high Bt density region, but this tendency was not found in low Bt density region. The data suggest that the planting of Bt cotton in high Bt density region was effective in controlling the population density of second-generation moths. Furthermore, the seasonal change of moth patterns associated with Bt cotton planting may regulate the regional occurrence and population development of this migratory insect.

The seasonal migration of the Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) over the Bohai Sea was observed with a searchlight trap and an entomological radar located on a small island in the center of the sea, and through a network of light-traps around the Bohai region. The H. armigera moths were observed to migrate over the sea at least as early as May and light trapping through a network suggested migration might start as early as April, as soon as the moths had emerged from overwintering pupae. H. armigera moths migrated toward the north in southerly winds during spring and summer, and returned south on nights with northerly winds, or at altitudes where the wind was northerly, during fall. The passage of a weather front (cold or warm) or trough at approximately 1700 hours provokes migration of H. armigera over the sea. The H. armigera generally flew at altitudes of below 1,500 m above sea level (asl) with layer concentrations at 200-500 m asl, where the wind direction, wind speed, and temperature were optimum. During fall migration, H. armigera tended to orient toward the southwest and was able to compensate for the wind drift by turning clockwise when the downwind direction was225 degrees but counterclockwise when it was225 degrees. The displacement speed measured with the radar was 24-41 km/h, the duration of flight was 8-11 h and the maximum migration rate was 1,894 moths per km per h.

Seven synthetic herbivore-induced plant volatiles (HIPVs) and a mixture of nonanal + (Z)-3-hexen-1-ol were field tested for their ability to attract beneficial insects in an open cotton field. Eleven species of the main natural enemies of insect pests in cotton fields were studied. Significantly greater numbers of the ladybird beetle Coccinella septempunctata were trapped on (Z)-3-hexenyl acetate-baited cards than on others that were HIPV baited or the control cards. Erigonidium graminicolum was attracted to traps baited with nonanal, (Z)-3-hexenyl acetate, and methyl salicylate (MeSA). The predatory bug Deraeocoris punctulatus was only attracted to traps baited with octanal. The predatory bug Orius similis responded to traps baited with 3,7-dimethyl,1,3,6-octatriene, nonanal, (Z)-3-hexenyl acetate, nonanal + (Z)-3-hexen-1-ol, and MeSA. Dimethyl octatriene, nonanal + (Z)-3-hexen-1-ol, and octanal significantly attracted the syrphid fly Paragus quadrifasciatus. The ladybird beetle Propylaea japonica, the green lacewing Chrysopa sinica, the bigeyed bug Geocoris pallidipennis, the syrphid fly Epistrophe balteata, and the parasitic wasp Campoletis chlorideae did not respond to any of the HIPVs tested. These results are discussed with regard to the potential of HIPVs as tools for recruiting natural enemies into cotton fields.

Various pesticides directly impact insect pest populations through acute toxicity (lethal) effects, but they also cause a range of indirect (sublethal) effects on pest physiology or behavior. In this study, we describe both lethal and sublethal effects of endosulfan on the survival, development, and fecundity of Apolygus lucorum Meyer-Dür (Hemiptera: Miridae), an important pest of Bacillus thuringiensis (Bt) cotton, Gossypium hirsutum L., in China. We quantify pesticide effects on adult mirids as well as on subsequent (F1, F2) generations. Toxicity bioassays determined the median lethal concentration (LC50) for adult A. lucorum as 0.58 ppm. To evaluate the sublethal effect of endosulfan, we used LC20 and LC40 concentrations at 0.30 ppm and 0.48 ppm, respectively. Sublethal concentrations of endosulfan significantly reduced longevity and fecundity of female adults, but slightly affected male longevity. We also found sublethal effects on the F1 generation, through a reduction of egg hatching rate, nymphal mortality, adult molting rate, and adult fecundity, and on the F2 generation, through a decrease of egg hatching rate. These results show that endosulfan has far greater effects on A. lucorum than estimated by acute toxicity tests, and the effects of this pesticide on surviving mirids can be comparatively long-lived. This pesticide likely causes severe disruptions of A. lucorum population dynamics, with single endosulfan application, invoking a great reduction of population growth of resident mirid populations. We discuss implications of this work for control programs for A. lucorum on Bt cotton.

Genetically modified insect-resistant rice lines containing the cry1Ac gene from Bacillus thuringiensis (Bt) or the CpTI (cowpea trypsin inhibitor) gene developed for the management of lepidopterous pests are highly resistant to the major target pests, Chilo suppressalis (Walker), Cnaphalocrocis medinalis (Guenée), and Scirpophaga incertulas (Walker), in the main rice-growing areas of China. However, the effects of these transgenic lines on Sesamia inferens (Walker), an important lepidopterous rice pest, are currently unknown. Because different insect species have varying susceptibility to Bt insecticidal proteins that may affect population dynamics, research into the effects of these transgenic rice lines on the population dynamics of S. inferens was conducted in Fuzhou, southern China, in 2005 and 2006. The results of laboratory, field cage, and field plot experiments show that S. inferens has comparatively high susceptibility to the transgenic line during the early growing season, with significant differences observed in larval density and infestation levels between transgenic and control lines. Because of a decrease in Cry1Ac levels in the plant as it ages, the transgenic line provided only a low potential for population suppression late in the growing season. There is a correlation between the changing expression of Cry1Ac and the impact of transgenic rice on the population dynamics of S. inferens during the season. These results indicate that S. inferens may become a major pest in fields of prospective commercially released transgenic rice, and more attention should be paid to developing an effective alternative management strategy.

Genetically modified cotton that produces a crystalline protein from Bacillus thuringiensis subsp. kurstaki (Berliner) (Bt) has been widely deployed to manage lepidopteran insect pests in cotton growing areas worldwide. However, susceptibility of different insect species to Bt protein varies, which may affect lepidopteran pest populations in the field. Studies on effects of two transgenic cotton lines (BG1560 and GK19) carrying a Cry1A gene on common cutworm Spodoptera litura F. (Lepidoptera: Noctuidae), were conducted during 2002-2005 in the cotton planting region of the Yangtze River valley of China. Results showed that common cutworm larvae had low susceptibility to Bt cotton. There was no significant difference in larval population densities in conventional and Bt cotton fields. However, the larval populations of the insect on conventional plants treated with chemical insecticides for control of target pest of Bt cotton were significantly lower than that in Bt cotton fields. These results indicated that the common cutworm was the potential to become a major and alarming pest in Bt cotton fields, and therefore efforts to develop an effective alternative management strategy are needed.

Transgenic cotton that has been engineered to produce insecticidal toxins from Bacillus thuringiensis (Bt) and so to resist the pest cotton bollworm ( Helicoverpa armigera ) has been widely planted in Asia. Analysis of the population dynamics of H. armigera from 1992 to 2007 in China indicated that a marked decrease in regional outbreaks of this pest in multiple crops was associated with the planting of Bt cotton. The study area included six provinces in northern China with an annual total of 3 million hectares of cotton and 22 million hectares of other crops (corn, peanuts, soybeans, and vegetables) grown by more than 10 million resource-poor farmers. Our data suggest that Bt cotton not only controls H. armigera on transgenic cotton designed to resist this pest but also may reduce its presence on other host crops and may decrease the need for insecticide sprays in general.

The soybean aphid, Aphis glycines (Matsumura), is native to eastern Asia and has recently invaded North America, where it is currently the most important insect pest of soybeans. The soybean aphid has spread rapidly within North America, presumably through a combination of active and passive (wind-aided) flight. Here, we studied the active flight potential of A. glycines under a range of environmental conditions using an aphid flight mill. Winged (alate) A. glycines were tested on a specially designed 32-channel, computer-monitored flight mill system. Aphids that were 12-24 h old exhibited the strongest flight behavior, with average flight durations of 3.3-4.1 h, which represented flight distances of 4.6-5.1 km. After the age of 72 h, A. glycines flight performance rapidly declined. The optimum temperature range for flight was 16-28 degrees C, whereas optimum relative humidity was 75%. Our findings show that A. glycines posseses a fairly strong active flight aptitude (ability and inclination) and point to the possibility of flight initiation under a broad range of environmental conditions. These results have the potential to aid forecasting and management protocols for A. glycines at the landscape level.

To clarify the environmental fate of the Cry1Ac protein from Bacillus thuringiensis subsp. kurstaki (Bt) contained in transgenic rice plant stubble after harvest, degradation was monitored under field conditions using an enzyme-linked immunosorbent assay. In stalks, Cry1Ac protein concentration decreased rapidly to 50% of the initial amount during the first month after harvest; subsequently, the degradation decreased gradually reaching 21.3% when the experiment was terminated after 7 mo. A similar degradation pattern of the Cry1Ac protein was observed in rice roots. However, when the temperature increased in April of the following spring, protein degradation resumed, and no protein could be detected by the end of the experiment. In addition, a laboratory experiment was conducted to study the persistence of Cry1Ac protein released from rice tissue in water and paddy soil. The protein released from leaves degraded rapidly in paddy soil under flooded conditions during the first 20 d and plateaued until the termination of this trial at 135 d, when 15.3% of the initial amount was still detectable. In water, the Cry1Ac protein degraded more slowly than in soil but never entered a relatively stable phase as in soil. The degradation rate of Cry1Ac protein was significantly faster in nonsterile water than in sterile water. These results indicate that the soil environment can increase the degradation of Bt protein contained in plant residues. Therefore, plowing a field immediately after harvest could be an effective method for decreasing the persistence of Bt protein in transgenic rice fields.

Lygus lucorum (Meyer-Dür) is a key pest of Bt cotton in China. This study reports on its flight potential examined by a flight-mill system. We found that 10-d-old mated females engaged in flight over the greatest distance (40.1 +/- 5.2 km) and duration (7.7 +/- 1.0 h) in 24-h flight assays in relation to age, sex, and mating status. Optimum temperature for flight was 20 degrees C, and optimum relative humidity was 75% RH. Flight potential of 10-d-old mated females under the optimum conditions (20 degrees C and 75% RH) was tested continuously for 48 h. Results showed that the flight distance amounted to 67.3 +/- 9.7 km, with a maximum distance of 151.3 km. This study shows that L. lucorum has the potential to undertake long-distance flight. The information will help in the development of the forecast and management of L. lucorum.

Transgenic cottons producing Cry toxins from Bacillus thuringiensis (Bt) provide for control of lepidopteran pests and were first commercially grown in Australia, Mexico and the USA in 1996. As of 2007, a total of six additional countries (Argentina, Brazil, China, Colombia, India, and South Africa) now grow Bt cotton on a total production area of 14 million hectares. The technology primarily provides highly selective and effective control of bollworms, which are the most damaging pests of cotton worldwide. It is estimated that between 1996 and 2005 the deployment of Bt cotton has reduced the volume of insecticide active ingredient used for pest control in cotton by 94.5 million kilograms and increased farm income through reduced costs and improved yields by US$7.5 billion, with most of the benefit accrued by farmers in developing nations. Reductions in insecticide use have broadened opportunities for biological control of all cotton pests but most other pest management tactics have remained largely unchanged by the use of Bt cotton. However, several non-target pests have become more problematic in Bt cotton fields in some countries largely due to reductions in insecticide use for target pests. After 11 years of Bt cotton cultivation, control failures due to resistance have not been detected under field conditions. This success can be largely credited to pre-emptive resistance management based on mandated refuges and monitoring programs as well as non-mandated refuge crops and natural refuges which collectively act to dilute any resistant alleles in pest populations. New products are in the pipeline to improve the effectiveness of genetically modified cotton cultivars for resistance to lepidopteran pests, and to address other pest problems in cotton. Debate over food and environmental safety, regulatory oversight, and farming community welfare are likely to continue as the technology moves forward with new crops and new adopting countries.

Bioassays and binding tests between Cry toxins (CrylAa, CrylAb, CrylAc, and Cry2Ab) and brush-border membrane vesicles (BBMVs) from larvae of a Bacillus thurningiensis (Bt)-susceptible (96S) and two CrylAc-resistant strains (BtR and LFR10) were conducted for investigating cross-resistance of CrylAc-resistant strains to Cry2Ab in Helicoverpa armigera (Hiibner) (Lepidoptera: Noctuidae). The resistance ratio (RR) values of the BtR and LFR10 strains to CrylAc and Cry2Ab were 2,971- and 1.1-fold and 253- and 1.0-fold, respectively, indicating that there was no cross-resistance to Cry2Ab. The binding experiments between Cry toxins and BBMVs from BtR, LFR10, and 96S larvae showed that all of the toxins could bind with these BBMVs, but the Cry2Ab could not displace 125I labeled CrylAc and CrylAb. The same results were observed in reciprocal binding tests, demonstrating that CrylA and Cry2Ab had different binding sites in H. armigera and providing a potential mechanism for the lack of cross-resistance between CrylA and Cry2Ab toxins. These results suggest that the transgenic cotton, Gossypium hirsutum L., expressing CrylAc and Cry2Ab genes may be deployed for management of CrylAc resistant H. armigera.

The onset of human lung cancer involves the loss of tumor suppressor genes and activation of tumor inducing genes. Current studies have proved EGFR, RAS, ALK as as key driver genes, however, in about 50% lung cancer samples there are no known genes to explain tumorigenesis, suggesting many novel pathways remain to be elucidated. Recent study on whole-genome and transcriptome sequencing NSCLC identified cell fate determinatiton gene DACH1 as novel alteration in genes involved in NSCLC. However, the role of DACH1 in NSCLC and its downstrem targets are largely unknown. We showed that expression of DACH1 was significantly decreased in human NSCLC tissues andits protein abundance was inversely correlated with the tumor stage and grade. Restoration of DACH1 function in NSCLC cells significantly reduced cellular proliferation, clone formation, migrasion and invasion in vitro, as well as tumor growth in vivo. By unbiased screen of secreting factors, we found out that CXCL5 was dramaticcaly repressed by DACH1. Furthermore, functional study suggested that CXCL5 was contribute to the DACH1-mediated repression of migrative in vitro and tumorigenesis in vivo. Consistingly, there was a inverse relationship between the mRNA level of DACH1 and CXCL5 in both lung cancer cell lines and huamn NSCLC tissues. Moreover, Kaplan-Meier analysis of human NSCLC samples demonstrated that high mRNA abundance of DACH1 was predicted favorate prognosis for both relaspse-free survivl and overall survival; in contrast, high CXCL5 expression predict worse prognosis for survival. To our understanding, this is the first study to demonstrate DACH1 regulated paracrine signaling in lung cancer and DACH1 could predict prognosis. In conclusion, DACH1 acts as a tumor suppressor gene through repression of paracrine signaling and is a potential biomarker for prognosis. Citation Format: Na Han, Xun Yuan, Hua Wu, Hanxiao Xu, Qian Chu, Mingzhou Guo, Shiying Yu, Yuan Chen, Kongming Wu. Cell fate determination factor DACH1 inhibits lung adenocarcinoma invasion and tumor growth through repression of CXCL5 signaling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 515. doi:10.1158/1538-7445.AM2015-515

Various studies have evaluated the significance of Notch1 expression in breast cancer, but the results have ever been disputed. By using 21 studies involving 3867 patients, this meta-analysis revealed that the expression of Notch1 was significantly higher in breast cancer than in normal tissues (OR=7.21; 95%CI, 4.7-11.07) and that higher Notch1 expression was associated with transition from ductal carcinoma in situ (DCIS) to invasive cancer (OR=3.75; 95% CI, 1.8-7.78). Higher Notch1 activity was observed in the basal subtype of breast cancer (OR=2.53; 95% CI, 1.18-5.43). Moreover, patients with Notch1 overexpression exhibited significantly worse overall and recurrence-free survival. Our meta-analysis suggests that Notch inhibitors may be useful in blocking the early progression of DCIS and that the outcomes of clinical trials for Notch1-targeting therapeutics could be improved by the molecular stratification of breast cancer patients.

Polygalacturonase (PG) is an enzyme in the salivary glands of piercing-sucking mirid bugs (Hemiptera: Miridae) that plays a key role in plant feeding and injury. By constructing a full-length cDNA library, we cloned and characterized 14 PG genes from the salivary glands of Apolygus lucorum, a pestiferous mirid bug in cotton, fruit trees and other crops in China. BLAST search analysis showed that the amino acid sequences deduced from transcripts of the PG genes were closely related to PGs from other mirid bugs. Phylogenetic analysis showed that the PGs of mirid bugs had six main branches, PG1-PG6 (Genbank accession numbers: KF881899~KF881912). We investigated the mRNA expression patterns of the A. lucorum PG genes using real-time PCR. All 14 PGs were expressed significantly higher in the salivary glands than in other tissues (head, thorax, abdomen, leg and wing). For eggs and nymphs, the expression levels of these PGs were much higher in the 5th instar stage than in the egg, and 1st and 3rd instar stages. The PG expression levels in 1-day-old adults were very low, and increased in 5, 20 and 30-day-old adults. Additionally, PG expression levels were generally similar between males and females. The possible physiological functions of PGs in A. lucorum were discussed.

Obstacles to the expansion of cells with proliferative potential include the induction of cell death, telomere-based senescence, and the pRb and p53 tumor suppressors. Not infrequently, the molecular pathways regulating oncogenesis recapitulate aberrations of processes governing embryogenesis. The genetic network, consisting of the dachshund (dac), eyes absent (eya), eyeless, and sine oculis (so) genes, regulates cell fate determination in metazoans, with dac serving as a cointegrator through a So DNA-binding factor. Here, DACH1 inhibited oncogene-mediated breast oncogenesis, blocking breast cancer epithelial cell DNA synthesis, colony formation, growth in Matrigel, and tumor growth in mice. Genetic deletion studies demonstrated a requirement for cyclin D1 in DACH1-mediated inhibition of DNA synthesis. DACH1 repressed cyclin D1 through a novel mechanism via a c-Jun DNA-binding partner, requiring the DACH1 alpha-helical DS domain which recruits corepressors to the local chromatin. Analysis of over 2,000 patients demonstrated increased nuclear DACH1 expression correlated inversely with cellular mitosis and predicted improved breast cancer patient survival. The cell fate determination factor, DACH1, arrests breast tumor proliferation and growth in vivo providing a new mechanistic and potential therapeutic insight into this common disease.

Ovarian cancer is resistant to the antiproliferative effects of transforming growth factor-β (TGF-β); however, the mechanism of this resistance remains unclear. We used oligonucleotide arrays to profile 37 undissected, 68 microdissected advanced-stage, and 14 microdissected early-stage papillary serous cancers to identify signaling pathways involved in ovarian cancer. A total of seven genes involved in TGF-β signaling were identified that had altered expression >1.5-fold (P < 0.001) in the ovarian cancer specimens compared with normal ovarian surface epithelium. The expression of these genes was coordinately altered: genes that inhibit TGF-β signaling (DACH1, BMP7, and EVI1) were up-regulated in advanced-stage ovarian cancers and, conversely, genes that enhance TGF-β signaling (PCAF, TFE3, TGFBRII, and SMAD4) were down-regulated compared with the normal samples. The microarray data for DACH1 and EVI1 were validated using quantitative real-time PCR on 22 microdissected ovarian cancer specimens. The EVI1 gene locus was amplified in 43% of the tumors, and there was a significant correlation (P = 0.029) between gene copy number and EVI1 gene expression. No amplification at the DACH1 locus was found in any of the samples. DACH1 and EVI1 inhibited TGF-β signaling in immortalized normal ovarian epithelial cells, and a dominant-negative DACH1, DACH1-ΔDS, partially restored signaling in an ovarian cancer cell line resistant to TGF-β. These results suggest that altered expression of these genes is responsible for disrupted TGF-β signaling in ovarian cancer and they may be useful as new and novel therapeutic targets for ovarian cancer. (Cancer Res 2006; 66(17): 8404-12)

Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin-like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect’s resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence, suggesting that the decrease of expression quantity of BtR-harm is one of the main reasons for H. armigera resistance to Cry1Ac.

Pax6 and c-Maf regulate multiple stages of mammalian lens development. Here, we identified novel distal control regions (DCRs) of the alphaA-crystallin gene, a marker of lens fiber cell differentiation induced by FGF-signaling. DCR1 stimulated reporter gene expression in primary lens explants treated with FGF2 linking FGF-signaling with alphaA-crystallin synthesis. A DCR1/alphaA-crystallin promoter (including DCR2) coupled with EGFP virtually recapitulated the expression pattern of alphaA-crystallin in lens epithelium and fibers. In contrast, the DCR3/alphaA/EGFP reporter was expressed only in 'late' lens fibers. Chromatin immunoprecipitations showed binding of Pax6 to DCR1 and the alphaA-crystallin promoter in lens chromatin and demonstrated that high levels of alphaA-crystallin expression correlate with increased binding of c-Maf and CREB to the promoter and of CREB to DCR3, a broad domain of histone H3K9-hyperacetylation extending from DCR1 to DCR3, and increased abundance of chromatin remodeling enzymes Brg1 and Snf2h at the alphaA-crystallin locus. Our data demonstrate a novel mechanism of Pax6, c-Maf and CREB function, through regulation of chromatin-remodeling enzymes, and suggest a multistage model for the activation of alphaA-crystallin during lens differentiation.