Aquaporins (AQPs) are integral membrane proteins that can transport water and small molecules across cell membranes in organisms.In a previous report, we identifi ed an AQP (SlAQP1) gene of the cotton leafworm, Spodoptera littoralis.In the current study, we identify and characterize two more SlAQP genes and their developmental and tissues expression.Predicted amino acid sequence and phylogenetic analysis revealed that SlAQP2 is a glycerol channel belonging to the insect specifi c Eglp subfamily, whereas SlAQP3 is a water specifi c channel belonging to the Prip subfamily.SlAQP2 expression was detected mainly in the fi rst four larval instars and adult males, whereas SlAQP3 was detected in all developmental stages.SlAQP2 gene expression was mainly detected in larval midgut and Malpighian tubules and adult male testes, whereas SlAQP3 was detected in all the tissues tested.RNAi mediated knockdown of each gene separately resulted in deleterious effects including larval and pupal mortality, deformed pupae and adults and prolonged development.Results were confi rmed using qRT-PCR, which revealed downregulation of both genes after injection of larvae with gene specifi c dsRNA.Our results confi rm the presence of Prip and Eglp AQPs in S. littoralis and suggest that Eglp has a role in male reproductive ability.

With increasing reports of resistance to traditional insecticides, there is a need for innovative ways for mosquito control. RNAi is a sequence-specific molecular biology technique for gene silencing through degradation of mRNA and prevention of protein translation. Some genes are essential for insect life and their silencing can lead to insect morbidity and/or mortality. Searching for lethal genes in Culex quinquefasciatus, we found dynamin, ROP, HMGR and JHAMT to be lethal targets for RNAi in initial screening through larval soaking in dsRNA solution. Two delivery methods, chitosan nanoparticles and genetically modified yeast cells, were used in this study and proved effective in inducing high larval mortality and low adult emergence. Adult emergence after chitosan nanoparticles/dsRNA treatment was 12.67% ± 1.76 (HMGR), 17.33% ± 1.76 (dynamin), 18.67% ± 0.67 (ROP), and 35.33% ± 0.67 (JHAMT). Genetically modified yeast increased mortalities as adult emergence was 8.33% ± 1.67 (HMGR), 13.33% ± 3.33 (dynamin), and 10% ± 2.89 (JHAMT and ROP). Chitosan nanoparticles retained 75% of its biological activity whereas yeast cells retained >95% of their activities after 7 days of incubation in water. In conclusion, our results showed that these four genes are good targets for C. quinquefasciatus control using RNAi packaged in either chitosan nanoparticles or genetically modified yeast cells.

Culex quinquefasciatus, a member of the Culex pipiens complex, is widespread in Saudi Arabia and other parts of the world. It is a vector for lymphatic filariasis, Rift Valley fever, and West Nile virus. Studies have shown the deleterious effect of RNA interference (RNAi)-mediated knockdown of various lethal genes in model and agricultural pest insects. RNAi was proposed as a tool for mosquito control with a focus on Aedes aegypti and Anopheles gambiae. In this study, we examined the effect of RNAi of selected target genes on both larval mortality and adult emergence of Cx. quinquefasciatus through two delivery methods: soaking and nanoparticles. Ten candidate genes were selected for RNAi based on their known lethal effect in other insects. Disruption of three genes, chitin synthase-1, inhibitor of apoptosis 1, and vacuolar adenosine triphosphatase, resulted in the highest mortality among the selected genes using the two treatment methods. Silencing the other seven genes resulted in a medium to low mortality in both assays. These three genes are also active against a wide range of insects and could be used for RNAi-based mosquito control in the future.

Jojoba has become a significant plant due to its precious seed oil with numerous applications. In an attempt to explore the genetic variation and the association with different traits, ISSR analysis was conducted for the selected Jojoba clones. In the present study, sixteen Jojoba plants (clones) including 1 male and 15 female clone were selected to conduct this experiment. Ten ISSR primers were used in this study. The analysis of oil content in the seeds revealed that one plant (GIADC2) contain up to 55.93% oil, which is quite high percentage. Increasing in oil content and seed weight is not always related to each other. ISSR analysis revealed 61 monomorphic bands, where, the maximum number of monomorphic bands (9) was generated by the ISSR-3, while the lowest (1) produced by the ISSR-9. However, the total number of polymorphic bands was 70. The percentage of polymorphism was ranged from 20% (ISSR-20) to 91% (ISSR-9) with average 52.6%. The dendrogram showed two main clusters; the first main cluster has grouped three Jojoba genotypes (5, 10 and 15). While the second main cluster was divided into two sub-clusters; one sub-cluster contain 10 clones (4, 6, 8, 12, 9, 14, male, 13, 1 and 11) and the other sub-cluster contained three Jojoba clones (2, 7 and 3). The association of ISSR marker with oil content revealed that ISSR-01, ISSR-08 and ISSR-19 were associated with oil content. On the hand, the association of ISSR marker with seed weight revealed that ISSR-01, ISSR-04, ISSR-05, ISSR-09, ISSR-19 and ISSR-20 primers were associated. Finally, molecular marker such as ISSR markers is important in trait marker association study for improving plant cultivars.

Ultraviolet radiation has negative effects on different living organisms. The effects of ultraviolet radiation have been widely studied on some insects, such as stored‐product pests. However, few studies have examined the effects of ultraviolet radiation on mosquitoes. In this study, we report the effect of ultraviolet radiation type C, or ultraviolet C, on the immature stages of Culex quinquefasciatus. Our results suggest that early larval instars were more susceptible to ultraviolet C than late instars, and longer ultraviolet C exposure times were more lethal to larvae regardless of age. Ultraviolet C caused 100% mortality in pupae after 11 minutes of exposure. No egg hatching was observed after 15 minutes of exposure. Mortality in immature stages occurred over time, especially during larval molting and adult emergence.

Mosquito-transmitted diseases pose a threat for a great portion of the world population. Chemical insecticides are the main tool for mosquito control. Heavy dependence on chemicals created several problems such as resistance development in many mosquito species, environmental effects, and human health issues. Other tools for mosquito control were developed and used in some parts of the world. Ribonucleic acid interference (RNAi) is a reverse genetic mechanism that was recently introduced as a new tool for pest control. Regarding mosquito, RNAi was used to study gene function and to discover genes that can be used as targets for control purposes. Several delivery methods are used to induce RNAi in mosquito larvae. Some methods such as injection and soaking are used routinely in RNAi research but have no application in the field. Other methods such as nanoparticles and microbes have some characteristics that make them good candidates for field application. In this report, we will focus on delivery methods for RNAi in mosquito larvae and will give examples for each method.

Precise identification of jojoba plant species using DNA barcode is very important because of the economic value of its seed oil. In this study, we selected 16 jojoba trees (clones) including one male and 15 female trees cultivated in the same farm to conduct this experiment. DNA samples were extracted from leaf tissues and subjected to PCR amplification using specific primers for rbcL gene fragment. The amplification was successful for all tested samples producing the same fragment with a size about 550 bp. Sequence analysis was performed for the purified products and consequently subjected to GenBank database analysis. The data analysis produced by BLASTn database revealed that the similarity search was from 98-100% with rbcL gene region form available DNA sequence in GenBank. Multiple sequence alignment was performed after confirmation of the DNA sequence. The alignment clearly indicated the high similarity between all DNA sequences for the tested samples. Furthermore, a phylogenetic tree was produced from the alignment process which revealed the close relationship between the tested samples. The tree can be divided into two main branches one containing samples 1-13 and 16 while the other branch contains samples 14 and 15. These results indicate that the rbcL region is a good option for molecular identification of jojoba species. Finally, the obtained sequence data were submitted to GenBank to provide the accession numbers for the nucleotide sequences.

BACKGROUND:Mosquito-borne pathogens are important causes of diseases in the Kingdom of Saudi Arabia. Knowledge of the mosquito fauna is needed for the appropriate control of the vectors that transmit the pathogens and prevent the diseases they cause. An important first step is to have an up-to-date list of the species known to be present in the country. Original occurrence records were obtained from published literature and critically scrutinized to compile a list of the mosquito species that occur within the borders of the Kingdom.RESULTS:Fifty-one species have been recorded in the Kingdom; however, the occurrence of two of these species is unlikely. Thus, the mosquito fauna of the Kingdom comprises 49 species that include 18 anophelines and 31 culicines. Published records are provided for each species. Problematic records based on misidentifications and inappropriate sources are discussed and annotated for clarity.CONCLUSION:Integrated morphological and molecular methods of identification are needed to refine the list of species and accurately document their distributions in the Kingdom.

Trypsin Modulating Oostatic Factor (TMOF) is a decapeptide hormone that inhibits the biosynthesis of digestive enzymes in the mosquito midgut. The hormone inhibits food digestion and ultimately leads to starvation and death. It has been used as a biological insecticide to control mosquitoes. In an attempt to increase the insecticidal activity of TMOF, a combination of CryIC (-endotoxin from Bacillus thuringiensis) and TMOF was determined. Eight recombinant proteins fused with GST (glutathione-S-transferase) were expressed in Escherichia coli cells. Their insecticidal activities were determined against Culex pipiens and Spodoptera littoralis larvae. Purified GST-TMOF and its analogue GST-YDPAS exhibited a moderate toxicity on C. pipiens larvae with LC50 of 145.9 and 339.9g/mL, respectively. Unexpectedly, no mortality was observed in first instar larvae of S. littoralis. Puirified GST-TMOF and GST-YDPAS together with Bt toxin showed a synergistic toxic effect on both Culex and Spodoptera larvae. In the presence of 100g/mL GST-TMOF and GST-YDPAS, the median lethal concentration of entomocidus on culex larvae decreased from 52.1 to 16.7 and 31.9g/mL, respectively. Likewise, GST-TMOF and GST-YDPAS incorporated with 0.07g/cm(2) of enotmocidus showed insecticidal activity against S. littoralis with LC50 of 16.4 and 21.9g/cm(2). The E. coli lysates containing GST-CryIC and its 3-truncated version showed low toxicity against the lepidopteran insect (10.8 and 16.6g/cm(2)) compared to 0.15g/cm(2) of the native crystalline form of CryIC. Similarly, the mosquitocidal activity of the recombinant Bt toxins was low.

Aquaporins (AQPs) are integral membrane proteins belong to the Major Intrinsic Proteins (MIP) superfamily identified in different tissues from all living organisms including mammals, plants, invertebrates and microorganisms. Only few AQPs isolated and studied from terrestrial insects. In the present study, we report the cloning of a partial cDNA sequence of AQP1 gene from Spodoptera littoralis. The cDNA is 1148bp with an open reading frame of 684bp encoding for 227 amino acids with similarity of 98% and 76% to Spodoptera litura and Grapholita molesta, respectively. Protein structure analysis showed the presence of conserved features found in all studied AQPs such as six transmembrane domains, two conserved motifs of NPA, and the aromatic/arginine (ar/R) selectivity filter. Our study revealed that AQP1 gene is expressed in all insect developmental stages (eggs, larvae, pupae, and adults including males and females). Furthermore, AQP1 expression was detected in tissues of the digestive and excretory system (foregut, midgut, hindgut and Malpighian tubules) of fourth, fifth and sixth larval instars. Gene expression was also detected in the reproductive system of adult females and males. Therefore, we suggest that this AQP1 gene has critical role in maintaining water balance in Spodoptera littoralis insects.

Chitin is a major component of insect exoskeleton, tracheal system and gut where it is synthesized by chitin synthase (CHS) enzymes. In this paper, we report the isolation and RNAi of chitin synthase A ( PhoCHSA ) from the potato tuber moth Phthorimaea operculella . The full-length cDNA of PhoCHSA is 5,627 bp with 4,689 bp open reading frame coding for 1,563 amino acids. Structural analysis of conceptual amino acid translation showed three distinct regions found in all known insect CHS proteins; N-terminus region having 9 transmembrane helices, middle catalytic region containing several conserved domains identified in insect CHS enzymes, and C-terminus region containing seven transmembrane spans. Phylogenetic analysis showed that PhoCHSA protein clustered with CHSA enzymes identified from insects from different insect orders. RNAi targeting three different regions of the gene showed different efficacy against potato tuber moth larvae and dsRNA targeting the 5′ region has the highest efficacy. Results were verified by qRT-PCR which showed that dsRNA targeting the 5′ region caused the highest reduction in PhoCHSA mRNA level. Our results show the importance of selecting the RNAi target region and that chitin synthase A can be a suitable RNAi target for the potato tuber moth control.

Juvenile hormone (JH) controls the growth, development, metamorphosis, and reproduction of insects. For many years, the general assumption has been that JH regulates tick and other acarine development and reproduction the same as in insects. Although researchers have not been able to find the common insect JHs in hard and soft tick species and JH applications appear to have no effect on tick development, it is difficult to prove the negative or to determine whether precursors to JH are made in ticks. The tick synganglion contains regions which are homologous to the corpora allata, the biosynthetic source for JH in insects. Next-gen sequencing of the tick synganglion transcriptome was conducted separately in adults of the American dog tick, Dermacentor variabilis, the deer tick, Ixodes scapularis, and the relapsing fever tick, Ornithodoros turicata as a new approach to determine whether ticks can make JH or a JH precursor. All of the enzymes that make up the mevalonate pathway from acetyl-CoA to farnesyl diphosphate (acetoacetyl-CoA thiolase, HMG-S, HMG-R, mevalonate kinase, phosphomevalonate kinase, diphosphomevalonate decarboxylase, and farnesyl diphosphate synthase) were found in at least one of the ticks studied but most were found in all three species. Sequence analysis of the last enzyme in the mevalonate pathway, farnesyl diphosphate synthase, demonstrated conservation of the seven prenyltransferase regions and the aspartate rich motifs within those regions typical of this enzyme. In the JH branch from farnesyl diphosphate to JH III, we found a putative farnesol oxidase used for the conversion of farnesol to farnesal in the synganglion transcriptome of I. scapularis and D. variabilis. Methyltransferases (MTs) that add a methyl group to farnesoic acid to make methyl farnesoate were present in all of the ticks studied with similarities as high as 36% at the amino acid level to insect JH acid methyltransferase (JHAMT). However, when the tick MTs were compared to the known insect JHAMTs from several insect species at the amino acid level, the former lacked the farnesoic acid binding motif typical in insects. The P450s shown in insects to add the C10,11 epoxide to methyl farnesoate, are in the CYP15 family; this family was absent in our tick transcriptomes and in the I. scapularis genome, the only tick genome available. These data suggest that ticks do not synthesize JH III but have the mevalonate pathway and may produce a JH III precursor.

Sugarcane plants of cv. Ph8013, G98/28, G98/24 and GT54-C9 were used for somaclonal variation induction using the spindle leaves as ex-plants. The used varieties differed according to the vegetative traits, juice quality and their similarity index. The effect of five concentrations from 2,4-D and three concentrations of coconut water were examined for the somatic embryogenesis induction. The 2, 4-D was applied at 0, 1, 2, 3, 4 and 5 mg/l in combination with 0, 5 and 10 g/l coconut water. Also, the effect of using NAA at five concentrations i.e. 0, 1, 3, 5 and 7 mg/l in combination with five concentrations of sucrose i.e. 20, 30, 40 50 and 60 g/l were examined to obtain the highest root and shoot elongation. The obtained results indicated the different responses of studied varieties with the different treatments of 2,4-D and coconut water. The varieties gave the highest callus induction percentage and somatic embryogenesis callus induction percentage with different concentrations of 2, 4-D in range between 2 - 4 mg/l. Lowering 2, 4-D to 1 mg/l enhanced the embryos development. On the other hand, all the varieties responded to 7 mg/l NAA but differ with the sucrose concentrations, they ranged between 40, 50 and 60 g/l. Molecular genetic studies were done to assess the somaclonal variation induced through tissue culture compared to the parental varieties. Five out of six primers succeeded in amplifying DNA fragments. Tissue culture was thus responsible for the generation of new variability, since an increase in the rate of molecular polymorphism was observed. Twenty eight out of 46 loci were found to be polymorphic in the regenerated genotypes. Similarity index (as percentage) based on RAPD-PCR analysis using UPGMA computer analysis between the regenerated varieties and their parental lines Phil. 8013, G98-28 and G98-24 were 84%, 85% and 88% respectively, which indicated the induction of mutation via somaclonal variation through tissue culture. The consensus tree was developed based on the banding patterns of the four varieties and their three regenerated genotypes using SPSS statistical analysis program to study the genetic relationships among these genotypes at the molecular level. The results confirmed that in vitro culture of the sugarcane is very efficient and has tremendous potential for producing novel and useful varieties.

Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing ∼57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick–host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host ‘questing’, prolonged feeding, cuticle synthesis, blood meal concentration, novel methods of haemoglobin digestion, haem detoxification, vitellogenesis and prolonged off-host survival. We identify proteins associated with the agent of human granulocytic anaplasmosis, an emerging disease, and the encephalitis-causing Langat virus, and a population structure correlated to life-history traits and transmission of the Lyme disease agent.

The first full-length message for vitellogenin (V g) was sequenced from ticks. The V g cDNA from the American dog tick, Dermacentor variabilis was 5744 nt in length (Genbank Accession number DQ285422) which coated for a protein of 1844 aa with a calculated molecular weight of 208 K. This protein had a 19 aa signal sequence, a single RXXR cleavage signal which predicts two subunits (49.5 and 157 Kin molecular weight), and an expected lipoprotein N-terminal and c~boxy von Willebrand factor type D domain. Tryptic digest MS analysis ofvitellin protein confirmed the function ofDQ285422 as the tick yolk protein. Apparently, vitellin in D. variabilis is oligomeric possibly dimeric and is comprised of a mixture of the uncleaved monomer and subunits that were predicted from the single RXXR cleavage signal. The highly conserved GL/ICG motif.close to the C-terminus in insect Vg genes was different in the tick Vg message, i.e., GLCS; this variant was confirmed for another partial sequence ofVg from Boophilus microp/us. A phylogenic analysis showed that the full length V g from D. variabilis and the partial sequence from B. mi crop/us was distinct from insects and Crustacea. The V g message was not found in whole body RNA from unfed or fed males or in unfed and partially fed (virgin) females as determined by Northern blotting. The message was found in replete (mated) pre-ovipositional females, increased to higher levels in ovipositing females and was absent after egg laying was complete. The endocrine regulation of the V g message is discussed. The tissue sources of the V g message are both the gut and fat body. Tryptic digest MS fingerprinting suggests that a second V g message might be present in the American dog tick, which needs further study. J'i j

Although eusociality evolved independently within several orders of insects, research into the molecular underpinnings of the transition towards social complexity has been confined primarily to Hymenoptera (for example, ants and bees). Here we sequence the genome and stage-specific transcriptomes of the dampwood termite Zootermopsis nevadensis (Blattodea) and compare them with similar data for eusocial Hymenoptera, to better identify commonalities and differences in achieving this significant transition. We show an expansion of genes related to male fertility, with upregulated gene expression in male reproductive individuals reflecting the profound differences in mating biology relative to the Hymenoptera. For several chemoreceptor families, we show divergent numbers of genes, which may correspond to the more claustral lifestyle of these termites. We also show similarities in the number and expression of genes related to caste determination mechanisms. Finally, patterns of DNA methylation and alternative splicing support a hypothesized epigenetic regulation of caste differentiation.

Co-feeding of aliphatic polyethylene glycol (PEG), phospholipase A2, anionic and ionic detergents, and amphipathic glycoside with bovine serum albumin (BSA) as a model protein to fourth stadium tobacco budworms, Heliothis virescens, did not affect the levels of BSA in the hemolymph. Covalent conjugation of small proteins like the decapeptide trypsin modulating oostatic factor (TMOF) to polyethylene glycol was previously shown to protect the peptide from protease attack and enhance its accumulation in the insect hemocoel. Whether this polymer chemistry could do the same for larger proteins was examined. The chemistry for the synthesis of polydispersed aliphatic PEG350-insulin and monodispersed aliphatic PEG333-insulin are described herein. Insulin was used for this synthesis and not BSA to better control conjugation among the available free amine groups. When PEGylated insulin or free insulin were fed in artificial diet to fifth stadium budworms, greater concentrations of insulin using the PEGylated variants were found in the hemolymph than when free insulin was used (a 6.7 and 7.3-fold increase for the PEG350 and PEG333 conjugates, respectively). When insulin is topically applied to the dorsum of H. virescens, no insulin is found in the hemolymph. However, after topical application of the PEGylated insulins, PEG350-insulin and PEG333-insulin were detected in the hemolymph. After injections of insulin into the hemocoel of fourth stadium H. virescens, insulin is completely cleared from the hemolymph in 120min. In comparison, PEG350-insulin and PEG333-insulin were present in the hemolymph for 300 and 240min after injection, respectively, translating to a 3.3 and 2.7-fold increase in the length of time insulin remains in the hemolymph after injection.

Development of embryogenic cultures having high regeneration efficiency from important, commercial varieties of banana is a prerequisite for genetic manipulation and for in vitro propagation. In the present study, we have studied the induction of somatic embryogenesis from young immature male inflorescences of the banana cultivar Williams (Musa spp. AAA group) via secondary somatic embryogenesis and cell suspension. Primary somatic embryos were produced when explants of immature male flower buds were cultured on Murashige and Skoog (MS) medium plus 1 mg/l biotin, 100 mg/l malt extract, 100 mg/l glutamine, 4 mg/l 2,4-dichlorophenoxyacetic acid, 1 mg/l indole-3-acetic acid (IAA), 1 mg/l alfa naphthaleneacetic acid, 30 g/l sucrose and 2.6 g/l Phytagel, pH 5.8 (M1 medium) and then transferred to M1 medium plus 200 mg/l casein hydrolysate and 2 mg/l proline (MM1). Secondary somatic embryogenesis (SE2) was developed when primary embryos were subculture on SK4 medium (MS medium supplemented with 10 ml of coconut milk). Embryos differentiated when sub-cultured on SK8 medium (MS medium supplemented with 5 mg/l BA and the embryos germinated when subculture on MS free hormone medium. Suspension cultures were initiated from SE2 embryogenic tissues from when placed in liquid medium supplemented with 2,4-D (1mg/l), biotin (1 mg/l), L-glutamate (100 mg/l), malt extract (100 mg/l), and sucrose (45 g/l), pH of the medium was adjusted to 5.3. The packed cell volume (PCV) of the suspension increased 2-5 fold with each monthly cycle. The somatic embryos were developed when suspension culture were aspirated on MS medium supplemented with biotin (1 mg/l), malt extract (100 mg/l), Glutamine (100mg/l), NAA (1mg/l), Kinetin (0.5 mg/l) Zeatin (0.2 mg/l), sucrose (45 g/l), and phytagel (2.6 g/l) (SK13). Differentiated embryos were transferred to MS medium supplemented with 5 mg/l 6-benzylaminopurine (BA) for development of mature somatic embryos, which were isolated and cultured on hormone-free MS medium for germination and development into plantlets. Approximately 55% of the somatic embryos germinated and developed into plantlets. Somatic embryogenesis via SE2 and cell suspension might be an excellent technique for mass propagation, developing a breeding strategy and genetic transformation of banana.

Whole-body transcriptomes for nymphs and adults of the green stink bug, Acrosternum hilare (Say), were sequenced on an Illumina® Genome Analyzer IIx sequencer. The insects were collected from sites in North Carolina and Virginia, USA. The cDNA library for each sample was sequenced on one lane of an eight-lane flow cell. Total read count was 25,211,411 and 23,601,951 for the nymph and adult libraries, respectively. Mean read length was 124 bps (base pairs) for nymph and 125 bps for adult. Total sequencing data were 3,127,036,121 bps for nymph cDNA and 2,956,794,438 bps for adult. Trinity® software was used to assemble the nymph and adult reads into 79,062 and 74,271 contigs (contiguous sequences), respectively. Blast2GO® software was used to align, map, and annotate the contigs. For the alignment step, the contigs were translated to peptides and compared to the GenBank nr (non-redundant) protein database using the BLASTx (Basic Local Alignment Search Tool) algorithm with E-value cut-off set at E-3 (10). The average length of BLASTx translated sequences was 1248 (range = 201 – 7968) amino acids for the nymph and 1320 (range = 201 – 7957) for the adult. Several nymph and adult sequences showed high homology (E-value ≤ 1.5 E-13) with database hormones, neuropeptides, neurotransmitters and/or their receptors. Seventeen nymph and adult sequences were identical (E-value = 0) to A. pisum, T. castaneum, B. germaninca, and P. humanus corporis hormones. After alignment of green stink bug contigs with GenBank nr database sequences, all BLAST hits (E-value ≤ E-3) were mapped and annotated with GO (Gene Ontology) terms that assigned the translated sequences to categories of putative protein function. Mapping and annotation steps yielded 13,226 nymph and 14,725 adult sequences assigned to 13 GO functional categories: Catalysis, binding, transport, signal transduction, structural molecule activity, enzyme regulation, electron carrier activity, antioxidant activity, metallochaperone, channel regulation, protein tag, translation regulation and nutrient reservoir. The goal of sequencing and of on-going bioinformatic analysis of these transcriptomes is to identify candidate RNAi gene targets with potential to be developed as insecticidal transgenes for control of stink bugs and other hemipteran pests of cotton. The resulting data can also be applied to other protein function and genetics studies.

454 Pyrosequencing was used to characterize the expressed genes from the synganglion and associated neurosecretory organs of unfed and partially fed virgin and mated replete females of the American dog tick, Dermacentor variabilis. A total of 14 881 contiguous sequences (contigs) was assembled, with an average size of 229 bp. Gene ontology terms for Level 2 biological processes were assigned to 4366 contigs. Seven acetylcholinesterases, a muscarinic acetylcholine (ACh) receptor, two nicotinic ACh receptor β-subunits, two ACh unc-18 regulators, two dopamine receptors, two gamma aminobutyric acid (GABA) receptors, two GABA transporters, two norepinephrine transporters and an octopamine receptor are described. Microarrays were conducted to examine global gene expression and quantitative real-time polymerase chain reaction was used to verify expression of selected neuropeptides. Hierarchical clustering of all differentially expressed transcripts grouped part-fed and replete ticks as being more similar in terms of differentially expressed genes with unfed ticks as the outgroup. Nine putative neuropeptides (allatostatin, bursicon-β, preprocorazonin, glycoprotein hormone α, insulin-like peptide, three orcokinins, preprosulphakinin) and a gonadotropin releasing hormone receptor were differentially expressed, and their developmental expression and role in reproduction was investigated. The presence of eclosion hormone, corazonin and bursicon in the synganglion, which in insects regulate behaviour and cuticle development associated with moulting, suggest that this system may be used in ticks to regulate blood feeding, cuticle expansion and development related to female reproduction; adult ticks do not moult.