Ovary-Specific Transcriptome and Essential Role of Nanos in Ovary Development in the Oriental Fruit Fly (Diptera: Tephritidae).

We used transcriptome analysis to research ovary development in Bactrocera dorsalis (Hendel). The ovary transcriptome of B. dorsalis yielded 66,463,710 clean reads that were assembled into 23,822 unigenes. After aligning to the Nr database in NCBI, 15,473 (64.95%) of the unigenes were matched to identified proteins. As determined by BLAST search, 11,043 (46.36%), 6,102 (25.61%), and 12,603 (52.90%) unigenes were each allocated to clusters via gene ontology, orthologous groups, and SwissProt, respectively. The Kyoto encyclopedia database of genes and genomes (KEGG) was further used to annotate these sequences, and 11,068 unigenes were mapped to 255 known pathways. Afterward, the genes that were possibly involved in oogenesis and ovary development were obtained from the transcriptome data and analyzed. Interestingly, seven ovary-specific genes were identified, including a Nanos gene that is involved in maintaining the primordial germ cells in many insects. Therefore, we further focused on the function of the BdNanos gene, and the gene was injected into B. dorsalis. As expected, the knocking down of Nanos gene expression led to significant inhibition of ovary development, suggesting an important role of this gene in the reproductive process of B. dorsalis. In summary, the present study provides an important reference for identifying the molecular mechanisms of oogenesis and ovary development in B. dorsalis. The BdNanos gene is crucial for ovary development in B. dorsalis and is therefore a potential new pest control target.

Transcriptome Profile Changes Associated With Heat Shock Reaction in the Entomopathogenic Nematode, Steinernema carpocapsae.

The entomopathogenic nematode Steinernema carpocapsae has been used for control of soil insects. However, S. carpocapse is sensitive to environmental factors, particularly temperature. We studied an S. carpocapse group that was shocked with high temperature. We also studied the transcriptome-level responses associated with temperature stress using a BGIseq sequencing platform. We de novo assembled the reads from the treatment and control groups into one transcriptome consisting of 43.9 and 42.9 million clean reads, respectively. Based on the genome database, we aligned the clean reads to the Nr, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases and analyzed the differentially expressed genes (DEGs). Compared with the control, the heat-shocked group had significant differential expression of the heat shock protein (HSP) family, antioxidase [glutathione S-transferases (GSTs) and superoxide dismutase (SOD)], monooxygenase (P450), and transcription factor genes (DAF-16 and DAF-2). These DEGs were demonstrated to be part of the Longevity pathway and insulin/insulin-like signaling pathway. The results revealed the potential mechanisms, at the transcriptional level, of S. carpocapsae under thermal stress.

PT-symmetric phonon laser under gain saturation effect.

As an analog of optical laser, phonon laser has potential applications in various areas. We study a type of phonon laser implemented by two coupled micro-cavities, one of which carries optical gain medium. The phonon laser operation is under a blue detuned external drive leading to dynamical instability. The saturation of the optical gain is considered, and its induced nonlinearity results in more complicated behaviors in stimulated phonon emission. To deal with such complex dynamics, we apply a composite numerical approach, in addition to a previously used dynamical approach, to the time evolution of the system. The workable phonon laser operation is found to be achievable by choosing the proper system parameters. Moreover, low threshold for the phonon laser operation is possible with the suitable coupling between the cavities and an optimum damping rate in one cavity.

Influence of various stressors on the expression of core genes of the small interfering RNA pathway in the oriental fruit fly, Bactrocera dorsalis.

RNA interference (RNAi)-based technology has emerged as a potential tool for controlling insect pests, however, previous studies found that the efficiency of RNAi in Bactrocera dorsalis was variable. In nature, insects often meet various challenges, such as pathogen infections, extreme temperatures, lack of nutrition and heavy metals. To better understand the association of the stressors with efficiency of RNAi, in the current study we tested the expression of three core genes, dicer2 (Bddcr2), r2d2 (Bdr2d2) and argonaute2 (Bdago2), of the small interfering RNA (siRNA) pathway of B. dorsalis upon various stressors. Our results showed that all three genes were upregulated by the infection of invertebrate iridescent virus 6, which suggested a function of the siRNA pathway against viral infection. The loading of FeCl3 could also increase the expression of Bddcr2. The treatments of Escherichia coli, extremely high (40°C) and low (0°C) temperatures, as well as starvation, could negatively influence the expression of Bddcr2 and/or Bdago2. In total, our results showed that various stressors could influence the expression of core components of B. dorsalis siRNA pathway. This highlights further speculation on the RNAi efficiency upon these stressors. Considering the complexity and variation of RNAi efficiency in different conditions, these results provide initial aspects in possible environmental stressors to influence the activity of the siRNA pathway, but the real impact of RNAi efficiency posed by these stressors requires further studies.

Molecular characterization and functional analysis of BdFoxO gene in the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae).

The forkhead box O transcription factor (FoxO) is an important downstream transcription factor in the well-conserved insulin signaling pathway, which regulates the body size and development of insects. In this study, the FoxO gene (BdFoxO) was identified from the oriental fruit fly, Bactrocera dorsalis (Hendel). The open reading frame of BdFoxO (2732 bp) encoded a 910 amino acid protein, and the sequence was well conserved with other insect species. The BdFoxO was highly expressed in larvae and pupae among different development stages, and the highest tissue-specific expression level was found in the fat bodies compared to the testis, ovary, head, thorax, midgut, and Malpighian tubules of adults. Interestingly, we found BdFoxO expression was also up-regulated by starvation, but down-regulated when re-fed. Moreover, the injection of BdFoxO double-stranded RNAs into third-instar larvae significantly reduced BdFoxO transcript levels, which in turn down-regulated the expression of other four genes in the insulin signaling pathway. The silencing of BdFoxO resulted in delayed pupation, and the insect body weight increased significantly compared with that of the control. These results suggested that BdFoxO plays an important role in body size and development in B. dorsalis.

Two Chitin Biosynthesis Pathway Genes in Bactrocera dorsalis (Diptera: Tephritidae): Molecular Characteristics, Expression Patterns, and Roles in Larval-Pupal Transition.

Glucose-6-phosphate isomerase (G6PI) and UDP-N-acetylglucosamine pyrophosphorylase (UAP), two key components in the chitin biosynthesis pathway, are critical for insect growth and metamorphosis. In this study, we identified the genes BdG6PI and BdUAP from the oriental fruit fly, Bactrocera dorsalis (Hendel). The open reading frames (ORFs) of BdG6PI (1,491 bp) and BdUAP (1,677 bp) encoded 496 and 558 amino acid residues, respectively. Multiple sequence alignments showed that BdG6PI and BdUAP had high amino acid sequence identity with other insect homologues. Quantitative real-time polymerase chain reaction (qPCR) analysis indicated that BdG6PI was mainly expressed in the early stages of third-instar larvae and adults, while significantly higher expression of BdUAP was observed in adults. Both transcripts were expressed highly in the Malpighian tubules, but only slightly in the tracheae. The expression of both BdG6PI and BdUAP was significantly up-regulated by 20-hydroxyecdysone exposure and down-regulated by starvation. Moreover, injection of double-stranded RNAs of BdG6PI and BdUAP into third-instar larvae significantly reduced the corresponding gene expressions. Additionally, silencing of BdUAP resulted in 65% death and abnormal phenotypes of larvae, while silencing of BdG6PI had a slight effect on insect molting. These findings provide some data on the roles of BdG6PI and BdUAP in B. dorsalis and demonstrate the potential role for BdUAP in larval-pupal transition.

Phenoloxidase and its zymogen are required for the larval-pupal transition in Bactrocera dorsalis (Diptera: Tephritidae).

Phenoloxidases (POs) play a key role in melanin production, are involved in invertebrate immune mechanisms, and are considered important enzymes in the insect development process. In the present study, we report the developmental stage and tissue-specific expression patterns of BdPPO1 and PO activity from Bactrocera dorsalis. The results showed that the activity of PO and its zymogen expression were closely related to the development of B. dorsalis during the larval-pupal transition, particularly in the integument. Additionally, biochemical characterization showed that PO from different developmental stages and tissues all had maximum activity at pH 7.5 and 37°C. After feeding a metal ion-containing artificial diet, the activity of PO and expression of BdPPO1 were significantly increased, indicating that PO was a metalloprotein and it could be activated by Zn2+, Mg2+, Ca2+, and Cu2+. The functional analysis showed that the expression of BdPPO1 could be regulated by 20-hydroxyecdysone (20E) after injection. Furthermore, injection of the double-stranded RNA of BdPPO1 into the 3rd instar larvae significantly reduced mRNA levels after 24 h and 48 h, and resulted in a lower pupation rate and abnormal phenotype. These results expand the understanding of the important role of PO and its zymogen in the growth of B. dorsalis.

De novo cloning and annotation of genes associated with immunity, detoxification and energy metabolism from the fat body of the oriental fruit fly, Bactrocera dorsalis.

The oriental fruit fly, Bactrocera dorsalis, is a destructive pest in tropical and subtropical areas. In this study, we performed transcriptome-wide analysis of the fat body of B. dorsalis and obtained more than 59 million sequencing reads, which were assembled into 27,787 unigenes with an average length of 591 bp. Among them, 17,442 (62.8%) unigenes matched known proteins in the NCBI database. The assembled sequences were further annotated with gene ontology, cluster of orthologous group terms, and Kyoto encyclopedia of genes and genomes. In depth analysis was performed to identify genes putatively involved in immunity, detoxification, and energy metabolism. Many new genes were identified including serpins, peptidoglycan recognition proteins and defensins, which were potentially linked to immune defense. Many detoxification genes were identified, including cytochrome P450s, glutathione S-transferases and ATP-binding cassette (ABC) transporters. Many new transcripts possibly involved in energy metabolism, including fatty acid desaturases, lipases, alpha amylases, and trehalose-6-phosphate synthases, were identified. Moreover, we randomly selected some genes to examine their expression patterns in different tissues by quantitative real-time PCR, which indicated that some genes exhibited fat body-specific expression in B. dorsalis. The identification of a numerous transcripts in the fat body of B. dorsalis laid the foundation for future studies on the functions of these genes.

Study on application of biological iron sulfide composites in treating vanadium-extraction wastewater containing chromium (VI) and chromium reclamation.

In this study, the Cr(VI)-resistant properties and regeneration characteristics of biological iron sulfide composites were investigated, which consist of sulfate reducing bacteria (SRB) and its in situ synthesized nanosized iron sulfides. Then the application of the composites in treating vanadium-extraction wastewater containing high concentration Cr(VI) and reclaiming Cr were performed. It was found that SRB in composites still survived after being used to treat vanadium-extraction wastewater, which could reduce reaction products Fe3+ and sulphur into Fe2+ and S2 by using them as the electron accepters and thus regenerating biological iron sulfide composites. The SRB also could be resistant to 600 mgl(-1) Cr(VI) and reduce it gradually. Based on the Cr(VI)-resistant properties and regeneration characteristics of the composites, a reduction-regeneration recirculation process for treating vanadium-extraction wastewater and reclamation of Cr was developed. The results indicated that the contaminants in effluent reached the Chinese discharge standard of pollutants for vanadium industry (GB 26452-2011), i.e. the concentration of total Cr(TCr) was less than 0.912 mgl(-1), Cr(VI) was less than 0.017 mgl(-1) and V was less than 0.260 mgl(-1). After 10 cycles of treatment, the Cr2O3 content in sludge reached 41.03%, and the ratio of Cr2O3/FeO was 7.35. The sludge reached the chemical and metallurgical (hydrometallurgy) grade of chromite ore and could be reclaimed.

Removal of Cu2+ ions from aqueous solution by the abandoned mushroom compost of Flammulina velutipes.

The abandoned mushroom compost of Flammulina velutipes, a cheap and easy by-product to get, was used as biosorbent for removing copper ions from aqueous solution. Batch experiments were carried out to investigate the effect of contact time, solution pH, biomass dosage, initial concentration of Cu2+ ions and temperature on biosorption efficiency. The maximum sorption capacity could be reached at pH 5.0 in 60 min. Langmuir, Freundlich, and Redlich- Peterson isotherm models were used to fit the experimental data and their model parameters were evaluated. The calculated qm based on Langmuir equation was 35.608 mg g(-1) at 288 K, 48.711 mg g(-1) at 298 K, and 42.330 mg g(-1) at 308 K, respectively. The kinetics were discussed by pseudo- first order and pseudo-second order models, and the result showed that the latter was more suitable. The thermodynamics of biosorption was also investigated, and the biosorption process was feasible, spontaneous and endothermic in nature.

[Property analysis of nanosized iron sulfide produced by sulfate reducing bacteria and its application in the treatment of wastewater containing high concentration of Cr (VI)].

The properties of novel nanosized iron sulphide produced by sulfate reducing bacteria (SRB) and its application to the removal of Cr (VI) from wastewater were studied. The structural properties were examined by transmission electron microscope (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The influences of pH, temperature and dosage on removal of Cr (VI) from wastewater by nanosized iron sulphide were investigated, and the effect comparison of nanosized iron sulphide and other traditional wastewater treatment was also evaluated. TEM result shows that the length of the nanosized iron sulphide is approximately 45-80 nm, and the ratio of length to width is 10-15. XPS and XRD results indicate that the Fe/S is 1.07-1.11, and the main composition of nanosized iron sulphide is amorphous FeS and mackinawite. While in treating wastewater containing Cr (V), pH, temperature and dosage of nanosized iron sulphide are the main factors to control the reaction. The Cr (VI) reduction rate increases with decrease of pH and increase of dosage and temperature. Among these factors, pH is the most significant influence factor. At 25 degrees C, pH 3, when the mole ratio of nanosized iron sulphide to chromium (VI) reaches 1.17/1, wastewater containing Cr (VI) 0.03 mol/L can be treated to meet the discharge standard within 10 min. Moreover, in treating wastewater containing Cr (VI), compared with commercial iron sulfide (AR.), nanosized iron sulphide has advantages of low dosage and short treatment time, and compared to conventional biological treatment by SRB, nanosized iron sulphide has advantages of low effluent COD and small amount of sludge.