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Aims: Spodoptera exigua is a serious pest of many agricultural crops. However, the bacterial communities of S. exigua are poorly studied, particularly over their entire life cycle. We aimed to study the biodiversity of the microbiota across the life cycle of S. exigua and to provide a better and obtain insight into new pest control strategies. Methods and Results: The bacterial diversity across the life cycle of S. exigua was studied using Illumina MiSeq sequencing of 16S rRNA genes. Spodoptera exigua is dominated by Proteobacteria and Firmicutes, with a total relative abundance of 90Á03%. Enterococcus (24Á6%), Pseudomonas (12Á2%) and Asaia (45Á9%) were abundant and active in eggs, while Methylobacterium (18Á7%) and Halomonas (16Á5%) dominated freshly eclosed larvae. The 3rd and 5th instar larvae were dominated by Enterococcus (76Á3 and 62Á0%). Pupal stages had the highest microbial diversity. There was no significant difference between newly emerged males and females. Symbionts of eggs were extremely similar and probably vertically transmitted by males during mating. Conclusions: The result showed that the bacterial community was affected by the host developmental stages. Our results also suggest that symbionts of egg mass are probably vertically transmitted control by male spawning adults. Significance and Impact of the Study: Our study documents the symbiont bacteria across the life cycle of S. exigua. Understanding the microbial symbionts may provide clues to develop potential biocontrol techniques against this pest.

BACKGROUND: Aphis gossypii, a polyphagous and recurrent pest induced by pesticides, causes tremendous loss crop yields each year. Previous studies on the mechanism of pesticide-induced sublethal effects mainly focus on the gene level. The symbiotic bacteria are also important participants of this mechanism, but their roles in hormesis are still unclear.RESULTS: In this study, life table parameters and 16S rRNA sequencing were applied to evaluate the sublethal and transgenerational effects of sulfoxaflor on adult A. gossypii after 24-h LC 20 (6.96 mg L −1 ) concentration exposure. The results indicated that the LC 20 of sulfoxaflor significantly reduced the finite rate of increase (⊗) and net reproductive rate (R 0 ) of parent generation (G0), and significantly increased mean generation time (T) of G1 and G2, but not of G3 and G4. Both reproductive period and fecundity of G1 and G2 were significantly higher than those of the control. Furthermore, our sequencing data revealed that more than 95% bacterial communities were dominated by the phylum Proteobacteria, in which the maximum proportion genus was the primary symbiont Buchnera and the facultative symbiont Arsenophonus. Compared to those of the control, the abundance and composition of symbiotic bacteria of A. gossypii for three successive generations (G0-G2) were changed after G0 A. gossypii was exposed to sulfoxaflor: the diversity of the bacterial community was decreased, but the abundance of Buchnera was increased (G0), while the abundance of Arsenophonus was decreased. Contrary to G0, G1 and G2 cotton aphid exhibited an increased relative abundance of Arsenophonus in the sublethal treatment group. CONCLUSION: Taken together, our results provide an insight into the interactions among pesticide resistance, aphids, and symbionts, which will eventually help to better manage the resurgence of A. gossypii.

Bacteria and insects have a mutually beneficial symbiotic relationship. Bacteria participate in several physiological processes such as reproduction, metabolism, and detoxification of the host. Adelphocoris suturalis is considered a pest by the agricultural industry and is now a major pest in cotton, posing a serious threat to agricultural production. As with many insects, various microbes live inside A. suturalis. However, the microbial composition and diversity of its life cycle have not been well-studied. To identify the species and community structure of symbiotic bacteria in A. suturalis, we used the HiSeq platform to perform high-throughput sequencing of the V3–V4 region in the 16S rRNA of symbiotic bacteria found in A. suturalis throughout its life stages. Our results demonstrated that younger nymphs (1st and 2nd instar nymphs) have higher species richness. Proteobacteria (87.06%) and Firmicutes (9.43%) were the dominant phyla of A. suturalis. At the genus level, Erwinia (28.98%), Staphylococcus (5.69%), and Acinetobacter (4.54%) were the dominant bacteria. We found that the relative abundance of Erwinia was very stable during the whole developmental stage. On the contrary, the relative abundance of Staphylococcus, Acinetobacter, Pseudomonas, and Corynebacterium showed significant dynamic changes at different developmental stages. Functional prediction of symbiotic bacteria mainly focuses on metabolic pathways. Our findings document symbiotic bacteria across the life cycle of A. suturalis, as well as differences in both the composition and richness in nymph and adult symbiotic bacteria. Our analysis of the bacteria in A. suturalis provides important information for the development of novel biological control strategies.

The optimal synchronous conditions to extract tea polysaccharides (TPS) and tea polyphenols (TPP) from Ya'an Tibetan tea were investigated, and the antioxidative capacity of TPS and TPP was measured, and the tea was analyzed to identify the polyphenol compounds it contained. On the basis of single-factor experiments, a Box-Behnken design and response surface methodology were applied to optimize the hot water extraction conditions. The optimal extraction technology was determined as extraction temperature of 83°C, time of 104 min, and liquid-to-material ratio of 41 ml/g, yielding TPP and TPS at 42.70 ± 2.38 mg/g and 53.86 ± 3.79 mg/g, respectively. The TPS and TPP in Ya'an Tibetan tea have high eliminating activities on DPPH and strong reducing power, with TPP showing a higher antioxidant activity than TPS. UHPLC-QqQ-MS/MS analysis identified EGCG, GCG, and ECG as major polyphenol components in Ya'an Tibetan tea. These findings might promote the application of Ya'an Tibetan tea in the food industry. K E Y W O R D Santioxidant activity, optimizing synchronous extraction, tea polyphenols, tea polyphenols compositions, tea polysaccharides, Ya'an Tibetan tea

Reference genes have been utilized in estimating gene expression levels using quantitative reverse transcriptase-quantitative polymerase chain reaction (qRT-PCR) analysis. Aphidius gifuensis Ashmaed is one of the most widely used biological control agents for aphids. The biological properties of this species have been studied in detail, and current investigations are focused on elucidating the regulatory mechanisms in its host However, the appropriate reference genes for target gene expression studies have not been identified. In this study, the expression profiles of 12 candidate reference genes were evaluated under different experimental conditions(development stage, sex, tissue type, diet) by using dedicated algorithms, including geNorm, Normfinder, BestKeeper, and ΔCt. In addition, RefFinder was used to rank the overall stability of the candidate genes. Finally, we recommend three optimal reference genes for the normalization of qRT-PCR data in the presence of specific variables, which include ACTB, RPL13, and PPI for different developmental stages; RPS18, ACTB, and RPL13 for sexes; RPL13, PRII3, and RPS18 in different tissue types; and RPL13, RPL27, and ACTB in diverse diets. The present study has identified optimal reference genes that could be used in estimating the expression levels of specific genes under these conditions following the Minimum Information for publication of Quantitative real-time PCR Experiments (MIQE) guidelines, which would facilitate in advancements in functional genomics research on A. gifuensis.

Magnetic dummy molecularly imprinted polymers (MDMIPs) were prepared by combining the surface imprinting technique with computer simulation for selective recognition of phthalate esters (PAEs). A computational study based on the density functional theory was performed to evaluate the template-monomer geometry and interaction energy in the prepolymerization mixture. The MDMIPs were characterized by transmission electron microscopy, scanning electron microscopy, vibrating sample magnetometry, X-ray diffraction, and Fourier transform infrared spectroscopy. They exhibited (a) high saturation magnetization of 53.14 emu g (leading to fast separation), and (b) large adsorption capacity, fast binding kinetics, and high selectivity for PAEs. Subsequently, a molecularly imprinted solid-phase extraction procedure followed by GC-MS was established for selective extraction and determination of 10 PAEs in food samples. Under the optimal experimental conditions, the response (peak area) was linear in the 0.5-100 ng mL concentration range. The limits of detection ranged from 0.15 to 1.64 ng g. The method was applied to the determination of PAEs in spiked real samples. The recoveries for 10 PAEs from various foods were in the range of 73.7%-98.1%, with relative standard deviations of 1.7%-10.2%. Graphical abstract Magnetic dummy molecularly imprinted polymers (MDMIPs) were prepared and successfully were applied as a special sorbent for the selective recognition and fast enrichment of 10 PAEs from different complex matrix.

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