The oriental armyworm, Mythimna separata, is a major insect pest in China and other Asian countries. Unfortunately, suitable reference genes for quantitative real-time PCR (qRT-PCR) have not been previously identified in M. separata for normalizing target gene expression. In this study, we evaluated the expression stability of eight candidate genes (18S, ACT, EF1-α, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative ΔCt method, BestKeeper, Normfinder geNorm and ReFinder, a comprehensive software platform. The results indicated that the appropriate reference gene varied depending on the experimental conditions. We found that ACTIN, EF1-α and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-α showed the most stable expresssion in different tissues; RPS13 and 18S were the best reference genes for monitoring expression under high temperature conditions; TUB, RPS13 and RPS7 exhibited the most stable expression under larval-crowding conditions; RPS7, EF1-α, RPL32 and GAPDH were the best for pesticide exposure experiments. This study provides tools for reliable normalization of qRT-PCR data and forms a foundation for functional studies of target gene expression in M. separata.

Superoxide dismutase (SOD) is an antioxidant metalloenzyme that catalyzes the dismutation of the superoxide anion O2− to O2 and H2O2. Many studies have focused on the role of SOD in response to abiotic stress, but its role during biotic stress, such as changes in organismal population density, has rarely been investigated. The oriental armyworm, Mythimna separata, is an economically important pest that exhibits phenotypic changes in response to population density. Solitary and gregarious phases occur at low and high population density, respectively. To examine the role of SODs in response to population density stress, we cloned two genes encoding SOD, MsCuZnSOD and MsMnSOD, and compared their expression in solitary and gregarious phases of M. separata. The MsCuZnSOD and MsMnSOD ORFs were 480 and 651 bp and encoded predicted protein products of 159 and 216 amino acids, respectively. The two SODs contained motifs that are typical of orthologous proteins. Real-time PCR indicated that the two SOD genes were expressed throughout developmental stages and were significantly upregulated in more mature stages of gregarious M. separata. Expression of the two SOD genes in various tissues of sixth-instar larvae was higher in gregarious versus solitary insects. Furthermore, expression of the SOD genes was significantly upregulated in response to crowding in solitary individuals, but suppressed in gregarious insects subjected to isolation. Collectively, these results suggest that population density may be key factor in the induction of SOD genes in M. separata.

Background: The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith), is known to cause large agricultural production losses. Emamectin benzoate is one of the most effective insecticides to control this pest; however, its effective time is not sufficiently long to control FAW. Therefore, it is important that new controlled insecticide formulations with new application methods are developed.Results: A series of emamectin benzoate polymer gel granules were prepared with sizes ranging from 0.95 to 1.5 mm. As the bentonite content increased, the release rate decreased. The cumulative release process of emamectin benzoate mainly depends on the cracks in the surface of the granules, and the release rate can be described by non-Fickian and Fickian diffusion, which are closely related to the water content. By spreading the developed polymer gel granules into maize leaf whorls, the control effect reached 83% after 21 days in field trials.Conclusion: A novel polymer gel granule was developed that can effectively regulate emamectin benzoate release. By broadcasting polymer gel granules into maize leaf whorls, significant control efficacy against FAW can be obtained, and this could potentially be used for the effective control of FAW.

Key indicatorsSingle-crystal X-ray study T = 298 K Mean (C-C) = 0.010 Å R factor = 0.080 wR factor = 0.154 Data-to-parameter ratio = 19.8For details of how these key indicators were automatically derived from the article, see

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