Locusta migratoria is a classic hemimetamorphosis insect and has caused widespread economic damage to crops as a migratory pest. Researches on the expression pattern of functional genes in L. ...migratoria have drawn focus in recent years, especially with the release of genome information. Real-time quantitative PCR is the most reproducible and sensitive approach for detecting transcript expression levels of target genes, but optimal internal standards are key factors for its accuracy and reliability. Therefore, it's necessary to provide a systematic stability assessment of internal control for well-performed tests of target gene expression profile. In this study, twelve candidate genes (Ach, Act, Cht2, EF1α, RPL32, Hsp70, Tub, RP49, SDH, GAPDH, 18S, and His) were analyzed with four statistical methods: the delta Ct approach, geNorm, Bestkeeper and NormFinder. The results from these analyses aimed to choose the best suitable reference gene across different experimental situations for gene profile study in L. migratoria. The result demonstrated that for different developmental stages, EF1α, Hsp70 and RPL32 exhibited the most stable expression status for all samples; EF1α and RPL32 were selected as the best reference genes for studies involving embryo and larvae stages, while SDH and RP49 were identified for adult stage. The best-ranked reference genes across different tissues are RPL32, Hsp70 and RP49. For abiotic treatments, the most appropriate genes we identified were as follows: Act and SDH for larvae subjected to different insecticides; RPL32 and Ach for larvae exposed to different temperature treatments; and Act and Ach for larvae suffering from starvation. The present report should facilitate future researches on gene expression in L. migratoria with accessibly optimal reference genes under different experimental contexts.
Recent studies have focused on determining functional genes and microRNAs in the pest Helicoverpa armigera (Lepidoptera: Noctuidae). Most of these studies used quantitative real-time PCR (qRT-PCR). ...Suitable reference genes are necessary to normalize gene expression data of qRT-PCR. However, a comprehensive study on the reference genes in H. armigera remains lacking.
Twelve candidate reference genes of H. armigera were selected and evaluated for their expression stability under different biotic and abiotic conditions. The comprehensive stability ranking of candidate reference genes was recommended by RefFinder and the optimal number of reference genes was calculated by geNorm. Two target genes, thioredoxin (TRX) and Cu/Zn superoxide dismutase (SOD), were used to validate the selection of reference genes. Results showed that the most suitable candidate combinations of reference genes were as follows: 28S and RPS15 for developmental stages; RPS15 and RPL13 for larvae tissues; EF and RPL27 for adult tissues; GAPDH, RPL27, and β-TUB for nuclear polyhedrosis virus infection; RPS15 and RPL32 for insecticide treatment; RPS15 and RPL27 for temperature treatment; and RPL32, RPS15, and RPL27 for all samples.
This study not only establishes an accurate method for normalizing qRT-PCR data in H. armigera but also serve as a reference for further study on gene transcription in H. armigera and other insects.
•Twelve reference genes from H. armigera were evaluated under different treatments.•28S and RPS15 for developmental stages; RPS15 and RPL13 for larvae tissues.•EF and RPL27 for adult tissues; GAPDH, RPL27 and β-TUB for NPV infection.•RPS15 and RPL32 for insecticide treatment; RPS15 and RPL27 for temperature treatment.
After injury many insects could regenerate lost limb. In this study, Helicoverpa armigera Hubner and Locusta migratoria manilensis (Meyen, 1835) were chosen to compare the regeneration potency of ...holometabolous and hemimetabolous insects. We employed the classical approach of surgical excision to verify the regeneration ability and to investigate the factors that affect the extent of regeneration. The results found that H. armigera could regenerate intact legs when the larval legs were excised at the first and second instar and that legs of adult H. armigera had a close relationship with their larval counterparts. However, the adult legs became malformed or disappeared when excised at other older instars. For the L. migratoria, we found the legs have weak partial regeneration ability when amputation was conducted at the joint of two segments. The regeneration potency might be stronger the more proximal the operation. Regeneration process had a negative impact on the larval development. This is the first report of complete leg regeneration capacity having a strong correlation with the instar but not with the position where amputation occurred for H. armigera, while for the L. migratoria, partial regenerative ability had a close relationship with the position where amputation occurred but not with instars.
Ecdysteroids play an important role in the growth, development, and reproduction of insects. The ecdysteroid signal is transduced through a heterodimeric complex of two nuclear receptors: the ...ecdysone receptor (EcR) and the ultraspiracle (USP). To investigate the role of USP during the development of Grapholita molesta, ultraspiracle (USP) cDNA was obtained from the transcriptome of G. molesta and verified by PCR. The deduced amino acid sequence of G. molesta USP (GmUSP) was highly homologous to those of other lepidopterans, especially the Manduca sexta USP-1 isoform. Relatively high expression levels of GmUSP were observed immediately prior to the last larval (L10) molt and during the early pupal stage (P1). Furthermore, high levels of GmUSP mRNA were found in the midgut during the last larval stage. Silencing of GmUSP could significantly down-regulate the transcriptional level of ecdysone-inducible genes and result in increased mortality during metamorphosis and a prolonged prepupal duration. In RNAi experiments, ecdysone-inducible genes were significantly down-regulated following GmUSP silencing, suggesting that they were under the control of GmUSP. In addition, the GmUSP knockdown resulted in increased mortality during metamorphosis and prolonged the prepupal period. Taken together, the present results indicate that GmUSP may directly or indirectly affect the development of G. molesta.
After injury many insects could regenerate lost limb. In this study, Helicoverpa armigera Hubner and Locusta migratoria manilensis (Meyen, 1835) were chosen to compare the regeneration potency of ...holometabolous and hemimetabolous insects. We employed the classical approach of surgical excision to verify the regeneration ability and to investigate the factors that affect the extent of regeneration. The results found that H. armigera could regenerate intact legs when the larval legs were excised at the first and second instar and that legs of adult H. armigera had a close relationship with their larval counterparts. However, the adult legs became malformed or disappeared when excised at other older instars. For the L. migratoria, we found the legs have weak partial regeneration ability when amputation was conducted at the joint of two segments. The regeneration potency might be stronger the more proximal the operation. Regeneration process had a negative impact on the larval development. This is the first report of complete leg regeneration capacity having a strong correlation with the instar but not with the position where amputation occurred for H. armigera, while for the L. migratoria, partial regenerative ability had a close relationship with the position where amputation occurred but not with instars.
More and more studies have focused on microRNAs (miRNAs) expression in the pest Helicoverpa armigera (Lepidoptera: Noctuidae) recently. Quantitative real-time PCR (qRT-PCR) is being widely used in ...miRNA expression studies. Suitable reference genes are necessary for the correct analysis of results. In this study, 10 candidate genes of H. armigera were selected and analyzed for their expression stability under different biotic and abiotic conditions with 3 statistical methods, including geNorm, NormFinder, and Bestkeeper. Combination the best number of reference genes was calculated by geNorm. One target gene, let-7, was used to validate the selection of reference genes. The suitable candidate reference genes were shown as follows: miR-9 and U6 snRNA for developmental stages, miR-100 and U6 snRNA for larval tissues, miR-100 and miR-305 for adult tissues, miR-9 and miR-279 for parasitic treatment, miR-998 and U6 snRNA for nuclear polyhedrosis virus infection, miR-9 and U6 snRNA for insecticide treatment, miR-92a, miR-100, and miR-279 for temperature treatment, miR-92a, miR-305, and miR-998 for starvation treatment, miR-9 and miR-279 for light treatment, miR-305 and miR-998 for hormone treatment, and there was not one reference gene suitable for all samples. This study could promote future research on miRNAs expression in H. armigera with optimal reference genes under different experimental conditions.
Apolygus lucorum (Meyer-Dür) (Hemiptera: Heteroptera: Miridae) is an important insect pest of cotton and other crops, but the species also feeds on some insect pests. In this study, the effect of 5 ...food condition treatments, including corn kernel, cotton leaves, eggs of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), a combination of corn + H. armigera eggs, and of cotton leaves + H. armigera eggs, on the biological parameters of A. lucorum was evaluated in the laboratory. Mixed foods (e.g., corn + H. armigera eggs) significantly increased nymphal survival, prolonged adult longevity, and improved female fecundity. The survival rate of nymphs with only H. armigera egg was the lowest, and development times with cotton leaves were the longest. Under mixed food conditions, nymphs preyed on more eggs in the combination with cotton leaves than with corn. These results indicate that A. lucorum does have a certain ability to prey on H. armigera eggs, but that during the growth and development of A. lucorum, plant food is still necessary for development.
Existing experimental results have shown that using a semi-log linear relationship between the permanent volumetric strain and cyclic number underestimates the volumetric deformation of rockfill ...materials with a large cyclic number, and that the error increases with the confining pressure. The existing permanent deformation models are not suitable for the seismic safety analysis of high dams during strong earthquakes. In this study, a series of large-scale triaxial cyclic loading tests of rockfill materials were performed, and a new permanent deformation model of rockfill materials was developed and validated with three kinds of rockfill materials. The results show that the proposed model can properly reflect the general features of the permanent deformation of rockfill materials. The main features of the model are as follows: (1) relations between the cyclic number and permanent volumetric/shear strain are described by hyperbolic functions, which can avoid underestimating the volumetric deformation occurring during strong earthquakes; (2) the model can capture the effect of the mean effective stress on the permanent volumetric strain, with greater confining pressure correlating to greater permanent volumetric deformation, and the permanent volumetric strain under low confining pressure near the dam crest can be well represented; and (3) the model can reflect the effect of the consolidation stress ratio on the permanent shear strain.
Locusta migratoria is a classic hemimetamorphosis insect and has caused widespread economic damage to crops as a migratory pest. Researches on the expression pattern of functional genes in L. ...migratoria have drawn focus in recent years, especially with the release of genome information. Real-time quantitative PCR is the most reproducible and sensitive approach for detecting transcript expression levels of target genes, but optimal internal standards are key factors for its accuracy and reliability. Therefore, it's necessary to provide a systematic stability assessment of internal control for well-performed tests of target gene expression profile. In this study, twelve candidate genes (Ach, Act, Cht2, EF1 alpha , RPL32, Hsp70, Tub, RP49, SDH, GAPDH, 18S, and His) were analyzed with four statistical methods: the delta Ct approach, geNorm, Bestkeeper and NormFinder. The results from these analyses aimed to choose the best suitable reference gene across different experimental situations for gene profile study in L. migratoria. The result demonstrated that for different developmental stages, EF1 alpha , Hsp70 and RPL32 exhibited the most stable expression status for all samples; EF1 alpha and RPL32 were selected as the best reference genes for studies involving embryo and larvae stages, while SDH and RP49 were identified for adult stage. The best-ranked reference genes across different tissues are RPL32, Hsp70 and RP49. For abiotic treatments, the most appropriate genes we identified were as follows: Act and SDH for larvae subjected to different insecticides; RPL32 and Ach for larvae exposed to different temperature treatments; and Act and Ach for larvae suffering from starvation. The present report should facilitate future researches on gene expression in L. migratoria with accessibly optimal reference genes under different experimental contexts.
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