Selection and evaluation of reference genes for qRT-PCR analysis of different developmental stages in Chilo suppressalis

He, Fang-Fang; Lu, Ming‐Xing; Du, Yu‐Zhou

doi:10.1016/j.aspen.2021.08.008

Journal of Asia-Pacific Entomology

2021

DOI: 10.1016/j.aspen.2021.08.008

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Selection and evaluation of reference genes for qRT-PCR analysis of different developmental stages in Chilo suppressalis

Fang-Fang He

Ming‐Xing Lu

Yu‐Zhou Du

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2023

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Cited by 3 publications

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“…Each reference gene has highly conservative and stable expression characteristics. However, recent studies have found that these commonly used reference genes are unstable, and their expression levels usually vary greatly in different types of cells and tissues, stages of development, and experimental treatments (Berruien et al 2021; He et al 2021; Odetti et al 2021). For this reason, specific reference genes must be rescreened in specific species.…”

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confidence: 99%

Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

Tan

Jin

Xiao

et al. 2023

Journal of Entomological Science

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A stable and specific heat shock protein 27.2 (HSP27.2) antibody was prepared and analyzed for protein level research in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). The full-length hsp27.2 was amplified from H. armigera larvae and constructed into the prokaryotic expression vector. The purified His-tag fused protein was used to immunize rabbits for the antibody preparation. Western blot analysis indicated that this antibody specifically recognized the HSP27.2 encoded by H. armigera and detected the HSP27.2 encoded by other noctuid larvae. Further analysis of HSP27.2 expression in H. armigera under infection by different pathogenic microorganisms and in different tissues showed that the expression of HSP27.2 is continually stable. The HSP27.2 antibody is efficient and capable as a reference antibody for functional studies involving genes and proteins in H. armigera and other lepidopteran insects.

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confidence: 99%

Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

Tan

Jin

Xiao

et al. 2023

Journal of Entomological Science

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Depending on different apoptosis pathways, the effector Cscaspase-3 in Chilo suppressalis exposed to temperature and parasitic stress was induced

Dong

Zhu

et al. 2023

International Journal of Biological Macromolecules

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Determination of suitable reference genes for RT-qPCR normalisation in Bombyx mori (Lepidoptera: Bombycidae) infected by the parasitoid Exorista sorbillans (Diptera, Tachinidae)

Liu,

Gu,

et al. 2023

Bull. Entomol. Res.

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The silkworm Bombyx mori (Lepidoptera: Bombycidae) is a lepidopteran model insect of great economic importance. The parasitoid Exorista sorbillans (Diptera, Tachinidae) is the major pest of B. mori and also a promising candidate for biological control. However, the molecular interactions between hosts and dipteran parasitoids have only partially been studied. Gene expression analysis by reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) is indispensable to characterise their interactions. Accurate normalisation of RT-qPCR-based gene expression requires the use of reference genes that are constantly expressed irrespective of experimental conditions. In this study, the expression stability of 13 traditionally used reference genes was estimated by five statistical algorithms (ΔCt, geNorm, Normfinder, BestKeeper, and RefFinder) to determine the best reference genes for gene expression studies in different tissues of B. mori under E. sorbillans parasitism. Specifically, TATA-box-binding protein was the best reference gene in epidermis and testis, while elongation factor 1α was the most stable gene in prothoracic gland and midgut. Elongation factor 1γ, ribosomal protein L3, actin A1, ribosomal protein L40, glyceraldehyde-3-phosphate dehydrogenase and eukaryotic translation initiation factor 4A were the most suitable genes in head, silk gland, fat body, haemolymph, Malpighian tubule and ovary, respectively. Our study offers a set of suitable reference genes for gene expression normalisation in B. mori under the parasitic stress of E. sorbillans, which will benefit the in-depth exploration of host-dipteran parasitoid interactions, and also provide insights for further improvements of B. mori resistance against parasitoids and biocontrol efficacy of dipteran parasitoids.

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Selection and evaluation of reference genes for qRT-PCR analysis of different developmental stages in Chilo suppressalis

Cited by 3 publications

References 43 publications

Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

Depending on different apoptosis pathways, the effector Cscaspase-3 in Chilo suppressalis exposed to temperature and parasitic stress was induced

Determination of suitable reference genes for RT-qPCR normalisation in Bombyx mori (Lepidoptera: Bombycidae) infected by the parasitoid Exorista sorbillans (Diptera, Tachinidae)

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