Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)

Bai, Yu; Lv, Yanan; Zeng, Mei; Jia, Peng; Lu, Hu-Na; Zhu, Yuanfeng; Li, Sheng; Cui, Yingying; Luan, Yigang

doi:10.3390/genes12010021

Cited by 20 publications

(11 citation statements)

References 40 publications

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“…More convincing results were obtained when two genes were used for expression normalization ( Figures 5A,B ). Our results corroborate the current notion of using two or three genes for target gene expression normalization to enhance accuracy (Arya et al, 2017 ; Li et al, 2018c ; Wei et al, 2020 ; Bai et al, 2021 ; Fu and Meyer-Rochow, 2021 ).…”

Section: Discussionsupporting

confidence: 90%

“…RPS3 also exhibited high stability under larval tissues in Lucilia sericata (Baumann et al, 2015 ). Similarly, RPL9 and RPL10 genes showed higher expression stability in different developmental stages and tissues of Sogatella furcifera (An et al, 2016 ), whereas RPS26 and RPL32 genes showed the same in Thermobia domestica (Bai et al, 2021 ). Furthermore, RPS18 and RPL13 genes showed the highest expression stability in Rhopalosiphum padi tissues (Li et al, 2021 ).…”

Section: Discussionmentioning

confidence: 98%

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Reference Gene Selection for Normalizing Gene Expression in Ips Sexdentatus (Coleoptera: Curculionidae: Scolytinae) Under Different Experimental Conditions

et al. 2021

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Ips sexdentatus (Coleoptera: Curculionidae: Scolytinae) is one of the most destructive and economically important forest pests. A better understanding of molecular mechanisms underlying its adaptation to toxic host compounds may unleash the potential for future management of this pest. Gene expression studies could be considered as one of the key experimental approaches for such purposes. A suitable reference gene selection is fundamental for quantitative gene expression analysis and functional genomics studies in I. sexdentatus. Twelve commonly used reference genes in Coleopterans were screened under different experimental conditions to obtain accurate and reliable normalization of gene expression data. The majority of the 12 reference genes showed a relatively stable expression pattern among developmental stages, tissue-specific, and sex-specific stages; however, some variabilities were observed during varied temperature incubation. Under developmental conditions, the Tubulin beta-1 chain (β-Tubulin) was the most stable reference gene, followed by translation elongation factor (eEF2) and ribosomal protein S3 (RPS3). In sex-specific conditions, RPS3, β-Tubulin, and eEF2 were the most stable reference genes. In contrast, different sets of genes were shown higher stability in terms of expression under tissue-specific conditions, i.e., RPS3 and eEF2 in head tissue, V-ATPase-A and eEF2 in the fat body, V-ATPase-A and eEF2 in the gut. Under varied temperatures, β-Tubulin and V-ATPase-A were most stable, whereas ubiquitin (UbiQ) and V-ATPase-A displayed the highest expression stability after Juvenile Hormone III treatment. The findings were validated further using real-time quantitative reverse transcription PCR (RT-qPCR)-based target gene expression analysis. Nevertheless, the present study delivers a catalog of reference genes under varied experimental conditions for the coleopteran forest pest I. sexdentatus and paves the way for future gene expression and functional genomic studies on this species.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 98%

Reference Gene Selection for Normalizing Gene Expression in Ips Sexdentatus (Coleoptera: Curculionidae: Scolytinae) Under Different Experimental Conditions

et al. 2021

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“…Some of the internal reference genes screened by us can be directly used in RNAi experiments after verification. For the tissues, the reference genes used by Thermobia domestica were RPS26 and RPL32 in the different tissues under dsRNA microinjection treatment [ 56 ]. RNAi studies of specific target genes usually also screen the suitable reference genes.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Reference Genes in Glenea cantor (Fabricius) by Using qRT-PCR

Huang

Qin

et al. 2021

Genes

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Kapok is the main host of Glenea cantor (Fabricius), which causes serious damage and is difficult to control. In severe cases, it often causes the kapok trees to die continuously, which seriously affects the results of urban landscaping. To provide reference for the functional research on related genes in G. cantor, we screened the stable expression of candidate reference genes at different developmental stages (i.e., eggs, larvae, pupae, and adults), in various adult tissues (i.e., head, thorax, abdomen, feet, antennae, and wings), and sexes (i.e., male pupae, female pupae, male adults, and female adults). In this study, 12 candidate reference genes (i.e., ACTINLIKE, ACTININ, TUB, RPL36, RPL32, RPS20, TBP, GAPDH, 18S rRNA, EF1A1, EF1A2, and UBQ) were evaluated using different adult tissues, developmental stages, and sexes. RefFinder, geNorm, NormFinder, and BestKeeper were used to evaluate and comprehensively analyze the stability of the expression of the candidate reference genes. The results show that RPL32 and EF1A1 were the most suitable reference genes in the different adult tissues, and RPL36 and EF1A1 were best at the different developmental stages. RPL36 and EF1A2 were the best fit for the qRT-PCR reference genes in the different sexes, while RPL36 and EF1A1 were the most appropriate qRT-PCR reference genes in all samples. Results from geNorm showed that the optimal number of reference genes was two. We also surveyed the expression of cellulase at the different developmental stages and in the different adult tissues. Results further verified the reliability of the reference genes, and confirmed the best reference genes under the different experimental conditions. This study provides a useful tool for molecular biological studies on G. cantor.

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“…It is also the most reliable method for detecting and quantifying transcriptional abundance [4]. When using RT-qPCR to study the expression levels of target genes, appropriate reference genes must be used to calibrate and standardize the experimental samples [5][6][7]. The reference genes should not be affected by external environments and should be stably expressed in various tissues or cells and under various experimental conditions [8].…”

Section: Introductionmentioning

confidence: 99%

“…Several genes have been confirmed as stable reference genes and are widely used when studying insects, such as glyceraldehyde 3-phosphate dehydrogenase (GAPDH), actin (ACT), translation elongation factor 1α (TEF-1α), phospholipase A2 (PLA2), arginine kinase (AK), tubulin (TUB), TATA binding protein (TBP), and ribosomal proteins [7,[16][17][18][19][20]. However, few reference genes are consistently expressed across all conditions (e.g., tissues, developmental stages, and experimental conditions) [19,[21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Screening Potential Reference Genes in Tuta absoluta with Real-Time Quantitative PCR Analysis under Different Experimental Conditions

Yang

Wang

Huang

et al. 2021

Genes

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Tuta absoluta is one of the most significant invasive pests affecting tomato plants worldwide. RT-qPCR has emerged as one of the most sensitive and accurate methods for detecting gene expression data. The screening of stable internal reference genes is the most critical step for studying the molecular mechanisms of environmental adaptability. The stable reference genes expressed in T. absoluta under specific experimental conditions have not yet been clarified. In this study, seven candidate reference genes (RPL27, RPS13, RPS15, EF1-α, TUB, TBP, and β-actin) and their optimal numbers were evaluated under biotic (developmental stages and adult tissues) and abiotic (insecticide, temperature, and plant VOC) conditions using four software programs. Our results identified the following reference genes and numbers as optimal: three genes (EF1-α, RPS13, and RPL27) for different developmental stages (egg, larva, pupa, unmated adult), two genes (RPS13 and TBP) for adult tissues (antenna, head, thorax, abdomen, leg), two genes (TBP and RPS13) for insecticides (Bacillus thuringiensis, chlorpyrifos, abamectin-aminomethyl, and chlorantraniliprole), two genes (RPL27 and TUB) for temperature-induced stresses (0, 25, and 40 °C), and two genes (RPS13 and TUB) for VOC-induced stresses (nonanal, α-phellandrene, and tomato leaves). Our results provide a reference for selecting appropriate reference genes for further study of the functional genes of T. absoluta under different experimental conditions.

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Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)

Cited by 20 publications

References 40 publications

Reference Gene Selection for Normalizing Gene Expression in Ips Sexdentatus (Coleoptera: Curculionidae: Scolytinae) Under Different Experimental Conditions

Reference Gene Selection for Normalizing Gene Expression in Ips Sexdentatus (Coleoptera: Curculionidae: Scolytinae) Under Different Experimental Conditions

Evaluation of Reference Genes in Glenea cantor (Fabricius) by Using qRT-PCR

Screening Potential Reference Genes in Tuta absoluta with Real-Time Quantitative PCR Analysis under Different Experimental Conditions

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