Evaluation of Reference Genes in (Fabricius) by Using QRT-PCR

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2021 Dec 24

PMID 34946935

Citations 6

Authors

Ran-Ran Su

Zhong-Yan Huang

Chao-Wei Qin

Xia-Lin Zheng

Wen Lu

Xiao-Yun Wang

Affiliations

Soon will be listed here.

Abstract

Kapok is the main host of (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 , 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., , , , , , , , , , , , and ) 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 and were the most suitable reference genes in the different adult tissues, and and were best at the different developmental stages. and were the best fit for the qRT-PCR reference genes in the different sexes, while and 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 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 .

Citing Articles

Cellulose degradation in Glenea cantor (Fabricius) (Coleoptera: Cerambycidae): functional characterization of GcEGaseZ7 and Cellulase reveals a novel enzymatic activity.

Su R, Lan T, Pan B, Zheng X, Lu W, Wang X J Insect Sci. 2024; 24(6).

PMID: 39576188 PMC: 11583219. DOI: 10.1093/jisesa/ieae101.

Transcriptome and Expression Analysis of Glycerol Biosynthesis-Related Genes in Fabricius (Cerambycidae: Lamiinae).

Lan T, Su R, Dong Z, Tong X, Zheng X, Wang X Int J Mol Sci. 2024; 25(21).

PMID: 39519387 PMC: 11545850. DOI: 10.3390/ijms252111834.

Characterization of bacterial diversity and screening of cellulose-degrading bacteria in the gut system of (Fabricius) larvae.

Su R, Pan B, Luo Y, Zheng X, Lu W, Wang X Front Bioeng Biotechnol. 2024; 12:1340168.

PMID: 38456003 PMC: 10919226. DOI: 10.3389/fbioe.2024.1340168.

Antennal Transcriptome Analysis and Identification of Olfactory Genes in Fabricius (Cerambycidae: Lamiinae).

Wu G, Su R, Ouyang H, Zheng X, Lu W, Wang X Insects. 2022; 13(6).

PMID: 35735890 PMC: 9224838. DOI: 10.3390/insects13060553.

Identification of neuropeptides and neuropeptide receptor genes in Phauda flammans (Walker).

Wu H, Wang X, Hu J, Su R, Lu W, Zheng X Sci Rep. 2022; 12(1):9892.

PMID: 35701459 PMC: 9198061. DOI: 10.1038/s41598-022-13590-7.

References

Yin J, Sun L, Zhang Q, Cao C . Screening and evaluation of the stability of expression of reference genes in Lymantria dispar (Lepidoptera: Erebidae) using qRT-PCR. Gene. 2020; 749:144712. DOI: 10.1016/j.gene.2020.144712. View

Guo C, Pan H, Zhang L, Ou D, Lu Z, Khan M . Comprehensive Assessment of Candidate Reference Genes for Gene Expression Studies Using RT-qPCR in , a Predominant Parasitoid of . Genes (Basel). 2020; 11(10). PMC: 7601638. DOI: 10.3390/genes11101178. View

Etschmann B, Wilcken B, Stoevesand K, von der Schulenburg A, Sterner-Kock A . Selection of reference genes for quantitative real-time PCR analysis in canine mammary tumors using the GeNorm algorithm. Vet Pathol. 2006; 43(6):934-42. DOI: 10.1354/vp.43-6-934. View

Hruz T, Wyss M, Docquier M, Pfaffl M, Masanetz S, Borghi L . RefGenes: identification of reliable and condition specific reference genes for RT-qPCR data normalization. BMC Genomics. 2011; 12:156. PMC: 3072958. DOI: 10.1186/1471-2164-12-156. View

Deng Y, Zhao H, Yang S, Zhang L, Zhang L, Hou C . Screening and Validation of Reference Genes for RT-qPCR Under Different Honey Bee Viral Infections and dsRNA Treatment. Front Microbiol. 2020; 11:1715. PMC: 7406718. DOI: 10.3389/fmicb.2020.01715. View

Zhang L, Jing X, Chen W, Bai J, Vasseur L, He W . Selection of reference genes for expression analysis of plant-derived microRNAs in Plutella xylostella using qRT-PCR and ddPCR. PLoS One. 2019; 14(8):e0220475. PMC: 6675394. DOI: 10.1371/journal.pone.0220475. View

Yang X, Pan H, Yuan L, Zhou X . Reference gene selection for RT-qPCR analysis in Harmonia axyridis, a global invasive lady beetle. Sci Rep. 2018; 8(1):2689. PMC: 5807316. DOI: 10.1038/s41598-018-20612-w. View

Jing D, Zhang T, Bai S, He K, Prabu S, Luan J . Sexual-biased gene expression of olfactory-related genes in the antennae of Conogethes pinicolalis (Lepidoptera: Crambidae). BMC Genomics. 2020; 21(1):244. PMC: 7081556. DOI: 10.1186/s12864-020-6648-3. View

Wang X, Zheng X, Lu W . The complete mitochondrial genome of an Asian longicorn beetle (Coleoptera: Cerambycidae: Lamiinae). Mitochondrial DNA B Resour. 2020; 4(2):2906-2907. PMC: 7706684. DOI: 10.1080/23802359.2019.1661295. View

10.

Pfaffl M, Tichopad A, Prgomet C, Neuvians T . Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper--Excel-based tool using pair-wise correlations. Biotechnol Lett. 2004; 26(6):509-15. DOI: 10.1023/b:bile.0000019559.84305.47. View

11.

Gao Z, Deng W, Zhu F . Reference gene selection for quantitative gene expression analysis in black soldier fly (Hermetia illucens). PLoS One. 2019; 14(8):e0221420. PMC: 6697329. DOI: 10.1371/journal.pone.0221420. View

12.

Bustin S . Developments in real-time PCR research and molecular diagnostics. Expert Rev Mol Diagn. 2010; 10(6):713-5. DOI: 10.1586/erm.10.65. View

13.

Lu W, Wang Q, Tian M, Xu J, Qin A, He L . Host selection and colonization strategies with evidence for a female-produced oviposition attractant in a longhorn beetle. Environ Entomol. 2012; 40(6):1487-93. DOI: 10.1603/EN10280. View

14.

Lu J, Chen S, Guo M, Ye C, Qiu B, Wu J . Selection and Validation of Reference Genes for RT-qPCR Analysis of the Ladybird Beetle . Front Physiol. 2018; 9:1614. PMC: 6243213. DOI: 10.3389/fphys.2018.01614. View

15.

Lu W, Wang Q, Tian M, Xu J, Lv J, Wei S . Reproductive traits of Glenea cantor (Coleoptera: Cerambycidae: Lamiinae). J Econ Entomol. 2013; 106(1):215-20. DOI: 10.1603/ec12251. View

16.

Dong Z, Yang Y, Dou F, Zhang Y, Huang H, Zheng X . Observations on the Ultrastructure of Antennal Sensilla of Adult Glenea cantor (Cerambycidae: Lamiinae). J Insect Sci. 2020; 20(2). PMC: 7082121. DOI: 10.1093/jisesa/ieaa013. View

17.

Li X, Gong P, Wang B, Wang C, Li M, Zhang Y . Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae). Sci Rep. 2020; 10(1):21951. PMC: 7738536. DOI: 10.1038/s41598-020-78974-z. View

18.

Fu X, Meyer-Rochow V . Selection and Validation of Suitable Reference Genes for RT-qPCR Analysis in the Rare Aquatic Firefly (Coleoptera: Lampyridae). Insects. 2021; 12(4). PMC: 8072524. DOI: 10.3390/insects12040359. View

19.

Andersen C, Jensen J, Orntoft T . Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 2004; 64(15):5245-50. DOI: 10.1158/0008-5472.CAN-04-0496. View

20.

Bustin S, Benes V, Garson J, Hellemans J, Huggett J, Kubista M . The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009; 55(4):611-22. DOI: 10.1373/clinchem.2008.112797. View