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Open AccessArticle

Screening the Reference Genes for Quantitative Gene Expression by RT-qPCR During SE Initial Dedifferentiation in Four Gossypium hirsutum Cultivars that Have Different SE Capability

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Key Laboratory of Xinjiang Phytomedicine Resource and Utilization of Ministry of Education, College of Life Sciences, Shihezi University, Shihezi 832003, China
2
Key Laboratory of Oasis Eco-Agriculture, Shihezi University, Shihezi 832003, China
*
Authors to whom correspondence should be addressed.
Genes 2019, 10(7), 497; https://doi.org/10.3390/genes10070497
Received: 3 June 2019 / Revised: 24 June 2019 / Accepted: 25 June 2019 / Published: 28 June 2019
(This article belongs to the Section Plant Genetics and Genomics)
RNA sequencing (RNA-Seq)-based gene expression analysis is applicable to a wide range of biological purposes in various species. Reverse transcription quantitative PCR (RT-qPCR) is also used to assess target gene expression utilizing stably expressed reference genes as internal control under a given set of conditions. However, investigations of the reference genes for RT-qPCR normalization in the process of somatic embryogenesis (SE) initial dedifferentiation in Gossypium hirsutum are rarely reported. In this study, on the basis of our previous transcriptome data of three different induction stages during SE initial dedifferentiation process in four G. hirsutum cultivars that have different SE capability, 15 candidate genes were selected during SE initial dedifferentiation process, and their expression stability was evaluated by geNorm, NormFinder, and BestKeeper. The results indicated that the two genes of endonuclease 4 (ENDO4) and 18S ribosomal RNA (18S rRNA) showed stable expression in the four different G. hirsutum cultivars, endowing them to be appropriate reference genes during three induction stages in the four cotton cultivars. In addition, the stability and reliability of the two reference genes of ENDO4 and 18S rRNA were further verified by comparing the expressions of auxin-responsive protein 22 (AUX22) and ethylene-responsive transcription factor 17 (ERF17) between RT-qPCR results and the RNA-seq data, which showed strong positive correlation coefficient (R2 = 0.8396–0.9984), validating again the steady expression of ENDO4 and 18S rRNA as the reliable reference genes. Our results provide effective reference genes for RT-qPCR normalization during SE process in different G. hirsutum cultivars. View Full-Text
Keywords: reference gene; quantitative gene expression; RT-qPCR; Gossypium hirsutum; somatic embryogenesis reference gene; quantitative gene expression; RT-qPCR; Gossypium hirsutum; somatic embryogenesis
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MDPI and ACS Style

Cao, A.; Shao, D.; Cui, B.; Tong, X.; Zheng, Y.; Sun, J.; Li, H. Screening the Reference Genes for Quantitative Gene Expression by RT-qPCR During SE Initial Dedifferentiation in Four Gossypium hirsutum Cultivars that Have Different SE Capability. Genes 2019, 10, 497. https://doi.org/10.3390/genes10070497

AMA Style

Cao A, Shao D, Cui B, Tong X, Zheng Y, Sun J, Li H. Screening the Reference Genes for Quantitative Gene Expression by RT-qPCR During SE Initial Dedifferentiation in Four Gossypium hirsutum Cultivars that Have Different SE Capability. Genes. 2019; 10(7):497. https://doi.org/10.3390/genes10070497

Chicago/Turabian Style

Cao, Aiping; Shao, Dongnan; Cui, Baiming; Tong, Xuecheng; Zheng, Yinying; Sun, Jie; Li, Hongbin. 2019. "Screening the Reference Genes for Quantitative Gene Expression by RT-qPCR During SE Initial Dedifferentiation in Four Gossypium hirsutum Cultivars that Have Different SE Capability" Genes 10, no. 7: 497. https://doi.org/10.3390/genes10070497

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