Expression Analysis of XTH in Stem Swelling of Stem Mustard and Selection of Reference Genes.

Mengyao Li,Ya Luo,Qing Chen,Fangjie Xie,Yong Zhang,Ronggao Gong,Jie Li,Fen Zhang,Haoru Tang,Jiali Liu,Bo Sun,Qi He,Yan Wang,Xiaorong Wang

doi:10.3390/genes11010113

Mengyao Li, Ya Luo + Show 12 more

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https://doi.org/10.3390/genes11010113

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Journal: Genes	Publication Date: Jan 20, 2020
Citations: 16	License type: CC BY 4.0

Affiliation: Sichuan Agricultural University, Institute of Pomology

Abstract

Accurate analysis of gene expression requires selection of appropriate reference genes. In this study, we report analysis of eight candidate reference genes (ACTIN, UBQ, EF-1α, UBC, IF-4α, TUB, PP2A, and HIS), which were screened from the genome and transcriptome data in Brassica juncea. Four statistical analysis softwares geNorm, NormFinder, BestKeeper, and RefFinder were used to test the reliability and stability of gene expression of the reference genes. To further validate the stability of reference genes, the expression levels of two CYCD3 genes (BjuB045330 and BjuA003219) were studied. In addition, all genes in the xyloglucan endotransglucosylase/hydrolase (XTH) family were identified in B. juncea and their patterns at different periods of stem enlargement were analyzed. Results indicated that UBC and TUB genes showed stable levels of expression and are recommended for future research. In addition, XTH genes were involved in regulation of stem enlargement expression. These results provide new insights for future research aiming at exploring important functional genes, their expression patterns and regulatory mechanisms for mustard development.

Highlights

Quantitative real-time PCR is considered an important method for detection and analysis of levels of gene expression
We found the value of E% in the eight reference genes to be between 94.2 and 108.7%, and the R2 was above 0.99
We found the value of E% in the eight reference genes to be between 94.2 and 108.7%, and the R2 was above 0.99 (Table 1 and Figure S3)

Summary

Introduction

Quantitative real-time PCR (qPCR) is considered an important method for detection and analysis of levels of gene expression. It has many advantages such as high accuracy, specificity, low cost, and reproducibility [1]. A number of reference genes have been identified including ACTIN, PP2A, and TUB and are commonly used in gene expression analyses [3]. These genes are mainly involved in maintaining basic cellular functions such as cell structure and primary metabolism

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