Selection and Validation of Reference Genes for qRT-PCR Gene Expression Analysis in Hydrangea macrophylla under Aluminum Treatment

doi:10.7668/hbnxb.20191716

Abstract

Abstract: In order to select the stable reference genes to ensure the reliability and accuracy in gene expression analysis of Hydrangea macrophylla under aluminum treatment, ten candidate reference genes including five traditional genes and five novel genes were detected by qRT-PCR in the different tissues and organs of Al³⁺-sensitive (Bailer) and insensitive (Ruby) cultivars based on transcriptome data. The expression stability of these genes was evaluated by geNorm,NormFinder,BestKeeper,ΔCt and RefFinder. The results showed that the expression levels of β-TUB (β-tubulin) and UPL7 (E3 ubiquitin protein ligase) were relatively high and relatively stable; The most suitable internal reference genes were UPL7, β-TUB, ACT7 and β-TUB, UBP15, UPL7 in Bailer and Ruby under Al₂(SO₄)₃ treatment, respectively. Simultaneously, the optimal internal reference genes were β-TUB, UBP15, UPL7 for different tissues, organs and all samples. Therefore, β-TUB and UPL7 were the most stable reference genes in different tissues of Hydrangea macrophylla under aluminum treatment. The relative expression levels of HmVALT were showed same expression pattern taking β-TUB, UPL7 and β-TUB + UPL7 as reference genes.

Key words: Hydrangea macrophylla, Aluminum treatment, Reference gene, Expression stability

CLC Number:

S682
Q78

CHEN Shuangshuang, QI Xiangyu, FENG Jing, CHEN Huijie, WANG Huadi, QIN Ziyi, DENG Yanming. Selection and Validation of Reference Genes for qRT-PCR Gene Expression Analysis in Hydrangea macrophylla under Aluminum Treatment[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 9-18. doi: 10.7668/hbnxb.20191716.

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