铝处理下绣球实时荧光定量PCR内参基因筛选及验证

doi:10.7668/hbnxb.20191716

摘要/Abstract

摘要： 为筛选适合铝处理下绣球中稳定表达的内参基因，在转录组数据分析的基础上，以Al₂（SO₄）₃处理下绣球铝敏感品种Bailer（商品名：Endless Summer^TM，中文名译为无尽夏）和不敏感品种Ruby（中文名译为红宝石）的不同组织为材料，分析10个候选内参基因（5个传统内参基因和5个新基因）的表达水平，并利用geNorm、NormFinder、BestKeeper、ΔCt及RefFinder 5个程序分析候选内参基因的表达稳定性。结果表明，β-TUB（β-微管蛋白）和UPL7（E3泛素蛋白连接酶）的表达水平较高，而且相对较稳定；表达稳定性分析结果显示，在Al₂（SO₄）₃处理下Bailer中最适内参基因为UPL7和β-TUB，其次是ACT7，Ruby中最适内参基因为β-TUB和UBP15，其次是UPL7；适用于不同组织器官以及所有样本的最适内参基因同样为β-TUB、UBP15和UPL7。因此，β-TUB和UPL7为铝处理下绣球实时荧光定量PCR最适内参基因；以β-TUB、UPL7和β-TUB+UPL7作为内参基因时，绣球铝转运蛋白基因HmVALT的表达模式一致。

关键词: 绣球, 铝处理, 内参基因, 表达稳定性

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

中图分类号:

S682
Q78

陈双双, 齐香玉, 冯景, 陈慧杰, 王华娣, 秦紫艺, 邓衍明. 铝处理下绣球实时荧光定量PCR内参基因筛选及验证[J]. 华北农学报, 2021, 36(2): 9-18. doi: 10.7668/hbnxb.20191716.

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.

http://www.hbnxb.net/CN/Y2021/V36/I2/9

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