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Identification and validation of reference genes for quantitative real-time PCR studies in alligatorweed (Alternanthera philoxeroides)

文献类型： 外文期刊

作者： Yin, Junliang 1 ; Hou, Lu 2 ; Jiang, Xinchen 3 ; Yang, Jie 3 ; He, Yang 3 ; Zhou, Xiaokang 3 ; Zhu, Xiongmeng 3 ; Gong, A 1 ;

作者机构： 1.Yangtze Univ, Hubei Key Lab Waterlogging Disaster & Agr Use Wet, Jingzhou, Hubei, Peoples R China

2.Qinghai Univ, Qinghai Prov State Key Lab Plateau Ecol & Agr, Key Lab Agr Integrated Pest Management, Qinghai Acad Agr & Forestry Sci, Xining, Qinghai, Peoples R China

3.Yangtze Univ, Coll Hort & Gardening, Jingzhou, Hubei, Peoples R China

4.Xinyang Normal Univ, Coll Life Sci, Xinyang, Henan, Peoples R China

5.Coll Hort & Gardening, Jingzhou, Hubei, Peoples R China

6.Yangtze Univ, Coll Hort & Gardening, Jingzhou 434000, Hubei, Peoples R China

关键词： Housekeeping gene; internal control; qRT-PCR; RefFinder

期刊名称：WEED SCIENCE （ 影响因子：2.713； 五年影响因子：2.868 ）

ISSN： 0043-1745

年卷期： 2021 年 69 卷 4 期

页码：

收录情况： SCI

摘要： Alligatorweed [Alternanthera philoxeroides (Mart.) Griseb.] is an invasive semiaquatic weed that poses a serious threat to agricultural production and ecological balance worldwide. However, information about genetic factors associated with the adaptation and invasion mechanisms of this species is limited. Screening for appropriate reference genes is important for gene expression and functional analysis research in A. philoxeroides. In this study, 30 candidate genes that showed stable expression in different A. philoxeroides tissues under various treatments in RNA-seq data were chosen to design quantitative real-time PCR (qRT-PCR) primers. After the amplification specificity validation, 25 candidates were selected and further evaluated in a diverse set of A. philoxeroides samples, including leaf, stem, and root tissues under drought, salinity, heat, chilling, five herbicides, and corresponding untreated controls using qRT-PCR. The delta-CT method, geNorm, NormFinder, BestKeeper, and RefFinder algorithms were used to identify stable reference genes from A. philoxeroides samples. Overall, CoA, RFI2, Tubby, SRP19, and V-ATPase were the top five ideal reference genes in all organs and conditions. Tubby and CoA were the most stable reference genes in the leaf/stem; and RFI2, ERprr, and SPR19 were suitable reference genes for the roots. This work provided a foundation for exploring gene expression profiling of A. philoxeroides, especially those adaptation- and invasion-related genes, which may help in management of this invasive weed.

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