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21 April 2021 Identification and Validation of Reference Genes for Quantitative Real-Time PCR Studies in Alligatorweed (Alternanthera philoxeroides)
Junliang Yin, Lu Hou, Xinchen Jiang, Jie Yang, Yang He, Xiaokang Zhou, Xiongmeng Zhu, Andong Gong, Yongxing Zhu, Zhongyi Chen
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Abstract

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.

© The Author(s), 2021. Published by Cambridge University Press on behalf of the Weed Science Society of America.
Junliang Yin, Lu Hou, Xinchen Jiang, Jie Yang, Yang He, Xiaokang Zhou, Xiongmeng Zhu, Andong Gong, Yongxing Zhu, and Zhongyi Chen "Identification and Validation of Reference Genes for Quantitative Real-Time PCR Studies in Alligatorweed (Alternanthera philoxeroides)," Weed Science 69(4), 404-411, (21 April 2021). https://doi.org/10.1017/wsc.2021.32
Received: 13 July 2020; Accepted: 8 April 2021; Published: 21 April 2021
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