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  • Open Access


    Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (Eriobotrya japonica)

    Shoukai Lin1,2,#, Shichang Xu1,#, Liyan Huang1, Fuxiang Qiu1, Yihong Zheng1, Qionghao Liu1, Shiwei Ma1,2, Bisha Wu1,2, Jincheng Wu1,2,*

    Phyton-International Journal of Experimental Botany, Vol.92, No.4, pp. 1185-1201, 2023, DOI:10.32604/phyton.2023.026752

    Abstract Loquat (Eriobotrya japonica Lindl.) is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components. Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression. In this study, eight candidate reference genes were selected from our previously published RNA-seq data, and primers for each candidate reference gene were designed and evaluated. The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples, including 12 subgroups of developing or abiotic-stressed tissues. Different combinations of stable reference genes were screened… More >

  • Open Access


    Selection and validation of reference genes for quantitative real-time polymerase chain reaction analyses of Serratia ureilytica DW2


    BIOCELL, Vol.47, No.3, pp. 647-656, 2023, DOI:10.32604/biocell.2023.024758

    Abstract Background: Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C. pilosula; nonetheless, until now, no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction (RT–qPCR) data. Methods: To screen stable reference genes of S. ureilytica DW2, the expression of its eight candidate reference genes (16S rRNA, ftsZ, ftsA, mreB, recA, slyD, thiC, and zipA) under different treatment conditions (pH, temperature, culture time, and salt content) was assayed by RT–qPCR. The expression stability of… More >

  • Open Access


    Selection of Stable Reference Genes for Quantitative Real-Time PCR on Herbaceous Peony (Paeonia lactiflora Pall.) in Response to Drought Stress

    Zhipeng Sheng, Yuting Luan, Cong Xu, Jun Tao*, Daqiu Zhao*

    Phyton-International Journal of Experimental Botany, Vol.92, No.3, pp. 801-814, 2023, DOI:10.32604/phyton.2023.024953

    Abstract Herbaceous peony (Paeonia lactiflora Pall.), as a high-end cut flower in the international market, has high ornamental and medicinal values. But in Northern China, drought is a major environmental factor influencing the growth and development of P. lactiflora. Quantitative real-time polymerase chain reaction (qRT-PCR) can evaluate gene expression levels under different stress conditions, and stable internal reference is the key for qRT-PCR. At present, there is no systematic screening of internal reference for correcting gene expressions of P. lactiflora in response to drought stress. In this study, 10 candidate genes [ubiquitin (UBQ2), UBQ1, elongation factor 1-α (EF-1α), Histidine (His), eukaryotic… More >

  • Open Access


    Selection and Verification of Reference Genes for qRT-PCR Analysis in Iris domestica under Drought

    Qiang Ai, Cuijing Liu*, Mei Han*, Limin Yang

    Phyton-International Journal of Experimental Botany, Vol.91, No.11, pp. 2537-2548, 2022, DOI:10.32604/phyton.2022.021889

    Abstract Iris domestica is a plant of the Iridaceae family and is drought-tolerant, but its drought-resistance mechanism is not yet clear. Analysing the gene expression changes of I. domestica by qRT-PCR is an important mean to understand its drought resistance characteristics. Nevertheless, a lack of reference genes greatly hinders investigation and research on the adaptation of I. domestica to drought at the molecular and genetic levels. In this study, we assessed the expression stability of 11 candidate gene in I. domestica under drought stress conditions and different tissues using geNorm, NormFinder, BestKeeper and RefFinder tools. The results showed that EF1β was… More >

  • Open Access


    Identification of Suitable Reference Genes for qRT-PCR Normalization in Tilia miqueliana Maxim

    Huanli Wang, Lingjun Yan, Xi Huang, Zhongwei Wang, Yuanhao Yue, Shijie Tang*

    Phyton-International Journal of Experimental Botany, Vol.91, No.10, pp. 2191-2210, 2022, DOI:10.32604/phyton.2022.020735

    Abstract Quantitative real-time polymerase chain reaction (qRT-PCR) is a rapid and effective approach toward detecting the expression patterns of target genes. The selection of a stable reference gene under specific test condition is essential for expressing levels of target genes accurately. Tilia miqueliana, considered endangered, is a prominent native ornamental and honey tree in East China. No study has evaluated the optimal endogenous reference gene for qRT-PCR analysis in T. miqueliana systematically. In this study, fifteen commonly used reference genes were selected as candidate genes, and the stabilities of their expressions were assessed using four algorithms (GeNorm, NormFiner, BestKeeper, and DeltaCt)… More >

  • Open Access


    Selection of Stable Reference Genes for Quantitative Real-Time PCR on Paeonia ostii T. Hong et J. X. Zhang Leaves Exposed to Different Drought Stress Conditions

    Yuting Luan1, Cong Xu1, Xiaoxiao Wang1, Daqiu Zhao1, Jun Tao1,2,*

    Phyton-International Journal of Experimental Botany, Vol.91, No.5, pp. 1045-1059, 2022, DOI:10.32604/phyton.2022.018767

    Abstract The definition of relatively stable expressed internal reference genes is essential in both traditional blotting quantification and as a modern data quantitative strategy. Appropriate internal reference genes can accurately standardize the expression abundance of target genes to avoid serious experimental errors. In this study, the expression profiles of ten candidate genes, ACT1, ACT2, GAPDH, eIF1, eIF2, α-TUB, β-TUB, TBP, RNA Pol II and RP II, were calculated for a suitable reference gene selection in Paeonia ostii T. Hong et J. X. Zhang leaves under various drought stress conditions. Data were processed by the four regularly used evaluation software. A comprehensive… More >

  • Open Access


    Reference Gene Selection for qRT-PCR Normalization in Iris germanica L.

    Yinjie Wang, Yongxia Zhang, Qingquan Liu, Liangqin Liu, Suzhen Huang, Haiyan Yuan*

    Phyton-International Journal of Experimental Botany, Vol.90, No.1, pp. 277-290, 2021, DOI:10.32604/phyton.2020.011545

    Abstract Quantitative real-time PCR (qPCR) is an effective and widely used method to analyze expression patterns of target genes. Selection of stable reference genes is a prerequisite for accurate normalization of target gene expression by qRT-PCR. In Iris germanica L., no studies have yet been published regarding the evaluation of potential reference genes. In this study, nine candidate reference genes were assessed at different flower developmental stages and in different tissues by four different algorithms (GeNorm, NormFinder, BestKeeper, and RefFinder). The results revealed that ACT11 (Actin 11) and EF1α (Elongation factor 1 alpha) were the most stable reference genes in different… More >

  • Open Access


    Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues

    Gang Li1, Yao Zhou2, Yaqi Zhao2, Yaxue Liu2, Yuwei Ke2, Xiaoqing Jin1, Haoli Ma1,2,*

    Phyton-International Journal of Experimental Botany, Vol.89, No.2, pp. 329-344, 2020, DOI:10.32604/phyton.2020.08874

    Abstract Quantitative real-time PCR (qRT-PCR) is widely used for investigating gene expression patterns and has many advantages, including its high sensitivity, fidelity, and specificity. Selecting a satisfactory internal reference gene is crucial for obtaining precise gene expression results in qRT-PCR analyses. In this study, the transcriptomic data of 2 potato varieties were screened for housekeeping genes with stable expression patterns. A total of 77 putative genes were selected, which were highly and stably expressed. Then, qRT-PCR analyses were performed to examine the expression levels of these 77 candidate reference genes in various potato tissues, including leaves, flowers, stolons, and tubers. Gene… More >

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