Phcog.Net logo
Submit

BROWSE ALL JOURNALS

    SEE ALL 6 JOURNALS
    Pharmacognosy MagazineNOW IN
    Article

    Selection of Suitable Reference Genes for Reverse Transcription‑quantitative Polymerase Chain Reaction Normalization in Artemisia annua L. Plants at Different Stages of Growth and Development

    Javed Ahmad1, Nazima Nasrullah1, Naved Quadri1, Umara Nissar1, Shashi Kumar2, Malik Zainul Abdin1★★ Corresponding author

    1. 1Centre for Transgenic Plant Development, Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, India.
    2. 2International Centre for Genetic Engineering and Biotechnology, New Delhi, India.

    CORRESPONDENCE

    Malik Zainul Abdin

    Centre for Transgenic Plant Development, Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, India.

    malikzabdin@gmail.com

    Received: 01-12-2018; Revised: 06-03-2019.

    Volume 15, Issue 66s · pp. S377–S385 · PUBLISHED 28 November 2019 · DOI: 10.4103/pm.pm_600_18

    View on Pharmacogn. Mag. original site ↗

    ABSTRACT

    Objectives: Our main aim for this study was to select the suitable reference gene for the normalization of reverse transcription‑quantitative polymerase chain reaction ( Reverse transcription RT‑qPCR) data in different tissues at various developmental stages in A. annua L. plants. Materials and Methods: Six candidate reference genes, namely; β‑actin (ACT), elongation factor 1‑alpha (EF1α), TAP‑42 interacting protein (TAP42), SAND family protein, β‑tubulin, and protein phosphatase 2A (PP2A) for their expression stability in the root, stem, leaf, and flower of A. annua L. plants at vegetative, preflowering, and flowering stages were analyzed using geNorm, NormFinder and BestKeeper, the Excel‑based research tools. Results: The genes ACT/PP2A, PP2A/TAP42, and EF1α/ PP2A were appropriate as reference genes in the leaf tissues at vegetative, preflowering, and flowering stages, respectively. In addition, EF1α/PP2A genes at vegetative and flowering stage, while EF1α/TAP42 gene at preflowering stage was found suitable reference genes for normalization of expression data in the stem. In the root samples, ACT/EF1α, EF1α/ PP2A, and ACT/TAP42 sets were found to be reliable reference genes at vegetative, preflowering, and flowering stages, respectively, whereas, PP2A/TAP42 gene set was found suitable for flower tissues at flowering stage. Conclusion: These results will be helpful in the normalization of expression data in RT‑qPCR to find the reliable outcome.

    KEYWORDS

    Open in new tab

    REFERENCES

    As published

    Showing references and in-text citations exactly as published.

      Cite this article

      SELECT FORMAT

      Ahmad, J., Nasrullah, N., Quadri, N., Nissar, U., Kumar, S., & Abdin, M. Z. (2019). Selection of Suitable Reference Genes for Reverse Transcription‑quantitative Polymerase Chain Reaction Normalization in Artemisia annua L. Plants at Different Stages of Growth and Development. Pharmacognosy Magazine, 15(66s), S377–S385. https://doi.org/10.4103/pm.pm_600_18