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Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
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Reference genes selection for transcript normalization in kenaf (Hibiscus cannabinus L.) under salinity and drought

Xiaoping Niu1, Jianmin Qi1, Meixia Chen2

  • 1Key Laboratory for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University , Fuzhou , China.

Peerj
|December 9, 2015
PubMed
Summary

Accurate gene expression analysis in kenaf under salinity and drought stress requires reliable reference genes. TUBα and 18S rRNA were identified as the most stable reference genes for normalizing gene expression in kenaf.

Keywords:
Gene expressionKenaf (Hibiscus cannabinus L.)Reference geneSalinity and drought stress

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Area of Science:

  • Plant Science
  • Molecular Biology
  • Agricultural Science

Background:

  • Kenaf (Hibiscus cannabinus) is a vital fiber crop susceptible to yield and quality reduction from salinity and drought stress.
  • Understanding kenaf's molecular response to these stresses is crucial for crop improvement.
  • Accurate gene expression normalization using stable reference genes is essential for transcriptomic studies.

Purpose of the Study:

  • To identify and validate the most stable reference genes for quantitative gene expression analysis in kenaf under salinity and drought stress.
  • To provide reliable normalization factors for future gene expression studies in kenaf.

Main Methods:

  • Ten candidate reference genes were selected from the kenaf transcriptome.
  • Gene expression stability was assessed using quantitative reverse transcription-PCR (qRT-PCR) on samples treated with NaCl (salinity) and PEG (drought).
  • Expression data were analyzed using geNorm, NormFinder, and BestKeeper algorithms.

Main Results:

  • TUBα and 18S rRNA were identified as the most stable reference genes under both salinity and drought stress conditions.
  • The expression of WRKY28 and WRKY32 genes was validated using TUBα and 18S rRNA as reference genes, confirming their reliability.
  • The study provides a validated set of reference genes for kenaf gene expression studies.

Conclusions:

  • TUBα and 18S rRNA are recommended as optimal reference genes for qRT-PCR normalization in kenaf under salinity and drought stress.
  • This finding will enhance the accuracy and reliability of future molecular research on kenaf stress responses.
  • The validated reference genes will contribute to a better understanding of kenaf's adaptation mechanisms to environmental challenges.