Abstract
Kenaf (Hibiscus cannabinus L.), an environmental friendly and economic fiber crop, has a certain tolerance to abiotic stresses. Identification of reliable reference genes for transcript normalization of stress responsive genes expression by quantitative real-time PCR (qRT-PCR) is important for exploring the molecular mechanisms of plants response to abiotic stresses. In this study, nine candidate reference genes were cloned, and their expression stabilities were assessed in 132 abiotic stress and hormonal stimuli samples of kenaf using geNorm, NormFinder, and BestKeeper algorithms. Results revealed that HcPP2A (Protein phosphatase 2A) and HcACT7 (Actin 7) were the optimum reference genes across all samples; HcUBC (Ubiquitin-conjugating enzyme like protein) was the worst reference gene for transcript normalization. The reliability of the selected reference genes was further confirmed by evaluating the expression profile of HcWRKY28 gene at different stress durations. This work will benefit future studies on discovery of stress-tolerance genes and stress-signaling pathways in this important fiber crop.
Highlights
Being sessile, plants have to adapt and survive in various unfavorable environmental conditions
Transcript abundances of 9 candidate reference genes were assessed by qRT-PCR in 396 samples, and the Ct values were examined under three groups including abiotic stresses, hormonal stimuli, and all samples
Transcriptomics and functional genomics approaches have been implemented for understanding the defense mechanisms of plant response to unfavorable environmental conditions. quantitative real-time PCR (qRTPCR) has become the most powerful method for detection of transcriptomics data and study the molecular mechanisms of plant stress responses
Summary
Plants have to adapt and survive in various unfavorable environmental conditions. Soil desertification and salinization are rapidly increasing on a global scale, declining average yields for most major crop plants by more than 50% (Bartels and Sunkar, 2005). In response to these stress factors, plants can perceive abiotic stresses, and develop diverse signaling pathways comprising the regulation network of protein-protein reactions, protein-DNA interaction to combat or tolerate stresses. Elucidating the molecular mechanism of plant tolerance to adverse conditions is of fundamental importance to crop improvement
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