1. Identification of Reference Genes for Quantitative Gene Expression Studies in a Non-Model Tree Pistachio (Pistacia vera L.)
- Author
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Seyed Mahdi Seyedi, Christopher J. Botanga, Maryam Moazzam Jazi, and Effat Ghadirzadeh Khorzoghi
- Subjects
0106 biological sciences ,0301 basic medicine ,Leaves ,Gene Expression ,lcsh:Medicine ,Artificial Gene Amplification and Extension ,Plant Science ,Sodium Chloride ,01 natural sciences ,Biochemistry ,Polymerase Chain Reaction ,Plant Roots ,Peptide Elongation Factor 1 ,Gene Expression Regulation, Plant ,Tubulin ,Reference genes ,Gene expression ,lcsh:Science ,Cytoskeleton ,Genetics ,Abiotic component ,Regulation of gene expression ,Multidisciplinary ,Genes, Essential ,Pistacia ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Plant Anatomy ,Salt Tolerance ,Reference Standards ,Adaptation, Physiological ,Droughts ,Nucleic acids ,Cold Temperature ,Ribosomal RNA ,RNA extraction ,Research Article ,Cell biology ,Computer and Information Sciences ,Cellular structures and organelles ,Nucleic acid synthesis ,Research and Analysis Methods ,Genes, Plant ,Computer Software ,03 medical and health sciences ,Extraction techniques ,Stress, Physiological ,Botany ,Chemical synthesis ,RNA synthesis ,Non-coding RNA ,Molecular Biology Techniques ,Gene ,Molecular Biology ,lcsh:R ,Biology and Life Sciences ,biology.organism_classification ,Actins ,Gene expression profiling ,Plant Leaves ,Biosynthetic techniques ,030104 developmental biology ,RNA ,lcsh:Q ,Ribosomes ,010606 plant biology & botany - Abstract
The tree species, Pistacia vera (P. vera) is an important commercial product that is salt-tolerant and long-lived, with a possible lifespan of over one thousand years. Gene expression analysis is an efficient method to explore the possible regulatory mechanisms underlying these characteristics. Therefore, having the most suitable set of reference genes is required for transcript level normalization under different conditions in P. vera. In the present study, we selected eight widely used reference genes, ACT, EF1α, α-TUB, β-TUB, GAPDH, CYP2, UBQ10, and 18S rRNA. Using qRT-PCR their expression was assessed in 54 different samples of three cultivars of P. vera. The samples were collected from different organs under various abiotic treatments (cold, drought, and salt) across three time points. Several statistical programs (geNorm, NormFinder, and BestKeeper) were applied to estimate the expression stability of candidate reference genes. Results obtained from the statistical analysis were then exposed to Rank aggregation package to generate a consensus gene rank. Based on our results, EF1α was found to be the superior reference gene in all samples under all abiotic treatments. In addition to EF1α, ACT and β-TUB were the second best reference genes for gene expression analysis in leaf and root. We recommended β-TUB as the second most stable gene for samples under the cold and drought treatments, while ACT holds the same position in samples analyzed under salt treatment. This report will benefit future research on the expression profiling of P. vera and other members of the Anacardiaceae family.
- Published
- 2016