Selection and validation of garlic reference genes for quantitative real-time PCR normalization
作者： Min Liu , Zhen Wu , Fangling Jiang
DOI: 10.1007/S11240-015-0780-9
关键词: Genetics 、 Polymerase chain reaction 、 18S ribosomal RNA 、 Ribosomal RNA 、 Gene expression 、 Reference genes 、 Genotype 、 Biology 、 Housekeeping gene 、 Gene
摘要: A stable reference gene is a prerequisite to obtaining reliable quantitative real-time PCR (qPCR) analysis results. Despite the global distribution and nutritional, medicinal economic value of garlic, genes for qPCR have not been reported. Eight garlic housekeeping genes, 18S ribosomal RNA (18S), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor (EF-1α), α-tubulin (TUA) β-tubulin (TUB), polyubiquitin (UBQ), cyclophilin (CYP), were chosen as candidate genes. Their stability was evaluated by GeNorm, NormFinder, BestKeeper, comparative ΔCt method, RefFinder. The results demonstrated that TUA, GAPDH, UBQ, CYP, ACT are most suitable eight different genotypes, organs, developmental stages, abiotic stresses media treatments, respectively. To further validate these expression levels superoxide dismutase (SOD) analyzed. relative quantification SOD varied according gene, thus highlighting importance using in qPCR. Our also provide molecular biological basis studying mechanisms development stress tolerance garlic.
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