Selection of suitable reference genes for qPCR normalization under abiotic stresses and hormone stimuli in carrot leaves.
Selection of suitable reference genes for qPCR normalization under abiotic stresses and hormone stimuli in carrot leaves.
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选择合适的参考基因用于胡萝卜叶非生物胁迫和激素刺激下的 qPCR 标准化。
DOI：
10.1371/journal.pone.0117569
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Xiong AS
中科院分区：
文献类型：
--
作者：
Tian C;Jiang Q;Wang F;Wang GL;Xu ZS;Xiong AS
胡萝卜是菊科二年生草本植物，是世界上最重要的蔬菜作物之一。本研究从胡萝卜中克隆了9个候选参考基因(GAPDH、肌动蛋白、eIF-4α、PP2A、SAND、TIP41、UBQ、EF-1α和TUB)。胡萝卜植株受到非生物胁迫(热、冷、盐和干旱)和激素刺激(赤霉素、水杨酸、茉莉酸甲酯和脱落酸)的影响。候选参考基因的表达谱在三个技术和生物学重复中进行了评估。实时qPCR数据分析使用三个常用的基于Excel的小程序，即BestKeeper、geNorm和NormFinder。肌动蛋白和TUB是所有样本组中确定的最稳定的基因，但个体分析显示它们的表达谱发生了变化。GAPDH对大多数单次应力表现出最大的稳定性。为了进一步验证本研究中确定的参考基因的适用性，本研究对拟南芥AtDREB-A1基因同源基因DcDREB-A1在胡萝卜中的表达谱进行了研究。选择了在不同实验条件下稳定表达的合适参比基因。
Carrot, a biennial herb of the Apiaceae family, is among the most important vegetable crops in the world. In this study, nine candidate reference genes (GAPDH, ACTIN, eIF-4α, PP2A, SAND, TIP41, UBQ, EF-1α, and TUB) were cloned from carrot. Carrot plants were subjected to abiotic stresses (heat, cold, salt, and drought) and hormone stimuli (gibberellin, salicylic acid, methyl jasmonate, and abscisic acid). The expression profiles of the candidate reference genes were evaluated in three technical and biological replicates. Real-time qPCR data analyses were performed using three commonly used Excel-based applets namely, BestKeeper, geNorm, and NormFinder. ACTIN and TUB were the most stable genes identified among all sample groups, but individual analysis revealed changes in their expression profiles. GAPDH displayed the maximum stability for most of single stresses. To further validate the suitability of the reference genes identified in this study, the expression profile of DcDREB-A1 gene (homolog of AtDREB-A1 gene of Arabidophsis) was studied in carrot. The appropriate reference genes were selected that showed stable expression under the different experimental conditions.
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影响因子：
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作者：
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通讯作者：
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