Validation of Reference Genes for Accurate Normalization of Gene Expression in Lilium davidii var. unicolor for Real Time Quantitative PCR.
Validation of Reference Genes for Accurate Normalization of Gene Expression in Lilium davidii var. unicolor for Real Time Quantitative PCR.
复制标题
百合基因表达准确标准化的参考基因验证。
DOI：
10.1371/journal.pone.0141323
复制
发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Sun H
中科院分区：
文献类型：
--
作者：
Li X;Cheng J;Zhang J;Teixeira da Silva JA;Wang C;Sun H
百合是重要的商品球茎花卉。qRT-PCR是一种非常重要的追踪基因表达水平的技术。对合适的内参基因进行规范化的需求日益显著和迫切。生物体内内控基因的表达在不同的实验条件下差异很大。对于具有重要经济意义的百合，目前报道的qRT-PCR内参基因数量有限。本研究评估了α-TUB、β-TUB、ACT、eIF、GAPDH、UBQ、UBC、18S、60S、AP4、FP和RH2等12个候选基因在代表不同发育过程、三种胁迫处理（冷、热、盐）和不同器官的29个样品中的表达稳定性。对于不同的器官，ACT、GAPDH和UBQ的组合是合适的，而ACT和AP4或ACT和GAPDH分别适用于不同发育阶段的叶片和鳞片的规范化。在胁迫处理下，叶片、鳞片和根中，FP、ACT和AP4的表达最稳定。本研究为不同实验条件下内参基因的选择提供了指导，也有利于今后百合多种基因型中更准确的基因表达研究。
Lilium is an important commercial market flower bulb. qRT-PCR is an extremely important technique to track gene expression levels. The requirement of suitable reference genes for normalization has become increasingly significant and exigent. The expression of internal control genes in living organisms varies considerably under different experimental conditions. For economically important Lilium, only a limited number of reference genes applied in qRT-PCR have been reported to date. In this study, the expression stability of 12 candidate genes including α-TUB, β-TUB, ACT, eIF, GAPDH, UBQ, UBC, 18S, 60S, AP4, FP, and RH2, in a diverse set of 29 samples representing different developmental processes, three stress treatments (cold, heat, and salt) and different organs, has been evaluated. For different organs, the combination of ACT, GAPDH, and UBQ is appropriate whereas ACT together with AP4, or ACT along with GAPDH is suitable for normalization of leaves and scales at different developmental stages, respectively. In leaves, scales and roots under stress treatments, FP, ACT and AP4, respectively showed the most stable expression. This study provides a guide for the selection of a reference gene under different experimental conditions, and will benefit future research on more accurate gene expression studies in a wide variety of Lilium genotypes.