Reference Gene Selection for RT-qPCR Analysis of Flower Development in Chrysanthemum morifolium and Chrysanthemum lavandulifolium.
Reference Gene Selection for RT-qPCR Analysis of Flower Development in Chrysanthemum morifolium and Chrysanthemum lavandulifolium.
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菊花和薰衣草花发育 RT-qPCR 分析参考基因的选择
DOI：
10.3389/fpls.2016.00287
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发表时间：
2016
影响因子：
5.6
通讯作者：
Dai S
中科院分区：
文献类型：
--
作者：
Qi S;Yang L;Wen X;Hong Y;Song X;Zhang M;Dai S
实时荧光定量PCR（qPCR）是一种用于了解花发育的分子机制的常用和强有力的工具。然而，这种方法的准确性取决于参考基因的稳定性。菊科的头状花序非常特殊，由辐状小花和盘状小花组成。两种类型小花在对称性、性别、组织结构和功能上存在明显差异。此外，伞辐小花具有各种形状。本研究的目的是鉴定菊花和熏衣草花发育过程中的稳定参考基因。本研究选择了9个候选参考基因，并对其在花发育过程中的表达稳定性进行了评价，并通过4种不同的算法（Bestkeeper，Normandy，GeNorm和Ref-finder）验证了它们的稳定性。SAND家族蛋白（SAND family protein，SAND家族蛋白）是在所有样品或不同组织中表达最稳定的基因。熏衣草发育SAND和PGK（磷酸甘油酸激酶）在中国大花菊花品种中表现最稳定，而PGK在盆栽菊花中表现最好。品种间遗传背景复杂，最佳参考基因存在差异。这些研究结果为分析菊花花发育相关基因的表达模式提供了参考基因。
Quantitative real-time PCR (qPCR) is a popular and powerful tool used to understand the molecular mechanisms of flower development. However, the accuracy of this approach depends on the stability of reference genes. The capitulum of chrysanthemums is very special, which is consisting of ray florets and disc florets. There are obvious differences between the two types of florets in symmetry, gender, histological structure, and function. Furthermore, the ray florets have various shapes. The objective of present study was to identify the stable reference genes in Chrysanthemum morifolium and Chrysanthemum lavandulifolium during the process of flower development. In this study, nine candidate reference genes were selected and evaluated for their expression stability acrosssamples during the process of flower development, and their stability was validated by four different algorithms (Bestkeeper, NormFinder, GeNorm, and Ref-finder). SAND (SAND family protein) was found to be the most stably expressed gene in all samples or different tissues during the process of C. lavandulifolium development. Both SAND and PGK (phosphoglycerate kinase) performed most stable in Chinese large-flowered chrysanthemum cultivars, and PGK was the best in potted chrysanthemums. There were differences in best reference genes among varieties as the genetic background of them were complex. These studies provide guidance for selecting reference genes for analyzing the expression pattern of floral development genes in chrysanthemums.