Reference gene selection for RT-qPCR analysis of Chrysanthemum lavandulifolium during its flowering stages
Reference gene selection for RT-qPCR analysis of Chrysanthemum lavandulifolium during its flowering stages
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薰衣菊花期 RT-qPCR 分析参考基因选择
DOI：
10.1007/s11032-012-9784-x
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Dai, Silan
中科院分区：
文献类型：
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作者：
Fu, Jianxin;Wang, Yi;Dai, Silan
实时定量RT-PCR（RT-qPCR）是一种可用于分析基因表达丰度的技术。选择在不同发育阶段和光周期处理下在不同组织中保持恒定水平的参考基因，分析开花时间基因和花发育基因的表达水平。利用数字化基因表达技术，筛选出9个在熏衣草幼叶期（CK 1）和现蕾期（W1）叶片中表达量适中的参考基因，作为候选参考基因。共采集115份C.分析了熏衣草属植物不同发育阶段的不同组织和不同光周期处理的叶片。9个参考基因的稳定性在样品中略有变化，但MTP、SKIP 16和PGK是总体上最稳定的基因。此外，分析了ClFT在具有成花能力的植物不同组织中的相对表达量，以验证本研究所选的参考基因。
Quantitative real-time RT-PCR (RT-qPCR) is a technology that can be used to analyze the abundance of gene expression. Reference genes, which are assumed to remain at constant levels in different tissues at various developmental stages and photoperiodic treatments, were selected to analyze the expression levels of flowering time genes and floral development genes. Using digital gene expression technology, nine reference genes with moderate expression in the leaves of Chrysanthemum lavandulifolium at the juvenile phase (CK1) and the squaring stage (W1) were selected as the candidate reference genes for further study. A total of 115 biological samples of C. lavandulifolium were analyzed, including different tissues under various developmental stages and leaves with varied photoperiodic treatments. The stability of the nine reference genes was slightly variable across the samples, but MTP, SKIP16 and PGK were the most stable genes overall. In addition, the relative expression level of ClFT in different tissues of plants with the competence to flower was analyzed to verify the reference genes selected in this study.