Reference Gene Selection for Quantitative Real-Time PCR in Chrysanthemum Subjected to Biotic and Abiotic Stress
Reference Gene Selection for Quantitative Real-Time PCR in Chrysanthemum Subjected to Biotic and Abiotic Stress
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生物和非生物胁迫下菊花实时定量 PCR 参考基因的选择
DOI：
10.1007/s12033-011-9394-6
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发表时间：
2011-10-01
影响因子：
2.6
通讯作者：
Chen, Fadi
中科院分区：
文献类型：
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作者：
Gu, Chunsun;Chen, Sumei;Chen, Fadi
定量实时PCR（RT-qPCR）是一种可靠的方法，用于评估基因表达，前提是包括合适的参考基因来标准化数据。八个潜在参考基因（即微管蛋白）的表达稳定性（α-2，4微管蛋白）、肌动蛋白、EF 1 α（延伸因子1 α），UBC（泛素C），GAPDH（甘油醛-3-磷酸脱氢酶）（代表光系统I的光合作用相关质体基因）、PP 2Acs（蛋白磷酸酶2A的催化亚基）和PGK使用geNorm软件在遭受蚜虫侵染、热胁迫或涝渍胁迫的菊花植物中评估磷酸甘油酸激酶（磷酸甘油酸激酶）。广泛使用的参考基因EF 1 α表现良好的蚜虫侵染的植物，但不好的水浸的。蛋白磷酸酶2A催化亚基（PP 2Acs）在高温和淹水胁迫下表现最好，而在蚜虫侵染下表现最差。常用的参考基因肌动蛋白通常是最不稳定的。没有一个基因适合单独进行标准化。参考基因的选择是基于RT-qPCR的基因表达研究中的重要因素。
Quantitative real-time PCR (RT-qPCR) is a reliable method for assessing gene expression, provided that suitable reference genes are included to normalize the data. The stability of expression of eight potential reference genes, namely, tubulin (alpha-2,4 tubulin), actin, EF1 alpha (elongation factor 1 alpha), UBC (ubiquitin C), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), psaA (photosynthesis-related plastid gene representing photosystem I), PP2Acs (catalytic subunit of protein phosphatase 2A), and PGK (phosphoglycerate kinase), was assessed in chrysanthemum plants subjected to aphid infestation, heat stress or waterlogging stress using geNorm software. The widely used reference gene EF1 alpha performed well for aphid infested plants but poorly for waterlogged ones. The catalytic subunit of protein phosphatase 2A (PP2Acs) was the best performing one during heat and waterlogging stress, but was the worst during aphid infestation. The commonly used reference gene actin was generally the least stable of the set. No single gene was suitable for normalization on its own. The choice of reference gene(s) is an important factor in gene expression studies based on RT-qPCR.