Validation of Reference Genes for Gene Expression Studies in Aphis glycines (Hemiptera: Aphididae)
Validation of Reference Genes for Gene Expression Studies in Aphis glycines (Hemiptera: Aphididae)
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DOI：
10.1603/ec12095
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发表时间：
2012-08-01
影响因子：
2.2
通讯作者：
Michel, Andy P.
中科院分区：
文献类型：
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作者：
Bansal, Raman;Mamidala, Praveen;Michel, Andy P.
定量实时聚合酶链反应（qRT-PCR）是用于精确定量mRNA转录物的常用且稳健的工具。为了使结果标准化，管家基因（[HKG]、参考基因或内源性对照基因）是必需的。大豆蚜（Aphis glycines Matsumura）是一种重要的大豆蚜虫，在我国大豆生产中具有重要的地位。我的天，然而，由于缺乏稳定的HKG，基因表达和功能基因组学研究受到阻碍。我们评估了七个潜在的香港公司，（SDFS，琥珀酸脱氢酶黄素蛋白亚基; EF 1a，延伸因子-1 α; HEL，解旋酶; GAPDH，甘油醛-3磷酸脱氢酶; RPS 9，核糖体蛋白S9; TBP，TATA盒结合蛋白;和UBQ，泛素缀合蛋白），以确定在组织，发育阶段，以及以敏感和抗宿主植物大豆为食的蚜虫。使用GeNorm和Normalt确定HKG稳定性。三种不同实验条件的结果显示，与分析样品中的其他HKG相比，TBP具有较高的稳定性。RPS 9在感病和抗病植株上的蚜虫中稳定表达，而EF 1a在组织和发育阶段中稳定表达。因此，我们建议TBP作为一个合适的HKG有效的治疗，组织和A的发育阶段之间的正常化。甘氨酸此外，RPS 9可用于宿主植物抗性实验，EF 1a可用于测试组织或发育阶段的差异表达。这些结果将使A.甘氨酸
Quantitative real-time polymerase chain reaction (qRT-PCR) is a common and robust tool for accurate quantification of mRNA transcripts. To normalize results, a housekeeping gene ([HKG], reference gene or endogenous control gene) is mandatory. Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is a significant soybean, Glycine max (L.) Merr., pest, yet gene expression and functional genomics studies are hindered by a lack of stable HKGs. We evaluated seven potential HKGs (SDFS, succinate dehydrogenase flavoprotein subunit; EF1a, elongation factor-1 alpha; HEL, helicase; GAPDH, glyceraldehyde-3 phosphate dehydrogenase; RPS9, ribosomal protein S9; TBP, TATA-box binding protein; and UBQ, ubiquitin-conjugating protein) to determine the most efficient HKGs that have stable expression among tissues, developmental stages, and aphids fed on susceptible and host plant-resistant soybean. HKG stability was determined using GeNorm and NormFinder. Results from three different experimental conditions revealed high stability of TBP compared with the other HKGs profiled across the samples assayed. RPS9 showed stable expression among aphids on susceptible and resistant plants, whereas EF1a showed stable expression in tissues and developmental stages. Therefore, we recommend the TBP as a suitable HKG for efficient normalization among treatments, tissues, and developmental stages of A. glycines. In addition, RPS9 may be used for host-plant resistance experiments and EF1a could be considered for testing differential expression across tissues or developmental stages. These results will enable a more accurate and reliable normalization of qRT-PCR data in A. glycines.