High-quality reference genes for quantifying the transcriptional responses of Oryza sativa L. (ssp indica and japonica) to abiotic stress conditions
High-quality reference genes for quantifying the transcriptional responses of Oryza sativa L. (ssp indica and japonica) to abiotic stress conditions
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DOI：
10.1007/s11434-013-5726-1
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发表时间：
2013-06-01
影响因子：
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通讯作者：
Selvaraj, Gopalan
中科院分区：
文献类型：
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作者：
Maksup, Sarunyaporn;Supaibulwatana, Kanyaratt;Selvaraj, Gopalan
水稻（Oryza saliva L.）对于粮食安全非常重要，也是众多谷类作物的优良模式植物。水稻功能基因组学研究包括通过定量实时 PCR (qPCR) 分析表征基因表达动态；这是开发能够应对不利气候变化的水稻品种的重要关键。 qPCR 分析需要使用适当的参考基因，以使任何定量解释有意义。在这里，新的潜在参考基因是从水稻微阵列实验的庞大公共数据库中选出的。通过geNorm(PLUS)和NormFinder软件验证了14个候选基因和4个常规内参基因的表达稳定性。七个候选基因优于 qPCR 中常规使用的参考基因，并且三个基因可以可靠地用于定量涉及非生物胁迫反应的基因的表达。这些高质量参考 EP (LOC_Os05g08980)、HNR (LOC_Os01g71770) 和 TBC (LOC_Os09g34040) 在三种籼稻基因型和一种粳稻基因型中效果良好。 KDML105 是泰国开发的籼稻基因型之一，其中包括茉莉香米，但迄今为止尚未报道其参考基因。
Rice (Oryza saliva L.) is important to food security and is also an excellent model plant for numerous cereal crops. A functional genomics study in rice includes characterization of the expression dynamics of genes by quantitative real-time PCR (qPCR) analysis; this is a significant key for developing rice varieties that perform well in the face of adverse climate change. The qPCR analysis requires the use of appropriate reference genes in order to make any quantitative interpretations meaningful. Here, the new potential reference genes were selected from a huge public database of rice microarray experiments. The expression stability of 14 candidates and 4 conventional reference genes was validated by geNorm(PLUS) and NormFinder software. Seven candidates are superior to the conventionally used reference genes in qPCR and three genes can be used reliably for quantitating the expression of genes involved in abiotic stress responses. These high-quality references EP (LOC_Os05g08980), HNR (LOC_Os01g71770), and TBC (LOC_Os09g34040) worked very well in three indica genotypes and one japonica genotype. One of indica genotypes including the Jasmine rice, KDML105 developed in Thailand for which no reference genes have been reported until now.