Selection of optimized candidate reference genes for qRT-PCR normalization in rice (Oryza sativa L.) during Magnaporthe oryzae infection and drought
Selection of optimized candidate reference genes for qRT-PCR normalization in rice (Oryza sativa L.) during Magnaporthe oryzae infection and drought
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DOI：
10.4238/2014.november.27.7
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发表时间：
2014-01-01
影响因子：
0.4
通讯作者：
Petrofeza, S.
中科院分区：
文献类型：
--
作者：
Bevitori, R.;Oliveira, M. B.;Petrofeza, S.
由稻瘟病菌引起的干旱和稻瘟病是对全球水稻生产最严重的两个威胁。为了探索逆境诱导水稻基因表达的机制，在过去的几年里，人们进行了涉及转录组分析的研究。因此，有一组可靠的参考基因来标准化受不同胁迫影响的水稻基因的表达水平是至关重要的。为了寻找水稻在稻瘟病菌侵染和干旱条件下目标基因差异表达的潜在参考基因，本研究对5个基因表达标准化的看家基因进行了评价。使用分析软件geNorm和NormFinder评估这些基因表达的稳定性。对于所分析的所有样品，稳定性等级为UBQ5>；GAPDH>；EIF-4α>；β-Tub>；18S rRNA。这些数据表明，UBQ5、GAPDH和eIF-4α基因是合适的、高效的参考基因，将在未来水稻真菌感染和干旱的表达研究中发挥重要作用。
Drought and rice blast disease caused by Magnaporthe oryzae are two of the most serious threats to global rice production. To explore the mechanisms underlying gene expression induced in rice by stresses, studies involving transcriptome analyses have been conducted over the past few years. Thus, it is crucial to have a reliable set of reference genes to normalize the expression levels of rice genes affected by different stresses. To identify potential reference genes for studies of the differential expression of target genes in rice under M. oryzae infection and drought conditions, the present study evaluated five housekeeping genes for the normalization of gene expression. The stability of the expression of these genes was assessed using the analytical software packages geNorm and NormFinder. For all samples analyzed, the stability rank was UBQ5 > GAPDH > eIF-4 alpha > beta-TUB > 18S rRNA. The data showed that the UBQ5, GAPDH, and eIF-4 alpha genes are appropriate, high-performing reference genes and will be highly useful in future expression studies of fungal infections and drought in rice.