白叶枯病菌侵染下的水稻内参基因稳定性

Selection of Optimized Candidate Reference Genes for Qrt-pcr Normalization in Rice during Xanthomonas Oryzae Pv. Oryzae Infection

水稻白叶枯病是由黄单胞菌水稻致病变种引起的一种细菌性枯萎病害,严重危害水稻产量。水稻受白叶枯侵染后,通过对相关基因表达变化进行检测,是研究水稻-白叶枯病菌分子互作机理的一种重要手段。为了筛选出稳定表达的内参基因,以确保后续基因表达分析结果的可靠性,本研究以接种了白叶枯病菌PXO61、PXO99的水稻叶片为研究材料,在侵染后不同时间点取样,对6个常用的水稻内参基因(TUB,EF1α,UBQ5,ACT,18Sr RNA和GAPDH)的表达进行qRT-PCR检测并用ge Norm、Norm Finder和Best Keeper三种软件对6对常用内参基因的稳定性进行了分析。结果发现:6个内参基因在两种白叶枯病菌侵染后的表达量变化均有差异;综合三种软件算法,受白叶枯PXO61和PXO99侵染后水稻叶片中表达最稳定的基因为EF1α、ACT和TUB,为水稻响应白叶枯病菌基因表达及其进一步的功能研究提供了参考和借鉴。

Xanthomonas oryzae pv. oryzae（Xoo）, which causes bacterial blight, is one of the devastating diseases of rice worldwide. The differential expression pattern of rice genes affected by Xoo will contribute to understand the molecular mechanisms in rice-Xoo interaction and further provide a tool for rice resistance breeding strategy.Thus, it is crucial to have a reliable set of reference genes to normalize the expression levels of rice genes affected by Xoo. To identify potential reference genes for studies of the differential expression of target genes in rice under Xoo infection, we assessed the gene expression stability of 6 frequently used housekeeping genes, including TUB,EF1α, UBQ5, ACT, 18 Sr RNA and GAPDH in a diverse set of 9 rice samples. The stability of the expression of these genes was assessed using the analytical software packages ge Norm, Norm Finder and Bestkeeper. Our data showed that EF1α,ACT and TUB were the most stable and high-performing reference genes and will be highly useful in future expression studies of rice-Xoo interaction.