Validation of reference genes for quantifying changes in gene expression in virus-infected tobacco
Validation of reference genes for quantifying changes in gene expression in virus-infected tobacco
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DOI：
10.1016/j.virol.2017.06.029
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发表时间：
2017-10-01
期刊：
影响因子：
3.7
通讯作者：
Palukaitis, Peter
中科院分区：
文献类型：
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作者：
Baek, Eseul;Yoon, Ju-Yeon;Palukaitis, Peter
为了便于对病毒感染烟草植株的基因表达变化进行量化，研究了8个管家基因在三种系统感染病毒（黄瓜花叶病毒、马铃薯病毒X、马铃薯病毒Y）或一种只在接种叶片上产生超敏反应诱导病毒（烟草花叶病毒；TMV）感染期间的表达稳定性。通过5个参考基因验证程序，确定了系统感染病毒最稳定的基因顺序为核糖体蛋白L25 > p -微管蛋白> Actin，最不稳定的基因为泛素结合酶(UCE) < PP2A < GAPDH。对于TMV局部感染，最稳定的基因为EFl α >半胱氨酸蛋白酶> Actin，最不稳定的基因为GAPDH < PP2A < UCE。利用两个最稳定和两个最不稳定的验证内参基因，检测了三个防御应答基因，比较了它们在每种病毒引起的基因表达的相对变化。
To facilitate quantification of gene expression changes in virus-infected tobacco plants, eight housekeeping genes were evaluated for their stability of expression during infection by one of three systemically-infecting viruses (cucumber mosaic virus, potato virus X, potato virus Y) or a hypersensitive-response-inducing virus (tobacco mosaic virus; TMV) limited to the inoculated leaf. Five reference-gene validation programs were used to establish the order of the most stable genes for the systemically-infecting viruses as ribosomal protein L25 > P-Tubulin > Actin, and the least stable genes Ubiquitin-conjugating enzyme (UCE) < PP2A < GAPDH. For local infection by TMV, the most stable genes were EFl alpha > Cysteine protease > Actin, and the least stable genes were GAPDH < PP2A < UCE. Using two of the most stable and the two least stable validated reference genes, three defense responsive genes were examined to compare their relative changes in gene expression caused by each virus.