离子对高效液相色谱法分析黄瓜花叶病毒侵染烟草的总RNA水平变化

Analysis of Total RNA in CMV Infected Tobacco by Ion Pair Reverse Phase High Performance Liquid Chromatography

为研究植物病毒在RNA水平上对寄主植物的基因表达产生的影响,采用离子对高效液相色谱法,对黄瓜花叶病毒侵染烟草造成的总RNA的组成成分变化进行研究。离子对液相色谱法,是近几年才应用于对RNA进行分离、纯化和分析的一种试验技术,具有操作简单、重复性好、所需时间短等优点。选择适宜的离子对试剂,并对选定的离子对试剂正己胺/1,1,1,3,3,3-六氟-2-丙醇进行了优化,达到了较好的总RNA分离效果,并观察到健康植株和染病植株分离峰之间的差异,有差异的RNA种类还需进一步试验来验证。为研究植物病毒的致病机制提供一种新的试验方法和途径。

To study the impact of plant virus on host plant at the RNA level gene expression, in this study differential expressing of total RNA induced by cucumher mosaic virus （ CMV ） infection in tobacco through ion pair reversed phase liquid chromatography （ RP-HPLC ） was analyzed. Ion pair RP-HPLC has been applied for RNA isolation, purification and analysis in recent years, and has the advantages of direct injection, good reproducibility, and rapid analysis. Suitable ion pair reagents as the hexylamine/1, 1, 1, 3, 3, 3-Hexafluoro-2-propanol （ HA/ HFIP ） system was optimized to achieve a acceptable total RNA separation, the separation peak differences between healthy and diseased plant were observed, but needed further experiments to verify. Therefore, this paper provided an alternative method for study the viral effects on gene expression of host plants at the RNA level and plant virus pathogenic mechanisms.