【摘要】 为阐明泡桐多倍体响应盐胁迫的分子机制,本试验以白花泡桐为材料,采用同位素相对标记与绝对定量技术(isobaric tags for relative and absolute quantitation,iTRAQ)结合液相色谱与串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)定量蛋白质组技术,研究盐处理前后白花泡桐四倍体和对应二倍体的蛋白质表达变化。同时,选择6个差异表达蛋白质进行实时荧光定量PCR(qRT-PCR)验证。通过比对分析,共鉴定到2 851个蛋白质,有32个是倍性相关的盐应答差异蛋白质。KEGG pathway分析发现,这些差异蛋白质主要参与光合作用,信号转导,植物病原相互作用及淀粉和糖代谢等。其中,2-酮戊二酸脱氢酶E1、过氧化物酶和氢离子转运ATP合酶表达差异显著,表明这些蛋白质可能是潜在的盐应答蛋白质。QRT-PCR验证结果表明,mRNA与蛋白质表达水平不一致。更多还原

【Abstract】 To clarify the mechanism of polyploidy response to salt stress,isobaric tags for relative and absolute quantitation( iTRAQ) technology and liquid chromatography-tandem mass spectrometry( LCMS / MS) were used to identify changes of proteins in response to salt stress in tetraploid Paulownia fortunei and its diploid. Meanwhile,quantitative real-time PCR( qRT-PCR) was used to validate the expression data of six randomly selected differentially abundant proteins. Through comparative analysis,2851 proteins were identified,of which,32 were differentially abundant proteins related to salt stress in diploid and tetraploid Paulownia,whose functions were mainly involved in photosynthesis,biosynthesis of starch and sugar,plant hormone signal transduction and plant-pathogen interaction. Among these differentially abundant proteins,2-oxoglutarate dehydrogenase E1,H+-transporting ATPase and peroxidase were differentially expressed,suggesting that these proteins were the potential target proteins in response to salt stress in Paulownia. QRT-PCR showed that the expression levels of differentially abundant proteins were not consistent with those of the quantitative mass spectrometry.更多还原