利用转录组及iTRAQ技术筛选高油酸油菜抗病相关基因

The Study of High Oleic Acid Rapeseed Disease Resistance Related Genes by Transcriptome and iTRAQ Analysis

高油酸油菜和低油酸油菜对菌核病的抗性存在显著差异,为了弄清其分子机理,以一组高油酸油菜近等基因系自交授粉后20-35 d的种子为材料,分别进行转录组和同位素相对标记与绝对定量技术分析。分析了与抗病相关的氧化磷酸化、植物激素信号转导通路和植物病原体互作3个分类,将注释的基因和蛋白进行关联,并对其中可能与抗病相关的基因进行定量PCR验证。结合前人研究发现,基因表达或蛋白表达发生显著变化的基因gi｜260505503（多聚半乳糖醛酸酶抑制蛋白）、gi｜226346102（HSR203J类蛋白）、gi｜470103214（钙调蛋白类）与抗病相关;而gi｜297843222（结合蛋白）、gi｜18397961（2-铁,2-硫-铁氧化还原类蛋白）、gi｜196052306（还原型烟酰胺腺嘌呤二核苷酸脱氢酶亚基）、gi｜18423437（还原型烟酰胺腺嘌呤二核苷酸脱氢酶（辅酶）1α-复形5）和gi｜297794581（激酶家族蛋白）等基因差异显著。

There are obvious differences between high oleic acid rapeseed and low oleic acid rapeseed on sclerotinia sclerotiorum resistance. In order to find out the molecule mechanism,the high oleic acid rapeseed inbred line seeds 20-35 d after pollination were used as material for transcriptome analysis and i TRAQ analysis respectively in this study. The classifications associated with disease resistance were oxidative phosphorylation,plant hormone signal transduction and plant-pathogen interaction were discussed,and the relationship between differential genes and the corresponding protein were investigated too. Then real-time quantitative PCR（ q PCR） analysis was used to verify the expression levels of differentially expressed genes which may be associated with disease resistance. Combined with previous study,the genes related with disease resistance were： gi ｜ 260505503（ polygalacturonase inhibitory protein）,gi ｜ 226346102（ HSR203J-like protein） and gi ｜ 470103214（ caltractin-like）. And the genes of gi ｜ 297843222（ binding protein）,gi ｜ 18397961（ 2Fe-2S ferredoxin-like protein）,gi ｜ 196052306（ NADH dehydrogenase subunit）,gi ｜18423437（ NADH dehydrogenase（ ubiquinone） 1alpha subcomplex 5） and gi ｜ 297794581（ kinase family protein）have obvious difference.