HrpN_(CSDS001)基因克隆及其表达产物诱导拟南芥基因表达谱变化的研究

Study of hrpN_(CSDS001) and the gene expression profile of Arabidopsis thaliana induced by Harpin_(CSDS001)

Erwinia carotovora subsp.carotovora CSDS001菌株具有可直接诱导烟草过敏反应特征,从构建的CSDS001菌株基因组文库,鉴定、克隆到hrpNCSDS001基因,GenBank登录号AY939927;构建的重组hrpNCSDS001基因工程菌株经IPTG诱导培养,获得的高效表达HarpinCSDS001蛋白,可诱导烟草发生过敏反应。30μg/mLHarpinCSDS001蛋白喷施拟南芥后,分析第3h、12h、24h、36h和48h拟南芥全基因谱表达动态变化,结果显示发生显著表达差异(logratio≤?1或≥1)的基因数分别为912、1787、2393、1833和1755。对被诱导发生显著表达差异的转录因子基因分析表明,有13个转录因子家族:ZIM、BES1、TCP、C2C2、AP2/EREBP、WRKY、bHLH、bZIP、GARP、MYB、NAC、HB、C2H2与HarpinCSDS001蛋白作用相关,这些转录因子家族主要参与调控植物抗性、光合作用、生长发育、开花等相关功能基因表达。

Erwinia carotovora subsp, carotovora CSDS001 elicits hypersensitive reaction （HR） in tobacco. From the genomic libraries of Erwinia carotovora subsp, carotovora CSDS001, the hrpNCSDS001 gene （GenBank number AY939927）, was isolated. The hrpNCSDS001 fusion protein was produced in Escherichia coli, and was used to induce HR by injecting into tobacco. We further examined the global regulation ofArabidopsis thaliana genes in response to HarpinCSDS001 at a concentration of 30 μg/mL. We indicated that 912, 1787, 2393, 1833 and 1,755 genes that were regulated significantly （log ratio ≤-1 or ≥1） at 3 h, 12 h, 24 h, 36 h and 48 h respectively after the treatment. Analysis of some transcription factors （TF） showed that 13 TF families responded to HarpinCSDS001 including ZIM, BES1, TCP, C2C2, AP2/EREBP, WRKY, bHLH, bZIP, GARP, MYB, NAC, HB, C2H2. These families mainly function in biological processes of plant defense, photosynthesis, development and flowering.