甘蓝型油菜BnEIN3基因克隆及菌核病诱导表达

目的]探讨甘蓝型油菜EIN3（BnEIN3）基因与菌核病抗性的关系。方法]根据GenBank中已公布的拟南芥EIN-3基因的保守区及甘蓝型油菜EST序列，设计特异性引物，分别进行基因组PCR和RT-PCR扩增，克隆BnEIN3基因。对菌核病不同抗性的3个油菜品种中BnEIN3的表达进行荧光定量PCR检测分析。结果]克隆得到1947bp的基因片段，与拟南芥EIN3一致性高达82%，有完整的开放阅码框，编码614个氨基酸，其中包含1个EIN3结构域，初步认定该片段为油菜BnEIN3基因。在高抗品种（D083）中，菌核病在侵染后期快速诱导BnEIN3上调表达，且显著高于中抗（中双9号）及高感品种（84039）中该基因的表达水平。在中抗及高感品种中，菌核病均抑制BnEIN3的表达，且中抗品种BnEIN3表达量一直大于高感品种。结论]BnEIN3可能与油菜对菌核病抗性相关。

Cloning of Brassica napus EIN3 Gene and Its Expression Induced by Sclerotinia sclerotiorum

Objective] The study was to investigate roles of Brassica napus EIN3 in resistance to Sclerotinia sclerotiorum.Methods] Genomic PCR and RT-PCR were carried out to isolate genomic DNA and cDNA sequences of BnEIN3 from oilseed rape,based on the highly conserved region of EIN3 gene from Arabidopsis thaliana and the homologous sequences of oilseed rape ESTs.Expression levels of BnEIN3 were detected in three varieties of oilseed rape inoculated with S.sclerotiorum by real-time quantitative PCR.Results]A 1947-bp DNA fragment was obtained from oilseed rape.The fragment shared 82% identity to A.thaliana EIN3,encoding 614 amino acids containing an EIN3 domain,and was named as BnEIN3.Real-time PCR results showed that expression patterns of BnEIN3 were drastically different in different varieties.In highly resistant oilseed rape variety D083,BnEIN3 expression level was significantly increased 72 h after S.sclerotiorum inoculation whereas in middle resistant and susceptible varieties Zhongshuang 9 and 84039,BnEIN3 expression was suppressed.Conclusion] BnEIN3 may play an important role in oilseed rape resistance to S.sclerotiorum.