摘要
以从小麦品种间杂交组合‘川35050／山农483’F7株系中分离出的感、抗小麦黄矮病的2个近等基因系为材料，利用DDRT—PCR技术和抗病基因保守结构域序列设计的7条简并或特异引物进行差异显示，PCR分析获得4条差异序列，并分别命名为：Rbdv1～Rbdv4（GenBank注册号：EU267934～EU267937）。以Rbdv1为靶序列，利用RACE对其3′末端进行PCR扩增，得到长778bp、末端有poly（A）尾巴的序列。用DNAMAN软件将3′RACE得到的序列与靶序列拼接得到1196bp长的片段，命名为A1（GenBank注册号：EU267938）。BlastN分析表明，A1与拟南芥、水稻中CDC48蛋白的同源性分别为81％和90％，其氨基酸序列中包含1个P—Loop，具有R基因的特征结构域，推测该片段很可能与抗小麦黄矮病基因相关。
Two near-isogenic lines for susceptible and resistant to WYDV （wheat yellow dwarf virus） derived from a F7 line of the cross between the wheat varieties ‘Chuan 35050＇ and ‘Shannong 483＇ were employed in this study. Seven specific or degenerated primers were designed on the basis of sequences of certain domains conversed to disease resistance genes （R gene）. Using these primers and DDRT-PCR （differential display reverse transcription PCR）, four expressed cDNA fragments Rbdvl-Rbdv4 （registered numbers in GenBank are EU267934-EU267937） were gained. Regarding Rbdvl as target sequence, 778 bp sequence of 3′ end was obtained using RACE （Rapid Amplification of cDNA Ends）. We assembled the sequences of the RACE product and Rbdvl under the software DNAMAN and obtained a 1 196 bp sequence named A1 （registered number in GenBank is EU267938）. And A1 showed close identity to CDC48 protein from Arabidopsis and rice （81% and 90% respectively） according to BlastN search in GenBank. Further more,the amino acid sequence of A1 included a P-Loop which was the conversed domain of R gene. There may be some relationships between the fragment A1 and the R gene of WYDV.
出处
《西北植物学报》
CAS
CSCD
北大核心
2008年第4期645-650,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家"十一五"科技支撑计划(2006BAD13B02)
