用cDNA-AFLP技术分析山核桃嫁接过程中的CcARF基因表达

Auxin response factor fragments cloned by cDNA-AFLP during Carya cathayensis grafting process

利用cDNA-AFLP(互补脱氧核糖核酸-扩增片段长度多态性)技术,在山核桃Carya carthayensis嫁接过程中分离得到1个与山核桃嫁接相关的cDNA片段。与美国国家生物技术信息中心(NCBI)同源性比较分析表明,该cDNA片段与小麦生长素响应因子部分区段有81%的同源性,命名为CcARF。荧光定量实时聚合酶链式反应(RT-PCR)分析表明:该基因在山核桃嫁接前砧木和接穗中都有强烈表达,但在嫁接后3d急剧下降,在这之后砧木中的表达增强不大,而接穗中嫁接后7d表达开始增强,到嫁接后14d表达明显,比嫁接后3d增强了4.7倍。该CcARF片段可能对山核桃嫁接起到基因表达调控的作用。

In order to improve survival of Carya cathayensis （Chinese hickory） grafted seedling, a cDNA fragment related to Chinese hickory graft was obtained by eDNA-amplified fragment length polymorphism （eDNA-AFLP） techniques during the grafting process. Analyses were conducted using the National Center for Bioteehnology Information （NCBI）homology comparison and Fluorescence Real Time Polymerase Chain Reaction （RT-PCR）. The NCBI homology comparison showed that a eDNA fragment, named CcARF, was 81% homologous with a wheat section of the auxin response factor （ARF）. Meanwhile, RT-PCR revealed that before grafting, gene expressions were strongly induced both in the rootstock and scion of Chinese hickory and declined sharply at 3 d after grafting. After 3 d, expression in rootstock was weakly in- duced. However, in the scion, 7 d after grafting, expression increased reaching a maximum 14 d after grafting; this was 4.7 times stronger than 3 d after grafting. Thus, when grafting Chinese hickory, the CcARFfragment may be involved in the regulation of gene expression. [Ch, 3 fig. 1 tab. 21 ref.]