论文部分内容阅读
利用电子克隆和RT-PCR技术,在本实验室前期构建的非亲和互作的SSH文库基础上,从条锈菌侵染的小麦水原11叶片中首次分离出一个含CBS结构域蛋白的基因,暂命名为TaCDCP1(Triticum aestivum CBS domain containing protein 1)。TaCDCP1包含一个完整的654 bp的开放阅读框,编码217个氨基酸。推测该基因拟编码的蛋白具有2个CBS保守结构域,不含跨膜区且无信号肽,定位在叶绿体基质内;经过同源比对,小麦TaCDCP1氨基酸序列与大麦、水稻和玉米等的同源序列的相似性较高;该基因表达量在小麦叶中显著高于在根和茎中;在小麦与条锈菌的非亲和、亲和组合中,TaCDCP1基因均受到条锈菌诱导,分别在接种后18 h和96 h达到表达高峰,非亲和组合表达量在侵染前期(接种后18~48 h)高于亲和组合,而在侵染后期(接种后96~120 h)低于亲和组合;外源植物激素脱落酸诱导该基因上调表达,苄基腺嘌呤,乙烯,赤霉素,茉莉酸甲酯和水杨酸处理后其表达量在不同程度上受到抑制;TaCDCP1在低温和干旱条件下表达量上升,在机械伤害和高盐处理下表达量无明显差异。表明TaCDCP1可能通过脱落酸等信号途径参与小麦对条锈菌的防御反应,同时参与低温和干旱环境下的信号转导途径。这些结果对于明确CBS结构域的功能以及CBS结构域蛋白尤其是TaCDCP1在小麦与条锈菌互作中的作用奠定了基础。
Based on the non-affinity interaction SSH library constructed in our lab, a CBS-domain-containing gene was first isolated from stripe rust-infected wheat Suwon 11 using electron cloning and RT-PCR , Temporarily named TaCDCP1 (Triticum aestivum CBS domain containing protein 1). TaCDCP1 contains a complete 654 bp open reading frame encoding 217 amino acids. It is speculated that the protein to be encoded by this gene has two CBS conserved domains, no transmembrane region and no signal peptide, and locates in the chloroplast matrix. After homologous comparison, the amino acid sequence of TaCDCP1 in wheat is homologous with the same in barley, rice and maize The gene sequence of TaCDCP1 gene was highly similar in root and stem in wheat leaves. In non-affinity and affinity combinations of wheat and stripe rust, the TaCDCP1 gene was induced by Stripe rust, The peak of expression reached the peak at 18 h and 96 h after inoculation, respectively. The expression level of non-affinity combination was higher than that of the affinity combination in the early stage of infection (18-48 h after inoculation), while in the late stage of infection (96-120 h after inoculation) Which was lower than the affinity combination. Exogenous plant hormone abscisic acid induced up-regulated expression of the gene, and the expression of the gene was inhibited to a certain extent by benzyladenine, ethylene, gibberellin, methyl jasmonate and salicylic acid; TaCDCP1 Under low temperature and drought conditions, the expression level increased, no significant difference in the expression of the mechanical injury and high salt treatment. This indicates that TaCDCP1 may participate in the defensive reaction of wheat stripe rust through signal pathway such as abscisic acid and participate in signal transduction pathways in low temperature and drought environment. These results lay the foundation for clarifying the function of the CBS domain and the role of CBS domain proteins, especially TaCDCP1, in the interaction between wheat and stripe rust.