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【目的】PPR(pentatricope ptide repeat)蛋白家族在植物的生长发育、细胞器形成、细胞质雄性不育的育性恢复、RNA的编辑加工、细胞核与细胞器之间信号传递、逆境防御等方面具有重要作用。研究旨在了解PPR蛋白家族SoDIPPR基因在马铃薯中的表达情况及其在干旱胁迫下的作用【。方法】以抗旱马铃薯二倍体品系H145为材料,利用cDNA-AFLP技术寻找与抗旱性密切相关的基因cDNA片段,利用RACE技术克隆其基因全长cDNA,并利用半定量RT-PCR和Northern杂交法研究其在干旱条件下的表达情况。【结果】从抗旱马铃薯二倍体品系H145中克隆了一个836bp全长cDNA序列,命名为SoDIPPR,该序列开放阅读框长为588bp,编码195个氨基酸序列。序列分析表明,SoDIPPR蛋白存在PPR结构域、激酶结合区、和C端SMR(small MutS related protein)区域。SoDIPPR蛋白序列与GenBank数据库中的其它PPR蛋白进行同源序列比对,构建系统进化树,发现SoDIPPR与其它14个高等植物的PPR蛋白同源性为57%～82%,与苜蓿DNA错配修复蛋白MuTs2、水稻盐诱导蛋白(Q9LS25)和叶绿体RNA结合蛋白(Q75IP8)的同源性达69%～82%。半定量RT-PCR和Northern杂交结果表明,SoDIPPR基因在干旱胁迫下叶片和根系里的表达量明显增加,说明SoDIPPR基因在马铃薯抗旱中起一定作用。在持续干旱条件下,SoDIPPR基因在抗旱品系H145与干旱敏感品系H214中表达模式不同。【结论】马铃薯SoDIPPR基因编码的蛋白与其它植物PPR家族蛋白同源性较高,SoDIPPR基因参与马铃薯对干旱胁迫的应答反应,可能对抵抗干旱胁迫有一定作用。
【Objective】 The pentatricope ptide repeat (PPR) protein family plays an important role in plant growth and development, organelle formation, fertility restoration of cytoplasmic male sterility, RNA editing and processing, signal transduction between nuclear and organelles, and defense against adverse reactions. This study aimed to understand the expression of SoPIPPR gene in potato and its role in drought stress. Methods The drought-resistant potato diploid line H145 was used as a material to search for cDNA fragments closely related to drought resistance by using cDNA-AFLP technique. The full-length cDNA was cloned by RACE technique. Semi-quantitative RT-PCR and Northern blotting Study its expression under drought conditions. 【Result】 The full-length cDNA sequence of 836bp was cloned from drought-resistant potato diploid H145 and named SoDIPPR. The open reading frame of this cDNA was 588bp and encoded a 195 amino acid sequence. Sequence analysis showed that the SoDIPPR protein has PPR domain, kinase binding region, and C-terminal SMR (small MutS related protein) region. SoDIPPR protein sequences were compared with other PPR proteins in GenBank database to construct phylogenetic tree. The results showed that SoPIPPR shared 57% -82% homology with PPR protein of other 14 higher plants, The homology of protein MuTs2, rice salt-inducible protein (Q9LS25) and chloroplast RNA-binding protein (Q75IP8) was 69% -82%. The results of semi-quantitative RT-PCR and Northern blot showed that the expression of SoDIPPR gene in leaves and roots significantly increased under drought stress, indicating that SoDIPPR gene plays a role in potato drought resistance. Under continuous drought conditions, the expression pattern of SoDIPPR was different between drought-resistant line H145 and drought-sensitive line H214. 【Conclusion】 The protein encoded by potato SoDIPPR gene shares high homology with PPR family proteins of other plants. SoDIPPR gene is involved in the response of potato to drought stress and may play a role in resisting drought stress.