State Key Laboratory of Crop Stress Biology for Arid Area, College of Agronomy, Northwest A&F University, Yangling, 712100, Shaanxi, China,
Mol Biol Rep. 2013 Nov;40(11):6245-53. doi: 10.1007/s11033-013-2736-7. Epub 2013 Sep 25.
The non-protein-coding genes have been reported as a critical control role in the regulation of gene expression in abiotic stress. We previously identified four expressed sequence tags numbered S18 (EL773024), S73 (EL773035), S106 (EL773041) and S108 (EL773042) from a SSH-cDNA library of bread wheat Shaanmai 139 infected with Puccinia striiformis f. sp. tritici (Pst). Here, we isolated four cDNA clones and referred them as TalncRNA18, TalncRNA73, TalncRNA106 and TalncRNA108 (GenBank: KC549675-KC549678). These cDNA separately consisted of 1,393, 667, 449 and 647 nucleotides but without any open reading frame. The alignment result showed that TalncRNA18 is a partial cDNA of E3 ubiquitin-protein ligase UPL1-like gene, TalncRNA73 is an antisense transcript of hypothetical protein, TalncRNA108 is a homolog to RRNA intron-encoded homing endonuclease, and lncRNA106 had no similarly sequence. Quantitative RT-PCR studies confirmed that these four lncRNAs were differentially expressed in three near isogenic lines. TalncRNA108 was significantly stepwise decreased at early stage of inoculation with Pst, while the others were upregulated, especially at 1 and 3 dpi (days post-inoculation). Using Chinese Spring nulli-tetrasomic lines and its ditelosomic lines, TalncRNA73 and TalncRNA108 were located to wheat chromosome 7A and the short arm of chromosome 4B, respectively, while TalncRNA18 and TalncRNA106 were located to chromosome 5B. Comparing the sequence of DNA and cDNA of four lncRNAs with polymerase chain reaction primers, the results showed that all of them have no introns. The kinetics analyses of lncRNAs expression as a result of pathogen challenge in immune resistant genotype indicated that they may play the roles of modulating or silencing the protein-coding gene into pathogen-defence response.
非蛋白编码基因在非生物胁迫下的基因表达调控中发挥着关键的控制作用。我们之前从感染小麦条锈菌的小麦 Shaanmai 139 的 SSH-cDNA 文库中鉴定了四个表达序列标签,编号为 S18(EL773024)、S73(EL773035)、S106(EL773041)和 S108(EL773042)。在这里,我们分离了四个 cDNA 克隆,并将其命名为 TalncRNA18、TalncRNA73、TalncRNA106 和 TalncRNA108(GenBank:KC549675-KC549678)。这些 cDNA 分别由 1393、667、449 和 647 个核苷酸组成,但没有开放阅读框。序列比对结果表明,TalncRNA18 是 E3 泛素蛋白连接酶 UPL1 样基因的部分 cDNA,TalncRNA73 是假定蛋白的反义转录本,TalncRNA108 是 rRNA 内含子编码的归巢内切酶的同源物,而 lncRNA106 没有相似的序列。定量 RT-PCR 研究证实,这四个 lncRNA 在三个近等基因系中差异表达。TalncRNA108 在接种小麦条锈菌早期阶段呈显著的逐步下降趋势,而其他基因则上调,特别是在 1dpi 和 3dpi。利用中国春缺体-四体系及其易位系,TalncRNA73 和 TalncRNA108 分别定位在小麦 7A 染色体和 4B 染色体短臂上,而 TalncRNA18 和 TalncRNA106 定位在 5B 染色体上。用聚合酶链反应引物比较四个 lncRNA 的 DNA 和 cDNA 序列,结果表明它们都没有内含子。在免疫抗性基因型中,病原体胁迫导致 lncRNA 表达的动力学分析表明,它们可能在调节或沉默蛋白编码基因对病原体防御反应中发挥作用。