Chen Z J, Cao Z M, Yu Z D
College of Forestry, Northwest A&F University, Yangling, Shaanxi, China.
College of Forestry, Northwest A&F University, Yangling, Shaanxi, China
Genet Mol Res. 2014 Mar 24;13(1):2082-93. doi: 10.4238/2014.March.24.12.
Poplars are extensively cultivated worldwide, and their susceptibility to the foliar rust fungus leads to considerable damages in plantations. To better understand the molecular basis of poplar responses to the foliar rust, suppression subtractive hybridization was used to identify the potential important or novel genes involved in the Populus szechuanica infection by Melampsora larici-populina. A total of 515 expressed sequence tags (ESTs) with high quality were obtained and clustered into 66 contigs and 75 singletons to give a set of 141 uniESTs. The nucleotide Basic Local Alignment Search Tool (BLASTn) program was used to search for homologous sequences of the uniESTs in the GenBank database. Among them, 92.9% showed homology to the poplar genome, and 2% showed similarity to the rust fungus genome. In addition, homology to known genes was analyzed by the BLASTx algorithm, and approximately 50% of the uniESTs were significantly homologous to genes encoding proteins with known functions. Based on a reverse transcription-polymerase chain reaction (PCR) and quantitative PCR approach, five uniESTs were analyzed, and the results showed that the expression level of the thaumatin-like gene was highest at 72 h post-inoculation, and the pathogenesis-related protein 1 gene was highest at 48 h post-inoculation. The information generated in this study provides new clues to aid in the understanding of incompatibility between poplar and the foliar rust.
杨树在全球广泛种植,其对叶锈菌的易感性导致人工林遭受相当大的损害。为了更好地理解杨树对叶锈病反应的分子基础,采用抑制性消减杂交技术来鉴定参与四川杨被落叶松杨栅锈菌感染过程的潜在重要或新基因。共获得了515个高质量的表达序列标签(EST),并聚类成66个重叠群和75个单拷贝序列,得到一组141个单一EST。使用核苷酸基本局部比对搜索工具(BLASTn)程序在GenBank数据库中搜索单一EST的同源序列。其中,92.9%与杨树基因组具有同源性,2%与锈菌基因组具有相似性。此外,通过BLASTx算法分析与已知基因的同源性,约50%的单一EST与编码具有已知功能蛋白质的基因具有显著同源性。基于逆转录-聚合酶链反应(PCR)和定量PCR方法,对5个单一EST进行了分析,结果表明,类thaumatin基因的表达水平在接种后72小时最高,病程相关蛋白1基因在接种后48小时最高。本研究产生的信息为帮助理解杨树与叶锈病之间的不相容性提供了新线索。