宿根矮化病菌诱导甘蔗差异表达基因的cDNA-SCoT分析

宿根矮化病是危害严重的世界性甘蔗主要病害之一，为了探究甘蔗在该病胁迫下的抗病分子机制，本研究以甘蔗品种新台糖22健康植株为材料，利用含有宿根矮化病病原菌的蔗汁诱导其抗性，分别在接种后2、4、6、8和10 d取样，构建宿根矮化病侵染处理与对照的RNA混合池，利用cDNA-SCoT法进行差异显示研究。结果表明，80条SCoT引物扩增出500多条带，长度在100~1800 bp之间。从中筛选出30个差异明显的条带，测序后序列拼接，获得22条高质量的EST序列。生物信息学分析显示，对甘蔗宿根矮化病的差异基因主要涉及能量代谢、防卫反应、信号转导、蛋白质类代谢等方面。进一步基因功能分析显示，油菜素内酯合成蛋白、NBS-LRR类抗性蛋白、茉莉酸诱导蛋白、α-微管蛋白、ABA胁迫成熟蛋白、富含脯氨酸蛋白、翻译起始因子eif-2b α亚族等可能参与了甘蔗宿根矮化病病原菌互作的过程。

cDNA-SCoT Analysis of Differentially Expressed Genes in Sugarcane Induced by Leifsonia xyli subsp.xyli

Ratoon stunting disease (RSD) is one of the major diseases, which is harmful to sugarcane in the world. To investigate the molecular mechanism of gene differential expression in sugarcane under ratoon stunting disease stress, we inoculated the stem of sugarcane variety ROC22 with cane juice with RSD pathogen Leifsonia xyli subsp. xyli to induce resistance, then took stem samples at 2, 4, 6, 8, and 10 days, respectively, after inoculation, and established different cDNA mixture pools transcribed from the stem RNA mixture pools of RSD treatment and control for cDNA-SCoT analysis to detect the gene differential expressions. More than 500 bands with length of 100–1800 bp were obtained using 80 SCoT primers. A total of 30 differentially expressed EST sequences were screened out, and 22 non-redundant ESTs with high quality were obtained by cluster analyses of the ESTs sequencing. The results of BlastN showed that the differentially expressed genes of ROC22 mainly related to energy metabolism, disease defense, signal transduction, and protein metabolism. Further analysis of gene function indicated that brassinosteroid biosynthesis-like proteins, NBS-LRR type disease resistance protein, jasmonic acid induced protein, α-tubulin, abscisic stress ripening protein, proline-rich protein, and translation initiation factor eif-2b α subunit may be involved in the process of the incompatible interaction between plants and Leifsonia xyli subsp. xyli.