宿根矮化病菌诱导甘蔗差异表达基因的cDNA-SCoT分析  

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

作　　者：陈明辉[1] 张保青[1] 宋修鹏[1] 陈虎 杨丽涛[1,3] 李杨瑞[1,3] 陈保善[1] 

机构地区：[1]广西大学/亚热带农业生物资源保护与利用国家重点实验室,广西南宁530005 [2]广西林业科学院,广西南宁530002 [3]中国农业科学院甘蔗研究中心/广西农业科学院甘蔗研究所/广西作物遗传改良生物技术重点开放实验室/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,广西南宁530007

出　　处：《作物学报》2013年第6期1119-1126,共8页Acta Agronomica Sinica

基　　金：广西自然科学基金重点项目(2012GXNSFDA053011);广西自然科学基金创新团队项目(2011GXNSFF018002);广西自然科学基金重点项目(2012GXNSFDA053);广西壮族自治区主席科技资金项目(11166-02);科技部国际合作项目(2008DFA30600,2009DFA30820和2013DFA31600);广西农业科学院创新团队项目(桂农科2011YT01);广西科学研究与技术开发计划项目(桂科产1123008-1和桂科能0815011)资助

摘　　要：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10d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100～1800bp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。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 bio- synthesis-like proteins, NBS-LRR type disease resistance protein, jasmonic acid induced protein, α-tubulin, abscisic stress ripen-ing protein, proline-rich protein, and translation initiation factor eif-2b α subunit may be involved in the process of the incompati- ble interaction between plants and Leifsonia xyli subsp, xyli.

关 键 词：甘蔗 宿根矮化病 cDNA-SCoT 差异基因 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]