枯萎病菌诱导感、抗陆地棉品种的基因表达谱分析

Expression Profiling Analysis between Resistant and Susceptible Cotton Cultivars(Gossypium hirsutum L.) in Response to Fusarium Wilt

选用陆地棉抗枯萎病品种中棉所12和感枯萎病品种新陆早7号,利用Solexa高通量测序技术分析棉花幼苗受到枯萎病菌侵染后3 h和6 h根部组织的基因表达谱。中棉所12受病菌侵染后3 h与0 h相比,6 h与0 h相比以及6 h与3 h相比,分别鉴定出4447、5481和2559个差异表达基因;新陆早7号分别鉴定出8615、6727和2078个差异表达基因;2个品种比较,在3 h和6 h分别鉴定出1879个和500个差异表达基因。基因功能分析表明,差异表达基因划分为生物学过程、细胞组分和分子功能注释本体,并进一步细分为48个功能类别。代谢通路分析显示,中棉所12和新陆早7号在枯萎病菌侵染后的6 h与0 h相比鉴定出的代谢通路(Pathway)最多,均有126条;在枯萎病菌侵染后6 h,2个品种相比鉴定出的代谢通路最少,仅有89条。各个比对组的代谢通路涉及的基因可被划分为次生代谢产物的生物合成、多糖的生物合成和代谢、环境适应和免疫系统等13类。在环境适应和免疫系统分类中存在着植物与病原菌相互作用代谢通路,涉及到996个已知功能的差异表达基因,有444个基因上调表达,552个基因下调表达。其中,差异表达基因最多的是WRKY转录因子家族基因,其次为丝氨酸/苏氨酸蛋白激酶基因,而DNA损伤修复/耐受性蛋白,JAZ1、RAR1和RPM1互作蛋白,S位点的特异性糖蛋白S6前体以及钙牵引蛋白等6类基因参与该代谢通路的数目最少。最后随机抽取6个基因进行实时荧光定量PCR验证,其结果与测序结果一致。

Fusarium Wilt resistant cotton cultivar Zhongmiansuo 12 and susceptible cultivar Xinluzao 7 were used to analyze the gene expression profiling of root tissues at three hours and six hours after the seedlings infection by Fusarium Wilt using Solexa sequencing technology. Compared with no infection, 4447 and 5481 differential genes after three hours and six hours infected by Fusarium Wilt were identified, and compared six hours with three hours, 2559 differential genes were identified in Zhongmiansuo 12; while 8615, 6727, and 2078 was respectively identified in Xinluzao 7. There were 1879 and 500 differential genes in three hours and six hours after infection when the two cultivars were compared. According to the Gene Ontology, these genes were divided into these groups of biological process, cellular component and molecular function; then further subdivided into 48 func-tional categories. By analyzing the pathways, the most of them were identified in 6 h/0 h after infection between Zhongmiansuo 12 and Xinluzao 7, which were 126 each in the two cultivars; the least of pathways were at 6 h after infection in two cultivars, which was 89 only. However, all the pathways in each comparison group could be classified into 13 categories, such as biosynthe-sis of other secondary metabolites, glycan biosynthesis and metabolism, environmental adaptation, and immune system. A path-way, named plant-pathogen interaction in the environmental adaptation and immune system category, was involved in 996 differ-ential genes; and the number of up regulated genes was 444 and that of down regulated genes was 552. The most differential genes were in WRKY transcription factor family, the serine/threonine kinase had the medium number of differential genes, while DNA damage-repair/toleration protein, JAZ1, RAR1, RPM1-interacting protein,S locus specific glycoprotein S6 precursor and caltractin had the least genes. Finally, six random genes were subjected to RT-PCR and the results validated the Solexa sequencing conclusions.