作　　者： ; ;

机构地区： 广州大学生命科学学院

出　　处： 《生物技术通讯》 2007年第4期581-586,共6页

摘　　要： 目的:采用基因表达谱分析方法,探讨小麦耐铝的分子机理。方法:利用抑制消减杂交(SSH)技术,以小麦的铝敏感品种Chisholm及其耐铝近等基因系Chisholm-T(其耐铝性来自小麦品种Atlas66)的根尖为材料,构建了2个铝胁迫后的SSHcDNA文库,共含有1628个表达序列标签(EST),利用这些EST制作了小麦根系的cDNA基因芯片。以cDNA基因芯片为平台,在铝胁迫后6h、1d、3d和7d,分别比较Chisholm和Chisholm-T之间的基因表达谱差异。结果:在各个时间点,耐铝和不耐铝小麦材料之间约有5%的EST表现出差异表达。对所有差异表达的EST进行测序分析,序列数据经Pipe-Online2.0进行毗连序列群(contig)拼接,发现只有8.3%的重复序列。结论:SSH是一种非常有效的差减和均一化的建库方法。对有功能注释的差异表达基因进行功能分类分析,表明这些基因参与了植物体内的电子传递、信号传导、植物保护和次生物质的代谢活动。 Objective： Aluminum（A1） is one of the most abundant metals in the earth and a major limiting factor to plant growth and development in acid soil. Although plant tolerance to A1 toxicity has been attractive to a growing number of researchers, the physiological, biochemical and molecular mechanisms of plant tolerance to A1 remain to be investigated. In help of understanding the molecular mechanism for A1 tolerance, expression profiles were conducted in this study. Methotis： Using suppression subtractive hybridization（SSH） technique, two subtractive libraries were constructed with expressed sequence tags （EST） of Al-stressed roots between Chisholm （an Al-sensitive wheat cultivar） and its near-isogenic line Chisholm-T in which wheat cultivar Atlas 66 was the donor of A1 tolerance. A total of 1 628 cDNA clones were isolated from the libraries and spotted on the glass slides. Expression patterns of those ESTs were examined at four time courses （6 h, 1 d, 3 d and 7 d after A1 stress）. Results： Approximately 5% of transcripts between Chisholm and Chisholm-T were differentially expressed in various time courses under A1 stress. Significantly up- or down-regulated ESTs were further examined by sequence analysis. Those EST sequences were assembled using PipeOnline 2.0 and showed only 8.3% redun- dancy, their putative functions were identified. Conclusion： This study indicats that SSH is an efficient method for library normalization. Function classification based on BLAST search results indicates that many genes with known function show differential expression and are involved in electron transportation, signal transduction, plant defense and secondary metabolism in the four time courses of A1 stress.

关 键 词： 小麦 基因芯片 耐铝性 表达谱

领　　域： [生物学] [生物学]