棉花幼苗盐胁迫条件下solexa转录组测序结果的研究及验证-study and verification of sequencing results of so lexa transcriptome under salt stress in cotton seedlings.docxVIP

棉花幼苗盐胁迫条件下 棉花幼苗盐胁迫条件下 Solexa 转录组测序结果的分析及验证 4 4 We confirmed the sequencing result by detected relative expression level of 22 genes by RT-PCR. Generally, the expression patterns of these genes showed agreement with the solexa data, which basically confirmed the reliability of our transcriptome analysis. Transcript levels of 7 LEA genes were determined by qRT-PCR, the result suggested the 4 LEA genes we got from solexa data were most responsive to salt stress. Then, expression analysis of the 4 LEA genes was performed in leaves and roots. The result showed that the relative expression level of M19406 is much higher than TA24166_3635 and X15086, and the 3 LEA genes exhibited non-specific expression pattern. M19379 was likely root specific expression, the accumulation of its transcripts in leaves was barely detected even upon NaCl treatments. Full-length of the 4 LEA genes were cloned by the means of RACE. We collected 4 LEAs from cotton, 18 LEAs from Arabidopsis and 14 LEAs from rice. Then, we performed a ClustalX alignment of these genes. The unrooted phylogeny tree result showed that the identified LEA groups are quite distinct from each other, and the 4 LEA genes in cotton were classified into different groups. We expressed the 4 LEA genes in yeast, then analyzed the stress tolerance in solid and liquid YPG. The result suggested transgenic yeast with the 4 LEA genes have an improved tolerance to salt、osmotic and oxidative stress, which indicated the role of the 4 LEA genes in the adaptation of cotton to stress conditions. These results provide useful basic data for more in-depth analysis in Arabidopsis. Key words: salt stress; cotton; solexa sequencing; qRT-PCR; LEA protein; yeast 关于学位论文原创性和使用授权的声明 本人所呈交的学位论文，是在导师指导下，独立进行科学研究所取得 的成果。对在论文研究期间给予指导、帮助和和做出重要贡献的个人或集 体，均在文中明确说明。本声明的法律责任由本人承担。 本人完全了解山东农业大学有关保留和使用学位论文的规定，同意学 校保留和按要求向国家有关部门或机构送交论文纸质本和电子版，允许论 文被查阅和借阅。本人授权山东农业大学可以将本学位论文的全部或部分 内容编入有关数据库进行检索，可以采用影印、缩印或其他复制手段保存 论文和汇编本学术论文。同时授权中国科学技术信息研究所将本学位论文 收录