分类号： S311 单位代码： 10335 密 级： 学 号：10716031 博 士 学 位 论 文 中文论文题目：中文论文题目：白菜型油菜同源四倍体与二倍体之间盐白菜型油菜同源四倍体与二倍体之间盐 和镉胁迫耐性的差异及其分子机理和镉胁迫耐性的差异及其分子机理 英文论文题目英文论文题目 Comparison between a tetraploid turnip and its diploid progenitor for their adaptation to salinity and cadmium stress and the underlying molecular mechanism 申请人姓名： 孟 华 兵 指导教师： 蒋 立 希 专业名称： 作 物 学 研究方向： 作 物 逆 境 分 子 生 理 所在学院： 农 业 与 生 物 技 术 学 院 论文提交日期论文提交日期 2010 年年 4 月月 26 日日 白菜型油菜同源四倍体与二倍体之间盐和镉胁迫耐白菜型油菜同源四倍体与二倍体之间盐和镉胁迫耐 性的差异及其分子机理性的差异及其分子机理 论文作者签名论文作者签名: 指导教师签名指导教师签名: 论文评阅人 1： 评阅人 2： 评阅人 3： 评阅人 4： 评阅人 5： 答辩委员会主席：张国平 教授 浙江大学农业与生物技术学院 委员 1：钱前 研究员 中国水稻研究所 委员 2：寿惠霞 教授 浙江大学生命科学学院 委员 3：邬飞波 教授 浙江大学农业与生物技术学院 委员 4：蒋立希 教授 浙江大学农业与生物技术学院 委员 5： 答辩日期： 2010 年 6 月 1 日 Comparison between a tetraploid turnip and its diploid progenitor for their adaptation to salinity and cadmium stress and the underlying molecular mechanism Authors signature: Supervisor s signature: External Reviewers: (姓名职称单位,下同) (隐名评阅学位论文省略) Examining Committee Chairperson: Guoping Zhang Professor CAB, ZJU Examining Committee Members: Qian Qian Professor CNNRI Huixia Shou Professor CLS, ZJU Feibo Wu Professor CAB, ZJU Lixi Jiang Professor CAB, ZJU Date of oral defence： June 1, 2010 Comparison between a tetraploid turnip and its diploid progenitor for their adaptation to salinity and cadmium stress and the underlying molecular mechanism A Dissertation Submitted to Zhejiang University in Partial Fulfillment of the Requirements for the Ph. D degree (Crop Science) 本研究受以下项目资助：本研究受以下项目资助： 国家自然基金（国家自然基金（30671339，30971700） 、浙江省钱江人才计 划 （ ） 、浙江省钱江人才计 划 （ 2006R10002 ）、 中 国 农 业 部 资 助 项 目 （ ）、 中 国 农 业 部 资 助 项 目 （2009ZX08009-076B） 、国家高技术研究发展计划（） 、国家高技术研究发展计划（863， 2006AA10A113） 、国家重点基础研究发展计划（） 、国家重点基础研究发展计划（973， 2006CB101602） 。） 。 This research work was supported by Natural Science Foundation (30671339, 30971700), The Science and Technology Bureau of Zhejiang province (2006R10002), The Agricultural Ministry of China (2009ZX08009-076B), National High-Tech R whereas the total methylaton ratio of the tetraploid in the control, salinity and cadmium stressed conditions were 20.3, 18.8, 22.7. The total methylaton level declined in both the diploid and tetraploid after salt treatment, but increased after cadmium treatment. In diploid the methylaton level decreased by 1.5 point, whereas in the tetraploid it decreased by 2.5 point after salt treatment. The full methylation ratio in diploid was 2.2 points higher than in tetraploid, while half methylation ratio in tetraploid was 1.6 points higher than in the diploid. 3. By using 96 primer pair combinations, about 2648, 2650, 2703, 2730, 2551 and 2580 cDNA fragments were generated respectively under the following treatments (1) tetraploid without stress; (2) diploid without stress; (3) tetraploid under salinity stress; (4) diploid under salinity stress; (5) tetraploid with cadmium toxicity; (6) diploid with cadmium toxicity. Salinity stress and cadmium toxicity led to 36 and 40 TDFs, respectively, which represented the functional genes that can be classified in to 3 or 4 categories, namely, transcriptional factors, stress responsive genes, genes relating to signal transduction and so on. Among the TDFs caused by salinity stress, 14 and 7 were repressed in the diploid and tetraploid, respectively; 5 and 10 were induced in the diploid and tetraploid, respectively. On the other hand, among the TDFs resulted from cadmium toxicity, 7 and 16 were repressed in the diploid and tetraploid, respectively; and 12 and 5 were induced in the diploid and tetraploid, respectively. Taken together, the tetraploid turnip had a better adaptation in the salinity and cadmium stressed environments. The MSAP and cDNA-AFLP analysis revealed the underlying genetic mechanisms that cause the morphological and physiological differences. Our work is a contribution to the advancement of our understanding on 浙江大学博士学位论文 摘要 vi the mechanism that an autopolyploid turnip deals with adverse environment. Key words: Brassica rapa; autotetraploid; tetraploid; diploid; salinity stress; cadmium stress 浙江大学博士学位论文 文中图、 表列表 vii 文中图、表列表文中图、表列表 图 1.0 胞嘧啶甲基化过程 图 2.1 不同的盐浓度胁迫下对二倍体白菜和四倍体白菜发芽率的影响 图2.2 不同的盐浓度在发芽期对四倍体白菜和二倍体白菜形态指标的影响 表 2.1 不同 NaCl 浓度下白菜发芽 7 d 后幼苗中 Na+含量 表 2.2 不同 NaCl 浓度下白菜发芽 7 d 后幼苗中 K+含量 表 2.3 不同 NaCl 浓度下白菜发芽 7 d 后幼苗中 Ca2+含量 表 2.4 不同 NaCl 浓度下白菜发芽 7 d 后幼苗中 K+/Na+比 表 2.5 不同 NaCl 浓度下白菜发芽 7 d 后幼苗中 Na+/Ca2+比 图 2.3 50 mM NaCl 处理对二倍体白菜和四倍体白菜的叶绿素、GSH、MDA 的影响 图 2.4 50 mM NaCl 处理对二倍体白菜和四倍体白菜的抗氧化酶的影响 图 2.5 50 mM NaCl 处理对二倍体白菜和四倍体白菜的钾钠离子含量分布的影响