菊芋幼苗叶片干旱胁迫的生理响应和蛋白质组学研究
文献类型：学位论文
| 作者 | 张美德 |
| 学位类别 | 博士 |
| 答辩日期 | 2010-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 沈世华 |
| 关键词 | 菊芋 干旱胁迫 蛋白质组 |
| 学位专业 | 生态学 |
| 中文摘要 | 菊芋（Helianthus tuberosus L.）是菊科向日葵属多年生植物，块茎中丰富的果糖可用来生产乙醇。由于菊芋具有很强的抗逆性，能在干旱、盐碱等恶劣环境生长，是一种理想的能源植物。然而由于菊芋不同品种间抗性差别较大，极大的影响其在生产实践中的推广应用。本文从生理、分子两方面着手，对菊芋两个不同耐旱性品种幼苗在20% PEG 诱导的干旱胁迫下的响应进行了生理、蛋白质组层面的剖析，取得了以下结果：干旱胁迫严重影响了菊芋的生理和生化过程。干旱胁迫造成了菊芋水势的降低，同时破坏细胞的膜系统，并积累了大量的活性氧(ROS)，对植株造成氧化胁迫伤害。和抗旱性较弱的品种“榆林”相比，“岫岩”有着较强的耐旱性，是由于“岫岩”在胁迫后能保持较高的水势、CAT 和 SOD 活性；另外，较高的游离脯氨酸积累也有助于“岫岩”耐旱性的增强。双向电泳结果表明，干旱胁迫后“岫岩”共有 69 个变化量在 1.5 倍以上的差异蛋白，“榆林”共有 63 个变化量在 1.5 倍以上的差异蛋白。经过 LC-MS/MS 分析，分别鉴定出 46 个差异蛋白（“岫岩”）和 34 个差异蛋白（“榆林”） 根据功。能不同，鉴定的蛋白质可分为光合作用相关蛋白、抗逆相关蛋白、信号转导类蛋白、碳代谢类蛋白、能量以及一些其他功能蛋白质等六大类。进一步的蛋白功能分析显示，两品种对干旱胁迫的蛋白调控有所不同。干旱胁迫导致多个光合相关的蛋白表达受到抑制，发生了下调。然而受干旱诱导，“岫岩”比“榆林”出现更多的光合相关的上调蛋白。如“榆林”的 Rubisco 大亚基的表达受干旱胁迫抑制发生下调，而“岫岩”的 Rubisco 大亚基的表达受干旱胁迫诱导发生上调。表明“岫岩”可能通过调节 Rubisco 表达量来其维持较高的光合速率，从而对干旱胁迫作出适应性反应。此外，胁迫后菊芋两品种都积极启动抗逆相关蛋白，以减轻逆境胁迫的影响。但是，更多的抗逆相关蛋白在“岫岩”中受诱导上调，如疾病、抵抗蛋白（disease resistance-like protein） Peroxiredoxin-2F 蛋白、乳酰谷胱苷肽裂解酶（lactoylglutathione lyase）。另外 ， 菊芋两品种在信号转导、碳代谢等方面的蛋白质表达也有差异。因此，干旱胁迫后，“岫岩”进行了更为积极有效的蛋白调控以应对逆境胁迫，这可能是菊芋两品种抗旱性差异的原因所在。综上所述，干旱胁迫后，菊芋体内抗氧化系统以及渗透调节能力对其抗旱性起着重要作用；而干旱响应蛋白(光合、抗逆、信号转导等方面蛋白)的有效调控也对菊芋的抗旱性起着重要作用。 |
| 英文摘要 | Jerusalem artichoke (Helianthus tuberosus L.) is a perennial herb belonging to Asteraceae family. Its tubers are rich in fructose which can be used to produce ethanol. Due to its high resistance to adverse conditions such as drought, saline, Jerusalem artichoke is regard as a promising candidate crop for the manufacture of bioethanol. But the resistance differs a lot between Jerusalem artichoke cultivars, which prevent its further practice and application. In this study, we investigated the physiological and proteomic responses of two different Jerusalem artichoke cultivars to polyethylene glycol (PEG) 6000 induced drought stress. Conclusions presented in this thesis can be summarized as follows:Drought stress influenced physiological and biochemical progress in Jerusalem artichoke. It caused the decrease in leaf water potential and membrane lipid peroxidation in Jerusalem artichoke. Drought also led to the accumulation of reactive oxygen species (ROS), which resulted in the oxidative damage. Compared to cv. Yulin, cv. Xiuyan was more tolerant to water stress-induced oxidative damage as evidenced by its relatively higher water status, CAT activity, and SOD activity. Moreover, relatively higher level of free proline accumulation in cv. Xiuyan was also associated with its better protection against water stress-induced oxidant damage.Two-dimensional electrophoresis (2-DE) results showed 69 protein spots in cv. Xiuyan changed in abundance more than 1.5-fold after drought treatments, while 63 protein spots in cv. Yulin changed in abundance more than 1.5-fold. 46 proteins in cv. Xiuyan and 34 proteins in cv. Yulin were identified through LC-MS/MS and NCBI database searching. The identified proteins could be sorted into 6 functional categories, including photosynthesis, defense, signal transduction, carbon metabolism, energy and others. Further protein function analysis revealed the different protein regulation in two cultivars. Drought stress inhibited expression of some photosynthetic proteins in two cultivars. However, more proteins related to photosynthesis up-regulated in cv. Xiuyan after stress as compared with cv. Yulin. For example, expression of protein Rubisco laege subunit in cv. Yulin was inhibited after stress, but it up-regulated in cv. Xiuyan. It may be explained that cv. Xiuyan increased expression of protein Rubisco laege subunit to maintain a relatively high photosynthetic rate fter drought stress. Drought stress also induced the expression of proteins related to defense in two cultivars. But more defense-related proteins appeared in cv. Xiuyan, such as disease resistance-like protein, Peroxiredoxin-2F, lactoylglutathione lyase and so on. In addition, expression of proteins related to signal transduction, carbon metabolism and other functional categories also differed in two cultivars. Above all, cv. Xiuyan occured more efficient protein regulations after drought stress, which resulted in the different drought resistance among two Jerusalem artichoke cultivars.In brief, the Jerusalem artichoke’s tolerance to drought stress is involved in ROS control and osmoprotective capacity. The effective regulation of some drought responsive proteins that relate to photosynthesis, defense, signal transduction, etc. also plays important roles in Jerusalem artichoke’s tolerance to drought stress. |
| 学科主题 | 能源植物资源 |
| 语种 | 中文 |
| 公开日期 | 2011-08-29 |
| 源URL | [http://ir.qibebt.ac.cn//handle/337004/322]  |
| 专题 | 青岛生物能源与过程研究所_植物代谢工程团队 |
| 推荐引用方式 GB/T 7714 | 张美德. 菊芋幼苗叶片干旱胁迫的生理响应和蛋白质组学研究[D]. 北京. 中国科学院研究生院. 2010. |
入库方式： OAI收割
来源：青岛生物能源与过程研究所
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