兰州大学机构库 >生命科学学院
蛋白磷酸酶PPSx参与拟南芥盐胁迫响应初步研究
Alternative TitleA Preliminary Study of Protein Phosphatase PPSx Involved on Salt Stress Response in Arabidopsis
沈杰
Thesis Advisor侯岁稳
2017-05-10
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword拟南芥 盐胁迫 PPSx PPSs SAO途径
Abstract

盐碱条件产生的渗透胁迫、离子毒害及营养失衡均会对细胞的生理活动和代谢平衡造成严重影响,如影响脂质代谢、蛋白质合成与修饰等一系列重要的生命活动过程。为应对不良的外界环境,植物进化出多种生理生化机制维持自身稳态,确保生长发育的正常进行。

本论文利用蛋白磷酸酶PPSx基因的一个点突变体ppsx-1和T-DNA插入突变体ppsx-3为材料进行研究,发现盐胁迫和渗透胁迫条件下ppsx-1和ppsx-3均出现更加敏感的应答缺陷表型,而对过氧化物、碱性条件及K+、Li+胁迫处理表现出与野生型相似的表型。pPPSx-GUS染色和定量PCR结果表明,同盐胁迫响应基因RD22和RD29B一样,PPSx也受盐胁迫诱导表达,由此证明PPSx参与了植物盐胁迫应答过程。进一步研究发现PPSx蛋白的C端结构域可以更好地恢复酵母AXT3K的盐敏感缺陷表型。二氨基联苯胺染色(DAB)和膜脂过氧化产物丙二醛(MDA)含量测定发现,盐胁迫能够引起ppsx-1和ppsx-3中活性氧(ROS)爆发和MDA含量积累,而且显著高于野生型。酵母双杂及pull-down实验结果表明,PPSx能够与SAO信号通路中的调控因子FI、FH3和FH4分别互作,暗示PPSx可能通过SAO信号通路调节植物耐盐过程。

为验证植物蛋白磷酸酶PPSs家族成员在盐胁迫调控过程中的功能,将9个成员转化盐敏感缺陷酵母AXT3K突变体,PPSe和PPSx可以较明显恢复酵母AXT3K的盐敏感缺陷表型,PPSg的恢复作用较弱,表明PPSs家族中PPSe、PPSx和PPSg可能均参与盐胁迫应答过程。酵母双杂实验结果显示PPSe、PPSx、PPSh、PPSi与FI存在互作,FH4只与PPSx存在互作。总之,我们的研究结果表明植物PPSs家族可能通过SAO信号通路参与盐胁迫应答过程。

Other Abstract

Osmotic stress, ion toxic, and nutrient deficiency cause severe effects on physical activity and metabolic balance in the cells. For example, they could affect a series of life activity processes, such as lipid metabolism, protein synthesis and modification. In order to resist the adverse external environment, plants have evolved different physiological and biochemical mechanisms to ensure their growth and development.

In this thesis, the results of physiological experiment showed that ppsx-1, a point mutant of PPSx, and ppsx-3, a T-DNA insertion mutant of PPSx, had defective response to salt and osmotic stress. However, the phenotype of ppsx-1 and ppsx-3 was similar to the phenotype of Col under the stress of peroxide, alkaline conditions, K+ and Li+. The results of GUS staining and qRT-PCR indicated that the expression of PPSx was induced under salt stress, as well as the action of the salt stress response genes RD22 and RD29B. These results indicated that PPSx was involved in the process of salt stress response in Arabidopsis. Furthermore, the C-terminal domain of PPSx could complement yeast AXT3K which showed strong salt sensitive phenotype. Reactive oxygen species (ROS) and malonaldehyde (MDA) content accumulated in ppsx-1 and ppsx-3 under salt stress. Yeast two hybrid experiment and pull-down assay showed that PPSx could interact with several regulatory factors of the SAO signal pathway, such as FI, FH3 and FH4. These results indicated that PPSx participated in the process of salt tolerance by regulating the SAO signal pathway.

To identify the function of PPSs in plants under salt stress, the nine members of PPSs were transformed into salt sensitive yeast AXT3K respectively. PPSe and PPSx could complement yeast AXT3K salt sensitive phenotype strongly. In addition, PPSg could complement yeast AXT3K weakly. These results suggested that PPSe, PPSx and PPSg may play an important role in salt stress response. Yeast two hybrid experiment results showed that PPSe, PPSx, PPSh and PPSi could interact with FI. In addition, FH4 could only interact with PPSx. In summary, our study demonstrated that PPSs participated in the process of salt tolerance by regulating the SAO signal pathway in Arabidopsis.

URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221590
Collection生命科学学院
Recommended Citation
GB/T 7714
沈杰. 蛋白磷酸酶PPSx参与拟南芥盐胁迫响应初步研究[D]. 兰州. 兰州大学,2017.
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