杜仲叶片干旱胁迫响应相关差异蛋白的筛选与鉴定

doi:10.7525/j.issn.1673-5102.2018.03.015

摘要/Abstract

摘要： 为研究干旱胁迫下杜仲幼苗生理生化及分子响应机制，利用盆栽试验，通过持续（3、6、9、12、15 d）干旱胁迫处理和复水处理，研究杜仲幼苗的生理响应特性。同时，通过研究对照与处理15 d后的杜仲幼苗差异蛋白质组，分析杜仲幼苗对干旱胁迫的分子响应机制。结果表明，随着干旱处理时间的延长，杜仲叶片的水分饱和亏逐渐增加；光合速率、蒸腾速率、胞间二氧化碳浓度、气孔导度均逐渐减小；SOD、POD、CAT活性呈先上升后降低的趋势；丙二醛含量则呈现先上升，然后下降，最后又上升的变化特点；脯氨酸和可溶性糖含量的变化趋势与SOD等活性变化一致，前期上升，后期下降。在复水后，杜仲叶片的所有指标均有所恢复，但未达到干旱处理之前的水平。表明干旱胁迫影响了杜仲叶片的正常生长代谢。通过对干旱处理15 d后杜仲叶片总蛋白进行双向电泳分离和MALDI-TOF-TOF生物质谱鉴定，成功鉴定出36个差异表达蛋白，其中22个上调表达，14个下调表达。对36个差异蛋白进行功能分析发现，这些差异蛋白主要涉及信号传导、光合作用、碳代谢、能量代谢、次级代谢物合成、抗氧化保护酶、氨基酸代谢和蛋白质代谢。推测杜仲为适应干旱胁迫，首先是感应干旱胁迫信号，并传导至细胞内，影响杜仲叶片中光合作用、次级代谢物合成和蛋白质的生物合成；同时，通过过氧化物保护酶的作用，将过多活性氧加以清除；另一方面，则是通过增强糖酵解，磷酸戊糖途径，产生能量供杜仲正常生长所需。从生理机制来看，杜仲叶片同过增加胞内脯氨酸、可溶性糖含量，降低胞内渗透势，减少叶片中水分损失，与氨基酸合成和糖代谢相关蛋白的表达量上升的结果一致。

关键词: 杜仲, 叶片, 干旱胁迫, 蛋白质组

Abstract: To uncover the underlying mechanism in leaves of Eucommia ulmoides in response to drought stress, we studied the physiological response characteristic in the leaves at the drought treatment time of 0, 3, 6, 9, 12, 15 d and 2 d of recovery by pot test. We used differential proteomic analysis between the leaves of 0 and 15 d to reveal the molecular mechanism. The results showed that with the stress time prolonged, the water saturation deficit was increased. While the photosynthesis rate, transpiration rate, intercellular CO₂ concentration and stomatal conductance were decreased. The activity of SOD, POD and CAT were firstly increased, then decreased. The content of MDA was firstly increased, then decreased, thereafter, increased again at the day of 15. The change profile of proline and soluble sugar content were in consistence with the change of SOD activity. After rehydration, all the parameters recovered to a certain level, but still could not recover to the control level. The total proteins extracted from the leaves that were collected at 0 and 15 d were separated by 2-DE, and the differentially expressed proteins were identified by MALDI-TOF-TOF analysis. About 36 protein spots were successfully identified, including 22 up-regulated and 14 down-regulated proteins. Further analysis of these differential expressed proteins revealed that these proteins were mainly involved in signal transduction, photosynthesis, carbon metabolism, energy metabolism, secondary metabolites synthesis, antioxidant protective enzyme, amino acid metabolism and protein metabolism. It was speculated that Eucommia ulmoides leaves firstly sensed the stress signal in response to drought, then affected photosynthesis, secondary metabolites synthesis and protein synthesis. Meanwhile, the excess ROS were eliminated by antioxidant protective enzyme. On the other hand, the glycolysis and pentose phosphate pathway were enhanced to produce the energy to supply demands of plant growth under drought stress. The physiological analysis results revealed that the proline and soluble sugar content were increased to reduce the water loss by decreasing the osmotic potential in plant cells. These results were in accordance with the change profiles of amino acid metabolism and sugar metabolism associated proteins.

Key words: Eucommia ulmoides, leaves, drought stress, proteomics

中图分类号:

Q949.751.5

赵欣, 白伟. 杜仲叶片干旱胁迫响应相关差异蛋白的筛选与鉴定[J]. 植物研究, 2018, 38(3): 422-432.

ZHAO Xin, BAI Wei. Screening and Identification of the Proteins Related to Drought Response in Leaves of Eucommia ulmoides[J]. Bulletin of Botanical Research, 2018, 38(3): 422-432.

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