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Examination of the leaf proteome during flooding stress and the induction of programmed cell death in maize

机译:淹水胁迫下叶片蛋白质组的检测和诱导的程序性细胞死亡

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Background Maize is a major economic crop worldwide, with substantial crop loss attributed to flooding. During a stress response, programmed cell death (PCD) can be an effective way for plants better adapt. To identify flooding stress related PCD proteins in maize leaves, proteomic analysis was performed using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrometry. Results Comparative proteomics was combined with physiological and biochemical analysis of maize leaves under flooding stress. Fv/Fm, qP, qN and relative water content (RWC) were found to be altered in response to flooding stress, with an increase in H2O2 content noted in vivo. Furthermore, DNA ladder detection indicated that PCD had occurred under flooding treatment. The maize leaf proteome was analyzed via 2D-DIGE gel, with a total of 32 differentially expressed spots isolated, 31 spots were successfully identified via MALDI-TOF/TOF MS which represent 28 proteins. The identified proteins were related to energy metabolism and photosynthesis, PCD, phytohormones and polyamines. To better characterize the role of translationally controlled tumor protein (TCTP) in PCD during a stress response, mRNA expression was examined in different plants by stress-induced PCD. These included heat stress induced rice protoplasts, Tobacco Mosaic Virus infected tobacco leaves and dark induced rice and Arabidopsis thaliana leaves, all of which showed active PCD, and TCTP expression was increased in different degrees. Moreover, S-adenosylmethionine synthase 2 (SAMS2) and S-adenosylmethionine decarboxylase (SAMDC) mRNA expression were also increased, but ACC synthase (ACS) and ACC oxidase (ACO) mRNA expression were not found in maize leaves following flooding. Lastly, ethylene and polyamine concentrations were increased in response to flooding treatment in maize leaves. Conclusions Following flooding stress, the photosynthetic systems were damaged, resulting in a disruption in energy metabolism, with the noted photosynthetic decline also possibly attributed to ROS production. The observed PCD could be regulated by TCTP with a possible role for H2O2 in TCTP induction under flooding stress. Additionally, increased SAMS2 expression was closely associated with an increased polyamine synthesis during flooding treatment.
机译:背景技术玉米是世界范围内的主要经济作物,由于洪灾造成大量作物损失。在胁迫反应期间,程序性细胞死亡(PCD)可能是植物更好适应的有效方法。为了鉴定玉米叶片中与洪水胁迫相关的PCD蛋白,使用二维荧光差异凝胶电泳(2D-DIGE)和质谱进行了蛋白质组学分析。结果比较蛋白质组学与水淹胁迫下玉米叶片的生理生化分析相结合。发现Fv / Fm,qP,qN和相对水含量(RWC)响应洪水压力而改变,体内的H2O2含量增加。此外,DNA阶梯检测表明在水淹处理下已经发生了PCD。通过2D-DIGE凝胶分析了玉米叶片的蛋白质组,共分离出32个差异表达的斑点,通过MALDI-TOF / TOF MS成功鉴定出31个斑点,代表28个蛋白质。鉴定出的蛋白质与能量代谢和光合作用,PCD,植物激素和多胺有关。为了更好地表征应激反应期间PCD中翻译控制的肿瘤蛋白(TCTP)的作用,通过胁迫诱导的PCD检测了不同植物中的mRNA表达。这些包括热胁迫诱导的水稻原生质体,烟草花叶病毒感染的烟叶以及深色诱导的水稻和拟南芥叶,所有这些均显示活性PCD,并且TCTP表达以不同程度增加。此外,淹水后玉米叶片中S-腺苷甲硫氨酸合酶2(SAMS2)和S-腺苷甲硫氨酸脱羧酶(SAMDC)的mRNA表达也增加,但未见ACC合酶(ACS)和ACC氧化酶(ACO)的mRNA表达。最后,响应于淹没处理,玉米叶片中的乙烯和多胺浓度增加。结论洪水胁迫后,光合作用系统受到破坏,导致能量代谢中断,其中明显的光合作用下降也可能归因于ROS的产生。 TCTP可以调节观测到的PCD,在洪水胁迫下H2O2可能在TCTP诱导中发挥作用。此外,SAMS2表达的增加与水淹处理期间多胺合成的增加密切相关。

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