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中国山核桃(Carya cathayensis)嫁接过程中琥珀酰化蛋白质组的定量分析。

Quantitative succinyl-proteome profiling of Chinese hickory (Carya cathayensis) during the grafting process.

机构信息

State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou, 311300, People's Republic of China.

Center for Cultivation of Subtropical Forest Resources (CCSFR, Zhejiang A&F University, Hangzhou, 311300, People's Republic of China.

出版信息

BMC Plant Biol. 2019 Nov 4;19(1):467. doi: 10.1186/s12870-019-2072-8.

Abstract

BACKGROUND

Chinese hickory (Carya cathayensis) is a popular nut plant having high economic value. Grafting is applied to accelerate the transition from vegetative phase to reproductive phase. Lysine succinylation occurs frequently in the proteins associated with metabolic pathways, which may participate in the regulation of the grafting process. However, the exact regulatory mechanism underlying grafting process in Chinese hickory has not been studied at post-translational modification level.

RESULTS

A comprehensive proteome-wide lysine succinylation profiling of Chinese hickory was explored by a newly developed method combining affinity enrichment and high-resolution LC-MS/MS. In total, 259 succinylation sites in 202 proteins were identified, representing the first comprehensive lysine succinylome in Chinese hickory. The succinylation was biased to occur in the cytosolic proteins of Chinese hickory. Moreover, four conserved succinylation motifs were identified in the succinylated peptides. Comparison of two grafting stages of Chinese hickory revealed that the differential expressed succinylated proteins were mainly involved in sugar metabolism, carbon fixation, amino acid metabolism and plant-pathogen interaction. Besides, seven heat shock proteins (HSPs) with 11 succinylation sites were also identified, all of which were observed to be up-regulated during the grafting process.

CONCLUSIONS

Succinylation of the proteins involved in amino acid biosynthesis might be required for a successful grafting. Succinylated HSPs might play a role in stress tolerance of the grafted Chinese hickory plants. Our results can be a good resource for functional validation of the succinylated proteins and a starting point for the investigation of molecular mechanisms during lysine succinylation occurring at grafting site.

摘要

背景

中国山核桃(Carya cathayensis)是一种具有较高经济价值的受欢迎的坚果植物。嫁接被应用于加速其从营养生长阶段向生殖生长阶段的转变。赖氨酸琥珀酰化在与代谢途径相关的蛋白质中经常发生,可能参与嫁接过程的调控。然而,在中国山核桃嫁接过程的精确调控机制尚未在翻译后修饰水平上进行研究。

结果

通过一种新开发的结合亲和富集和高分辨率 LC-MS/MS 的方法,对中国山核桃进行了全面的蛋白质组赖氨酸琥珀酰化谱分析。共鉴定了 202 个蛋白质中的 259 个琥珀酰化位点,代表了中国山核桃中第一个全面的赖氨酸琥珀酰化组。琥珀酰化偏向于发生在细胞质中的蛋白质。此外,在琥珀酰化肽中鉴定出四个保守的琥珀酰化基序。对中国山核桃的两个嫁接阶段进行比较发现,差异表达的琥珀酰化蛋白主要参与糖代谢、碳固定、氨基酸代谢和植物-病原体相互作用。此外,还鉴定了七个具有 11 个琥珀酰化位点的热休克蛋白(HSPs),它们在嫁接过程中均被观察到上调。

结论

参与氨基酸生物合成的蛋白质的琥珀酰化可能是嫁接成功所必需的。琥珀酰化的 HSPs 可能在嫁接中国山核桃植物的胁迫耐受中发挥作用。我们的结果可以为琥珀酰化蛋白的功能验证提供良好的资源,并为研究赖氨酸琥珀酰化发生在嫁接部位时的分子机制提供起点。

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