短日照处理诱导油松容器苗针叶差异表达蛋白质的功能分析

蒋乐; 刘勇; 李国雷; 林平; 史文辉; 万芳芳

doi:10.3969/j.issn.1000-2006.201605005

蒋乐,刘勇,李国雷,林平,史文辉,万芳芳

南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (02) : 17-24.

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南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (02) : 17-24. DOI: 10.3969/j.issn.1000-2006.201605005

研究论文

短日照处理诱导油松容器苗针叶差异表达蛋白质的功能分析

蒋乐,刘勇^*,李国雷,林平,史文辉,万芳芳

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Analysis of differentially expressed protein functions of Pinus tabulaeformis seedling leaves induced by short-day treatment

JIANG Le, LIU Yong^*, LI Guolei, LIN Ping, SHI Wenhui, WAN Fangfang

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摘要

【目的】基于蛋白质水平探讨短日照处理调控油松(Pinus tabulaeformis)容器苗基础代谢和抗逆性机理的研究,完善油松容器苗的育苗和造林技术,为短日照处理育苗技术在我国北方地区困难立地造林中推广应用提供理论依据。【方法】以经日照长度为10 h、持续3 周短日照处理的油松容器苗针叶为研究对象,采用改良的酚法提取针叶蛋白。应用双向电泳结合二级质谱分析的蛋白质组学技术,研究短日照处理诱导油松容器苗针叶蛋白质表达的变化,分析差异表达蛋白质的鉴定和功能。【结果】成功获取5个短日照处理诱导表达差异显著的蛋白质,上调表达的是叶绿体放氧增强蛋白1(oxygen-evolving enhancer protein 1, OEE1)(蛋白点404)、甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)(蛋白点539)、延伸因子-Tu(elongation factor-Tu, EF-Tu)(蛋白点654)、核酮糖-1,5-二磷酸羧化酶/加氧酶活化酶(ribulose bisphosphate carboxylase/oxygenase activase, RCA)(蛋白点681); 下调表达的是磷酸甘油酸激酶1(phosphoglycerate kinase 1, PGK1)(蛋白点641)。这些蛋白质的功能主要涉及光合代谢、糖代谢和胁迫防御。【结论】短日照处理诱导油松容器苗蛋白质差异表达,对油松容器苗的基础代谢和抗逆性产生重要影响。

Abstract

【Objective】 Determining basic metabolism and stress resistance response to short-day treatment at the protein level for Pinus tabulaeformis Carr. container seedling and providing theoretical basis for applying the short-day treatment on seedlings used in the difficult site afforestation in Northern China.【Method】Short-day treatment was applied to P. tabulaeformis container seedlings by artificially reduced day length to 10 hours for 21 days. Seedling leaves were sampled and leaf proteins were extracted by improved phenol method. A proteomic analysis using two-dimensional electrophoresis(2-DE)in combination with MALDI-TOF/TOF MS analysis was performed to investigate changes in protein expression. The MS/MS data were submitted to Mascot search engine to search the NCBInr and Swiss Prot database for protein identification. Functional information of the identified proteins was found in UniProt.【Result】We obtained five proteins expressed differentially(P < 0.05)induced by short-day treatment. They were up-regulated expression of oxygen-evolving enhancer protein 1(OEE1)(spot 404), glyceraldehyde-3-phosphate dehydrogenase(GAPDH)(spot 539), elongation factor Tu(EF-Tu)(spot 654), ribulose bisphosphate carboxylase/oxygenase activase(RCA)(spot 681), and down-regulated expression of phosphoglycerate kinase 1(PGK1)(spot 641). They are mainly involved in plant photosynthetic metabolism, sugar metabolism and stress defense.【Conclusion】The different expression of P. tabulaeformis seedling leaf proteins can be induced by short-day treatment. These proteins are mainly involved in plant primary metabolism and stress response, suggesting that short-day treatment could play important roles in the development of basic metabolism and stress resistance of seedling.

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蒋乐,刘勇,李国雷,林平,史文辉,万芳芳. 短日照处理诱导油松容器苗针叶差异表达蛋白质的功能分析[J]. 南京林业大学学报（自然科学版）. 2018, 42(02): 17-24 https://doi.org/10.3969/j.issn.1000-2006.201605005

JIANG Le, LIU Yong, LI Guolei, LIN Ping, SHI Wenhui, WAN Fangfang. Analysis of differentially expressed protein functions of Pinus tabulaeformis seedling leaves induced by short-day treatment[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2018, 42(02): 17-24 https://doi.org/10.3969/j.issn.1000-2006.201605005

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