低温胁迫下克恩氏冬青幼苗叶片差异蛋白质分析

周余华; 梁有旺; 彭方仁

doi:10.3969/j.issn.1000-2006.201703027

周余华,梁有旺,彭方仁

南京林业大学学报（自然科学版） ›› 2017, Vol. 41 ›› Issue (06) : 187-192.

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南京林业大学学报（自然科学版） ›› 2017, Vol. 41 ›› Issue (06) : 187-192. DOI: 10.3969/j.issn.1000-2006.201703027

研究简报

低温胁迫下克恩氏冬青幼苗叶片差异蛋白质分析

周余华^{1, 2}, 梁有旺¹, 彭方仁^1*

作者信息 +

Proteomic analysis of Ilex seedling leaf response to low temperature

ZHOU Yuhua^{1, 2}, LIANG Youwang¹, PENG Fangren^1*

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文章历史 +

摘要

【目的】研究克恩氏冬青叶片应答低温胁迫的蛋白表达情况,探讨克恩氏冬青耐寒的作用机理。【方法】以2年生幼苗为试材,对其进行-8 ℃和-16 ℃低温胁迫处理,运用双向电泳技术(2-DE)结合质谱技术(MALDI-TOF/TOF MS)检测并分析叶片差异表达蛋白,并对差异蛋白进行了生物信息学分析。【结果】在低温胁迫下,克恩氏冬青幼苗叶片中蛋白表达发生了明显变化,胁迫前后发现共有31个显著差异的蛋白质点,经质谱检测及数据库检索,成功鉴定了其中23个差异表达蛋白点。对其中的20个蛋白成功进行了功能分类,主要涉及蛋白质和氨基酸代谢、光合作用、氧化还原平衡、防御作用、碳水化合物代谢、脂类代谢、氮素代谢等生物学过程。【结论】克恩氏冬青应答低温胁迫是多种蛋白质共同作用的结果,主要通过调节多种代谢过程中的相关蛋白表达来发挥作用。

Abstract

【Objective】Study the molecular mechanism of Ilex×koehneana ‘Emily Bruner' in response to low temperature stress.【Mothed】 We analyzed the proteomic differences in leaves among different temperature treatment(0 ℃, -8 ℃ and -16 ℃)by using 2-DE coupled with MALDI-TOF/TOF MS.【Result】 The 2-DE profile of the total protein in the leaves demonstrated that more than 1 000 protein spots were detected with good reproducibility in the range of molecular masses of 15-120 ku and pH 4-7. A total of 31 protein spots in low temperature treatments exhibited significant changes in abundance of less than 0.66-fold or more than 1.5-fold(P<0.05)compared to CK. After MALDI-TOF/TOF MS analysis, a total of 23 differentially accumulated protein spots were successfully identified. Among the 23 identified proteins, 20 proteins had assigned functions, and the other 3 were identified as novel with no assigned function. The 20 proteins with assigned functions were classified into 8 metabolic pathways and cellular processes, including protein and amino acid metabolism, carbohydrate metabolism, carbon fixation in photosynthetic organisms, lipid metabolism, photosynthesis, redox homeostasis, defense response, and nitrogen metabolism. The identifications of these differentially accumulated proteins indicates the presence of a specific different metabolic network in Ilex in response to cold stress.【Conclusions】These studies as a first step towards understanding the molecular mechanisms of low temperature resistance of Ilex, will provide the research basis for cultivating and screening relatively cold resistant varieties.

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周余华,梁有旺,彭方仁. 低温胁迫下克恩氏冬青幼苗叶片差异蛋白质分析[J]. 南京林业大学学报（自然科学版）. 2017, 41(06): 187-192 https://doi.org/10.3969/j.issn.1000-2006.201703027

ZHOU Yuhua, LIANG Youwang, PENG Fangren. Proteomic analysis of Ilex seedling leaf response to low temperature[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2017, 41(06): 187-192 https://doi.org/10.3969/j.issn.1000-2006.201703027

中图分类号： S718 Q945.78

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