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基于iTRAQ的发育和成熟的圆叶葡萄浆果定量蛋白质组学研究

iTRAQ-based quantitative proteomics of developing and ripening muscadine grape berry.

作者信息

Kambiranda Devaiah, Katam Ramesh, Basha Sheikh M, Siebert Shalom

机构信息

Plant Biotechnology Laboratory, Center for Viticulture and Small Fruit Research, Florida A&M University , 6505 Mahan Drive, Tallahassee, Florida 32317, United States.

出版信息

J Proteome Res. 2014 Feb 7;13(2):555-69. doi: 10.1021/pr400731p. Epub 2013 Dec 6.

Abstract

Grapes are among the widely cultivated fruit crops in the world. Grape berries like other nonclimacteric fruits undergo a complex set of dynamic, physical, physiological, and biochemical changes during ripening. Muscadine grapes are widely cultivated in the southern United States for fresh fruit and wine. To date, changes in the metabolites composition of muscadine grapes have been well documented; however, the molecular changes during berry development and ripening are not fully known. The aim of this study was to investigate changes in the berry proteome during ripening in muscadine grape cv. Noble. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS was used to detect statistically significant changes in the berry proteome. A total of 674 proteins were detected, and 76 were differentially expressed across four time points in muscadine berry. Proteins obtained were further analyzed to provide information about its potential functions during ripening. Several proteins involved in abiotic and biotic stimuli and sucrose and hexose metabolism were upregulated during berry ripening. Quantitative real-time PCR analysis validated the protein expression results for nine proteins. Identification of vicilin-like antimicrobial peptides indicates additional disease tolerance proteins are present in muscadines for berry protection during ripening. The results provide new information for characterization and understanding muscadine berry proteome and grape ripening.

摘要

葡萄是世界上广泛种植的水果作物之一。葡萄浆果与其他非跃变型果实一样,在成熟过程中会经历一系列复杂的动态、物理、生理和生化变化。麝香葡萄在美国南部广泛种植,用于生产新鲜水果和葡萄酒。迄今为止,麝香葡萄代谢物组成的变化已有充分记录;然而,浆果发育和成熟过程中的分子变化尚不完全清楚。本研究的目的是调查麝香葡萄品种诺布尔在成熟过程中浆果蛋白质组的变化。采用相对和绝对定量等压标签(iTRAQ)串联质谱法检测浆果蛋白质组中有统计学意义的变化。共检测到674种蛋白质,其中76种在麝香葡萄浆果的四个时间点上差异表达。对获得的蛋白质进行进一步分析,以提供其在成熟过程中潜在功能的信息。在浆果成熟过程中,几种参与非生物和生物刺激以及蔗糖和己糖代谢的蛋白质上调。实时定量PCR分析验证了9种蛋白质的表达结果。类豌豆球蛋白抗菌肽的鉴定表明,麝香葡萄中存在额外的抗病蛋白,以在成熟过程中保护浆果。这些结果为表征和理解麝香葡萄浆果蛋白质组和葡萄成熟提供了新的信息。

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