Pedreschi Romina, Vanstreels Els, Carpentier Sebastien, Hertog Maarten, Lammertyn Jeroen, Robben Johan, Noben Jean-Paul, Swennen Rony, Vanderleyden Jozef, Nicolaï Bart M
BIOSYST-MeBioS Division, Katholieke Universiteit Leuven, Belgium.
Proteomics. 2007 Jun;7(12):2083-99. doi: 10.1002/pmic.200600723.
2-DE was applied to study core breakdown disorder in controlled atmosphere stored 'Conference' pears. This physiological disorder is characterized by internal browning of the fruit tissue and the development of cavities. Suitable protein phenol extraction/ammonium acetate-methanol precipitation and 2-DE protocols for a wide pH range were established for pear tissue. The protein expression profiles of healthy, sound (intact tissue of pears with core breakdown) and brown tissue were analyzed with the univariate non-parametric Kolmogorov-Smirnov test and multivariate statistical techniques such as principal component analysis and partial least square discriminant analysis. Both statistical approaches revealed interesting differentially expressed proteins between healthy and disordered pears. LC-ESI-MS/MS identification of differentially expressed proteins between healthy and sound tissue revealed their participation in the energy metabolism, the antioxidant system and ethylene biosynthesis. Up-regulated characteristic proteins in brown tissue were mainly involved in energy metabolism and defense mechanisms. Proteomics coupled to univariate and multivariate statistical techniques seems to be an efficient approach to get a better insight into the different mechanisms and pathways leading to the core breakdown disorder.
采用双向电泳（2-DE）技术研究气调贮藏‘康佛伦斯’梨的果心褐变失调问题。这种生理失调的特征是果实组织内部褐变和空洞形成。针对梨组织，建立了适用于宽pH范围的蛋白质酚提取/醋酸铵-甲醇沉淀及双向电泳方案。采用单变量非参数柯尔莫哥洛夫-斯米尔诺夫检验以及主成分分析和偏最小二乘判别分析等多变量统计技术，对健康、完好（有果心褐变的梨的完整组织）和褐变组织的蛋白质表达谱进行了分析。两种统计方法均揭示了健康梨和失调梨之间有趣的差异表达蛋白质。液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）对健康组织和完好组织之间差异表达蛋白质的鉴定表明，它们参与能量代谢、抗氧化系统和乙烯生物合成。褐变组织中上调的特征蛋白主要参与能量代谢和防御机制。蛋白质组学与单变量和多变量统计技术相结合似乎是一种有效的方法，可更好地洞察导致果心褐变失调的不同机制和途径。
