Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, 350002 Fuzhou, China.
Planta. 2010 Mar;231(4):847-60. doi: 10.1007/s00425-009-1093-1. Epub 2010 Jan 5.
Two-dimensional gel electrophoresis (2-DE), coupled with mass spectroscopy, was used to study seed abortion in Dimocarpus longan Lour. (cv. Minjiao 64-1) by comparing normal and aborted seeds at three developmental stages. More than 1,000 protein spots were reproducibly detected in 2-DE gels, with 43 protein spots being significantly altered in their intensity between normal and aborted seeds at least at one stage. Thirty-five proteins were identified by matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS) analysis and protein database searching. Most of the identified proteins were associated with a variety of functions, including energy and metabolism (30%), programed cell death (9%), antioxidative processes (14%), chaperonin (23%), cell division, amino acid metabolism, secondary metabolism, and other functional classes. Furthermore, the expression patterns of HSP70 and cytosolic ascorbate peroxidase (cAPX) were validated by immunoblotting analysis. This study provides a novel, global insight into proteomic differences between normal and aborted seeds in longan. We anticipate that identification of the differentially expressed proteins may lead to a better understanding of the molecular basis for seed abortion in longan.
双向电泳(2-DE)结合质谱技术,通过比较三个发育阶段正常和败育种子,研究了龙眼(Dimocarpus longan Lour. cv. Minjiao 64-1)种子败育。在 2-DE 凝胶中可重复性地检测到 1000 多个蛋白质斑点,其中至少有一个阶段在正常和败育种子之间,有 43 个蛋白质斑点的强度有明显变化。通过基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF-MS/MS)分析和蛋白质数据库搜索鉴定了 35 种蛋白质。大多数鉴定出的蛋白质与多种功能相关,包括能量和代谢(30%)、程序性细胞死亡(9%)、抗氧化过程(14%)、伴侣蛋白(23%)、细胞分裂、氨基酸代谢、次生代谢和其他功能类别。此外,通过免疫印迹分析验证了 HSP70 和胞质抗坏血酸过氧化物酶(cAPX)的表达模式。本研究为龙眼正常和败育种子之间的蛋白质组差异提供了全新的、全面的见解。我们预计,差异表达蛋白的鉴定可能有助于更好地理解龙眼种子败育的分子基础。