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为从蛋白质的角度揭示自交不亲和甘蓝亲和授粉早期花粉与柱头相互作用,将蛋白质组学方法应用于自交不亲和性甘蓝柱头与亲和花粉互作早期差异蛋白质的研究。结果表明,亲和授粉后 3 ～ 5min 与 1 h 对比,有 116 个差异蛋白质点,而授粉后 1 h 与 2 h 差异不显著。按照特异表达与变化量大的蛋白质点优先原则,初步选取差异点中 71 个点做质谱分析,鉴定出 31 个可信差异蛋白质。与亲和授粉后 3 ～ 5 min 组相比,1 h 组中特异表达蛋白质有 19 个,上调表达 9 个,下调表达 3 个。31 种蛋白质的生物信息学分析表明,有两种蛋白与前人发现的与花粉管在柱头中生长发育相关蛋白相同,其他 29 种参与包括胁迫与防御应答在内多种生理过程。对早期差异蛋白中这种“既有促进花粉管发育的蛋白,也有与防御相关蛋白”现象分析表明,花粉管生长性侵入柱头的过程有可能伴随着柱头抗性的提高。
In order to reveal the interaction between early pollen and stigma in self-incompatible cabbage affinity pollen from the perspective of protein, proteomic methods were applied to study the early differential proteins between self-incompatibility cabbage stigma and affinity pollen. The results showed that there were 116 differential protein spots at 3 ~ 5 min after 1 h and 1 h after pollination, but no significant difference at 1 h and 2 h after pollination. According to the principle of preferential expression of protein spots with large amount of changes, we selected 31 of the difference points for mass spectrometry analysis, and identified 31 differential proteins. Compared with 3 ~ 5 min after in vitro fertilization, 19 proteins were specifically expressed in 1 h group, 9 up-regulated and 3 down-regulated. Bioinformatics analysis of 31 kinds of proteins showed that there are two kinds of proteins which are the same as those found in the stigma of the pollen tube. The other 29 kinds involved in various physiological processes including stress and defense responses. The analysis of the early differential proteins in this “both the promotion of pollen tube development and defense-related proteins” shows that the process of pollen tube growth invasion into the stigma may be accompanied by stigma resistance.