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为了研究玉米异交不亲和的蛋白表达机制,以玉米(Zea mays L.)异交不亲和基因Gal-S的一对近等基因系W22(GG)与w22(gg)为材料,组配自交(GG×GG)、正交(gg×GG)与反交(GG×gg)3个组合。首先采用荧光显微技术,观察比较了3个组合中花粉管在花丝中的生长过程;其次采用IEF/SDS-PAGE双向凝胶电泳与基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,比较研究了授粉后10h自交与反交母本W22(GG)花丝蛋白质组的特异表达差异。结果表明,授粉后10h,自交与正交花粉管伸长可达花丝基部,而反交花丝基部未观察到花粉管;自交与反交母本W22(GG)花丝蛋白质组共检测到25个差异蛋白质点,其中15个在自交中特异表达,10个在反交中特异表达。通过MALDI-TOF-MS质谱测序和MASCOT序列分析,注释了12个蛋白质点,其中自交中特异表达的蛋白质点11、12、14和异交中特异表达的蛋白质点18、22、24可能与玉米异交不亲和密切相关。
In order to study the protein expression mechanism of out-of-phase incompatibility in maize, a pair of near-isogenic lines W22 (GG) and w22 (gg) of the Gal-S gene of maize (Zea mays L.) There were 3 combinations of selfing (GG × GG), orthogonal (gg × GG) and reciprocal (GG × gg). Firstly, fluorescence microscopy was used to observe the growth of pollen tube in the filament in three combinations. Secondly, using IEF / SDS-PAGE two-dimensional gel electrophoresis and matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF- Technology to compare the differences in the specific expression of the W22 (GG) filiform proteome between the selfed and the offspring at 10 h after pollination. The results showed that the elongation of selfed and orthogonal pollen tubes reached the basal part of the pollen at 10h after pollination but no pollen tube was observed at the base of the opposite crossed filament. Among them, 15 were differentially expressed in inbreds and 10 were specifically expressed in reciprocal crosses. 12 protein spots were annotated by MALDI-TOF-MS mass spectrometry and MASCOT sequence analysis, wherein protein spots 11,12,14 specifically expressed in the inbred and protein spots 18,22,24 specifically expressed in the outcross may be related to Maize is not closely related to the other.