Vítámvás Pavel, Saalbach Gerhard, Prásil Iija Tom, Capková Vera, Opatrná Jana, Ahmed Jahoor
Department of Genetics and Plant Breeding, Research Institute of Crop Production, Drnovská 507, 161 06, Prague, Czech Republic.
J Plant Physiol. 2007 Sep;164(9):1197-207. doi: 10.1016/j.jplph.2006.06.011. Epub 2006 Sep 11.
The amount of proteins soluble upon boiling (especially WCS120 proteins) and the ability to develop frost tolerance (FT) after cold acclimation was studied in two frost-tolerant winter wheat cultivars, Mironovskaya 808 and Bezostaya 1. Protein gel blot analysis, mass spectrometry (MS) and image analysis of two-dimensional gel electrophoresis (2-DE) gels were used to identify and/or quantify the differences in protein patterns before (non-acclimated, NA) and after 3 weeks of cold acclimation (CA) of the wheats, when FT increased from -4 degrees C (lethal temperature (LT(50)), for both cultivars) to -18.6 degrees C in Bezostaya 1 and -20.8 degrees C in Mironovskaya 808. Only WCS120 protein was visible in NA leaves while all five WCS120 proteins were induced in the CA leaves. Mironovskaya 808 had higher accumulation of three members of WCS120 proteins (WCS120, WCS66 and WCS40) than Bezostaya 1. MS analysis of total sample of proteins soluble upon boiling showed seven COR proteins in the CA samples and only three COR proteins in the NA samples of cultivar Mironovskaya 808 (MIR). In conclusion, the level of the accumulation of WCS120, WCS66 and WCS40 distinguished our two frost-tolerant winter wheat cultivars. Moreover, the differences of CA and NA samples of the MIR were shown by liquid chromatography (LC)-tandem mass spectrometry (MS/MS).
研究了两个抗冻冬小麦品种米罗诺夫斯卡亚808(Mironovskaya 808)和别佐斯塔亚1号(Bezostaya 1)中煮沸后可溶的蛋白质数量(尤其是WCS120蛋白)以及冷驯化后形成抗冻性(FT)的能力。利用蛋白质凝胶印迹分析、质谱(MS)和二维凝胶电泳(2-DE)凝胶的图像分析来鉴定和/或量化小麦在冷驯化3周前(未驯化,NA)和冷驯化后(CA)蛋白质模式的差异,此时两个品种的抗冻性从-4℃(致死温度(LT(50)))分别提高到别佐斯塔亚1号的-18.6℃和米罗诺夫斯卡亚808的-20.8℃。在未驯化的叶片中仅可见WCS120蛋白,而在冷驯化的叶片中所有五种WCS120蛋白均被诱导。米罗诺夫斯卡亚808中WCS120蛋白的三个成员(WCS120、WCS66和WCS40)的积累量高于别佐斯塔亚1号。对煮沸后可溶的蛋白质总样品进行MS分析表明,米罗诺夫斯卡亚808品种(MIR)冷驯化样品中有七种COR蛋白,未驯化样品中只有三种COR蛋白。总之,WCS120、WCS66和WCS40的积累水平区分了我们的两个抗冻冬小麦品种。此外,通过液相色谱(LC)-串联质谱(MS/MS)显示了米罗诺夫斯卡亚808品种冷驯化和未驯化样品之间的差异。