Seedling proteome of Arabidopsis thaliana pux10-3 mutant

description: Lipid droplets (LD) of seedlings lacking PUX10 stay significantly smaller during germination than the LDs of wild type seedlings. The observation that the removal of a LD-associated protein from a plant affects LD morphology has been observed previously {Siloto et al., 2006}{Gidda et al., 2016}. Especially the effect of the knock-out of the major structural LD protein Oleosin1 leading to a strongly increased size of LDs described by Siloto et al. might indicate that the abundance of structural LD proteins influences the LD size. Thus, the influence of the absence of PUX10 on the abundance of these proteins was investigated via a proteomic approach. Here, proteins isolated from homogenized seedlings of different lines (Ws-4 and pux10-3) and different developmental stages (wet seeds, and 1 to 3 days after imbibition (dai)) were subjected to tryptic digest and LC-MS/MS analysis.

缺失PUX10的幼苗的脂滴（Lipid droplets, LD）在萌发过程中尺寸显著小于野生型幼苗的脂滴。此前已有研究观察到，从植物中移除脂滴相关蛋白会影响脂滴形态[Siloto等，2006；Gidda等，2016]。其中，Siloto等所报道的主要结构脂滴蛋白油质蛋白1（Oleosin1）的敲除会导致脂滴尺寸显著增大的效应，或提示结构脂滴蛋白的丰度会影响脂滴尺寸。因此，本研究通过蛋白质组学方法探究了PUX10缺失对这类蛋白丰度的影响。本研究对不同遗传株系（Ws-4和pux10-3）、不同发育阶段（湿种子，以及吸胀后1至3天（dai））的匀浆幼苗中分离得到的蛋白进行胰蛋白酶酶解，并开展液相色谱-串联质谱（LC-MS/MS）分析。