Liu Shengxuan, Liu Tengfei, Wang Enshuang, Cheng Yunxia, Liu Tiantian, Chen Guogang, Guo Minrui, Song Botao
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China.
School of Food Science and Technology, Shihezi University, Shihezi 832003, China.
Plants (Basel). 2022 Jul 24;11(15):1915. doi: 10.3390/plants11151915.
The chloroplast, the energy organelle unique to plants and green algae, performs many functions, including photosynthesis and biosynthesis of metabolites. However, as the most critical tuber crop worldwide, the chloroplast proteome of potato () has not been explored. Here, we use Percoll density gradient centrifugation to isolate intact chloroplasts from leaves of potato cultivar E3 and establish a reference proteome map of potato chloroplast by bottom-up proteomics. A total of 1834 non-redundant proteins were identified in the chloroplast proteome, including 51 proteins encoded by the chloroplast genome. Extensive sequence-based localization prediction revealed over 62% of proteins to be chloroplast resident by at least one algorithm. Sixteen proteins were selected to evaluate the prediction result by transient fluorescence assay, which confirmed that 14 were distributed in distinct internal compartments of the chloroplast. In addition, we identified 136 phosphorylation sites in 61 proteins encoded by chloroplast proteome. Furthermore, we reconstruct the snapshots along starch metabolic pathways in the two different types of plastids by a comparative analysis between chloroplast and previously reported amyloplast proteomes. Altogether, our results establish a comprehensive proteome map with post-translationally modified sites of potato chloroplast, which would provide the theoretical principle for the research of the photosynthesis pathway and starch metabolism.
叶绿体是植物和绿藻特有的能量细胞器,具有多种功能,包括光合作用和代谢物的生物合成。然而,作为全球最重要的块茎作物,马铃薯()的叶绿体蛋白质组尚未得到研究。在这里,我们使用Percoll密度梯度离心法从马铃薯品种E3的叶片中分离完整的叶绿体,并通过自下而上的蛋白质组学方法建立马铃薯叶绿体的参考蛋白质组图谱。在叶绿体蛋白质组中总共鉴定出1834个非冗余蛋白质,其中包括51个由叶绿体基因组编码的蛋白质。广泛的基于序列的定位预测显示,至少有一种算法表明超过62%的蛋白质定位于叶绿体。选择了16种蛋白质通过瞬时荧光测定法评估预测结果,证实其中14种分布在叶绿体的不同内部区域。此外,我们在叶绿体蛋白质组编码的61种蛋白质中鉴定出136个磷酸化位点。此外,通过对叶绿体和先前报道的造粉体蛋白质组进行比较分析,我们重建了两种不同类型质体中淀粉代谢途径的快照。总之,我们的结果建立了一个包含马铃薯叶绿体翻译后修饰位点的综合蛋白质组图谱,这将为光合作用途径和淀粉代谢的研究提供理论依据。