Biotechnology Research Institute, Chinese Agricultural and Academic Sciences, Beijing, PR China.
College of Life Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
J Sci Food Agric. 2021 Mar 30;101(5):1869-1878. doi: 10.1002/jsfa.10802. Epub 2020 Sep 30.
An isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis was employed to study the seeds of two genetically modified (GM) rice lines, T2A-1 and T1C-19, and their nontransgenic isogenic variety, MH63, to investigate the unintended effects of genetic modification.
A total of 3398 proteins were quantitatively identified. Seventy-seven differentially abundant proteins (DAPs) were identified in the T2A-1/MH63 comparison, and 70 and 7 of these DAPs were upregulated and downregulated, respectively. A pathway enrichment analysis showed that most of these DAPs participated in metabolic pathways and protein processing in endoplasmic reticulum and were ribosome components. Some 181 DAPs were identified from the T1C-19/MH63 comparison, and these included 115 upregulated proteins and 66 downregulated proteins. The subsequent pathway enrichment analysis showed that these DAPs mainly participated in protein processing in endoplasmic reticulum and carbon fixation in photosynthetic organisms and were ribosome components. None of these DAPs were identified as new unintended toxins or allergens, and only changes in abundance were detected. Fifty-four co-DAPs were identified in the seeds of the two GM rice lines, and protein-protein interaction analysis of these co-DAPs demonstrated that some interacting proteins were involved in protein processing in endoplasmic reticulum and metabolic pathways, whereas others were identified as ribosome components. Representative co-DAPs and proteins related to nutrients were analyzed using qRT-PCR to determine their transcriptional levels.
The results suggested that the seed proteomic profiles of the two GM rice lines studied were not substantially altered from those of their natural isogenic control. © 2020 Society of Chemical Industry.
采用同位素标记相对和绝对定量(iTRAQ)蛋白质组学分析方法研究了两种转基因(GM)水稻品系 T2A-1 和 T1C-19 及其非转基因同系品种 MH63 的种子,以研究遗传修饰的意外影响。
共定量鉴定了 3398 种蛋白质。在 T2A-1/MH63 比较中鉴定出 77 个差异丰度蛋白(DAP),其中 70 个和 7 个分别上调和下调。途径富集分析表明,这些 DAP 大多数参与代谢途径和内质网蛋白质加工,是核糖体成分。从 T1C-19/MH63 比较中鉴定出 181 个 DAP,包括 115 个上调蛋白和 66 个下调蛋白。随后的途径富集分析表明,这些 DAP 主要参与内质网蛋白质加工和光合生物中的碳固定,是核糖体成分。这些 DAP 中没有一个被鉴定为新的非预期毒素或过敏原,只检测到丰度的变化。在这两种 GM 水稻种子中鉴定出 54 个共同 DAP,这些共同 DAP 的蛋白质-蛋白质相互作用分析表明,一些相互作用蛋白参与内质网蛋白质加工和代谢途径,而另一些则被鉴定为核糖体成分。使用 qRT-PCR 分析了代表性的共同 DAP 和与营养有关的蛋白质,以确定它们的转录水平。
研究结果表明,所研究的两种 GM 水稻品系的种子蛋白质组图谱与天然同系对照品没有明显改变。© 2020 化学工业协会。