State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou, China.
Crops Research Institute, Guangdong Academy of Agricultural Sciences (GAAS), South China Peanut Sub-Center of National Center of Oilseed Crops Improvement, Guangdong Key Laboratory for Crops Genetic Improvement, Guangzhou, China.
PLoS One. 2020 Dec 7;15(12):e0243132. doi: 10.1371/journal.pone.0243132. eCollection 2020.
In order to obtain more valuable insights into the protein dynamics and accumulation of allergens in seeds during underground development, we performed a proteomic study on developing peanut seeds at seven different stages. A total of 264 proteins with altered abundance and contained at least one unique peptide was detected by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). All identified proteins were classified into five functional categories as level 1 and 20 secondary functional categories as level 2. Among them, 88 identified proteins (IPs) were related to carbohydrate/ amino acid/ lipid transport and metabolism, indicating that carbohydrate/amino acid/ lipid metabolism played a key role in the underground development of peanut seeds. Hierarchical cluster analysis showed that all IPs could be classified into eight cluster groups according to the abundance profiles, suggesting that the modulatory patterns of these identified proteins were complicated during seed development. The largest group contained 41 IPs, the expression of which decreased at R 2 and reached a maximum at R3 but gradually decreased from R4. A total of 14 IPs were identified as allergen-like proteins by BLAST with A genome (Arachis duranensis) or B genome (Arachis ipaensis) translated allergen sequences. Abundance profile analysis of 14 identified allergens showed that the expression of all allergen proteins was low or undetectable by 2-DE at the early stages (R1 to R4), and began to accumulate from the R5 stage and gradually increased. Network analysis showed that most of the significant proteins were involved in active metabolic pathways in early development. Real time RT-PCR analysis revealed that transcriptional regulation was approximately consistent with expression at the protein level for 8 selected identified proteins. In addition, some amino acid sequences that may be associated with new allergens were also discussed.
为了更深入地了解过敏原蛋白在花生种子地下发育过程中的动态变化和积累情况，我们对七个不同发育阶段的花生种子进行了蛋白质组学研究。通过基质辅助激光解吸电离飞行时间/飞行时间质谱（MALDI-TOF/TOF MS）共检测到 264 种丰度发生变化的蛋白，其中至少含有一个独特肽段。所有鉴定到的蛋白均被分为 5 个一级功能分类和 20 个二级功能分类。其中，88 个鉴定蛋白（IP）与碳水化合物/氨基酸/脂质的运输和代谢有关，表明碳水化合物/氨基酸/脂质代谢在花生种子的地下发育过程中起着关键作用。系统聚类分析显示，根据丰度图谱，所有 IP 可分为 8 个聚类组，表明这些鉴定蛋白的调控模式在种子发育过程中较为复杂。最大的聚类组包含 41 个 IP，其表达在 R2 时下降，在 R3 时达到最大值，但从 R4 开始逐渐下降。共有 14 个 IP 通过与 A 基因组（Arachis duranensis）或 B 基因组（Arachis ipaensis）翻译的过敏原序列的 BLAST 鉴定为过敏原样蛋白。对 14 种鉴定过敏原的丰度谱分析表明，所有过敏原蛋白在早期（R1 到 R4）阶段通过 2-DE 检测表达量低或无法检测到，从 R5 阶段开始积累，并逐渐增加。网络分析表明，大多数显著蛋白都参与了早期发育的活跃代谢途径。实时 RT-PCR 分析表明，8 个选定的鉴定蛋白在转录水平上的调控与蛋白表达水平大致一致。此外，还讨论了一些可能与新过敏原相关的氨基酸序列。
