Table1_Systematic characterization of extracellular vesicles from potato (Solanum tuberosum cv. Laura) roots and peels: biophysical properties and proteomic profiling.docx

IntroductionExtracellular vesicles (EVs) facilitate inter and intra-species/kingdom communication through biomolecule transfer, including proteins and small RNAs. Plant-derived EVs, a hot topic in the field, hold immense capability both as a potential biomarker to study plant physiology and as a biomaterial that can be mass-produced to be used in various industries ranging from cosmetics and food additives to biological pesticides. However, a systematic characterization of plant EVs is required to establish a foundation for further applications and studies.
MethodsIn this study, EVs were enriched from hydroponically cultivated potato plants (Solanum tuberosum, cv. Laura). We isolated EVs from root exudates and the apoplastic wash of potato peels using vacuum infiltration. These EVs were then systematically characterized for their biophysical and chemical properties to compare with standard EV characteristics and to explore their roles in plant physiology.
ResultsBiophysical and chemical analyses revealed morphological similarities between potato root and peel-derived EVs. The average diameter of root-derived EVs (164.6 ± 7.3 nm) was significantly larger than that of peel-derived EVs (132.2 ± 2.0 nm, p < 0.004). Liquid chromatography–mass spectrometry (LC-MS) demonstrated substantial protein enrichment in purified EVs compared to crude samples, with a 42% enrichment for root EVs and 25% for peel EVs. Only 11.8% of the identified proteins were common between root and peel EVs, with just 2% of significantly enriched proteins shared. Enriched pathways in both EV proteomes were associated with responses to biotic and abiotic stress, suggesting a defensive role of EVs in plants.
DiscussionWith further experimentation to elucidate the specific methods of communication, these findings increase the details known about plant EVs in terms of their physical and chemical characteristics and their potential functions, aiding in sustainable agricultural waste utilization for large-scale EV production, aligning with the concept of “valorization”.

引言
细胞外囊泡（Extracellular Vesicles, EVs）可通过转运蛋白质、小RNA等生物分子，实现跨物种/界以及物种内、界内的信息交流。植物源细胞外囊泡是当前该领域的研究热点，其兼具巨大应用潜力：既可作为研究植物生理学的潜在生物标志物，又可作为生物材料实现规模化生产，应用于化妆品、食品添加剂、生物农药等多个行业。然而，要为其后续应用与研究奠定基础，仍需对植物源细胞外囊泡进行系统的表征分析。

方法
本研究从水培栽培的马铃薯（Solanum tuberosum，cv. Laura）中富集细胞外囊泡。我们采用真空浸润法，分别从马铃薯根系分泌物及马铃薯果皮的质外体冲洗液中分离细胞外囊泡。随后，对这些细胞外囊泡的生物物理与化学性质进行系统表征，一方面与标准细胞外囊泡特征进行比对，另一方面探究其在植物生理学中的功能。

结果
生物物理与化学分析结果显示，马铃薯根系来源与果皮来源的细胞外囊泡在形态上具有相似性。根系来源细胞外囊泡的平均直径为(164.6±7.3)nm，显著大于果皮来源细胞外囊泡的(132.2±2.0)nm（p<0.004）。液相色谱-质谱联用法（Liquid Chromatography-Mass Spectrometry, LC-MS）分析显示，与粗样品相比，纯化后的细胞外囊泡中蛋白质富集程度显著提升：根系来源细胞外囊泡的蛋白质富集率为42%，果皮来源者为25%。根系与果皮来源的细胞外囊泡中，仅11.8%的鉴定蛋白质为共有蛋白，且显著富集的蛋白质中仅有2%存在重叠。两种来源细胞外囊泡的蛋白质组中，富集的通路均与生物及非生物胁迫响应相关，这表明细胞外囊泡在植物中发挥防御功能。

讨论
若通过进一步实验阐明其具体的信息交流机制，本研究结果将丰富人们对植物源细胞外囊泡物理化学性质及潜在功能的认知，有助于实现农业废弃物的可持续利用以规模化生产细胞外囊泡，契合“资源化增值”的理念。