Ekanayake Gayandi, Piibor Johanna, Midekessa Getnet, Godakumara Kasun, Dissanayake Keerthie, Andronowska Aneta, Bhat Rajeev, Fazeli Alireza
Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu, Estonia.
Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
Front Plant Sci. 2024 Nov 15;15:1477614. doi: 10.3389/fpls.2024.1477614. eCollection 2024.
Extracellular 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.
In this study, EVs were enriched from hydroponically cultivated potato plants (, 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.
Biophysical 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, < 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.
With 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".
细胞外囊泡（EVs）通过生物分子转移促进种间和种内/界间通讯，这些生物分子包括蛋白质和小RNA。植物来源的细胞外囊泡是该领域的一个热门话题，它既具有作为研究植物生理学潜在生物标志物的巨大潜力，又具有作为一种生物材料的巨大潜力，这种生物材料可以大量生产，用于从化妆品、食品添加剂到生物农药等各种行业。然而，需要对植物细胞外囊泡进行系统表征，以便为进一步的应用和研究奠定基础。
在本研究中，从水培马铃薯植株（品种为Laura）中富集细胞外囊泡。我们通过真空渗透从马铃薯根分泌物和马铃薯皮的质外体洗液中分离细胞外囊泡。然后对这些细胞外囊泡的生物物理和化学性质进行系统表征，以与标准细胞外囊泡特征进行比较，并探索它们在植物生理学中的作用。
生物物理和化学分析揭示了马铃薯根和皮来源的细胞外囊泡在形态上的相似性。根来源的细胞外囊泡的平均直径（164.6±7.3纳米）显著大于皮来源的细胞外囊泡（132.2±2.0纳米，P<0.004）。液相色谱-质谱联用（LC-MS）表明，与粗样品相比，纯化后的细胞外囊泡中蛋白质大量富集，根细胞外囊泡的富集率为42%，皮细胞外囊泡的富集率为25%。在根和皮细胞外囊泡中鉴定出的蛋白质只有11.8%是相同的，显著富集的蛋白质只有2%是共享的。两个细胞外囊泡蛋白质组中富集的途径都与对生物和非生物胁迫的反应有关，这表明细胞外囊泡在植物中具有防御作用。
通过进一步的实验来阐明具体的通讯方式，这些发现增加了我们对植物细胞外囊泡在物理和化学特征及其潜在功能方面的了解，有助于将农业废弃物可持续利用以大规模生产细胞外囊泡，符合“增值”的概念。
