Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Garzón 780, Montevideo 12900, Uruguay.
Departamento de Biociencias, Facultad de Química, Universidad de la República, General Flores 2124, Montevideo 11800, Uruguay.
Biochimie. 2021 Jan;180:229-242. doi: 10.1016/j.biochi.2020.11.005. Epub 2020 Nov 13.
Antimicrobial peptides (AMPs) play an essential role in plant defense against invading pathogens. Due to their biological properties, these molecules have been considered useful for drug development, as novel agents in disease therapeutics, applicable to both agriculture and medicine. New technologies of massive sequencing open opportunities to discover novel AMP encoding genes in wild plant species. This work aimed to identify cysteine-rich AMPs from Peltophorum dubium, a legume tree from South America. We performed whole-transcriptome sequencing of P. dubium seedlings followed by de novo transcriptome assembly, uncovering 78 AMP transcripts classified into five families: hevein-like, lipid-transfer proteins (LTPs), alpha hairpinins, defensins, and snakin/GASA (Giberellic Acid Stimulated in Arabidopsis) peptides. No transcripts with similarity to cyclotide or thionin genes were identified. Genomic DNA analysis by PCR confirmed the presence of 18 genes encoding six putative defensins and 12 snakin/GASA peptides and allowed the characterization of their exon-intron structure. The present work demonstrates that AMP prediction from a wild species is possible using RNA sequencing and de novo transcriptome assembly, regarding a starting point for studies focused on AMP gene evolution and expression. Moreover, this study allowed the detection of strong AMP candidates for drug development and novel biotechnological products.
抗菌肽 (AMPs) 在植物抵御入侵病原体的防御中发挥着重要作用。由于其生物学特性,这些分子被认为可用于药物开发,作为疾病治疗的新型药物,适用于农业和医学。大规模测序的新技术为在野生植物物种中发现新的 AMP 编码基因提供了机会。本工作旨在从南美洲的豆科植物 Peltophorum dubium 中鉴定富含半胱氨酸的 AMP。我们对 P. dubium 幼苗进行了全转录组测序,然后进行从头转录组组装,共发现 78 种 AMP 转录本,分为五类:hevein 样、脂质转移蛋白 (LTPs)、α发夹素、防御素和 snakin/GASA (拟南芥中赤霉素刺激) 肽。未鉴定出与环肽或硫素基因具有相似性的转录本。通过 PCR 对基因组 DNA 进行分析证实了存在编码 6 种假定防御素和 12 种 snakin/GASA 肽的 18 个基因,并对其外显子-内含子结构进行了表征。本工作表明,使用 RNA 测序和从头转录组组装可以从野生物种中预测 AMP,为 AMP 基因进化和表达的研究提供了起点。此外,本研究还检测到了用于药物开发和新型生物技术产品的强有力的 AMP 候选物。