Demi̇rel Serap, Usta Mustafa, Korkmaz Gülüstan, Kurt Zeynelabidin
Molecular Biology and Genetic Department of Science Faculty, Van Yüzüncü Yıl University, Van, Türkiye.
Plant Protection Department of Agriculture Faculty, Van Yüzüncü Yıl University, Van, Türkiye.
BMC Plant Biol. 2025 Jul 29;25(1):978. doi: 10.1186/s12870-025-07080-x.
Ribosome-inactivating proteins (RIPs) are a group of proteins known to inhibit protein synthesis and contribute to plant defense responses. Although the antiviral properties of various RIPs have been demonstrated, the antiviral potential of tritin, a type 1 RIP from bread wheat (Triticum aestivum L.), remains unexplored. This study aimed to investigate the antiviral activity of recombinant tritin against zucchini yellow mosaic virus (ZYMV) in zucchini (Cucurbita pepo L.) plants.
The tritin gene was isolated from the wheat cultivar Kutluk-94, cloned into the pETDuet-1 expression vector, and expressed in Escherichia coli BL21 (DE3) cells. Following induction, recombinant tritin was purified using Ni-NTA affinity chromatography. Antiviral activity was assessed by measuring morphological parameters, disease severity, and the expression levels of the ZYMV Coat Protein (CP) gene and the Pathogenesis-Related 1 (PR1) through quantitative real-time PCR.
Tritin-treated plants exhibited significantly lower ZYMV-CP gene expression compared to virus-inoculated controls at 3- and 15- days post-inoculation. Furthermore, PR1 gene expression was upregulated in response to tritin application, suggesting the activation of systemic defense pathways. Morphological assessments revealed dose-dependent phytotoxic effects, including reductions in chlorophyll content and plant growth at higher tritin concentrations.
This study represents the first report in demonstrating that recombinant tritin exhibits antiviral activity against ZYMV, reducing viral replication and enhancing defense gene expression in zucchini plants. However, the phytotoxic effects observed at higher concentrations suggest the need for dose optimization before agricultural application. These findings provide a promising basis for the development of RIP-based antiviral strategies to improve crop tolerance.
核糖体失活蛋白（RIPs）是一类已知能抑制蛋白质合成并参与植物防御反应的蛋白质。尽管各种RIPs的抗病毒特性已得到证实，但来自面包小麦（Triticum aestivum L.）的1型RIP——小麦核糖体失活蛋白（tritin）的抗病毒潜力仍未得到探索。本研究旨在调查重组小麦核糖体失活蛋白对西葫芦（Cucurbita pepo L.）植株中西葫芦黄花叶病毒（ZYMV）的抗病毒活性。
从小麦品种Kutluk - 94中分离出小麦核糖体失活蛋白基因，克隆到pETDuet - 1表达载体中，并在大肠杆菌BL21（DE3）细胞中表达。诱导后，使用镍 - 氮三乙酸（Ni - NTA）亲和层析法纯化重组小麦核糖体失活蛋白。通过测量形态学参数、病害严重程度以及通过定量实时PCR检测ZYMV外壳蛋白（CP）基因和病程相关蛋白1（PR1）的表达水平来评估抗病毒活性。
在接种后3天和15天，与接种病毒的对照相比，用小麦核糖体失活蛋白处理的植株中ZYMV - CP基因表达显著降低。此外，响应于小麦核糖体失活蛋白的施用，PR1基因表达上调，表明系统防御途径被激活。形态学评估显示出剂量依赖性的植物毒性效应，包括在较高小麦核糖体失活蛋白浓度下叶绿素含量降低和植株生长受抑制。
本研究首次表明重组小麦核糖体失活蛋白对ZYMV具有抗病毒活性，可减少西葫芦植株中的病毒复制并增强防御基因表达。然而，在较高浓度下观察到的植物毒性效应表明在农业应用前需要进行剂量优化。这些发现为开发基于RIP的抗病毒策略以提高作物耐受性提供了有前景的基础。
