Department of Plant Protection, Faculty of Agriculture, Bolu Abant İzzet Baysal University, Bolu 14030, Türkiye.
Central Research Laboratory Application and Research Center, Çankırı Karatekin University, Çankırı 18100, Türkiye.
Viruses. 2024 Jul 26;16(8):1203. doi: 10.3390/v16081203.
Some mycoviruses can be considered as effective biocontrol agents, mitigating the impact of phytopathogenic fungi and consequently reducing disease outbreaks while promoting plant health. , the causal agent of chestnut blight and a highly destructive pathogen, experienced a notable decrease in its virulence with the identification of cryphonectria hypovirus 1 (CHV1), a naturally occurring biocontrol agent. In this study, two innovative diagnostic protocols designed for the accurate and efficient detection of CHV1 are introduced. The ORF A and ORF B regions of CHV1 are targeted by these techniques, which employ colorimetric loop-mediated isothermal amplification (LAMP) with 2 Colorimetric LAMP Master Mix and real-time quantitative PCR (qPCR) with SYBR Green chemistry, respectively. The LAMP assay presents a discernible color transition, changing from pink to yellow after a 35 min incubation period. Comparative analysis, when assessed against two established reverse transcription-PCR (RT-PCR) techniques, reveals a significant enhancement in sensitivity for both the LAMP approach, which offers a tenfold increase, and the qPCR method, which showcases a remarkable 100-fold sensitivity improvement. Throughout the comparison phase, it was evident that the RT-PCR, LAMP, and qPCR procedures displayed superior performance compared to the Bavendamm test, relying on phenol oxidase activity, effectively distinguishing hypovirulent strains. Consequently, this study introduces two pioneer diagnostic assays for highly sensitive CHV1 detection, representing a substantial advancement in the realm of CHV1 surveillance techniques. These methodologies hold significant promise for enhancing research endeavors in the domain of the biological control of . .
一些真菌病毒可以被视为有效的生物防治剂，减轻植物病原菌的影响，从而减少疾病爆发，同时促进植物健康。栗疫病的病原体栗疫病栗疫病菌是一种高度破坏性的病原体，随着 Cryphonectria hypovirus 1（CHV1）的发现，一种天然的生物防治剂，其毒力显著降低。在这项研究中，引入了两种用于准确和高效检测 CHV1 的创新诊断方案。这些技术针对 CHV1 的 ORF A 和 ORF B 区域，分别采用具有 2 种颜色的比色环介导等温扩增（LAMP）和 SYBR Green 化学实时定量 PCR（qPCR）。LAMP 分析在 35 分钟孵育后呈现明显的颜色转变，从粉红色变为黄色。与两种已建立的逆转录聚合酶链反应（RT-PCR）技术进行比较分析，发现 LAMP 方法的灵敏度显著提高，增加了十倍，qPCR 方法的灵敏度提高了 100 倍。在整个比较阶段，事实证明，RT-PCR、LAMP 和 qPCR 程序的性能优于依赖酚氧化酶活性的 Bavendamm 测试，有效地区分了弱毒菌株。因此，本研究引入了两种用于高度敏感 CHV1 检测的开创性诊断检测方法，这是 CHV1 监测技术领域的重大进展。这些方法为栗疫病的生物防治研究提供了巨大的潜力。
