Abstract：
Cotton root rot caused by Macrophomina phaseolina pose a significant threat to cotton production, leading to substantial yield and quality losses. Early and accurate diagnosis of this pathogen in soil is crucial for effective disease management. This study presents a pioneering investigation into the utilization of the nit gene encoding nitrilase for the development of a molecular diagnostic assay aimed at the rapid detection of M. phaseolina in field soils. The methodology involved the design and validation of primers targeting the Nit gene sequence, followed by the optimization of PCR conditions for efficient amplification. Leveraging state-of-the-art molecular techniques, the assay offers a novel protocol to accurately identify the presence of M. phaseolina in soil with high sensitivity and specificity. The specificity of the designed primers was confirmed through PCR amplification using DNA from M. phaseolina and other related fungi. Sensitivity tests demonstrated that the PCR assay reliably detected M. phaseolina DNA at concentrations as low as 1 ng. Furthermore, the performance of the diagnostic assay was rigorously evaluated using field soil samples with a known status of M. phaseolina infection, demonstrating its reliability and efficacy in real-world scenarios. This study introduces a novel molecular marker for the detection of M. phaseolina and offers a rapid and efficient means for screening M. phaseolina in large soil samples with minimal time and manpower.
摘要：
由巨噬细胞引起的棉花根腐病对棉花生产构成重大威胁，导致大量的产量和质量损失。土壤中这种病原体的早期和准确诊断对于有效的疾病管理至关重要。这项研究为利用编码腈水解酶的nit基因开发旨在快速检测田间土壤中的菜豆的分子诊断测定法提供了开创性的研究。该方法涉及针对Nit基因序列的引物的设计和验证，然后优化PCR条件进行高效扩增。利用最先进的分子技术，该检测方法提供了一种新的方案,能够以高灵敏度和特异性准确地鉴定土壤中的胡须谷杆菌.所设计的引物的特异性通过PCR扩增确认,所述PCR扩增使用来自菜豆属和其他相关真菌的DNA。灵敏度测试表明，PCR测定法可以可靠地检测到浓度低至1ng的菜豆分枝杆菌DNA。此外，诊断试验的性能进行了严格的评估，使用田间土壤样品与已知状态的豆科植物感染，在现实场景中展示其可靠性和有效性。这项研究介绍了一种用于检测菜豆的新型分子标记，并提供了一种快速有效的方法，以最少的时间和人力在大型土壤样品中筛选菜豆。
