The fig Bionectria canker, caused by , is rapidly spreading in fruit trees, posing a significant threat to the horticultural industry. Early and specific detection of this pathogen is crucial for effective management. We developed a PCR-based assay using species-specific primers designed by exploiting sequence differences in the translation elongation factor 1-alpha () gene for early, rapid, and reliable diagnostics of this pathogen. The primers successfully amplified the target pathogen and did not react with other non-target fig canker-causing agents in direct and nested PCR. The designated primer pair, TEF-Sb3, with a 577 bp amplicon, demonstrated its efficacy by successfully detecting the pathogen with the lowest detectable concentration of 1 × 10 and 1 × 10 g.L in direct and nested PCR, respectively. Furthermore, the selected primer pair had high performance in detecting the target pathogen in naturally infected samples with evident canker symptoms in nested PCR. Moreover, our molecular tool has effectively identified latent infection of in asymptomatic samples from fig and walnut trees from three different regions of Fars Province. This is a notable advancement, as traditional culture-based methods have failed to detect these infections. Our results suggested that the designed species-specific PCR assays were sufficiently sensitive, specific, and reliable for identifying from pure cultures and detecting the pathogen in symptomatic and asymptomatic trees. This assay can be a rapid and cost-effective tool for monitoring and managing Bionectria canker caused by in natural and potential host trees.

中文翻译：

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开发基于 PCR 的检测方法，用于检测果树溃疡病病原体 Stilbocrea banihashemiana 的明显和潜在感染

由 引起的无花果溃疡病正在果树中迅速蔓延，对园艺业构成重大威胁。对该病原体的早期和特异性检测对于有效管理至关重要。我们开发了一种基于 PCR 的检测方法，使用物种特异性引物，通过利用翻译延伸因子 1-α () 基因中的序列差异设计，以便对该病原体进行早期、快速和可靠的诊断。引物成功扩增了目标病原体，并且在直接和巢式 PCR 中不与其他非目标无花果溃疡病致病菌发生反应。指定的引物对 TEF-Sb3 具有 577 bp 的扩增子，通过在直接 PCR 和巢式 PCR 中分别成功检测出最低可检测浓度为 1 × 10 gL 和 1 × 10 gL 的病原体，证明了其功效。此外，所选择的引物对在巢式PCR中检测具有明显溃疡症状的自然感染样品中的目标病原体方面具有高性能。此外，我们的分子工具还有效地识别了来自法尔斯省三个不同地区的无花果树和核桃树的无症状样本中的潜在感染。这是一个显着的进步，因为传统的基于培养的方法无法检测到这些感染。我们的结果表明，所设计的物种特异性 PCR 检测方法对于从纯培养物中进行识别以及检测有症状和无症状树木中的病原体具有足够的敏感性、特异性和可靠性。该测定可以成为一种快速且经济高效的工具，用于监测和管理由天然和潜在寄主树引起的生物克氏菌溃疡病。