Data_Sheet_1_High throughput screening of fungal phytopathogens caught in Australian forestry insect surveillance traps.PDF

Post-border surveillance for forestry’s high priority pests and pathogens is conducted routinely through established programs focused on the main points-of-entry and across the major plantation growing regions. Currently, most diagnostic protocols used to identify fungal phytopathogens sampled during these surveys rely on traditional methods, such as morphological examination and DNA barcoding techniques. This stepwise process from isolation to species identification is often regarded as slow, expensive, and limited due to the need for disease manifestation and/or comprehensive expertise for rapid and accurate detection. In this study, we applied a recently validated high-throughput, dual-marker amplicon sequencing approach on insect surveillance traps from across Australia to assess its performance for the targeted surveillance of the Ophiostomatales, an order of fungi comprising notable phytopathogens which are vectored by bark beetles. By using a recently validated assay we were able to confidently characterize a range of Ophiostomatalean taxa known to be present in Australia, while reporting eight first detections from environmental DNA. Our study demonstrates the value of targeted multi-barcode amplicon sequencing for high-throughput screening of fungi caught in post-border surveillance traps, in addition to emphasizing research priorities that require further investigation before such methods can be implemented routinely for biosecurity.

针对林业高优先级害虫和病原体的边境后监测工作，通常通过针对主要入境点和主要种植区域的既定计划进行。目前，用于识别在此次调查中采集的真菌性植物病原体的诊断方案大多依赖传统方法，如形态学检查和DNA条形码技术。从分离到物种鉴定这一逐步过程，往往因疾病的表现和/或对快速准确检测所需的全面专业知识而显得缓慢、昂贵且受限。在本研究中，我们采用了一种最近得到验证的高通量、双标记扩增子测序方法，对来自澳大利亚各地的昆虫监测陷阱进行测试，以评估其在针对 Ophiostomatales 级真菌（由树皮甲虫传播的显著植物病原体组成）的针对性监测中的性能。通过使用最近得到验证的检测方法，我们能够自信地鉴定出已知存在于澳大利亚的多种 Ophiostomatalean 分类群，并从环境DNA中报告了八种首次发现。我们的研究证明了在边境后监测陷阱中捕获的真菌进行针对性多条形码扩增子测序的高通量筛选的价值，同时强调了在将此类方法常规应用于生物安全之前，需要进一步研究的科研优先事项。