Institute of Plant Genetics, Polish Academy of Sciences, Poznan, Poland.
J Appl Genet. 2009;50(4):411-9. doi: 10.1007/BF03195702.
Spores of many fungal pathogens are dispersed by wind. Detection of these airborne inocula is important in forecasting both the onset and the risk of epiphytotics. Species-specific primers targeted at the internal transcribed spacer (ITS) region of Leptosphaeria maculans and L. biglobosa - the causal organisms of phoma stem canker and stem lesions of Brassica spp., including oilseed rape - were used to detect DNA extracted from particles deposited on tapes obtained from a spore trap operated in Rarwino (northwest Poland) from September to November in 2004 and 2006. The quantities of DNA assessed by traditional end-point PCR and quantitative real-time PCR were compared to microscopic counts of airborne ascospores. Results of this study showed that fluctuations in timing of ascospore release corresponded to the dynamics of combined concentrations of DNA from L. maculans and L. biglobosa, with significant positive correlations between ascospore number and DNA yield. Thus the utilization of PCR-based molecular diagnostic techniques enabled the detection, identification, and accurate quantification of airborne inoculum at the species level. Moreover, real-time PCR was more sensitive than traditional PCR, especially in years with low ascospore numbers.
许多真菌病原体的孢子通过风传播。检测这些空气传播的接种物对于预测流行病的爆发和风险非常重要。针对 Leptosphaeria maculans 和 L. biglobosa（引起油菜黑胫病和十字花科植物茎部损伤的病原菌）内部转录间隔区（ITS）区域的物种特异性引物被用于检测从 2004 年和 2006 年 9 月至 11 月在波兰西北部 Rarwino 运行的孢子陷阱中获得的带材上沉积的颗粒中提取的 DNA。通过传统终点 PCR 和定量实时 PCR 评估的 DNA 量与空气中子囊孢子的显微镜计数进行了比较。这项研究的结果表明，子囊孢子释放时间的波动与 L. maculans 和 L. biglobosa 组合 DNA 浓度的动态变化相对应，子囊孢子数量与 DNA 产量之间存在显著的正相关关系。因此，基于 PCR 的分子诊断技术的利用能够在物种水平上进行空气传播接种物的检测、鉴定和准确定量。此外，实时 PCR 比传统 PCR 更敏感，尤其是在子囊孢子数量较低的年份。
