Species-specific detection and identification of fusarium species complex, the causal agent of sugarcane pokkah boeng in China.
Species-specific detection and identification of fusarium species complex, the causal agent of sugarcane pokkah boeng in China.
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中国甘蔗白粉病病原菌镰刀菌种群的物种特异性检测与鉴定。
DOI：
10.1371/journal.pone.0104195
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Zhang M
中科院分区：
文献类型：
--
作者：
Lin Z;Xu S;Que Y;Wang J;Comstock JC;Wei J;McCord PH;Chen B;Chen R;Zhang M
由镰刀菌复合体引起的 Pokkah boeng 病会导致甘蔗产量严重损失。因此，迫切需要对病原体进行快速、准确的检测和鉴定，以管理和防止甘蔗瘟疫的蔓延。 2012年和2013年从中国5个主要甘蔗产区采集的pokkah boeng样品中共回收到101株分离菌株。通过形态学观察、致病性测试和基于真菌保守的rDNA-ITS的系统发育分析，鉴定了病原菌。开发了物种特异性 TaqMan 实时 PCR 和常规 PCR 方法，用于快速准确地检测甘蔗 pokkah boeng 的致病因子。还对来自中国的总共 84 株镰孢菌分离株以及来自其他真菌病原体 Sporisorium scitamineum 和 Phoma sp. 的几种分离株进行了 PCR 测定的特异性和敏感性评估。和甘蔗内生菌 Acremium sp.通过形态学观察、致病性测试和系统发育分析，鉴定出引起甘蔗腐烂病的两个镰刀菌属（F. verticillioides 和 F. proliferatum）。物种特异性 TaqMan PCR 和常规 PCR 经过设计和优化，以靶向其 rDNA-ITS 区域。 TaqMan PCR 的灵敏度约为 10 pg 真菌 DNA 输入，是传统 PCR 的 1,000 倍，并成功检测到田间种植的甘蔗中的 pokkah boeng。这项研究首次鉴定了两种 F. verticillioides 和 F. proliferatum，它们是中国甘蔗 pokkah boeng 的致病病原体。它还描述了一种物种特异性 PCR 检测方法的开发，用于检测和确认中国大陆甘蔗植物中的这些病原体。这种方法对于广泛的研究工作以及甘蔗 pokkah boeng 的监管反应和管理非常有用。
Pokkah boeng disease caused by the Fusarium species complex results in significant yield losses in sugarcane. Thus, the rapid and accurate detection and identification of the pathogen is urgently required to manage and prevent the spreading of sugarcane pokkah boeng. A total of 101 isolates were recovered from the pokkah boeng samples collected from five major sugarcane production areas in China throughout 2012 and 2013. The causal pathogen was identified by morphological observation, pathogenicity test, and phylogenetic analysis based on the fungus-conserved rDNA-ITS. Species-specific TaqMan real-time PCR and conventional PCR methods were developed for rapid and accurate detection of the causal agent of sugarcane pokkah boeng. The specificity and sensitivity of PCR assay were also evaluated on a total of 84 isolates of Fusarium from China and several isolates from other fungal pathogens of Sporisorium scitamineum and Phoma sp. and sugarcane endophyte of Acremonium sp. Two Fusarium species (F. verticillioides and F. proliferatum) that caused sugarcane pokahh boeng were identified by morphological observation, pathogenicity test, and phylogenetic analysis. Species-specific TaqMan PCR and conventional PCR were designed and optimized to target their rDNA-ITS regions. The sensitivity of the TaqMan PCR was approximately 10 pg of fungal DNA input, which was 1,000-fold over conventional PCR, and successfully detected pokkah boeng in the field-grown sugarcane. This study was the first to identify two species, F. verticillioides and F. proliferatum, that were causal pathogens of sugarcane pokkah boeng in China. It also described the development of a species-specific PCR assay to detect and confirm these pathogens in sugarcane plants from mainland China. This method will be very useful for a broad range of research endeavors as well as the regulatory response and management of sugarcane pokkah boeng.
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