Root and crown rot (RCR) of dry bean is a disease complex caused by soil-borne pathogens that can limit bean production in Zambia. Identifying the primary pathogen in the complex would facilitate the control as well as breeding for resistance to the disease. Plant tissue samples and DNA embedded FTA ® Cards were collected to isolate, detect and molecularly identify RCR pathogens using genus/species specific primers as well as high throughput deep sequencing with illumina Miseq using universal eukaryote bar coded primers Euk7. A total of 204 isolates were characterized using morphological features and Sanger sequencing. Analysis of reads and operational taxonomic units from illumina sequencing, detection frequencies from polymerized chain reaction as well as morphological characterization revealed Fusarium species complex as the main component of RCR complex of dry bean in Zambia, primarily F. oxysporum , followed by F. solani . A sensitivity chi-square test comparing detection methods of the four major RCR pathogens showed statistical differences. A highly significant positive correlation ( p < 0.001) confirmed by a good agreement (kappa (k) 0.5 to 0.6, p < 0.05) was found between FTA ® Card and direct plant tissue. Use of FTA ® Cards can facilitate sampling, storing and analysis of DNA for RCR pathogen identification.

中文翻译：

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使用培养和分子方法鉴定和表征赞比亚干豆真菌根腐病病原体

干豆的根冠腐烂 (RCR) 是一种由土传病原体引起的疾病复合体，可限制赞比亚的豆类生产。确定复合体中的主要病原体将有助于控制和培育对疾病的抗性。收集植物组织样本和 DNA 嵌入的 FTA ® 卡，以使用属/种特异性引物分离、检测和分子鉴定 RCR 病原体，以及使用通用真核生物条码引物 Euk7 使用illumina Miseq 进行高通量深度测序。使用形态特征和 Sanger 测序对总共 204 个分离株进行了表征。分析来自照明测序的读数和操作分类单元，聚合链反应的检测频率以及形态特征表明，镰刀菌属复合体是赞比亚干豆 RCR 复合体的主要成分，主要是尖孢镰刀菌，其次是茄病镰刀菌。比较四种主要 RCR 病原体的检测方法的敏感性卡方检验显示出统计学差异。在 FTA ® Card 和直接植物组织之间发现了高度显着的正相关 (p < 0.001)，并得到了良好的一致性（kappa (k) 0.5 到 0.6，p < 0.05）的证实。使用 FTA ® 卡可以促进 DNA 的采样、存储和分析，以进行 RCR 病原体鉴定。在 FTA ® Card 和直接植物组织之间发现了高度显着的正相关 (p < 0.001)，并得到了良好的一致性（kappa (k) 0.5 到 0.6，p < 0.05）的证实。使用 FTA ® 卡可以促进 DNA 的采样、存储和分析，以识别 RCR 病原体。在 FTA ® Card 和直接植物组织之间发现了高度显着的正相关 (p < 0.001)，并得到了良好的一致性（kappa (k) 0.5 到 0.6，p < 0.05）的证实。使用 FTA ® 卡可以促进 DNA 的采样、存储和分析，以进行 RCR 病原体鉴定。