Liu Na, Jiang Shijun, Feng Songli, Shang Wenyan, Xing Guozhen, Qiu Rui, Li Chengjun, Li Shujun, Zheng Wenming
College of Life Sciences, Henan Agricultural University, Zhengzhou 450002, China.
College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China.
Plant Pathol J. 2019 Apr;35(2):172-177. doi: 10.5423/PPJ.OA.09.2018.0173. Epub 2019 Apr 1.
A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens and . The specific primers for were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, gene and gene, while the specific primers for were designed based on its ITS regions and tubulin gene. The specificity of the primers was determined using isolates of , and control samples. The results showed that the target pathogens could be detected from diseased tobacco plants by a combination of the specific primers. The sensitivity limitation was 100 fg/μl of pure genomic DNA of the pathogens. This new assay can be applied to screen out target pathogens rapidly and reliably in one PCR and will be an important tool for the identification and precise early prediction of these two destructive diseases of tobacco.
开发了一种双重PCR方法，用于同时检测和鉴定烟草根腐病病原体。基于核糖体基因的内部转录间隔区（ITS）区域、基因和基因开发了针对的特异性引物，而基于其ITS区域和微管蛋白基因设计了针对的特异性引物。使用、的分离株和对照样品确定引物的特异性。结果表明，通过特异性引物的组合可以从患病烟草植株中检测到目标病原体。灵敏度限制为病原体纯基因组DNA的100 fg/μl。这种新方法可用于在一次PCR中快速可靠地筛选出目标病原体，将成为这两种烟草毁灭性病害鉴定和精确早期预测的重要工具。
