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A Duplex PCR Assay for Rapid Detection of Phytophthora nicotianae and Thielaviopsis basicola
Liu, Na,Jiang, Shijun,Feng, Songli,Shang, Wenyan,Xing, Guozhen,Qiu, Rui,Li, Chengjun,Li, Shujun,Zheng, Wenming The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.2
A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens Phytophthora nicotianae and Thielaviopsis basicola. The specific primers for P. nicotianae were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, ras gene and hgd gene, while the specific primers for T. basicola were designed based on its ITS regions and ${\beta}$-tubulin gene. The specificity of the primers was determined using isolates of P. nicotianae, T. basicola 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 $100fg/{\mu}l$ 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.
A Duplex PCR Assay for Rapid Detection of Phytophthora nicotianae and Thielaviopsis basicola
Na Liu,Shijun Jiang,Songli Feng,Wenyan Shang,Guozhen Xing,Rui Qiu,Chengjun Li,Shujun Li,Wenming Zheng 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.2
A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens Phytophthora nicotianae and Thielaviopsis basicola. The specific primers for P. nicotianae were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, ras gene and hgd gene, while the specific primers for T. basicola were designed based on its ITS regions and β-tubulin gene. The specificity of the primers was determined using isolates of P. nicotianae, T. basicola 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.