Detection of Stemphylium lycopersici from tobacco seeds using PCR with specific primers
2016
Kurose, D. (CABI Europe-UK, Surrey (UK)) | Hoang, L.H. | Furuya, N. | Matsumoto, Y. | Takeshita, M. | Tsushima, S. | Tsuchiya, K.
Stemphylium lycopersici is a common pathogen on vegetable crops causing severe disease in the world. In 2009, a preemergence damping-off caused by S. lycopersici was observed on tobacco seeds (Nicotiana tabacum L.) in Fukuoka, Japan. However, the etiology and mode of infection of this pathogen in tobacco seeds have not been clarified yet. In the present study, a primer pair was designed for direct, rapid and specific detection of S. lycopersici based on the sequences of the internal transcribed spacer regions including the 5.8S rDNA. A PCR product of approximately 449 bp was obtained only when the DNA extracted from pure cultures of S. lycopersici was used. No amplification was observed for other Pleospora / Stemphylium species or other genera of fungi. The PCR amplified as little as 100 fg of genomic DNA of S. lycopersici. Furthermore, S. lycopersici was detected from resultant tobacco seeds after the flowers were inoculated with this pathogen. Therefore, the designed primer pair is useful for the specific detection of S. lycopersici in tobacco seeds.
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