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对番茄细菌性溃疡病菌进行种子带菌的模拟检测试验。对用菌液处理过的种子进行简单抽提和纯化,不经过DNA提取而以抽提和纯化的病原菌为模板直接进行一步法PCR检测,对纯菌液D irect-PCR最低检出限为2 600个细菌/m l,而N ested-PCR最低检出限可达到13个细菌/m l;对种子提取液,D irect-PCR不能检出,而N ested-PCR最低检出限可达到3×105个细菌/m l。一步法N ested-PCR可在12 h内对番茄种子携带的番茄溃疡病菌进行准确的定性鉴定。并且方法方便快速,成本低,灵敏度高,适用于种子携带番茄溃疡病菌的快速鉴定。
Simulated test on tomato bacteriocidal bacteria with seed carrier. The simple extraction and purification of the seed treated with the bacterium solution was carried out by one-step PCR directly using the extracted and purified pathogenic bacteria as a template without any DNA extraction. The detection limit of the pure-liquid D irect-PCR was 2 600 bacteria / ml, and N ested-PCR detection limit of up to 13 bacteria / ml; Seeds extract, Direct-PCR can not be detected, and N ested-PCR detection limit of up to 3 × 105 Bacteria / ml. One-step N ested-PCR can accurately identify tomato-resistant tomato canker in 12 days. The method is convenient, rapid, low in cost and high in sensitivity, and is suitable for the rapid identification of seeds carrying tomato canker.