摘要
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。
By simply extracting and purifying, without DNA extraction, we used PCR directly to detect C. michiganensis subsp, miehiganensis carried by tomato seeds, and found that limit detection in pure bacterial suspension by Direct-PCR was 2 600 bacteria/ml, but reached 13 bacteria/ml by Nested-PCR. For seed extracts, Direct-PCR had no sensitivity to detect bacteria, while limit detection by Nested-PCR was 3 × 10^5 bacteria/ml. We can exactly detect C. michiganensis subsp, michiganensis on tomato seeds within 12hr by one-step Nested PCR, which is characterized as convenient, low cost, hypersensitive for fast detection of these bacteria.
出处
《甘肃农业科技》
2006年第5期12-15,共4页
Gansu Agricultural Science and Technology
关键词
番茄溃疡病
Nested—PCR
快速检测
Clavibacter michiganensis subsp, michiganensis
Nested-PCR
Fast detection
