摘要
利用番茄溃疡病菌特异性引物对纯菌液、模拟带菌种子及自然感染种子提取液分别进行Direct-PCR和Nested-PCR检测。结果在纯菌液中,Direct-PCR的检测灵敏度为105cfu/mL,Nested-PCR为102cfu/mL;在模拟带菌种子提取液中,Direct-PCR的检测灵敏度为107cfu/mL,Nested-PCR为104cfu/mL;在自然感染种子提取液中,稀释104倍后Nested-PCR仍然可以检出。建立的番茄种子Nested-PCR检测技术,无需经过核酸提取步骤,可以在8 h内完成整个检测过程,方便快捷,成本低廉,可作为番茄种子带菌的常规检测方法。
Tomato bacterial canker is the most important bacterial diseases which caused substantial economic losses in the worldwide.Seeds were the main pathway of long-distance spread and seed-borne Clavibacter michiganensis subsp.michiganensis（Cmm） were difficult to detect by current methods because of they were low rate of infected.Direct-PCR and Nested-PCR were performed for the detection of Cmm in bacterial suspensions,artificial contaminated seed extracts and naturally infected seed extracts,respectively.The detection sensitivity of Direct-PCR is 105 cfu/mL and 107 cfu/mL in bacterial suspensions and artificial contaminated seed extracts,respectively.But the sensitivity of Nested-PCR was 102 cfu/mL and 104 cfu/mL respectively.And Nested-PCR could provide positive result in 104 fold dilution in naturally infected seed extracts.The Nested-PCR developed in our study was convenient and low-cost,which could be used a routine method for the detection of tomato bacterial canker.
出处
《西北农业学报》
CAS
CSCD
北大核心
2011年第7期28-31,共4页
Acta Agriculturae Boreali-occidentalis Sinica
基金
甘肃省科技厅中小企业创新基金计划(0802NCCA028)
国家质检总局项目(2007IK259,2009IK276)
