摘要： 为快速检测番茄种子表面的带菌情况,以便及时有效防控番茄溃疡病,从而减少病害损失。试验用细菌基因组DNA试剂盒提取番茄溃疡病的总基因组,用通用引物进行PCR反应。以番茄溃疡病菌的DNA为模板,使用专一性引物进行PCR扩增,通过对专一性引物的筛选,最终确定了ClaF1-ClaF2为快速检测番茄溃疡病菌PCR的特异性引物。该检测方法特异性强、灵敏度高,模拟种子带菌,对浸种水的检测灵敏度达1×105 CFU/mL。经PCR快速检测,22个番茄品种中只有黑贝番茄、粉红番茄和保罗塔带有番茄溃疡病菌,其余19个品种均未带菌。该方法直接对种子进行检测,不需进行病原菌分离培养,快速简便。
关键词:
番茄溃疡病菌,
PCR快速检测,
种子带菌
Abstract: Bacterial canker is the quarantine tomato seed-borne disease, it is rapid to detect the pathogen from the seed so that the disease can be controlled timely and effectivly and reduced the disease losses.The total genome DNA of Clavibacter michiganensis subsp. Michiganensis was extracted by Bacterial Genomic DNA Extraction Kit, then used for PCR with universal primer.A pair of primers ClaF1-ClaF2 was finally chose by screening specific PCR primers as rapid detection of Clavibacter michiganensis subsp. Michiganensis.The PCR assay was proved to be sensitive for detection the bacteria in the water of soaking seed which was carried bacterial artificially sufficiently reached to 1×105 CFU/mL.It was a specificity and high sensitivity detection method.With the PCR rapid detection method, the result showed that Heibei tomato, Fenhong tomato and Baoluota of 22 tomato varieties carried with Clavibacter michiganensis subsp. Michiganensis, others were not detected.This method was used for directly detecting the seeds, without isolating pathogens, fast and conveniently.
Key words:
Bacterial canker of tomato,
Rapid detection of PCR,
Seed-borne
中图分类号:
田晓燕, 张庆萍, 王燕春, 席先梅, 白海. 内蒙古地区番茄溃疡病菌PCR快速检测技术及22个番茄品种种子表面带菌检测[J]. 华北农学报, 2015, 30(1): 150-153. doi: 10.7668/hbnxb.2015.01.024.
TIAN Xiao-yan, ZHANG Qing-ping, WANG Yan-chun, XI Xian-mei, BAI Hai. Rapid Detection of Clavibacter michiganensis subsp. Michiganensis in Inner Mongolia Area Using PCR and Testing of Seed Surface Borne Bacteria of 22 Tomato Varieties[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(1): 150-153. doi: 10.7668/hbnxb.2015.01.024.