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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。
Direct-PCR and Nested-PCR were used to detect pure fungi, simulated fungi seeds and naturally infected seed extracts using tomato canker-specific primers. Results The detection sensitivity of Direct-PCR was 105cfu / mL and that of Nested-PCR was 102cfu / mL. The detection sensitivity of direct-PCR was 107cfu / mL and the value of Nested-PCR was 104cfu / mL; Nested-PCR can still be detected after 104-fold dilution in naturally infected seed extract. The establishment of tomato seed Nested-PCR detection technology, without nucleic acid extraction steps, the entire detection process can be completed within 8 h, convenient and fast, low cost, can be used as a routine detection of tomato seed carrier.