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针对进境商用苹果果实携带梨火疫病菌Erwinia amylovora数量有限的特点,选取源于病菌pEA29质粒的2对引物P29A/P29B和PEANT1/PEANT2配对组合成套式PCR,其检测灵敏度可达0.15 pg菌体DNA,检测灵敏度高于EPPO推荐的单管套式PCR方法和常规PCR方法。分别利用这3种PCR检测方法对美国、新西兰、日本和智利等国进境的166批苹果样品进行检测,3种检测方法的样品阳性率分别为53.6%、38.0%和8.4%,试验结果表明此套式PCR检测方法可用于进境商用苹果的梨火疫病菌快速检测。进境样品的检测结果证实了进境商用苹果果实中存在梨火疫病菌的可能性。
Aiming at the limited quantity of Erwinia amylovora carrying commercial apple fruit in the market, two pairs of primer P29A / P29B and PEANT1 / PEANT2 from the germ plasmid pEA29 were selected and used for nested PCR. The detection sensitivity was up to 0.15 pg DNA, detection sensitivity higher than EPPO recommended single-tube PCR method and conventional PCR method. 166 batches of apple samples entering the United States, New Zealand, Japan and Chile were detected by these three PCR methods. The positive rates of the three methods were 53.6%, 38.0% and 8.4% respectively. The results showed that This nested PCR detection method can be used for the rapid detection of imported pathogenic apple. The test results of the incoming samples confirmed the possibility of the presence of R. so on in the fruits of commercial apples entering the country.