CAJ | 학술논문

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.
침대진경상용평과과실휴대리화역병균Erwinia amylovora수량유한적특점,선취원우병균pEA29질립적2대인물P29A/P29B화PEANT1/PEANT2배대조합성투식PCR,기검측령민도가체0.15 Pg균체DNA,검측령민도고우EPPO추천적단관투식PCR방법화상규PCR방법.분별이용저3충PCR검측방법대미국、신서란、일본화지리등국진경적166비평과양품진행검측,3충검측방법적양품양성솔분별위53.6%、38.0%화8.4%,시험결과표명차투식PCR검측방법가용우진경상용평과적리화역병균쾌속검측.진경양품적검측결과증실료진경상용평과과실중존재리화역병균적가능성.
The specific primers P29A/P29B and PEANT1/PEANT2 derived from plasmid pEA29 of Erwinia amylovora were selected to combine nested PCR method for the detection of trace E. amylovora in imported apple fruits with a limit as low as 0. 15 pg of total DNA. From 166 samples of imported apple fruits from USA, Japan, New Zealand and Chile, this detection method achieved better results than nested PCR in a sin-gle closed tube that EPPO recommended and standard PCR with 53.6% , 38.0% and 8. 4% of positive sam-ples, respectively. This nested PCR method was proposed to be used for the detection of E. amylovora in im-ported commercial apple fruits. The results of detection verified the possibility of imported commercial apple fruits harbouring E. amylovora in the calyxes and fruit stalks.