| 进境苹果果实中梨火疫病菌的套式PCR检测 |
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| 引用本文： | 贾平乔,周国梁,吴杏霞,张卫东,杨晓君,徐殿胜,易建平.进境苹果果实中梨火疫病菌的套式PCR检测[J].植物病理学报,2009,39(5):449-457. |
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| 作者姓名： | 贾平乔 周国梁 吴杏霞 张卫东 杨晓君 徐殿胜 易建平 |
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| 作者单位： | 1 华东理工大学生物工程学院, 上海 200237;2 上海出入境检验检疫局, 上海 200135;3 国家质检总局标准法规中心, 北京 100088 |
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| 基金项目： | 科技部国家"十一五"科技支撑计划项目，国家质检总局科研项目 |
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| 摘 要： | 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。
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| 关 键 词： | 梨火疫病菌 苹果果实 套式PCR 检测 |
Detection of Erwinia amylovora in imported apple fruit with nested PCR |
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| JIA Ping-qiao,ZHOU Guo-liang,WU Xing-xia,ZHANG Wei-dong,YANG Xiao-jun,XU Dian-sheng,YI Jian-ping.Detection of Erwinia amylovora in imported apple fruit with nested PCR[J].Acta Phytopathologica Sinica,2009,39(5):449-457. |
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| Authors: | JIA Ping-qiao ZHOU Guo-liang WU Xing-xia ZHANG Wei-dong YANG Xiao-jun XU Dian-sheng YI Jian-ping |
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| Institution: | 1 East China University of Science and Technology, Shanghai 200237, China;2 Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China;3 International standard and regulation research center, General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Beijing 100088, China |
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| Abstract: | 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. |
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| Keywords: | Erwinia amylovora apple fruit nested PCR detection |
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