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园艺学报 ›› 2011, Vol. 38 ›› Issue (10): 2017-2022.

• 新技术与新方法 • 上一篇    下一篇

安祖花细菌性疫病的Nested-PCR检测

孟 鹤1,金茂勇2,肖橘清2,张宝珠2,明 军1,袁素霞1,刘 春1,*,张 宁3   

  1. (1中国农业科学院蔬菜花卉研究所,北京 100081;2北京市大兴区苗圃,北京 102601;3华中农业大学园艺林学学院,武汉 430070)
  • 收稿日期:2011-05-30 修回日期:2011-09-29 出版日期:2011-10-25 发布日期:2011-10-25
  • 通讯作者: 刘 春1,*,

Detection of Xanthomonas campestris pv. dieffenbachiae on Anthurium andraeanum by Nested-PCR

MENG He1,JIN Mao-yong2,XIAO Ju-qing2,ZHANG Bao-zhu2,MING Jun1,YUAN Su-xia1,LIU Chun1,*,and ZHANG Ning3   

  1. (1Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2Beijing Daxing Nursery,Beijing 102601,China;3College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,China)
  • Received:2011-05-30 Revised:2011-09-29 Online:2011-10-25 Published:2011-10-25
  • Contact: LIU Chun1,*,

摘要: 根据基因库中已发表的安祖花细菌性疫病致病菌粉黛黄单胞杆菌(Xanthomonas campestris pv. dieffenbachiae)基因序列设计引物XcF1/XcR1、XcF2/XcR2,通过扩增条件优化,建立了安祖花细菌性疫病的Nested-PCR检测方法,可以直接从感染细菌性疫病的安祖花组织DNA中扩增出1条343 bp的特异性条带。分析表明该序列完全存在于GenBank登录的Xanthomonas campestris pv. dieffenbachiae序列X70380.1中。通过对安祖花植株进行Nested-PCR扩增,在植株不同部位均检测到了343 bp的特异性条带,且病原菌在植株中分布不均匀,其中老叶叶柄中扩增强度最高,根茎连接处、老叶、嫩叶柄、嫩叶、佛焰苞柄扩增强度次之,根、佛焰苞和花序中扩增最弱。

关键词: 安祖花, 细菌性疫病, 粉黛黄单胞杆菌, Nested-PCR

Abstract: A Nested-PCR method was developed to detect the bacterial pathogens of Anthurium andraeanum by using two sets of specific primers designed according to Xanthomonas campestris pv. dieffenbachiae gene sequences. All samples infected by Xanthomonas campestris pv. dieffenbachiae could be amplified by Nested-PCR,yielding one special band with size of 343 bp. The amplified sequences were presented in the Xanthomonas campestris pv. dieffenbachiae sequence X70380.1 in GenBank. Xanthomonas campestris pv. dieffenbachiae was detected by Nested-PCR from different parts of Anthurium andraeanum,and the distribution of pathogens in plants was uneven. Remarkably high amplification existed in old petioles,followed by stem-root joints,old leaves,young petioles,young leaves and spathe petiole,and the weakest amplification was in root,spathes and inflorescences.

Key words: Anthurium andraeanum, bacterial blight, Xanthomonas campestris pv. dieffenbachiae, Nested-PCR

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