应用多重PCR技术同步检测柑橘4种重要嫁接传播病原(英文)  

Simultaneous and Rapid Detection of Four Important Graft-transmissible Pathogens of Citrus by Multiplex PCR

作　　者：刘金香[1,2] 周常勇[1,2] 唐科志[1,2] 李玲娣[1,2] 刘科宏[1,2] 田晓[1,2] 李中安[1,2] 

机构地区：[1]西南大学柑橘研究所 [2]中国农业科学院柑橘研究所,重庆400712

出　　处：《植物病理学报》2014年第3期295-301,共7页Acta Phytopathologica Sinica

基　　金：Chang-jiang Scholars and Innovative Research Team in University(PCSIRT,IRT0976);MOA's Special Fund for Agro-Scientific Research in the Public Interest(201203076);the National Natural Science Foundation of China(30600419);the Fundamental Research Funds for the Central Universities(XDJK2009C133)

摘　　要：柑橘衰退病毒(Citrus tristeza virus,CTV),柑橘碎叶病毒(Citrus tatter-leaf virus,CTLV),柑橘裂皮病类病毒(Citrus exocortis viroid,CEVd)和柑橘黄龙病(Huanglongbing,HLB)亚洲种病原(Candidatus liberobacter asiaticus)是重要的柑橘嫁接传播病原。本文建立了同时检测HLB病菌、CTV、CEVd和CTLV 4种柑橘嫁接病原的一步法、双温多重PCR检测技术体系,同时在体系中设置内参基因。应用该体系快速评价了4种嫁接传播病原在田问侵染情况,结果表明28个田间样品CTV、CEVd、CTLV和HLB感染率分别为89.3%、17.9%、10.7%和28.6%,接近半数样品为混合感染。并且将该方法应用于快速评价茎尖嫁接苗病毒的脱除情况。Citrus tristeza virus （CTV）, Citrus tatter leaf virus （CTLV）, Citrus exocortis viroid （CEVd）, and Candidatus Liberibacter asiaticus （Ca. L. asiaticus） associated with Huanglongbing （HLB） are important graft-transmissible pathogens of citrus. A rapid multiplex PCR protocol with an internal control was developed for the detection of these citrus pathogens using a timesaving two-temperature cycling for PCR amplification. The assay was applied for rapid detection of those pathogens in field-grown trees. The results indicated that 89.3%, 10.7%, 17.9% and 28.6% of the 28 field-grown citrus samples were infected with CTV, CTLV, CEVd and Ca. L. asiaticus, respectively, and approximately half of the samples were co-infected with more than one pathogen. In addition, the absence of multiple pathogens in shoot-tip grafted budlings was evaluated with the multiplex PCR assay.

关 键 词：柑橘衰退病毒 柑橘碎叶病毒 柑橘裂皮病类病毒 柑橘黄龙病菌 多重PCR 

分 类 号：S432.4[农业科学—植物病理学]