| 柑橘黄龙病、裂皮病和衰退病病原的多重RT-PCR检测 |
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| 引用本文： | 丁芳,曹庆,王国平,易干军,钟云.柑橘黄龙病、裂皮病和衰退病病原的多重RT-PCR检测[J].园艺学报,2006,33(5):947-952. |
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| 作者姓名： | 丁芳 曹庆 王国平 易干军 钟云 |
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| 作者单位： | (华中农业大学植物科学技术学院, 湖北武汉430070, 广东省农业科学院果树研究所, 广东广州510640) |
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| 基金项目： | 国家863课题(2001AA241142)，广东省科技攻关资助项目(2003B21601)，湖北省攻关计划(2006AA203B05) |
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| 摘 要： | 建立了一种同时检测柑橘黄龙病病原类细菌(Candidatus L iberibacter asiaticus, HLB) 、柑橘裂皮类病毒(Citrus exocortis viroid, CEVd) 、柑橘衰退病病毒(Citrus tristeza virus, CTV) 的多重RT-PCR技术体系。运用根据3 种病原核苷酸保守区序列设计的特异性引物, 成功的对同一样品中的HLB、CEVd、CTV进行多重RT2PCR扩增, 得到1 160 bp、371 bp、273 bp 3条特异性大小与试验设计相符的条带。就影响多重PCR 的主要因素引物浓度和退火温度进行了一系列优化, 建立了能同时检测HLB、CEVd、CTV的多重RT2PCR技术体系。该体系最低能从10 pg总RNA中检测出黄龙病类细菌和柑橘裂皮类病毒, 从1 pg总RNA中检测到柑橘衰退病毒。对采自田间37份材料的实际检测表明: 存在两种或3种病原的复合侵染。
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| 关 键 词： | 柑橘 多重RT-PCR 黄龙病 裂皮病 衰退病 检测 |
| 文章编号： | 0513-353X（2006）05-0947-06 |
| 收稿时间： | 2006-01-23 |
| 修稿时间： | 2006-01-232006-03-09 |
Studies on the Simultaneous Detection of Citrus Huanglongbing Pathogen, Citrus exocortis viroid, Citrus tristeza virus by Multiplex RT-PCR |
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| Ding Fang,Cao Qing,Wang Guoping,Yi Ganjun,Zhong Yun.Studies on the Simultaneous Detection of Citrus Huanglongbing Pathogen, Citrus exocortis viroid, Citrus tristeza virus by Multiplex RT-PCR[J].Acta Horticulturae Sinica,2006,33(5):947-952. |
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| Authors: | Ding Fang Cao Qing Wang Guoping Yi Ganjun Zhong Yun |
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| Institution: | (Plant Science and Technology Academy, Huazhong Agricultural University, Wuhan, Hubei 430070, China;Insititute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong 510640, China) |
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| Abstract: | A multiplex RT-PCR was successfully established to simultaneously detect Citrus Huanglongbing pathogen (Candidatus Liberibacter asiaticus, HLB), Citrus exocortis viroid(CEVd) and Citrus tristeza virus (CTV). Using three sets of specific primers designed according to the converted sequences of these three pathogens respectively, expected fragments of 1 160 bp (HLB), 371 bp (CEVd) and 273 bp (CTV) were successfully amplified by this multiplex RT-PCR systerm. To optimize the multiplex RT-PCR, the concentration of primers and annealing temperature were repeatedly tried. Final results showed that a multiplex RT-PCR system to simultaneously detect HLB, CEVd and CTV was firstly established successfully. Sensibility test showed: it could detect out HLB, CEVd in 10 pg, and CTV in 1 pg total RNA respectively. To confirm the utilization of the multiplex RT-PCR system, 37 samples collected from field were tested. The results showed: two or three pathogens mix-infection was in existence in fields. |
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| Keywords: | Citrus Multiplex RT-PCR Huanglongbing Citrus exocortis viroid Citrus tristeza virus Detection |
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