| 卢占军,丁鹏.三种提取单头柑橘木虱体内黄龙病菌DNA方法的比较研究[J].中国南方果树,2016,45(6): |
| 三种提取单头柑橘木虱体内黄龙病菌DNA方法的比较研究 |
| Three Methods of DNA Extraction for Candidatus Liberibacter asiaticus from Individual Adult Asian citrus psyllid |
| 投稿时间：2016-08-15 修订日期：2016-08-15 |
| DOI： |
| 中文关键词: 柑橘黄龙病菌 柑橘木虱 核酸提取 |
| 英文关键词:Candidatus Liberibacter asiaticus Citrus psyllid Nucleic acid extraction |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 摘要点击次数: 2001 |
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| 中文摘要: |
| 对饲养在温室内的感染柑橘黄龙病甜橙树上的柑橘木虱采用三种方法进行单头核酸提取，通过超微量核酸蛋白分析仪和细菌通用引物27F1492R常规PCR检测核酸质量，利用柑橘黄龙病菌特异性引物OI1/OI2c常规PCR和以r-rplLAs/f-rplLAs引物进行荧光定量qPCR检测。研究结果表明，三种方法均可以成功提取到单头柑橘木虱总核酸且均能检测到黄龙病菌，李菁方法时间短，步骤简便，提取的核酸浓度较高，但核酸质量较低；氯仿-异戊醇抽提方法步骤较多、历时长，提取核酸浓度较低，但核酸质量较高；试剂盒方法简便快速，提取的核酸浓度高，核酸质量一般，成本较高。三种方法各有优缺点，可根据后续分子实验具体要求选择合适的提取方法。本文为有效的检测柑橘木虱体内黄龙病菌提供了技术基础，也可以推广到其他微小昆虫的内生细菌等相关基因研究领域。 |
| 英文摘要: |
| Three methods were taken to extract DNA of individual adult Asian citrus psyllid which was cultivated on sweet orange with Candidatus Liberibacter asiaticus (CLa.) infected in greenhouse. The efficiency of different DNA extraction methods were evaluated by comparing the concentration, purity, PCR products of DNA, respectively. The results showed that three methods can extract DNA of CLa. and citrus psyllid. Li Jing’s method was simple, rapid and the concentration of DNA sample was higher, while the purity was lower; chloroform-isoamylol was complex and the concentration of DNA sample was lower, but the purity was higher; Animal Tissue Direct PCR Kit was simpler, the concentration was higher, while the costs was higher. Three methods have advantages and shortcoming respectively. Researcher can choose the appropriate extraction method according to the subsequent molecular experiment. This paper provides technical basis for effective detection CLa. from individual citrus psyllid, and it can be extended to related research about endophytic bacteria and other small insects. |
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