摘要
为找出较优的单头柑桔木虱黄龙病菌核酸提取方法,以饲养于温室内感染柑桔黄龙病的甜橙树上的柑桔木虱为试材,采用3种方法提取单头柑桔木虱的总核酸,对提取的核酸样品进行质量检测,并利用柑桔黄龙病菌特异性引物OI1/OI2c以及r-rplLAs/f-rplLAs分别进行了常规PCR和qPCR检测。结果表明,3种方法均可以成功提取到单头柑桔木虱总核酸且均能检测到黄龙病菌。3种方法各有优缺点:李菁的方法步骤简单、历时短,提取的核酸浓度较高,但核酸质量较低;氯仿-异戊醇法步骤较多、历时长,提取核酸浓度较低,但核酸质量较高;动物组织直接PCR试剂盒法简便快速,提取的核酸浓度高,核酸质量一般,成本较高。
DNA of individual adult Asian citrus psyllid cultured on sweet orange infected with Candidatus Liberibacter asiaticus（CLa.）in greenhouse was extracted by three methods.Quality of the DNA was detected.PCR and qPCR detection were performed with special primers of OI1/OI2 cand r-LrplLAs/f-LrplLAs,respectively.The results showed that three methods can extract DNA of CLa.from citrus psyllid.Li Jing’s method was simple,rapid and the concentration of DNA was higher,but the purity was low.The method of chloroform-Lisoamylol was complex and the concentration of DNA was low,but the purity was high.The method of Animal Tissue Direct PCR Kit was the simplest,the concentration was high,but the costs was high.
出处
《中国南方果树》
北大核心
2016年第6期1-5,共5页
South China Fruits
基金
国家自然科学基金(31560602)
江西省科技计划重大项目(20152ACF60003)
江西省科技计划重大项目(20161ACF60016)
江西省科技计划重大项目(20143ACF60012)资助
江西省2016年度研究生创新专项资金项目(YC2016-S408)
江西省科技厅青年科学基金计划(20133BAB21001)
江西省自然科学基金(20122BAB204024)
关键词
柑桔黄龙病菌
柑桔木虱
核酸提取
Candidatus Liberibacter asiaticus
citrus psyllid
nucleic acid extraction
