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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。
In order to find a better method for nucleic acid extraction of the yellow-headed pathogen of single-headed citrus psylla, three methods were used to extract single-headed orange The total nucleic acid of the orange lice, the nucleic acid samples were extracted for quality testing, and the use of Citrus Huanglongye specific primers OI1 / OI2c and r-rplLAs / f-rplLAs respectively by conventional PCR and qPCR detection. The results showed that all of the three methods could successfully extract the total nucleic acid of single-headed citrus psilocybe and could detect yellow dragon’s disease. The three methods have their own advantages and disadvantages: the method has the advantages of simple steps and short duration, high concentration of nucleic acid extracted, low quality of nucleic acid, more steps of chloroform-isopentyl alcohol method, longer extraction time and lower concentration of nucleic acid extracted Nucleic acid quality higher; animal tissue direct PCR kit method is simple and fast, high concentration of nucleic acid extraction, nucleic acid quality in general, higher cost.