[ Objective] This study aimed to develop a simple and effective DNA extraction method for citrus Huanglongbing pathogen detection. [ Method] From aspects of preparing procedure, prepare time and the quality of DNA, ad...[ Objective] This study aimed to develop a simple and effective DNA extraction method for citrus Huanglongbing pathogen detection. [ Method] From aspects of preparing procedure, prepare time and the quality of DNA, advantages and disadvantages of three sample preparation methods were compared, include two Direet-PCR extraction methods and one universal genomic DNA extraction kit method. In addition, PCR amplification effect on specific primers for 16S rDNA of ＂Candidatua Libefibacter asiaticus＂ （CLas） had also been evaluated. [ Result] The results showed that RT-qPCR detected CLas by using DNA obtained from one of Direct-PCR extraction method as templates. Under improved Direct-PCR extraction method, 16S rDNA of CLas could also be amplified by routine PCR. [ Conclusion] A simple and effective DNA extraction method of citrus Huanglongbing pathogen have been established, which provides technical supports for prepa- ration of large number of samples for detection of CLas.展开更多
基金Supported by Most-USDA Cooperation Program in Agriculture Science and Technology(10-8100-1452-CA)National Natural Science Foundation of China(31071712)
文摘[ Objective] This study aimed to develop a simple and effective DNA extraction method for citrus Huanglongbing pathogen detection. [ Method] From aspects of preparing procedure, prepare time and the quality of DNA, advantages and disadvantages of three sample preparation methods were compared, include two Direet-PCR extraction methods and one universal genomic DNA extraction kit method. In addition, PCR amplification effect on specific primers for 16S rDNA of ＂Candidatua Libefibacter asiaticus＂ （CLas） had also been evaluated. [ Result] The results showed that RT-qPCR detected CLas by using DNA obtained from one of Direct-PCR extraction method as templates. Under improved Direct-PCR extraction method, 16S rDNA of CLas could also be amplified by routine PCR. [ Conclusion] A simple and effective DNA extraction method of citrus Huanglongbing pathogen have been established, which provides technical supports for prepa- ration of large number of samples for detection of CLas.
