This study aimed to achieve rapid detection of Pseudomonas syringae pv.tabaci,the pathogen of tobacco wildfire disease.The specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as...This study aimed to achieve rapid detection of Pseudomonas syringae pv.tabaci,the pathogen of tobacco wildfire disease.The specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as the target gene.RAA was then combined with the lateral flow dipstick(LFD)to establish a LFD-RAA-based rapid detection system for the pathogen.Furthermore,the detection performance of the established method was tested.The results showed that the LFDRAA method had high specificity.The amplification could be completed after 25 min of reaction at 39℃.The sensitivity of the established method reached 0.0001 ng/μL,which was superior to that of PCR detection.Moreover,the LFD-RAA method could quickly detect P.syringae pv.tabaci from tobacco leaves,demonstrating field applicability.To sum up,the LFD-RAA method established in this study can be applied in the rapid detection and early diagnosis of tobacco wildfire disease.展开更多
Using genetic recombinant techniques, the tabtoxin resistant gene (trc gene) was to ligated a pBI121 vector under the CaMV 35S promoter to form the binary vector pBITR. Then the vector was introduced into Agrobacteriu...Using genetic recombinant techniques, the tabtoxin resistant gene (trc gene) was to ligated a pBI121 vector under the CaMV 35S promoter to form the binary vector pBITR. Then the vector was introduced into Agrobacterium tumefaciens by triparental mating. After transforming the tobacco, many transgenic plants were obtained. PCR tests indicated that the trc gene was inserted in the genome DNA of tobacco. Biological activity analysis also demonstrated that the transgenic plant alleviated the symptom caused by the pathogen of tobacco wildfire disease.展开更多
文摘This study aimed to achieve rapid detection of Pseudomonas syringae pv.tabaci,the pathogen of tobacco wildfire disease.The specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as the target gene.RAA was then combined with the lateral flow dipstick(LFD)to establish a LFD-RAA-based rapid detection system for the pathogen.Furthermore,the detection performance of the established method was tested.The results showed that the LFDRAA method had high specificity.The amplification could be completed after 25 min of reaction at 39℃.The sensitivity of the established method reached 0.0001 ng/μL,which was superior to that of PCR detection.Moreover,the LFD-RAA method could quickly detect P.syringae pv.tabaci from tobacco leaves,demonstrating field applicability.To sum up,the LFD-RAA method established in this study can be applied in the rapid detection and early diagnosis of tobacco wildfire disease.
文摘Using genetic recombinant techniques, the tabtoxin resistant gene (trc gene) was to ligated a pBI121 vector under the CaMV 35S promoter to form the binary vector pBITR. Then the vector was introduced into Agrobacterium tumefaciens by triparental mating. After transforming the tobacco, many transgenic plants were obtained. PCR tests indicated that the trc gene was inserted in the genome DNA of tobacco. Biological activity analysis also demonstrated that the transgenic plant alleviated the symptom caused by the pathogen of tobacco wildfire disease.
