The Study On Biological Characteristics And Molecular Detection Technique Of Alternaria Panax Whetz

To providing a theory basis for comprehensivetreatment of Alternaria panax Whetz, this study exploit biological characteristics,genetic diversity and molecule marker technology of Alternaria panax Whetz, the results are as follows:1. The result of Alternaria panax Whetz culture showed that:the optimum growth temperature of hypha was 20～25℃and the optimum pH was 6; the optimum germinate temperature of conidium was 25～35℃and the optimum pH was 5. sucrose was the optimal carbon source and starch was the worst;Peptone was the optimal nitrogen source and urea was the worst;PDA was the most suitable medium of all. The fatal temperature of hypha was 55℃when the fatal temperature of conidium was 50℃.2. Alternaria panax Whetz PCR reaction system was as follows:the optimize system was 20 ng·μL-1 DNA,150μmol·L-1 dNTP,10 pmol·L-1 primer, Mg2+1.5mol·L-1,1U Taq polymerase,10×Buffer 2.5μL in 25μL reaction system,and the lack part complement with DW. The reaction system was as follows:pre-denature at 94℃for 5 min, and then 94℃1 min,40℃1 min,72℃2min, for 40 cycles, finally extended at 72℃for 7mins.3. The genetic relationship among Alternaria panax Whetz from 6 areas were analysed with RAPD.With 6 primers amplified 68 bands and 35 bands were polymorphic, the ratio was 52.2%.Cluster analysis results showed that: at the level of genetic distance 0.20, Alternaria panax Whetz of Songjianghe Fusong and Quanyang Fusong were relatived.Cluster analysis results showed that the level of genetic similarity coefficient of 0.34 can be divided into three categories, namely West Side Changbai Mountain (Songjianghe Fusong, Quanyang Fusong and Antu),East Side Changbai Mountain(Toudao Helong),and Southwest Side Changbai Mountain (Cuocaogou Tonghua and Linjiang).The average genetic distance was 0.54.4. The total DNA from young leaves of Alternaria panax Whetz was extracted by CTAB method.Specific primers have been designed by the known sequence and all of the tested strains were amplified with PCR methods.Consistent with the predictions,the target bands were about 475bp,which could be used for specific molecular marking.Zhou extrating method was the best one in the three DNA extraction methods,Alternaria panax Whetz was successfully extracted.Alternaria panax Whetz was detected by PCR amplification in seven areas.