The Research On The Population Of Harmonia Axyridis Pallas With SSCP In The Area Of Maoershan

Harmonia axyridis Pallas has pleomorphism. There are many mottlings in their proalacrustacea,and some taxonomists classed them according to the mottlings.This thesis has analysed the COⅠof 13 phenotypes of Harmonia axyridis Pallas havingdifferent mottlings,using the method of PCR-SSCP, to study the kinship of them. According tothe COⅠ,optimize the experiment condition to obtain a stable and suitable condition for theexperiment,then check and analyse the results with DNA sequencing.The results:First,extract the genomic DNA of one Harmonia axyridis successfully with the method ofCTAB and it is suit for the experiment. Compared with the traditionary method for extractingthe DNA of insect using homogenate approach, this method is simpler and save more time andmoney.Second, optimize the electrophoresis condition of PAGE, and the suitable condition is: gelstrength: 5%, degree of cross linking:29:1, voltage:150V(15h), glycerol: none, electrophoresisbuffer: 1×TBE, the ratio between the sample and denaturing agent:3:3. The ssDNA can beseparated clearly under this condition. But the conformation of ssDNA change little betweenthe 13 Harmonia axyridis. The sequence of COⅠgene has about 685bp,and about 600bp ofthem are very conservative. During the 13 phenotypes of Harmonia axyridis, Harmoniaaxyridis ab. 1347-octosigna ta Yuan, ab.falca ta Hem,ab. 12345678-sedecimsi-gna ta Mls andab. circumscrip ta Hem have base mutation. The homology between the 13 Harmonia axyridisare 94%-100%. The genetic similarity between Harmonia axyridis ab. 12345678-sedecimsi-gna taMls and ab. circumscrip ta Hem is 100%, and that between Harmonia axyrisdisab.1/2123456789-undevigintisigna ta Fald and ab. succinea Hope is 100% too. This implys thattheir kinship are nearest in the 13 phenotypes of Harmonia axyridis.