Abstract:

To establish and apply of AFLP analysis system for genetic diversity analysis of Liparis japonica. The genomic DNA in the fresh leaves of Liparis japonica was extracted using the traditional SDS method, and an AFLP silver-stained analysis system suitable for L. japonica was established by optimizing several main factors and then applied for genetic diversity analysis of L. japonica populations in northeast China. 500 ng genomic DNA was digested for 3 h with EcoRⅠand MseⅠrestriction enzymes, ligated to adaptors for 12 h at 16 ℃, pre-amplified using EcoRⅠ+1 and MseⅠ+1 primers and then amplified using EcoRⅠ+3 and MseⅠ+3 primers. The amplified products were resolved in denaturing ployacrylamide gels and stained with silver, and then the clear ploymorphic fingerprints were obtained. AFLP analysis of 185 individuals of L. japonica using three pairs of primers generated a total of 221 bands, of which 195 were polymorphic and the PPB value was 88.24%. The AFLP analysis system we established could be used in the research of genetic diversity of L. japonica species.