AFLP Fingerprinting Analysis Of Pleurotus Ostreatus And Sequence Identification Of The Polymorphic AFLP Fragments

Pleurotus belongs to white-rot fungi. In addition to its value on edible, Pleurotus can secrete ligninolytic enzymes, laccases to oxidatively cleave lignin and aromatic pollutants. These enzymes are greatly potential in bio-pulping, bio-bleaching of pulp and treatment of environmental pollutants, etc. Pleurotus ostreatus which delegate the main characteristic of genus Pleurotus is the mode of the genus. Research in our laboratory have screened out some Pleurotus strains with high yield of ligninolytic enzymes, analyzed the activity and output of ligninolytic enzymes.Studies were carried out in the following 5 aspects and corresponding results were obtained. Firstly, the optimal submerged culture and growth conditions for P. ostreatus were screened. It was shown that the best medium for P. ostreatus is CMA culture and the temperature is 25℃. Secondly, conditions of the AFLP fingerprinting analysis for P. ostreatus were tested and then 14 strains of P. ostreatus from different areas were analyzed with AFLP method. The results showed that the primer pairs E-3/M-3 especially E-AGC/M-CAT and E-AGC/M-ACC could give more amplified DNA fragments than others. Thirdly, from the fingerprinting map of the primer pair E-AGC/M-CAT, 184 clear and stable DNA bands were demonstrated, including 101 polymorphous bands that are accounted for 54.89%. Fourthly, the fragments of the strain P. ostreatus 8101 withAFLP fingerprinting analysis were successfully purified and cloned in pMD18-T Vector. Then, two purified fragments were sequenced. Finally, The genetic distance of 14 strains was calculated and its dendrogram was performed based on AFLP data from 10 E-3/M-3 primer pairs.