Preparation of AFLP Mediated-Molecular Certificate for 12 Bred Strains of the Button Mushroom, Agaricus bisporus

Document Type : Research Article



As hybrid spawn is propagated asexually in edible mushrooms, producers may propagate it without the permission of the owner. For the protection of the owner right, there is a great need for a molecular certificate for each commercial line. The high-resolution genotyping method of amplified fragment length polymorphism (AFLP) analysis was used to prepare molecular certifications and fingerprinting of 12 lines of Agaricus bisporus. AFLP templates were prepared by digestion of Agaricus DNA with EcoRI and Tru9I restriction endonucleases and subsequent ligation of corresponding site specific adaptors. A total of 54 polymorphic bands were obtained using eight primer combinations. The largest polymorphic bands (12) were produced by using (E-CT/t-TC) primer combination. Molecular certificates were adjusted through presence (1) and non-presence (0) of polymorphic bands. The 12 Button Mushroom lines were distinctly allocated into three clusters with UPGMA method. These results indicated that AFLP‌ is a fast, highly discriminating and reproducible DNA fingerprinting method in distinguishing bred lines of white button mushroom.

Key words: Button Mushroom, Bred Lines, Primer Combinations, AFLP, Molecular Certificates