Genetic discrimination of button mushroom Agaricus bisporus by DNA fingerprinting using RAPD marker

Abstract

The investigation was conducted for DNA fingerprinting and genetic diversity analysis of five button mushroom strains (Agaricus bisporus-6, Agaricus bisporus-2, Agaricus bisporus-3, Agaricus bisporus-8 and Agaricus bisporus-5) using eight decamer RAPD primers (OPA01, OPA02, OPA03, OPA04, OPA07, OPB17, OPB12 and OPA10). A modified CTAB technique was used to isolate DNA. RAPD analysis resolved 152 scorable bands, 89 bands were monomorphic and 63 bands were polymorphic. The primer OPB17 amplified the highest number of band (30) and OPA01 amplified the lowest number of bands (09). The primer OPA04 produced the 8 polymorphic bands. Thus, it showed higher level of polymorphism (80%). Dendrogram based on linkage distance using Unweighted Pair Group Method of Arithmetic Means (UPGMA) indicated segregated of five button mushroom strains into two main clusters. Only Agaricus bisporus-6 (V1) grouped in cluster 1 (C1) while the major cluster C2 included all other strains, as well as V2 (Agaricus bisporus-2), V3 (Agaricus bisporus-3), V4 (Agaricus bisporus-8) and V5 (Agaricus bisporus-5). At the linkage distance of 50.5, the major cluster C2 formed sub-cluster C2A (Agaricus bisporus-3) and C2B (Agaricus bisporus-2, A. bisporus-8 & A. bisporus-5). Overall results from the dendrogram indicated that the V1 (Agaricus bisporus-6) button mushroom was shown to be outliers in the dendrogram, that is different from other strains because of its unique genetic makeup and high yields of this starin. The present work revealed that the DNA fingerprinting of five button mushroom strains based on RAPD showed significant different among the strains.