National Research Centre for Mushroom, Chambaghat, Solan 173 213 (H.P.), India
*Corresponding author. E-mail: sksingh1111@hotmail.com
Abbreviations: ARDRA, Amplified ribosomal DNA restriction analysis; ITS, Internal transcribed spacers; rDNA, Ribosomal DNA; RFLP, Restriction fragment length polymorphism; rRNA, Ribosomal RNA; UPGMA, Unweighed pair group method using arithmetic averages.
Thirty-six cultures representing eight Morchella and related genera, namely, Morchella esculenta, M. crassipes, M. spongiola, M. vulgaris, M. angusticeps, M. conica, Mitrophora semilibera and Verpa conica were subjected to restriction analysis of ITS1-5.8S-ITS2 region of rDNA. Six restriction endonuclease enzymes viz TaqI, EcoRl, Mspl, Rsal, Hinfl and BsuRl were used to generate restriction fragments and analysis of phylogenetic relationships among morels. The Amplified Ribosomal DNA Restriction Analysis (ARDRA) not only distinguished yellow morels from black morels but also separated related genera Mitrophora semilibera and Verpa conica from true morels. Simultaneously, each morel species could be separated from each other exhibiting considerable phylogenetic distances. The unique restriction fragment profiles generated by the restriction endonucleases enabled us to identify marker fragments to distinguish each species within and amongst the morel group. Since no intra-specific variation in restriction profiles by the six restriction endonucleases could be visualized among monospores, the technique could be used for rapid identification of wild morel specimens as a cheap alternative to direct sequencing for germplasm cataloguing.
Morchellaceae, Morchella, Verpa conica, Mitrophora semilibera, RFLP, ARDRA, 5.8S rRNA gene, phylogeny
