Identification of medicinal mushroom species based on nuclear large subunit rDNA sequences.
The purpose of this study was to develop molecular identification method for medical mushrooms and their preparations based on the nucleotide sequences of nuclear large subunit (LSU) rDNA. Four specimens were collected of each of the three representative medicinal mushrooms used in Korea: Ganoderma lucidum, Coriolus versicolor, and Fomes fomentarius. Fungal material used in these experiments included two different mycelial cultures and two different fruiting bodies from wild or cultivated mushrooms. The genomic DNA of mushrooms were extracted and 3 nuclear LSU rDNA fragments were amplified: set 1 for the 1.1-kb DNA fragment in the upstream region, set 2 for the 1.2-kb fragment in the middle, and set 3 for the 1.3-kb fragment downstream. The amplified gene products of nuclear large subunit rDNA from 3 different mushrooms were cloned into E. coli vector and subjected to nucleotide sequence determination. The sequence thus determined revealed that the gene sequences of the same medicinal mushroom species were more than 99.48% homologous, and the consensus sequences of 3 different medicinal mushrooms were more than 97.80% homologous. Restriction analysis revealed no useful restriction sites for 6-bp recognition enzymes for distinguishing the 3 sequences from one another, but some distinctive restriction patterns were recognized by the 4-bp recognition enzymes AccII and HhaI. This analysis was also confirmed by PCR-RFLP experiments on medicinal mushrooms.
Lee
Ji Seon
JS
Institute of Biotechnology, College of Pharmacy, Yeungnam University, Gyongsan 712-749, Republic of Korea.
Lim
Mi Ok
MO
Cho
Kyoung Yeh
KY
Cho
Jung Hee
JH
Chang
Seung Yeup
SY
Nam
Doo Hyun
DH
D004271:DNA, Fungal / Q000032:analysis; D004275:DNA, Ribosomal / Q000235:genetics*; D032902:Ganoderma / Q000145:classification* / Q000235:genetics / Q000254:growth & development; D007723:Korea; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques; D010946:Plants, Medicinal / Q000145:classification* / Q000235:genetics / Q000254:growth & development; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020072:Polyporales / Q000145:classification* / Q000235:genetics / Q000254:growth & development; D017422:Sequence Analysis, DNA
D004271:DNA, Fungal; D004275:DNA, Ribosomal