Mycobiology.  2003 Mar;31(1):23-31. 10.4489/MYCO.2003.31.1.023.

ITS Primers with Enhanced Specificity to Detect the Ectomycorrhizal Fungi in the Roots of Wood Plants

• Kim DH
• Chung HC
• Ohga S
• Lee SS

• KMID: 2052976
• DOI: http://doi.org/10.4489/MYCO.2003.31.1.023

Abstract

With universal primer ITS1-F, the specific DHJ2 primer was developed to detect the Ectomycorrhizal (ECM) root tips in soil and to identify the species of ECM fungi, as based on DNA sequences of rDNA stored in GeneBank of NCBI. This primer was designed with the common sites of rDNA of Amanita and Boletus, and was also designed with several DNA programs provided by NCBI. The DNA fragments synthesized by PCR were calculated to be 1,000 to 1,200 bps of DNA located to 18s to 28s rDNA to contain two variable sites of ITS, indicating much diversities for specific species or ecotypes of ECM fungi. The primer DHJ2 reacted with the genomic DNA's extracted from the tissues of basidiocarp at the rate of 73 of 80 fungi collected produced single bands with a 1,100 bps length. The DNA fragment synthesized with the genomic DNA that extracted from eight ECM tips of Pinus densiflora was confirmed and analysized to the rDNAs of ECM in full sequences, and informed to be a ECM fungal species in the forest.

Keyword

Amanita; Boletus; Ectomycorrhizal (ECM) fungal species; ECM root tips; Specific primer

MeSH Terms

Amanita
Base Sequence
DNA
DNA, Ribosomal
Ecotype
Fruiting Bodies, Fungal
Fungi*
Meristem
Pinus
Polymerase Chain Reaction
Sensitivity and Specificity*
Soil
Wood*
DNA
DNA, Ribosomal
Soil