Mycobiology.  2003 Jun;31(2):68-73.

Restriction Analyses of PCR Amplified Partial SSU Ribosomal DNA to Distinguish Arbuscular Mycorrhizal Fungi from Other Fungi Colonizing Plant Roots

• Lee JK
• Tae MS
• Eom AH
• Lee SS

Abstract

Roots of Glycine max and Miscanthus sinensis and soil samples were collected from various field sites at Goesan, Chungbuk in Korea. Microscopic observations of the roots indicated high colonization rates of both arbuscular mycorrhizal fungi (AMF) and other fungi. The partial small subunit of ribosomal DNA genes were amplified with the genomic DNA extracted from their roots by nested polymerase chain reaction (PCR) with universal primer NS1 and fungal specific primers AM1. Restriction fragment length polymorphism (RFLP) was analyzed using the combinations of three restriction enzymes, HinfI, AluI and AsuC21. Nucleotides sequence analysis revealed that ten sequences from Miscanthus sinensis and one sequence from Glycine max were close to those of arbuscular mycorrhizal fungi. Also, 33% of total clones amplified with NS31-AM1 primers from M. sinensis and 97% from G. max were close to Fusarium oxysporum or other pathogenic fungi, and they were successfully distinguished from AMF. Results suggested that these techniques could help to distinguish arbuscular mycorrhizal fungi from root pathogenic fungi in the plant roots. Especially, DNA amplified by these primers showed distinct polymorphisms between AMF and plant pathogenic species of Fusarium when digested with AsuC21.

Keyword

Arbuscular mycorrhizal fungi; AsuC21; Fusarium; Glomales; Glycine max; Miscanthus sinensis

MeSH Terms

Chungcheongbuk-do
Clone Cells
Colon*
DNA
DNA, Ribosomal*
Fungi*
Fusarium
Korea
Nucleotides
Plant Roots*
Plants*
Polymerase Chain Reaction*
Polymorphism, Restriction Fragment Length
Sequence Analysis
Soil
Soybeans
DNA
DNA, Ribosomal
Nucleotides
Soil