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Development and amplification of multiple co-dominant genetic markers from single spores of arbuscular mycorrhizal fungi by nested multiplex PCR
Ist Teil von
Fungal genetics and biology, 2005, Vol.42 (1), p.73-80
Ort / Verlag
United States: Elsevier Inc
Erscheinungsjahr
2005
Quelle
MEDLINE
Beschreibungen/Notizen
Multiple co-dominant genetic markers from single spores of the arbuscular mycorrhizal (AM) fungi
Glomus mosseae,
Glomus caledonium, and
Glomus geosporum were amplified by nested multiplex PCR using a combination of primers for simultaneous amplification of five loci in one PCR. Subsequently, each marker was amplified separately in nested PCR using specific primers. Polymorphic loci within the three putative single copy genes
GmFOX2,
GmTOR2, and
GmGIN1 were characterized by sequencing and single strand conformation polymorphisms (SSCP). Primers specific for the LSU rDNA D2 region were included in the multiplex PCR to ensure correct identification of the
Glomus spp. spores. Single AM fungal spores were characterized as multilocus genotypes by combining alleles of each amplified locus. Only one copy of each putative single copy gene could be amplified from each spore, indicating that spores are homokaryotic. All isolates of
G. mosseae had unique genotypes. The amplification of multiple co-dominant genetic markers from single spores by the nested multiplex PCR approach provides an important tool for future studies of AM fungi population genetics and evolution.