Open AccessTaxon‐specific oligonucleotide primers for detection of two ancient endomycorrhizal fungi, Glomus occultum and Glomus brasilianum
Author(s) -
Millner P.D.,
Mulbry W.W.,
Reynolds S.L.
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10559.x
Subject(s) - biology , glomus , primer (cosmetics) , ribosomal dna , strain (injury) , polymerase chain reaction , botany , genetics , phylogenetics , gene , chemistry , anatomy , spore , organic chemistry
A unique oligonucleotide pair, GOCC56:GOCC427, was designed that correctly primed specific amplification of a ∼370‐bp sequence spanning the ITS and 5.8S rDNA regions of Glomus occultum and Glomus brasilianum . In addition, this primer pair successfully detected G. occultum and G. brasilianum DNA in nested PCR using a primary PCR product amplified from highly diluted extracts of colonized corn ( Zea mays ) roots using modified ITS1:ITS4 primers. A second primer pair, GBRAS86:GBRAS388, primed specific amplification of a ∼200‐bp sequence spanning the ITS and 5.8S rDNA regions present only in G. brasilianum and Glomus strain GR582. Combined use of both primer pairs provides the means to detect and differentiate two ancient endomycorrhizal species, G. occultum and G. brasilianum , undetectable by standard root staining procedures. Sequence analysis showed that the purported G. occultum strain GR582 is likely a strain of G. brasilianum .