Open AccessArbuscular mycorrhizal fungi and bacteria: how to construct prokaryotic DNA‐free genomic libraries from the Glomales
Author(s) -
Hosny M.,
Tuinen D.,
Jacquin F.,
Füller P.,
Zhao B.,
GianinazziPearson V.,
Franken P.
Publication year - 1999
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.1574-6968.1999.tb13404.x
Subject(s) - spore , bacteria , biology , genomic dna , 16s ribosomal rna , glomus , microbiology and biotechnology , dna , ribosomal rna , polymerase chain reaction , gene , botany , genetics
Spores of various arbuscular mycorrhizal fungal isolates were analyzed for DNA of prokaryotic origin by amplification of the 16S rRNA. This shows that the presence of bacteria is not restricted to certain taxa within the Glomales, but distributed over all genera. Further experiments revealed, however, that, although single Glomus mosseae spores did not contain bacteria, samples of a number of spores were still contaminated with prokaryotes. In order to obtain genomic libraries from two arbuscular mycorrhizal fungi nearly free of clones of prokaryotic sequences, DNA extracted from spores was purified on CsCl gradients and used for library construction. Polymerase chain reaction analysis with primers for rRNA genes showed that the libraries contained if at all, only very low amounts of clones originated from bacteria.