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Typing method for N 2 ‐fixing bacteria based on PCR‐F — application to the characterization of Frankia strains
Author(s) -
JAMANN S.,
FERNANDEZ M. P.,
NORMAND P.
Publication year - 1993
Publication title -
molecular ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.619
H-Index - 225
eISSN - 1365-294X
pISSN - 0962-1083
DOI - 10.1111/j.1365-294x.1993.tb00095.x
Subject(s) - frankia , biology , actinorhizal plant , intergenic region , polymerase chain reaction , restriction enzyme , genetics , typing , microbiology and biotechnology , gene , genome , bacteria , nitrogen fixation , root nodule
DNA sequences of an intergenic spacer (IGS) and parts of genes in the nif cluster were amplified by the polymerase chain reaction (PCR) using two primers derived from nifD ‐and nifK ‐conserved sequences. The PCR products were cleaved by ten 4–base cutting restriction enzymes and the restriction patterns were used as fingerprints to type Frankia strains. The feasability of this PCR‐RFLP method for typing Frankia strains was investigated on Frankia reference strains belonging mainly to the Elaeagnaceae infectivity group but also on new Frankia isolates and on other N 2 ‐fixing microorganisms. By modulating the stringency of the amplifications, we showed the method allowed to target either Frankia strains or the whole N 2 ‐fixing microbial community. DNA digestion patterns were used to estimate the sequence divergence between the Frankia nifD‐K fragment. The estimated relationships deduced from these genotypic data correlated well with established Frankia taxonomic schemes.