Biologie et diversité génétique des souches deFrankiaassociées aux Casuarinacées

Absence of Cospeciation between the UnculturedFrankiaMicrosymbionts and the Disjunct ActinorhizalCoriariaSpecies

“…Individual lobes were selected, surface-sterilized in 30% (vol/vol) H 2 O 2 , and rinsed several times with distilled sterile water. The DNA extraction from single nodule lobes was performed as previously described by Rouvier et al [ 26 ]. Nodule lobes were crushed with sterile plastic mortars and pestles in 300 μ L of extraction buffer (100 mM Tris (pH 8), 20 mM EDTA, 1.4 M NaCl, 2% (wt/vol) CTAB (cetyltrimethyl ammonium bromide), and 1% (wt/vol) PVPP (polyvinyl polypyrrolidone)).…”

High-resolution phylogenetic analysis of NO2--oxidizing Nitrobacter species using the rrs-rrl IGS sequence and rrl genes.

“…It is at least twice as variable as the rrl gene and four times as variable as the rrs gene. It has been used for identification purposes in a number of bacteria (East & Collins, 1993 ;Honeycutt et al, 1995 ;Minnick et al, 1994 ;Rouvier et al, 1996 ;Riffard et al, 1998). The rrs-rrl IGS region of members of the genus Nitrobacter has also been used profitably for the characterization of natural populations using PCR-RFLP analysis (Navarro et al, 1992b) and has been found to be highly discriminating, even between strains with rrs sequences differing by less than 15 positions (99 % similarity ; Orso et al, 1994).…”