GENETIC CHARACTERIZATION OF STRAINS OF CYANOBACTERIA USING PCR‐RFLP OF THE cpcBA INTERGENIC SPACER AND FLANKING REGIONS¹

Abstract

Oligonucleotide primers, specific for conserved regions of the genes encoding the β‐ and α‐phycocyanin subunits of phycobilisomes (cpcB and cpcA) of cyanobacteria, were used to amplify a DNA fragment containing the intervening intergenic spacer region (cpcBA‐IGS) of 19 strains of three morphospecies of cyanobacteria. Six Australian strains were identified as Anabaena circinalis Rabenhorst, six strains were identified as Microcystis aeruginosa Kützing, and seven strains were identified as Nodularia spumigena Mertens. Restriction enzyme digestion of the amplification products from the strains revealed restriction fragment length polymorphism (RFLP) within all three morphospecies. Strains corresponding to M. aeruginosa were highly polymorphic: 11 of the 14 restriction enzymes used displayed RFLPs. The A. circinalis and N. spumigena strains were less variable: three of 14 enzymes and seven of 14 enzymes, respectively, showed RFLPs. The presence of genetic variation between strains within these three divergent morphospecies, which span two orders of cyanobacteria (Chroococcales Wettstein and Nostocales (Borzi) Geitler), show that the cpcBA‐ IGS fragment has broad application as a molecular marker for intrageneric studies of cyanobacteria systematics and genetics.

Characterization of Nodularia strains, cyanobacteria from brackish waters, by genotypic and phenotypic methods.

“…In this program, bootstrap analysis was used to evaluate the tree topologies by performing 1000 resamplings. Bolch et al (1996) ; 2, Bolch et al (1999) ;3, Camm & Stein (1974) ;4, Castenholz (1989) ;5, Fujii et al (1997a) ;6, Fujii et al (1997b) ;7, Giovannoni et al (1988) ;8, Herdman et al (1979a) ;9, Herdman et al (1979b) ;10, Koma! rek et al (1993) ; 11, Kononen et al (1993) ;12, Lachance (1981) ;13, Lehtima$ ki et al (1994) ;14, Lehtima$ ki et al (1997) ;15, Lyra et al (1997) ;16, Moisander et al (1996) ; 17, Neilan (1996) ; 18, Neilan et al (1995) ;19, Neilan et al (1997b) ;20, Neilan et al (1999) ;21, Nelissen et al (1994) ;22, Nordin & Stein (1980) ; 23, Rinne & Tarkiainen (1978) ; 24, Rippka et al (1979) ; 25, Sivonen et al (1989a) ;26, Sivonen et al (1989b) ; 27, S ) marda et al 28, Wilmotte (1994) ; 29, Wilmotte et al (1994).…”

Phylogenetic relationships of Arthrospira strains inferred from 16S rRNA gene and cpcBA-IGS sequences

“…Manen and Falquet (2002) suggested that 16S rRNA gene data and cpcBA-IGS data were often in agreement. Furthermore, another study investigated the use of the cpcBA-IGS region of the phycocyanin operon, flanked by cpcB and cpcA, as a marker for the subgenus characterization of cyanobacteria (Neilan et al 1995, Bolch et al 1996.…”

Phylogenetic analyses of Synechococcus strains (cyanobacteria) using sequences of 16S rDNA and part of the phycocyanin operon reveal multiple evolutionary lines and reflect phycobilin content.

“…In addition to analysis of 16S rRNA gene sequences (Giovannoni et al, 1988 ;Honda et al, 1999 ;Neilan et al, 1997 ;Otsuka et al, 1998 ;Turner et al, 1999 ;Urbach et al, 1998 ;Wilmotte, 1994), genes encoding the major light-harvesting accessory pigment proteins, particularly the phycocyanin (PC) operon (cpc), including the intergenic spacer (IGS) between cpcB and cpcA and the corresponding flanking regions (cpcBA-IGS), have been targeted for phylogenetic studies of cyanobacteria (Barker et al, 1999 ;Bolch et al, 1996 ;Hayes & Barker, 1997 ;Neilan et al, 1995).…”