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spread but not ubiquitous occurrence of norspermidine were observed as the major cellular polyamines within eukaryotic algae belonging to the phyla Chlorophyta, Rhodophyta, Heterokonta and Euglenozoa (Hamana and Matsuzaki, 1982, 1985; Hamana et al., 1988, 1990; Hegewald and Kneifel, 1981, 1982; Kneifel and Hegewald, 1980). Norspermine was sporadically found within the phyla. Norspermidine and norspermine have not been detected in the phylum Glaucophyta. The phylum Chlorophyta, consisting unicellular and multicellular green algae, is divided into the classes Chlorophyceae, Pedinophyceae, Picocystophyceae, Trebouxiophyceae, Ulvophyceae and Prasinophyceae, and the phylum Charophyta corresponds to a former class Charophyceae of the phylum Chlorophyta (Ali et al., 2001; Cavalier-Smith, 1993; NCBI home page, 2004). Recently it has been reported that non-photosynthetic achlorophyllous (achlorophyllic), animal-pathogenic, unicellular Helicosporidium and Prototheca species phylogenetically belong to Trebouxiophyceae (Boucies et al., 2001; Nedelcu, 2001; Tartar et al., 2002, 2003; Ueno et al, 2003). Helicosporidium spp. have been identified as invertebrate pathogens, whereas Prototheca spp. are known to be pathogenic to vertebrates (Galan et al., 1997; Mohabeer et al., 1997). Another non-photosynthetic unicellular green algal group consisting of the Polytoma and Polytomella species belongs to Chlorophyceae (Nedelcu, 2001; Tartar et al., 2003; Ueno et al., 2003). They have degenerate chloroplasts and grow heterotrophically in the dark whereas other green algae ordinarily grow phototrophically in the light. Their novel characteristics make it possible to effect their cellular polyamine profiles. In addition to established polyamine catalogues of Chlorophyta and Charophyta, cellular polyamines of 41 micro green algae including all four achlorophyllous genera were analyzed. Achlorophyllous Polytoma, Polytomella, Prototheca and Helicosporidium species were heterotrophically cultivated in JCM medium No. 31 broth (malt extract, glucose and peptone), No. 31 agar (2%) slant, Sabouraud glucose broth (SG) (glucose and enzymatic digest of casein) (DIFCO, Detroit, USA), ATCC 351 medium (Hutner’s medium), ATCC 1900 medium (soil extract salts medium) and/or IAM A-7 medium (sodium acetate, beef extract, acid hydrolysate of casein and yeast extract), at 25°C in the dark. A yellow mutant (IAM C-425) and a white mutant (IAM C-539) of Chlorella vulgaris (Schwarze and Frandsen, 1960) were heterotrophically grown in IAM A-5 medium containing glucose and proteose peptone and nutrient agar (NA) (Nissui Pharmaceutical Co., Tokyo, Japan) Distribution of norspermidine as a cellular polyamine within micro green algae including non-photosynthetic achlorophyllous Polytoma, Polytomella, Prototheca and Helicosporidium

journal of general and applied microbiology

Distribution of norspermidine as a cellular polyamine within micro green algae including non-photosynthetic achlorophyllous Polytoma, Polytomella, Prototheca and Helicosporidium