FEMS Microbiology Letters 218 (2003) 359^364

www.fems-microbiology.org

Evidence for limited species diversity of bacteriochlorophyll

b-containing purple nonsulfur anoxygenic phototrophs in
freshwater habitats

Gerrit J. Hoogewerf, Deborah O. Jung, Michael T. Madigan
Department of Microbiology and Center for Systematic Biology, Southern Illinois University, Carbondale, IL 62901-6508 USA

Received 8 October 2002; received in revised form 26 November 2002; accepted 26 November 2002

First published online 7 January 2003

Abstract

Thirteen new isolates of bacteriochlorophyll b-containing purple nonsulfur bacteria were isolated from four freshwater habitats using
specific enrichment methods including the use of long wavelength filters and extincting dilution of the inoculum. The new isolates were
compared with the type strain of Blastochloris viridis, strain DSM 133T , as regards pigments, morphology, carbon nutrition, and
phylogeny. All new isolates were budding bacteria, and phototrophic mass cultures were green, brown, or brown^green in color. The
pattern of carbon sources photocatabolized were similar in all strains; however, sugars, both mono- and disaccharides, were widely used
by the new isolates while they did not support growth of strain DSM 133T . Phylogenetic analysis showed all new strains to cluster tightly
with the type strain with the exception of one brown-colored strain and a mildly thermophilic strain. The results suggest that in contrast
to purple nonsulfur bacteria containing bacteriochlorophyll a, those containing bacteriochlorophyll b may not be morphologically or
phylogenetically diverse, and group into a tight phylogenetic clade distinct from all other anoxygenic phototrophs.
2 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Keywords : Anoxygenic phototrophic bacteria ; Purple bacteria; Blastochloris ; Bacteriochlorophyll b

1. Introduction than currently recognized but that either inherently low

Over 50 species of purple anoxygenic phototrophs have
been described and the majority produce bacteriochloro-
phyll (Bchl) a as their chlorophyll pigment [1]. Within the
purple nonsulfur bacteria, only species of the genus Blas-
tochloris (formerly Rhodopseudomonas) [2], contain Bchl b
[3^6]. The genus Blastochloris consists of two species,
B. viridis and B. sulfoviridis, both nutritionally diverse
phototrophs [4,7,8] and excellent nitrogen-¢xers [9,10]. In
addition to Blastochloris, four species of purple sulfur bac-
teria, Halorhodospira halochloris [11], Halorhodospira ab-
delmaleki [12], Thiococcus pfennigii [13,14], and Thioalka-
licoccus limnaeus [15], also produce Bchl b. Thus, of
cultured purple bacteria, those containing Bchl b are a
distinct minority.
We hypothesized that diversity among Bchl b-contain-
ing purple nonsulfur bacteria might actually be greater
* Corresponding author. Tel./Fax : +1 (618) 453 5130.
E-mail address : madigan@micro.siu.edu (M.T. Madigan).
0378-1097 / 03 / $22.00 2 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
doi:10.1016/S0378-1097(02)01195-3
population numbers or an inability to compete with Bchl
a-containing species bias their development in enrichment
cultures. To test this hypothesis we set up a series of en-
richment cultures using a light ¢lter that eliminated Bchl
a-containing phototrophs. The results of our study, based
on the morphology, pigments, carbon nutrition, and phy-
logeny of 13 new isolates of freshwater Bchl b-containing
purple nonsulfur bacteria, indicate that species diversity
within this group may indeed be quite low, and that cul-
turable representatives form a tight phylogenetic clade
within the alpha Proteobacteria.
2. Materials and methods
2.1. Enrichment and isolation
All new isolates of Bchl b-containing phototrophs were
obtained from enrichment cultures that employed a light
¢lter (Kodak Wratten Filter 87A) that transmitted less
than 1% of wavelengths shorter than 900 nm (Bchl a ab-

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360 G.J. Hoogewerf et al. / FEMS Microbiology Letters 218 (2003) 359^364

sorbs strongly near 800 and 850 nm) [16]. Water or mud^
water slurries were suspended in dilute phosphate bu¡er
and transferred to tubes of medium Enrich 1; inoculated
tubes were incubated in a light tight plastic tray under the
¢lter (32‡C). Illumination was from an incandescent spot-
light bulb providing ca. 1500 lux at the ¢lter surface. Pure
cultures from liquid enrichments were obtained on Petri
plates incubated phototrophically in un¢ltered incandes-
cent light.
Medium Enrich 1 contained the following (per liter of
distilled water): MgSO4 W7H2 O, 200 mg; CaCl2 W2H2 O, 50
mg; NaCl, 0.4 g; NH4 Cl, 0.5 g; sodium succinate, 0.75 g;
sodium acetate, 0.75 g; yeast extract, 0.1 g; sodium lac-
tate; 0.05%; KH2 PO4 , 0.2 g; K2 HPO4 , 0.3 g; trace ele-
ments [17], 1 ml. The ¢nal pH was adjusted to 7 and the
medium sterilized by autoclaving.
2.2. Characterizational studies
All isolates were characterized as to their morphology,
pigments, carbon nutrition, and phylogeny. Microscopy
was performed on an Olympus B-Max 60 photomicro-
scope. Absorption spectra were obtained from intact cells
suspended in 30% bovine serum albumin. Carbon nutri-
tion was determined using medium Enrich 1 in which in-
dividual carbon sources were substituted for succinate/ace-
tate/lactate at the concentrations indicated in Table 2.
Growth was monitored turbidimetrically using a Klett-
Summerson colorimeter (66, red ¢lter) versus no substrate
negative controls.
The phylogeny of newly isolated Bchl b-containing pho-
totrophs was determined by 16S ribosomal RNA gene
sequencing. Sequencing methods were basically as previ-
ously described [18] and approximately 1400 nucleotides
were used to generate the phylogenetic tree. Sequence
alignments were veri¢ed against the secondary structure
of the 16S rRNA gene of B. viridis DSM133T , exported
into PAUP [19], and a phylogenetic distance tree gener-
ated using the Jukes^Cantor correction and a heuristic
search pattern. All sequences have been deposited in Gen-
Bank under the accession numbers listed in Fig. 2.
3. Results
3.1. Enrichment, morphology, and pigments
Enrichment cultures for purple nonsulfur bacteria em-
ploying a ¢lter to exclude Bchl a-containing species readily
yielded new isolates containing Bchl b from four fresh-
water habitats (Table 1). Some isolates were obtained
from undiluted inocula while others were obtained from
extinction culturing of the original inoculum. However,
the highest dilution that yielded positive cultures was
only 1032 , indicating that populations of Bchl b-contain-
ing purple nonsulfur bacteria, at least those capable of
growth under our enrichment conditions, were low, 102
per ml or fewer.
A distinct variation in color was obvious in phototroph-
ically grown colonies of the di¡erent isolates. Colonies and
subsequent phototrophic liquid cultures varied from
brown or brown^green to distinctly olive-green in color.
These color di¡erences formed the basis for strain desig-
nations (G, green; B, brown, see Table 1). In addition to
pigmentation, cell morphology varied somewhat between
strains. All strains showed budding cell division, typical of

Table 1
Strains of B. viridis used in this study
Strains Origin Description In vivo Abs. maxima (nm)
Previous isolates
DSM 133T (type strain) Freiburg, Germany Green colonies, budding rods 1014, 833, 603, 483, 452, 424, 403
UN Campus Lake Carbondale, IL, USA Green colonies, budding rods 1014, 833, 604, 483, 452, 423, 403
GI Soda Dam hot spring Santa Fe National Green colonies, budding rods; thermophile 1012, 840, 605, 432, 450, 400
Forest, New Mexico
New isolatesa
G2 (ud) Crystal Lake Park, Urbana, IL, USA Brownish-green colonies, budding rods 1014, 832, 604, 481, 449, 397
G3 (ud) Crystal Lake Park, Urbana, IL, USA Brownish-green colonies, budding rods 1015, 833, 604, 483, 452, 425, 400
G5 (ud) Ramsey Lake, Ramsey, IL, USA Brownish-green colonies, budding rods 1016, 833, 604, 483, 452, 425, 403
G6 (1031 ) Allerton Pond, Monticello, IL, USA Dark green colonies, budding rods 1015, 832, 604, 483, 452, 427
G7 (1031 ) Allerton Pond, Monticello, IL, USA Light green colonies, medium sized budding 1015, 830, 603, 482, 451, 422, 401
rods
G8 (ud) Crystal Lake Park, Urbana, IL, USA Dark green colonies, budding rods 1015, 832, 604, 483, 452, 425, 404
G10 (1032 ) Ramsey Lake, Ramsey, IL, USA Dark green colonies, budding cocci to rods 1015, 831, 604, 483, 452, 428
G11 (ud) Campus Lake, Carbondale, IL, USA Green colonies, budding rods 1016, 832, 603, 483, 452, 423, 402
G12 (ud) Allerton Pond, Monticello, IL, USA Dark green colonies, budding rods 1015, 833, 604, 483, 452, 424, 401
B4 (1031 ) Ramsey Lake, Ramsey, IL, USA Brown colonies, budding rods 1015, 833, 604, 483, 452, 427
B5 (1031 ) Ramsey Lake, Ramsey, IL, USA Brown colonies, budding rods 1014, 830, 604, 482, 451, 422, 400
B7 (ud) Campus Lake, Carbondale, IL, USA Dark brown colonies, budding rods 1015, 832, 604, 483, 452, 428
B9 (1031 ) Campus Lake, Carbondale, IL, USA Light brown colonies, budding rods 1016, 830, 604, 483, 452, 423, 401
a
Numbers in parentheses indicate the dilution of the original inoculum in which liquid enrichments yielded a positive culture (that is, higher dilutions
did not yield positive enrichments over 6 weeks of incubation) ; ud, undiluted inoculum.

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 G.J. Hoogewerf et al. / FEMS Microbiology Letters 218 (2003) 359^364 361

near 480, 450 and 425 nm. The latter peaks indicated that
all strains contained the carotenoid neurosporene, charac-
teristic of B. viridis [3,6]. However, subtle di¡erences in
carotenoid absorption maxima and the ratio of these max-
ima were observed in distinctly di¡erent colored strains
(Fig. 1). A summary of the source, cell morphology, and
absorption properties of the new Bchl b-containing isolates
is shown in Table 1.

3.2. Carbon nutrition

In tests of carbon nutrition it was readily apparent that

Fig. 1. Absorption spectra of intact cells of B. viridis strain DSM 133T
and Bchl b-containing strain B5. For recording spectra, cells were sus-
pended in 30% bovine serum albumin.
Blastochloris species [6,7]. However, although cells of
green-colored strains were typically small and coccus to
egg-shaped, like those of the type strain, DSM 133T [6],
cells of brown or brown^green strains were somewhat
larger and more irregularly rod-shaped (Table 1). Other
morphologies, such as non-budding rods or spirilla, were
not observed in cultures of the new isolates.
Despite visible di¡erences in color among strains, their
in vivo absorption spectra were quite similar (Table 1 and
Fig. 1). Absorption maxima were obtained between 1012
and 1016 nm, due to Bchl b, and in the carotenoid region
the new Bchl b-containing isolates were nutritionally ver-
satile. Three previously characterized strains of B. viridis
including the type strain were also tested in this connec-
tion and the results are summarized in Table 2. A variety
of alcohols and short chain fatty and organic acids sup-
ported photoheterotrophic growth of the new isolates.
Particularly good growth was obtained on malate, while
one-carbon substrates (methanol, formate) were not used
by any strain (Table 2). Surprisingly, however, a number
of substrates supported growth of the new isolates that
were not used by the type strain of B. viridis. Propionate
and lactate, for example, supported good growth of all
newly isolated strains but not of the type strain (Table
2). Even more surprising were the results on sugars.
Although none of the sugars tested supported growth of
the type strain, the vast majority of green and brown-col-
ored strains photocatabolized sugars, including glucose,
fructose, sucrose, lactose and ribose; sucrose and lactose

Table 2
Carbon nutrition of new and established strains of B. viridisa;b
Strain Glucose Fructose Sucrose Lactose Ribose Lactate Propionate Mannitol
(10 mM) (10 mM) (10 mM) (10 mM) (10 mM) (20 mM) (10 mM) (10 mM)
DSM133T 3 3 3 3 3 3 3 3
UN 3 3 3 3 3 + + 3
G2 3 3 3 3 3 + ++ +
G3 ++ ++ ++ ++ ++ ++ ++ ++
G5 ++ ++ ++ ++ + +++ + ++
G6 ++ ++ ++ ++ + ++ ++ ++
G7 ++ ++ +++ +++ ++ ++ + ++
G8 + ++ ++ +++ 3 +++ ++ ++
G10 + + + + + ++ ++ ++
G11 +++ ++ ++ +++ 3 ++ + +++
G12 3 3 ++ 3 + + + 3
B4 + 3 ++ + ++ +++ + ++
B5 + 3 3 3 3 + ++ ++
B7 ++ + + +++ + +++ ++ ++
B9 ++ ++ ++ ++ + +++ + ++
GI + ++ n.d. n.d. n.d. n.d. n.d. n.d.
a
Phototrophic growth in all cases in medium Enrich 1 containing the substrate indicated as sole carbon source. Growth was scored after 10 days as fol-
lows: 3, 0^50 photometer units (p.u.) ; +, 51^150 p.u.; ++, 151^250 p.u.; +++, 251^350 p.u.; n.d., not determined. For strain DSM133T , 100 p.u. was
equivalent to approximately 0.2 mg bacterial dry weight ml31 of culture. Negative controls lacking a substrate were less than 50 p.u. in all cases.
b
The following substrates supported growth of each strain to approximately the same extent; ethanol (10 mM, ++); propanol (10 mM, +); butanol
(10 mM,+); pentanol (5 mM, +); acetate or butyrate (10 mM, ++), valerate (5 mM, ++); caproate (5 mM, ++); malate or succinate (20 mM, +++);
fumarate, pyruvate, or K-ketoglutarate (20 mM, ++); glutamate (20 mM, +). Strains UN, G3, and G11 grew weakly (+) on benzoate (5 mM). Strains
G15 and B9 grew weakly (+) on citrate. Methanol (10 mM) and formate (20 mM) did not support growth of any strain. Aspartate (20 mM) supported
weak growth (+) of strains G3, G11, and B9 only.

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were particularly well used (Table 2). Only three newly
isolated strains showed a similar pattern concerning
growth on sugars as did strain DSM 133T (Table 2). How-
ever, only strain G2 showed the same negative pattern on
all sugars as did strain DSM 133T (Table 2). The sugar
alcohol mannitol also supported good growth of most new
strains but not of the type strain (Table 2). Interestingly,
the aromatic compound benzoate, whose photocatabolism
is restricted to only a few purple nonsulfur bacteria [20],
was used by a few of the new strains of B. viridis but not
by the type strain (Table 2).
3.3. Phylogeny
The phylogeny of the new isolates was determined by
16S rRNA gene sequencing. The 16S rRNA gene of strain
DSM 133T previously had been sequenced [2] but was
sequenced here as an internal control on our methods.
The sequence of strain DSM133T generated herein was
identical to that of the published sequence, thus con¢rm-
ing the previous sequence and indicating that our sequenc-
ing error rate was very low.
Phylogenetic analysis using evolutionary distance
showed the new isolates to group tightly with the B. viridis
type strain (Fig. 2). In fact, the 16S rRNA gene sequences
of the new strains were nearly identical to that of the type
strain, varying by at most, ¢ve base substitutions over the
approximately 1400 bases analyzed for each isolate (Fig.
2). Green and brown strains were phylogenetically indis-
tinguishable from the type strain, with the exception of the
brown strain B9 (Fig. 2); however, the latter was still
99.5% identical to the type strain. It thus appears that
Fig. 2. Phylogenetic distance tree of Bchl b-containing purple nonsulfur
bacteria and closely related phototrophic species containing Bchl a.
GenBank accession numbers are listed in parentheses following each
strain designation. Rhodopseudomonas acidophila (Rhodoblastus acidophi-
lus), Rhodospirillum rubrum, and Rhodobacter capsulatus contain Bchl a.
FEMSLE 10814 23-1-03
the new isolates, both brown and green, are strains of
the species B. viridis. A more distant relative was strain
GI, a moderately thermophilic hot spring isolate [21] that
di¡ered from both B. sulfoviridis and B. viridis by about
2% in 16S rRNA gene sequence (Fig. 2). Phylogenetic
analysis using parsimony (data not shown) showed the
same branching order and tight grouping of new strains
as did the distance analyses, and also showed strains B9
and GI to be distinct from the main clade of B. viridis
isolates.
4. Discussion
The major objective of this study was to test the hypoth-
esis that hidden diversity existed among culturable Bchl
b-containing purple nonsulfur bacteria from freshwater
habitats. However, the results suggest that the diversity
of these phototrophs is indeed quite limited, perhaps just
to the species known at present. Our results certainly do
not preclude the existence of other genera or species of
Bchl b-containing anoxygenic phototrophs. But if such
species exist, they were either absent in the habitats we
sampled, unable to grow in the media employed, or were
unable to compete with B. viridis-type organisms under
our enrichment conditions. However, our data, coupled
with the fact that even metabolically unusual Bchl b-con-
taining purple nonsulfur bacteria, such as those capable of
photocatabolizing toluene and crude oil, still cluster
tightly within the Blastochloris clade [22], suggests that
phylogenetic diversity is not a hallmark of this group.
Color variation among strains of B. viridis has been
previously observed [3] but the basis for this is unknown.
Slight di¡erences in absorption maxima and clear di¡er-
ences in the ratios of spectral components in olive green
versus brown strains (Fig. 1) suggest that pigment di¡er-
ences may indeed exist between these strains. However,
only detailed analyses of the carotenoid content of these
strains could resolve this question. The di¡erences in ab-
sorption properties that we observed are unlikely to be due
to growth conditions, since for spectral analyses care was
taken to grow all strains at the same light intensity and in
the same medium ; cultures were also grown to approxi-
mately the same densities (late exponential phase) in each
case. Therefore, further work on the carotenoid composi-
tion of di¡erent colored B. viridis strains could be enlight-
ening.
A major surprise emerging from this study was how the
carbon nutritional properties of the new isolates were so
distinct from that of the type strain. For example, as re-
gards sugar utilization in B. viridis, the pattern displayed
by the type strain (DSM 133T ) [7] (Table 2) is clearly the
exception rather than the rule. In addition, our results that
showed growth of the type strain on C4 ^C6 fatty acids are
contrary to published reports [4,7,23]. The reason for the
latter discrepancy is unclear, but in the present work sub-
 G.J. Hoogewerf et al. / FEMS Microbiology Letters 218 (2003) 359^364
strate concentrations were kept quite low (Table 2) and
this may be why growth on these fatty acids was observed
here but not previously. At any rate, a major lesson to be
learned here is that physiological conclusions based on the
results obtained from a single isolate can be quite mislead-
ing. With inclusion of the present data, for example, the
spectrum of carbon sources photocatabolized by B. viridis
expands dramatically beyond that listed for this species in
Bergey’s Manual [7] and elsewhere [23]. Indeed, the excel-
lent growth of the new strains of B. viridis on sugars and
medium-chain-length fatty acids, capacities previously un-
recognized in this species [7,23], suggests that these may be
important substrates for B. viridis in nature.
It was hoped that phylogenetic analyses would reveal
that some of the budding strains isolated here were new
species. However, comparative sequencing supported the
phenotypic studies and showed the majority of the new
isolates to cluster tightly with B. viridis DSM 133T . The
only exceptions were B. sulfoviridis, whose phylogenetic
position relative to that of B. viridis was previously recog-
nized [2], the mildly thermophilic Blastochloris sp. strain
GI [21], and one brown-colored isolate, strain B9. As re-
gards strain GI, establishment of a new taxon for this
isolate may be warranted but will require further charac-
terization, in particular DNA:DNA hybridization, to bet-
ter establish its genomic relationships to B. viridis and
B. sulfoviridis.
The results of this study are the ¢rst experimental sup-
port of the current picture of biodiversity among purple
nonsulfur bacteria containing Bchl b. If cocci, spirilla, or
other morphotypes that contain Bchl b exist in freshwater
habitats, they have escaped our detection. If such organ-
isms do not exist, then the diversity of Bchl b-containing
phototrophs contrasts sharply with that of Bchl a-contain-
ing species, where freshwater habitats support a phyloge-
netically broad species diversity [1,7]. A possible approach
for extending our work would be to inspect the rRNA
gene sequences generated here for signature sequences
characteristic of this group and which are absent from
all known Bchl a-containing species. If exclusive signatures
exist, one could integrate them with FISH technology to
examine natural samples or enrichment cultures. Fluores-
cent labeling of only budding morphotypes in such studies
would con¢rm the enrichment and isolation results re-
ported here, and strengthen even further the contention
that Bchl b-containing purple nonsulfur bacteria inhabit-
ing freshwaters lack signi¢cant species diversity.
Acknowledgements
This research was supported in part by a grant from the
Illinois Council on Food and Agricultural Research (C-
FAR) Competitive Grants Program. We thank Laurie A.
Achenbach and Daniel L. Nickrent for help with DNA
sequencing and analysis.
FEMSLE 10814 23-1-03
363
References
[1] Imho¡, J.F. (1995) Taxonomy and physiology of phototrophic purple
bacteria and green sulfur bacteria. In: Anoxygenic Photosynthetic
Bacteria (Blankenship, R.E., Madigan, M.T. and Bauer, C.E.,
Eds.), pp. 1^15. Kluwer Academic Publishers, Dordrecht.
[2] Haraishi, A. (1997) Transfer of the bacteriochlorophyll b-containing
phototrophic bacteria Rhodopseudomonas viridis and Rhodopseudomo-
nas sulfoviridis to the genus Blastochloris gen. nov. Intl. J. Syst. Bac-
teriol. 47, 217^219.
[3] Biebl, H. and Drews, G. (1969) Das in-vivo speckrum als taxono-
misches merkmal bei untersuchungen zur Veirbreitung von Athiorho-
daceae. Zentralbl. Bakteriol. Parasiten. Infektionskr Hyg. Abt. II.
123, 425^452.
[4] Drews, G. and Giesbrecht, P. (1966) Rhodopseudomonas viridis, nov.
spec. ein neu isoliertes, obligat phototrophes Bakterium. Arch. Mi-
krobiol. 53, 255^262.
[5] Tru«per, H.G. and Imho¡, J.F. (1989) Genus Rhodopseudomonas. In:
Bergey’s Manual of Systematic Bacteriology (Staley, J.T., Bryant,
M.P., Pfennig, N. and Holt, J.G., Eds.), pp. 1672^1677. Williams
and Wilkins, Baltimore, MD.
[6] Whittenbury, R. and McLee, A.G. (1967) Rhodopseudomonas palus-
tris and Rh. viridis-photosynthetic budding bacteria. Arch. Mikro-
biol. 59, 324^334.
[7] Imho¡, J.F. and Tru«per, H.G. (1989) Purple Nonsulfur Bacteria. In:
Bergey’s Manual of Systematic Bacteriology (Staley, J.T., Bryant,
M.P., Pfennig, N. and Holt J.G., Eds.), pp. 1658^1661. Williams
and Wilkins, Baltimore, MD.
[8] Keppen, O.I. and Gorlenko, V.M. (1975) A new species of purple
budding bacteria containing bacteriochlorophyll b. Mikrobiologia 44,
224^229.
[9] Howard, K.S., Hales, B.J. and Socolofsky, M.D. (1983) Nitrogen
¢xation and ammonia switch-o¡ in the photosynthetic bacterium
Rhodopseudomonas viridis. J. Bacteriol. 155, 107^112.
[10] Madigan, M., Cox, S.S. and Stegeman, R.A. (1984) Nitrogen ¢xation
and nitrogenase activities in members of the family Rhodospirillaceae.
J. Bacteriol. 157, 73^78.
[11] Imho¡, J.F. and Tru«per, H.G. (1977) Ectothiorhodospira halochloris
sp. nov., a new extremely halophilic phototrophic bacterium contain-
ing bacteriochlorophyll b. Arch. Microbiol. 114, 115^121.
[12] Imho¡, J.F. and Tru«per, H.G. (1981) Ectothiorhodospira abdelmalekii
sp. nov., a new halophilic and alkaliphilic phototrophic bacterium.
Zentralbl. Bakteriol. 2, 228^234.
[13] Eimhjellen, K.E. (1970) Thiocapsa pfennigii sp. nov. a new species of
the phototrophic sulfur bacteria. Arch. Mikrobiol. 73, 193^194.
[14] Eimhjellen, K.E., Steensland, H. and Traetteberg, J. (1967) A Thio-
coccus sp. gen. nov., its pigments and internal membrane system.
Arch. Mikrobiol. 59, 82^92.
[15] Bryantseva, I.A., Gorlenko, V.M., Kompantseva, E.I. and Imho¡,
J.F. (2000) Thioalkalicococcus limnaeus gen. nov., sp. nov., a new
alkaliphilic purple sulfur bacterium with bacteriochlorophyll b. Intl.
J. Syst. Bacteriol. 50, 2157^2163.
[16] Biebl, H. and Pfennig, N. (1981) Isolation of members of the family
Rhodospirillaceae. In: The Prokaryotes (Starr, M.P., Stolp, H.,
Tru«per, H.G., Balows, A. and Schlegel, H.G., Eds.), pp. 267^273.
Springer-Verlag, New York.
[17] Wahlund, T.M., Castenholz, R.W., Woese, C.R. and Madigan, M.T.
(1991) A thermophilic green sulfur bacterium from New Zealand hot
springs. Chlorobium tepidum, nov. sp. Arch. Microbiol. 156, 81^91.
[18] Bryantseva, I.A., Gorlenko, V.M., Kompantseva, E.I., Achenbach,
L.A. and Madigan, M.T. (1999) Heliorestis daurensis, gen. nov. sp.
nov., an alkaliphilic rod-to-coiled-shaped phototrophic heliobacte-
rium from a Siberian soda lake. Arch. Microbiol. 172, 167^174.
[19] Swoford, D.L (2000) PAUP*. Phylogenetic Analysis Using Parsi-
mony (*and other methods), 4th edn. Sinauer, Sunderland, MA.
[20] Gibson, J. and Harwood, C.S. (1995) Degradation of aromatic com-
364 G.J. Hoogewerf et al. / FEMS Microbiology Letters 218 (2003) 359^364

pounds by nonsulfur purple bacteria. In: Anoxygenic Photosynthetic
Bacteria (Blankenship, R.E., Madigan, M.T. and Bauer, C.E., Eds.),
pp. 991^1003. Kluwer Academic Publishers, Dordrecht.
[21] Resnick, S.M. and Madigan, M.T. (1989) Isolation and character-
ization of a mildly thermophilic nonsulfur purple bacterium contain-
ing bacteriochlorophyll b. FEMS Microbiol. Lett. 65, 165^170.
[22] Zengler, K., Heider, J., Rossello¤-Mora, R. and Widdel, F. (1999)
Phototrophic utilization of toluene under anoxic conditions by a
new strain of Blastochloris sulfoviridis. Arch. Microbiol. 172, 204^
212.
[23] Imho¡, J.F., Tru«per, H.G. (1992) The genus Rhodospirillum and
related genera. In: The Prokaryotes, 2nd ed. (Balows, A., Tru«per,
H.G., Dworkin, M., Harder, W., and Schleifer, K.-H., Eds.), pp.
2141^2155. Springer-Verlag, New York.

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