Algological Studies 126 21–36 Stuttgart, April 2008

Leptolyngbya species from selected seep

walls in the Great Smoky Mountains National
Park
JEFFERY R. JOHANSEN1, CATHERINE E. OLSEN1, REX L. LOWE2,
KAROLINA FUČÍKOVÁ1 and DALE A. CASAMATTA3

1 Department of Biology, John Carroll University, Ohio, U.S.A.

2 Department of Biological Sciences, Bowling Green State University, Ohio,
U.S.A.
3 Department of Biology, University of North Florida, Florida, U.S.A.

With 4 figures and 2 tables

Abstract: The All Taxa Biodiversity Inventory (ATBI) began in 1997 with the goal
of inventorying and providing a database of information for every species located
in the Great Smoky Mountains National Park (GSMNP). In this study, algal sam-
ples were collected from three wet rock seeps within GSMNP. Seven Leptolyngbya
(Oscillatoriales, Cyanobacteria) species were identified from the three sites. Two
species are new to science (L. appalachiana, L. badia), two are reported as new
records to the park (L. angustissima, L. subtilissima), and three others do not fit
any previously circumscribed taxa, but we currently do not have molecular data on
these morphospecies and thus postpone description as new taxa.

Key words: All Taxa Biodiversity Inventory, cyanobacteria, Great Smoky Moun-
tains, Leptolyngbya, Pseudophormidium, 16S-23S Intergenic Spacer

Introduction
The Great Smoky Mountains National Park (GSMNP) has a rich store of
biodiversity and has been designated as an International Biosphere Re-
serve and as a world heritage site by the United Nations (SHARKEY 2001,
WELCH et al. 2002). Recent study of the algal communities of GSMNP has
yielded many new records to the park and possible new records to science.
GOMEZ et al. (2003) examined the non-diatom algal flora of wet walls in
GSMNP. They found that 91.5 % of species identified were new records
to the park, with 39 % percent of these representing possible new species.
JOHANSEN et al. (2004) reported a total of 584 algal taxa from the park, with
108 of those representing cyanobacterial records. Rexia erecta CASAMATTA

DOI: 10.1127/1864-1318/2008/0126-0021 1864-1318/08/0126-021 $ 4.00

© 2008 E. Schweizerbart’scheVerlagsbuchhandlung, D-70176 Stuttgart

Johansen_K.indd 21 30.04.2008 10:53:14 Uhr

 22 J. F. JOHANSEN et al.

et al. and Capsosira lowei CASAMATTA et al. were recently described from
the park (CASAMATTA et al. 2006), and new species in other phyla are in the
process of being published (JOHANSEN & LOWE 2007, unpublished manu-
scripts).
The present report concerns the discovery and description of three new
species of the cyanobacterial genus Leptolyngbya ANAGN. et KOM., along
with the characterization of four other species of Leptolyngbya, which could
either be assigned to existing species or were too rare to name at present.

Materials and methods

Samples were collected from three wet rock seeps within Great Smoky
Mountains National Park (Table 1). The rocks were scraped and organic
material and silt were put into Whirl-Pak bags. If the collection site was
wet enough, Whatman pH papers (range 4.0–7.0) were used to estimate the
pH of the rock face. In addition, latitude, longitude, and elevation were re-
corded at each site using a Garmin GPS unit.
All of the samples were refrigerated. A portion of the material was pre-
served, with the remainder of the material being left living for examination.
The portions of the samples that were to be preserved were put into poly-
carbonate bottles, and a 2 % glutaraldehyde solution was added to each of
the bottles.
Taxa identification and photography were performed using an Olympus
BH2 photomicroscope with high resolution Nomarski DIC optics. Taxonomic
identifications were made using standard references. For cyanobacteria,
GEITLER (1932), DESIKACHARY (1959), ANAGNOSTIDIS & KOMÁREK (1988),
KOMÁREK & ANAGNOSTIDIS (2005), and STARMACH (1966) were used.
When making taxonomic decisions, the ecological preference of previ-
ously described species was taken into consideration. It has been recog-
nized that ecological preference is a taxonomically significant characteristic
in bacteria (COHAN 2001, 2002), this being especially true in cyanobacteria
(KOMÁREK & ANAGNOSTIDIS 1999, 2005). Indeed, most of the keys in the
new revisions of the cyanobacteria of Europe have frequent couplets with
ecological dichotomies (KOMÁREK & ANAGNOSTIDIS 1999, 2005). Therefore,
a specimen with characteristics similar to a previously described species,
but with distinctly different ecological preference, was deemed to be a dif-
ferent species. This practice is consistent with the Ecotypic Species Concept
(COHAN 2001, 2002).
After the distinctiveness of the Leptolyngbya flora was discovered, we
returned to the park to the same localities and made efforts to isolate the
new Leptolyngbya taxa. We were successful in isolating L. appalachiana
and L. badia into Z8 culture medium. Descriptions and illustrations in this
paper for these two taxa are based on both field material and culture mate-
rial. Photographs are based on cultures.

Johansen_K.indd 22 20.04.2008 20:28:18 Uhr

Johansen_K.indd 23
Table 1. Sample descriptions. There were 3 samples from Cataloochie Road site 1, two samples from Cataloochie Road site 2,
and 4 samples from Hen Wallow Falls site 1.
Cataloochie Cataloochie Cataloochie Cataloochie Cataloochie Hen Hen Hen Hen
Rd. 1.1 Rd. 1.2 Rd. 1.3 Rd. 2.6 Rd. 2.7 Wallow Wallow Wallow Wallow
Falls 1.1 Falls 1.2 Falls 1.3 Falls 1.7
Date 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/5/02 5/5/02 5/5/02 5/5/02
Collected
Geology Longarm Longarm Longarm Basement Basement Roaring Roaring Roaring Roaring
quartzite quartzite quartzite complex complex Fork Sand- Fork Sand- Fork Sand- Fork Sand-
stone stone stone stone
pH ND ND 4.7 ND 5.0 ND 4.7 ND 4.7
Altitude (m) 762 762 762 548 548 791 791 791 791
Latitude N35.663° N35.663° N35.663° N35.641° N35.641° N35.7628° N35.7628° N35.7628° N35.7628°
Longitude W83.072° W83.072° W83.072° W83.060° W83.060° W83.2326° W83.2326° W83.2326° W83.2326°
Leptolyngbya from the Great Smoky Mountains
23

20.04.2008 20:28:18 Uhr

 24 J. F. JOHANSEN et al.

The 16S rRNA gene and associated 16S–23S ITS region were sequenced
in L. appalachiana, L. badia and the other Leptolyngbya strains follow-
ing the protocols commonly used in JOHANSEN’S laboratory (BOYER et al.
2001, 2002, FLECHTNER et al. 2002, CASAMATTA et al. 2005). Outgroup taxa
were obtained from GenBank (http://www.ncbi.nlm.nih.gov) and other se-
quenced taxa from the JOHANSEN laboratory (see bolded taxa in Fig. 2).
GenBank accession numbers are given for all sequences used (Fig. 3). Max-
imum parsimony trees were generated using a heuristic search constrained
by random sequence addition (1000), steepest descent, and tree-bisection-
reconnection branch swapping using PAUP v4.02b (SWOFFORD 1998). Boot-
strap values were obtained from 1000 replicates with one random sequence
addition. Distance analysis using the HKY85 distance method and assum-
ing a ti/tv ratio of two was also performed, and bootstrap scores (1000 rep-
licates) obtained.
Secondary structure of the 16S–23S ITS was determined using Mfold
version 2.3 (ZUKER 2003). Structures were determined by folding and iden-
tifying each conserved helix separately first, and then constraining the se-
quence to produce the entire structure. Apart from setting draw mode to
untangle with loop fix, default conditions were in all cases used.
Designated holotype and paratype materials (as preserved samples)
were deposited in the Herbarium for Nonvascular Cryptogams, Monte L.
Bean Museum, Brigham Young University, Provo, Utah, USA. Reference
strains were deposited in the UTEX (Austin, Texas) and UTCC (Toronto,
Ontario) culture collections.

Results
Several Leptolyngbya species were found that were either new to GSMNP
or possible new records to science (Table 2). Of the five species found that
could not be matched with a previously described species, two (Leptolyn-
gbya appalachiana sp. nov. and Leptolyngbya badia sp. nov.) have been
named as new species following the requirements of the International
Code of Botanical Nomenclature. Two others (Leptolyngbya cf. catarac-
tarum and Leptolyngbya sp. 9) are possibly new species to science (they fit
no described species), but not enough material was found at this time to de-
scribe them as new. Leptolyngbya angustissima (W. et G. S. WEST) ANAGN.
et KOM. and Leptolyngbya subtilissima (KÜTZING) KOM. have both been
inventoried as new records to GSMNP.

Species descriptions

Leptolyngbya appalachiana JOHANSEN et OLSEN sp. nov. (Figs 1A, 2 A, B)

D i a g n o s i s : Fila solitaria, 1.2–2.5 µm lata. Vagina incolorata, tenuis, inter-
dum absens. Trichomae constricta, non descresentes, cellulis apicalibus obtuse

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Johansen_K.indd 25
Table 2. Comparison of Leptolyngbya species
L. appalachiana L. badia L. angustissima L. subtilissima L. cataractarum L. sp. 1 L. sp.9
Filament 1.2-2.5 1.2-2.5 1.0-1.8 1.2-1.5 2.5 0.8-1.0 2.5
width [µm]
Trichome 1.2-1.7 1.0-1.3 0.8-1.0 1.2-1.5 2.5 0.8-1.0 2.0
width [µm]
Cell length 1.5-4.0 2.0 1.5-5 1.3-1.7 4.5-5.5 2-4 2-4
[µm]
Sheath charac- Not evident, or Firm, wide, Fine, open, Not evident Not evident Not evident, or Wide, fine,
teristics thin and color- smooth, open, distinctly wider very fine and colorless, open
less sooty brown than trichome thin
End cell shape Bluntly rounded Rounded Rounded Bluntly Bluntly Rounded Rounded
rounded rounded
Trichome Constricted Constricted Constricted Constricted Unconstricted Unconstricted Unconstricted
Cell contents Nongranular or Nongranular Nongranular or Nongranular Granular Nongranular Nongranular
with an occa- with an occa-
sional granule at sional minute
the crosswall granule at the
crosswall
Leptolyngbya from the Great Smoky Mountains
25

20.04.2008 20:28:18 Uhr

 26 J. F. JOHANSEN et al.

rotundis, 1.2–1.7 µm latae. Cellulae nongranulatae vel subinde granula singu-

laria ad septa, plerumque longiores quam latae, thylacoidibus peripheralibus
interdum visibilibus secus parietes externos, 1.5-4.0 µm longae.
Filaments solitary, 1.2–2.5 µm wide. Sheaths colorless, thin, sometimes
absent. Trichomes constricted, not tapering, with bluntly rounded end cells,
1.2–1.7 µm wide. Cells usually longer than broad, non-granular or with an
occasional granule at the crosswalls, with peripheral thylakoids sometimes
visible along the outside walls, 1.5–4.0 µm long.
Type collected 6 May 2002 from quartzite seep wall on Cataloochie Road
(Site 2.6), 35° 38.5' North Latitude, 83° 3.6' West Longitude, Great Smoky
Mountains National Park, Haywood County, North Carolina. Holotype
here designated: BRY C 37679, deposited in the Herbarium for Nonvascu-
lar Cryptogams, Brigham Young University, Provo, Utah, USA. Paratype
material also here designated: Hen Wallow Falls site 1.2 (BRY C 37680),
Cataloochie Road site 1.2 (BRY C 37681), Cataloochie Road site 1.3 (BRY
C 37682), Cataloochie Road site 2.7 (BRY C 37683).
Reference strain: GSM-SFF-MF60 deposited at University of Toronto
Culture Collection, Toronto, Ontario, Canada and University of Texas Cul-
ture Collection, Austin, Texas, USA.
L. appalachiana is named for the Appalachian Mountain chain.
This taxon is most similar to L. bijugata (KONGISSER) ANAGN. et KOM.,
but that taxon has thickenings between the cells and different habitat re-
quirements (muddy substrate, stagnant water). It bears a resemblance to
the broadly circumscribed soil taxon L. tenuis (GOM.) ANAGN. et KOM., but
lacks the tapered end cells of that taxon.
KOMÁREK (1999) has indicated that L. tenuis has been used as a junk
taxon, and that it needs revision. Rather than include our material in this
problematic taxon, we choose to describe it as new.
Leptolyngbya badia JOHANSEN et LOWE sp. nov. (Figs 1D, 2C, D)
D i a g n o s i s : Fila badia, dense intricata, trichomis plerumque una vel raro
duabus, usque ad 2.5 µm lata. Vagina fulginosa, firma, laevis, aperta, fulgi-
nosa. Trichoma constricta ad septa, non descrescentes, cellulis apicalibus
obtuse rotundis, 1.0–1.3 µm latae. Cellulae nongranulatae, subaeruginosae,
2.0 µm longae.
Filaments chocolate-brown, densely entangled, with trichomes usually
one or rarely two per sheath, up to 2.5 µm wide. Sheath sooty brown to
blackish, firm, smooth, open. Trichomes constricted at the crosswalls, unta-
pered, with bluntly rounded end cells, 1.0–1.3 µm wide. Cells nongranular,
pale blue-green, 2.0 µm long.
Type collected 6 May 2002 from quartzite seep wall on Cataloochie
Road (Site 2.6), 35° 38.5' North Latitude 83° 3.6' West Longitude, Great
Smoky Mountains National Park, Haywood County, North Carolina. Holo-
type here designated: BRY C 37684, deposited in the Herbarium for Non-
vascular Cryptogams, Brigham Young University, Provo, Utah, USA.

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 Leptolyngbya from the Great Smoky Mountains 27

Fig. 1. Leptolyngbya species found in the Great Smoky Mountains National Park.
A. Leptolyngbya appalachiana. Note the occasional presence of granules at the
crosswalls (left) and peripheral thylakoids (right). B. Leptolyngbya angustissima.
C. Leptolyngbya subtilissima. D. Leptolyngbya badia. E. Leptolyngbya noncon-
stricta. F. Leptolyngbya cf. cataractarum. G. Leptolyngbya sp. 9. [Scale bar =10 µm.]

Johansen_K.indd 27 20.04.2008 20:28:18 Uhr

 28 J. F. JOHANSEN et al.

L. badia was chosen as a name for this species for its chocolate color.
No other previously described species sharing similar ecological preference
exhibits a blackish chocolate-brown sheath.
The appearance of two trichomes in a common sheath is characteristic
of Pseudophormidium, a genus that has recently been separated from Plec-
tonema. This genus currently is in the Phormidiaceae, and has mostly taxa
with trichomes that are wider than 3 µm wide, including the type of the
genus, P. phormidioides (HANSG. ex FORTI) ANAGN. et KOM. We suspect that
the three taxa (P. purpureum (GOM.) ANAGN. et KOM., P. hollerbachianum
(ELENKIN) ANAGN., P. spelaeoides (ČADO) ANAGN.) with thin trichomes
likely belong to a genus in the Pseudanabaenaceae. Thus, we are hesitant
to describe L. badia, which clearly belongs in the Pseudanabaenales in the
potentially polyphyletic genus Pseudophormidium, whose type species is in
the Phormidiaceae.
Despite the occasional multiple trichomes per filament, we feel it is very
likely that L. badia belongs in the genus Leptolyngbya as it is currently de-
fined. Preliminary molecular studies on terrestrial Leptolyngbya taxa indi-
cate this genus will likely be split into several genera in an eventual revision
(CASAMATTA et al. 2005).
The blackish brown sheath color is highly unusual in the Pseudanabae-
nales. It is recorded in just two taxa in Leptolyngbya (L. nostocorum (BOR-
NET ex GOM.) ANAGN. et KOM., L. edaphica (HOLLERB. ex ELENKIN) AN-
AGN. et KOM.), and a few taxa in Schizothrix (S. simplicior SKUJA, S. heufleri
GRUN. ex GRUN., S. funalis W. et G. S. WEST, S. longearticulata (GEITLER)
ANAGN., S. braunii GOM., and S. incrustans (ERCEGOVIC) ANAGN. Of these
taxa, L. edaphica appears to be the closest in morphology of the filaments
(firm wide sheaths, one or rarely two trichomes per filament), but it dif-
fers in filament, trichome, and cell dimensions, as well as habitat. S. incrus-
tans possesses similar dimensions, but grows on calcareous rocks and is
incrusted with calcium carbonate. It is possible that these taxa with wide
brownish-black to bluish-black, firm sheaths actually form a cluster that
could eventually be placed in a genus separate both from Leptolyngbya and
Schizothrix. All other taxa in these two genera have colorless sheaths, or
less frequently sheaths with yellow to yellowish-brown coloration, or even
more rarely with reddish coloration.
The culture of L. badia from which the sequences and photomicrographs
were obtained died after the second transfer. Thus, no reference strain ex-
ists for this taxon.
Leptolyngbya angustissima (W. et G. S. WEST) ANAGN. et KOM. (Fig. 1B)
B a s i o n y m : Phormidium angustissimum W. et G. S. WEST 1897
Filaments 1.0–1.2 µm wide. Sheaths thin, colorless. Trichomes constricted,
untapered, with rounded end cells, 0.8-1.0 µm wide. Cells non-granular or
with an occasional minute granule at the crosswalls, thylakoid structure not
discernible in LM, longer than wide, 1.5–5 µm long.

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 Leptolyngbya from the Great Smoky Mountains 29

Collected from Cataloochie Road, site 2.6, Great Smoky Mountains Na-
tional Park.
This taxon has been repeatedly reported in floras (as P. angustissimum),
but not illustrated (GEITLER 1932, ELENKIN 1936–1949, DESIKACHARY
1959, STARMACH 1966, COMPÈRE 1986, VINOGRADOVA et al. 2000). Our
form is distinctly constricted at the crosswalls, whereas European material
is slightly or indistinctly constricted according to KOMÁREK. WHITFORD &
SCHUMACHER (1984) illustrated a nonconstricted taxon with very elongated
cells that is quite distinct from our specimens, and actually more similar to
Leptolyngbya sp. 1. This taxon is taxonomically and morphologically sepa-
rate from Jaaginema angustissimum (W. et G. S. WEST) ANAGN. et KOM. (ba-
sionym: Oscillatoria angustissima W. et G. S. WEST).

Leptolyngbya subtilissima (KÜTZING) KOMÁREK in ANAGN. (Fig. 1C)

B a s i o n y m : Leptothrix subtilissima KÜTZING
Filaments lacking sheaths, 1.2–1.5 µm wide. Trichomes distinctly con-
stricted at the crosswalls, with bluntly rounded end cells, 1.2–1.5 µm wide.
Cells non-granular, isodiametric, 1.3–1.7 µm long.
Collected from Hen Wallow Falls sites 1.1, Cataloochie Road site 2.6,
Great Smoky Mountains National Park.
Our specimens are a perfect morphological and ecological match to this
subaerophytic species. This taxon was reported previously from the park as
Leptolyngbya cf. subtilissima (JOHANSEN et al. 2004).

Leptolyngbya cf. cataractarum (RABENH. ex HANSG.) KOM. in ANAGN.

(Fig. 1F)
Trichome not constricted at the crosswalls, without evident sheath, bluntly
rounded end cells 2.5 µm wide. Cells granular, with thylakoid structure not
evident, 4.5–5.5 µm long.
Collected from Cataloochie Road, site 1.1, Great Smoky Mountains Na-
tional Park.
The ecology and cell size are similar to L. cataractarum, but our speci-
mens were found in solitary trichomes, rather than the mats in which L.
cataractarum is normally found. In addition, L. cataractarum has a more
evident sheath than our specimens. Leptolyngbya cf. cataractarum was con-
sidered too rare to justify description as a new species.

Leptolyngbya species 1 (Fig. 1E)

Filaments 0.8–1.0 µm wide. Sheath colorless, thin, rarely extending past the
apices of the trichomes, sometimes absent. Trichomes unconstricted at the
crosswalls, untapered, with rounded end cells, 0.8–1.0 µm wide. Cells non-
granular, longer than wide, 2–4 µm long.
Collected from seep wall on Cataloochie Road, Site 2.6, Great Smoky
Mountains National Park, Haywood County, North Carolina.
This taxon is clearly a new species to science based on morphological

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 30 J. F. JOHANSEN et al.

and ecological criteria, but we were not able to isolate the taxon into cul-
ture or obtain sequence data, and reviewers of this manuscript were unani-
mous in recommending it not be named at present. Cell sizes are similar
to Leptolyngbya angustissima, but this form is clearly unconstricted and
easily differentiated in our material (no intergradations were seen). It does
resemble the specimens illustrated as Phormidium angustissimum W. et G.
S. WEST (syn. L. angustissima) by WHITFORD & SCHUMACHER (1984), but
their specimens do not correspond with the circumscription of that taxon
(KOMÁREK & ANAGNOSTIDIS 2005). Cell sizes and habitat type are similar
to the unconstricted trichomes of L. truncata (LEMM.) ANAGN. et KOM., but
this form consistently has 1 to many granules per cell. This taxon also bears
a resemblance to Jaaginema angustissimum (W. et G. S. WEST) ANAGN. et
KOM. Our taxon differs from J. angustissimum in having longer cells and
different habitat requirements (J. angustissimum is planktic).

Leptolyngbya sp. 9 (Fig. 1G)

Filaments solitary and entangled among other algae, with a single trichome
per filament, without false branching, 2.5 µm wide. Sheath colorless, thin,
not lamellated, and open. Trichomes not evidently constricted at the cross-
walls, no necridia, not tapering, with rounded end cells, 2.0 µm wide. Cells
longer than wide, non-granular, pale blue-green, with thylakoids not visible
in the light microscope, 2–4 µm long.
Collected from Cataloochie Road, site 2.7, Great Smoky Mountains Na-
tional Park
The dimensions of our specimen were similar to L. benthonica (SKUJA)
ANAGN., but the specimen did not have tapering ends, and also did not have
a prominent aerotope at either side of the crosswalls. This taxon was also
similar to L. gracillima (ZOPF ex HANSG.) ANAGN. et KOM., but our speci-
men did not have gradually tapering ends. In addition, while fitting within
the description of L. gracillima, our specimen had considerably narrower
dimensions. Not enough material was found to describe Leptolyngbya sp. 9
as a new species.

Molecular characterization and phylogeny

Phylogenetic analysis of L. appalachiana and L. badia demonstrated that

they were very distinct from the type species of the genus, L. boryana
(GOM.) ANAGN. et KOM. (Fig. 3). Leptolyngbya appalachiana was sister
to the clade containing Leptolyngbya sensu stricto, those taxa with cells
shorter than wide to isodiametric, with constricted walls, necridia, a single
trichome per sheath, and false branching. Leptolyngbya badia fell into an
even more distant clade. We consider Leptolyngbya to be polyphyletic and
in need of revision. However, until sequences of most or all genera in the
Pseudanabaenales have been sequenced (particularly the generitypes), this

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 Leptolyngbya from the Great Smoky Mountains 31

Fig. 2. Light micrographs of Leptolyngbya species new to science. A–B. L. appala-

chiana, strain GSM-SFF-MK60. C–D. L. badia, strain CRS-1.
[Scale bars = 10 µm]

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 32 J. F. JOHANSEN et al.

Fig. 3. Distance tree of selected Pseudanabaenaceae. Rooted with outgroup taxon

Gloeobacter PCC8105. Bootstrap values for distance analysis given above the node,
bootstrap value from parsimony analysis below the node (or slash mark).

revision will be difficult to undertake because we cannot be sure that the

lineages currently called “Leptolyngbya” do not belong to established but
misunderstood taxa. We think it is probable based on this phylogeny that
when the genus is revised, L. badia will be transferred to a different genus
given its phylogenetic placement.

Johansen_K.indd 32 20.04.2008 20:28:24 Uhr

 Leptolyngbya from the Great Smoky Mountains 33

The ITS regions were also very distinct for both taxa. A detailed com-
parison of ITS structures in Leptolyngbya is reported elsewhere (JOHANSEN
et al. in review). Here we only report the structures for the two new taxa
(Fig. 4). Only operons containing both tRNALeu and tRNAAla genes in
the 16S-23S ITS were recovered from the Great Smoky Mountains taxa.
While only one operon was recovered from L. appalachiana, three distinct
operons were sequenced from L. badia. Two L. badia operons were identi-
cal in sequence and structure except for the region between the two tRNA
genes, commonly containing the V2 helix (ITEMAN et al. 2001). This helix
differed in length for the two operons (Fig. 4 J, K). The third operon (like
the L. appalachiana operon) lacked a V2 helix, and had distinctly different
D1-D1', Box B, and V3 helices than the other two operons from L. badia
(Fig. 4, compare these pairs of structures: A,B; E, F; and H,I). L. appala-
chiana was distinctive among all Leptolyngbya taxa for which we have
obtained ITS structure in that the helices were exceptionally long (Fig. 4
C, D, G). Typically the ITS region in L. boryana for those operons having
both tRNA genes is 479 nucleotides, but for L. appalachiana the total was

Fig. 4. Helices of the 16S-23S ITS regions of L. appalachiana and L. badia. A, B.

L. badia D1-D1' helices. C. L. appalachiana D1-D1' helix. D. L. appalachiana Box-
B helix. E, F. L. badia Box-B helices. G. L. appalachiana V3 helix. H, I. L. badia V3
helices. J, K. L. badia V2 helices.

Johansen_K.indd 33 20.04.2008 20:28:26 Uhr

 34 J. F. JOHANSEN et al.

602 nucleotides. L. badia ribosomal operons possessed ITS regions of vary-

ing lengths: 518, 504, and 497 nucleotides, respectively.

Conclusions
The material from GSMNP was extremely rich in cyanobacterial species. Of
the taxa identified 84 % had not previously been reported from GSMNP. In
addition, almost half of the cyanobacterial species seen were possible new
records to science. The diversity found in this limited number of samples
indicates that the genus Leptolyngbya is still poorly understood and de-
scribed. Eleven species of Leptolyngbya (mostly as Phormidium species)
have been reported from North America (KOMÁREK et al. 2003). Our find-
ings here and those of our research group (GOMEZ et al. 2003, JOHANSEN et
al. 2004) indicate that the Smoky Mountains likely contain many endemic
taxa yet to be discovered and described.
Many cyanobacterial taxa appear to be widely distributed (DESIKACH-
ARY 1959, GEITLER 1932). This is true particularly for lake-inhabiting spe-
cies. We suspect this may be true at least in part to the regular movement
of migrating waterfowl, which likely act as repeated and frequent vectors
for lentic species. Humans, with introductions of both plants and fish to
lakes, have also likely contributed to the introduction of European algal
species to the lakes of North America. The wet walls and headwater moun-
tain streams, which are the dominant aquatic environments found within
the park, do not attract migrating waterfowl, nor do they sustain fish popu-
lations. The absence of animal vectors capable of long-distance travel be-
tween first order streams and the high gradient of the streams present in
the mountains likely contribute to geographic isolation of algae within the
Smoky Mountains. We suspect the distinctive biotopes in combination with
geographic isolation have contributed to the development of endemic taxa.
The likelihood of our GSMNP taxa being conspecific with previously de-
scribed European species seems low to us. With the combination of mor-
phological and ecological data, it is possible to recognize the cyanobacterial
biodiversity of this region, and we intend to continue our studies in this
fascinating locality.

Acknowledgements

SHANNON GOMEZ assisted with collection of original material. MARGARET FITZ-

PATRICK assisted in collection and isolation of L. appalachiana and L. badia. This
work was supported in part with a grant from Discover Life in America and funds
from the National Science Foundation (DEB 0206360).

Johansen_K.indd 34 20.04.2008 20:28:27 Uhr

 Leptolyngbya from the Great Smoky Mountains 35

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Manuscript received February 16, 2007, accepted September 15, 2007.

Authors’ addresses:

JEFFREY R. JOHANSEN
Department of Biology
John Carroll University
University Heights, OH 44118, U.S.A.
E-mail address: johansen@jcu.ed

CATHERINE E. OLSEN
Department of Biology
John Carroll University
University Heights, OH 44118, U.S.A.

REX L. LOWE
Department of Biological Sciences
Bowling Green State University
Bowling Green, OH 43403, U.S.A.

KAROLINA FUČÍKOVÁ
Department of Ecology and Evolutionary Biology
University of Connecticut
75 North Eagleville Rd.
Storrs, CT 06269, U.S.A.

DALE A. CASAMATTA
Department of Biology
University of North Florida
Jacksonville, FL 32224, U.S.A.

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