International Journal of Systematic and Evolutionary Microbiology (2006), 56, 477–486 DOI 10.1099/ijs.0.

63980-0

Novel Neosartorya species isolated from soil in

Korea
Seung-Beom Hong,1 Hye-Sun Cho,1 Hyeon-Dong Shin,2 Jens C. Frisvad3
and Robert A. Samson4
1
Correspondence Korean Agricultural Culture Collection, NIAB, Suwon 441-707, Korea
Robert A. Samson 2
Division of Environmental Science and Ecological Engineering, College of Life and
[email protected] Environmental Science, Korea University, Seoul 136-701, Korea
3
Center for Microbial Biotechnology, Biocentrum-DTU, Technical University of Denmark,
Building 221, DK-2800 Kgs. Lyngby, Denmark
4
Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands

Phenotypic and genotypic characters of strains of Neosartorya spinosa and related taxa were
analysed. N. spinosa, Neosartorya botucatensis and Neosartorya paulistensis had identical partial
b-tubulin and calmodulin gene sequences and could not be differentiated on macro- and
micro-morphological characteristics, including by scanning electron microscopy. Based on partial
b-tubulin and calmodulin gene sequences and ascospore morphology, two separate groups
are distinguished and are proposed as novel species. Neosartorya laciniosa sp. nov. has
microtuberculate ascospores with two bent crests and two distinct equatorial rings of small
projections. Neosartorya coreana sp. nov. has rugose to weak reticulate ascospores with two often
bent crests, but without the equatorial rings of small projections. The type strain of N. laciniosa
is CBS 117721T (=NRRL 35589T=KACC 41657T) and the type strain of N. coreana is CBS
117059T (=NRRL 35590T=KACC 41659T).

INTRODUCTION based on secondary metabolite profiles (Samson et al., 1990)

and phenotypic and genotypic characteristics (Rinyu et al.,
Ascospore ornamentation is an important morphological
2000).
character for distinguishing species of the ascomycete genus
Neosartorya (Kozakiewicz, 1989). Based on ascospore orna- During a survey of soil-borne Aspergillus and Penicillium in
mentation, Horie et al. (2003) distinguished homothallic Korea, many Neosartorya strains were encountered and, in
Neosartorya species with white to pale-yellow ascomata and order to examine the taxonomic position of our isolates, we
ascospores with two distinct equatorial crests into six species. have re-examined N. spinosa, N. glabra and related taxa. We
In this group, Neosartorya spinosa and Neosartorya glabra have used a polyphasic taxonomic approach in order to
have been separated on the basis of smooth (for N. glabra) determine the delimitation and variability of the taxon.
or echinulate (for N. spinosa) ascospores, but many isolates For phenotypic analyses, morphology was examined using
possess intermediate patterns of ornamentation. Further- macro- and micro-morphology and scanning electron
more, N. glabra was also reported as highly polymorphic microscopy (SEM) methods and growth temperatures
were studied. For the phylogenetic analyses, b-tubulin and
Published online ahead of print on 21 October 2005 as DOI 10.1099/ calmodulin gene sequences were used, which have proved to
ijs.0.63980-0. be useful for Aspergillus taxonomy (Feibelman et al., 1998;
Abbreviations: NJ, neighbour-joining; SEM, scanning electron micro- Geiser et al., 1998; Ito et al., 2001; Varga et al., 2000a, b).
scopy.
The GenBank/EMBL/DDBJ accession numbers for the b-tubulin and METHODS
calmodulin gene sequences of the strains examined in this study are
shown in Table 1. Strains examined are listed in Table 1. The strains were maintained
A NJ phylogenetic tree based on calmodulin gene sequences and SEM on malt extract agar slants at 15 uC.
images of ascospores of Neosartorya species are available as supple- Morphology and extrolites. For macro-morphological observa-
mentary figures in IJSEM Online.
tions, Czapek yeast autolysate (CYA), malt extract autolysate (MEA)
The Mycobank accession numbers for Neosartorya laciniosa sp. nov. agar and Czapek agar (CZA) were used (Samson et al., 2004). Yeast
and Neosartorya coreana sp. nov. are MB 500233 and MB 500234, extract sucrose (YES) agar, oatmeal agar (OA) and creatine sucrose
respectively (http://www.mycobank.org). agar (CREA) were also used (Frisvad & Samson, 2004). The isolates

63980 G 2006 IUMS Printed in Great Britain 477

S.-B. Hong and others

Table 1. List of strains used in this study

CBS, Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands; NRRL, Agricultural Research Service Culture Collection, Peoria, USA;
IBT, Institute for Biotechnology, Lyngby, Technical University of Denmark; KACC, Korean Agricultural Culture Collection, Suwon, Korea.

Isolate Source Accession number

b-Tubulin Calmodulin

Aspergillus fumigatus CBS 545.65 Unknown source, K. B. Raper AY685151 AY689335

Aspergillus lentulus KACC 41391 Maize field soil, Yeongi, Korea AY685170 AY689354
Neosartorya aurata CBS 466.65T Jungle soil, Brunei AF057318* AY870685
Neosartorya botucatensis CBS 114221T Soil, Brazil AY870763 AY870723
Neosartorya coreana
CBS 117057 (=IBT 22051) Strawberries, Sydney, Australia AY870757 AY870717
CBS 117059T (=KACC 41659T=NRRL 35590T) Tomato field soil, Buyeo, Korea AY870758 AY870718
Neosartorya fennelliae CBS 411.89 Marine sludge, Japan AY870727 AY870686
Neosartorya fischeri
CBS 544.65T Canned apples AF057322* AY689370
CBS 317.89 Desert sand, Namibia AY870728 AY870687
CBS 420.96 Maize field, Nepal AY870729 AY870688
KACC 41668 Soil, Dominican Republic AY870730 AY870689
KACC 41673 Soil, Dominican Republic AY870731 AY870690
KACC 41674 Soil, Dominican Republic AY870732 AY870691
KACC 41664 Sesame field soil, Yeongi, Korea AY870733 AY870692
KACC 41665 Pepper field soil, Daejeon, Korea – –
Neosartorya glabra sensu stricto
CBS 111.55T Rubber scrap from old tyre, Iowa, USA AY870734 AY870693
CBS 165.63 Soil, Netherlands AY870735 AY870694
CBS 582.90 Raspberry pulp, Netherlands AY870736 AY870695
KACC 41654 Tomato field soil, Buyeo, Korea AY870737 AY870696
KACC 41655 Tomato field soil, Buyeo, Korea – –
KACC 41656 Tomato field soil, Buyeo, Korea – –
Neosartorya hiratsukae CBS 294.93T Pasteurized aloe juice, Japan AF057324* AY870699
Neosartorya laciniosa
KACC 41652 Perilla field soil, Daejeon, Korea AY870747 AY870707
CBS 117065 (=IBT 6660) Soil, Surinam AY870748 AY870708
KACC 41644 Soil, Dominican Republic AY870749 AY870709
KACC 41645 Soil, Dominican Republic AY870750 AY870710
KACC 41646 Soil, Dominican Republic AY870751 AY870711
KACC 41648 Soil, Kenya, AY870752 AY870712
CBS 206.92 Strawberry pulp, Netherlands AY870753 AY870713
CBS 315.89 Soil, Pakistan AY870754 AY870714
CBS 585.90 Vineyard, USA AY870755 AY870715
CBS 117059T (=KACC 41657T=NRRL 35589T) Tomato field soil, Buyeo, Korea AY870756 AY870716
KACC 41658 Tomato field soil, Buyeo, Korea – –
KACC 41660 Pepper field soil Daejeon, Korea – –-
Neosartorya paulistensis CBS 114216T Soil, Brazil AY870764 AY870724
Neosartorya pseudofischeri
KACC 41653 Tomato field soil, Buyeo, Korea AY870740 AY870700
KACC 41661 Perilla field soil, Daejeon, Korea AY870741 AY870701
CBS 208.92T Human vertebrae, USA AY870742 AY870702
CBS 100504 Pectin, dairy food, Netherlands AY870743 AY870703
CBS 110899 Human blood, Estonia AY870744 AY870704
CBS 117063 (=IBT 6472) Cherry filling, USA AY870745 AY870705
CBS 117073 (=IBT 3002) Soil, Denmark AY870746 AY870706
Neosartorya spinosa
CBS 483.65T Soil, Nicaragua AF057329 AY689371
CBS 117058 (=IBT 3001) Pineapple, Denmark AY870759 AY870719

478 International Journal of Systematic and Evolutionary Microbiology 56

 Two novel Neosartorya species

Table 1. cont.

Isolate Source Accession number

b-Tubulin Calmodulin

KACC 41647 Soil, Dominican Republic AY870760 AY870720

KACC 41650 Soil, Kenya AY870761 AY870721
KACC 41651 Soil, Kenya AY870762 AY870722
KACC 41662 Tomato field soil, Buyeo, Korea AY870765 AY870725
KACC 41663 Pepper field soil, Daejeon, Korea – –
Neosartorya stramenia CBS 498.65T Soil from maple–ash–elm forest, USA AY870766 AY870726
Neosartorya sp. CBS 841.96 Bark of Podocarpus sp., Papua, New Guinea AY870738 AY870697
Neosartorya sp. CBS 112.55 Garden soil, Australia AY870739 AY870698

*Sequence obtained from GenBank.

were inoculated at three points on each plate of each medium and strains were inferred from the NJ analysis and the resulting
incubated at 25 uC in the dark for 7 days and additionally at 37 uC phylogenetic tree is presented in Fig. 1.
on CYA. For micro-morphological observations, microscopic mounts
were made in lactic acid from MEA colonies and a drop of alcohol The five strains of N. spinosa, including the ex-type strain
was added to remove air bubbles and excess conidia. SEM was per-
CBS 483.65T, have identical partial b-tubulin gene sequences.
formed for every strain of Neosartorya used in this study using a
Hitachi S570 microscope. For SEM preparation, mature cleistothecia Strain CBS 117058 differs by only two bases from the other
were transferred to aluminium stubs using double-sided adhesive N. spinosa strains, while Neosartorya botucatensis CBS
tape. A small drop of 10 mM ACES buffer containing 0?05 % Tween 114221T and Neosartorya paulistensis CBS 114216T have
80 was added and the cleistothecia were crushed. The suspension identical sequences to N. spinosa. N. spinosa showed some
was air-dried and coated with platinum. gene sequence similarity to N. glabra, but showed stronger
Extrolites were analysed by HPLC using alkylphenone retention indices similarity to the two novel taxa, which are proposed as Neo-
and diode array UV-VIS detection as described by Frisvad & Thrane sartorya coreana sp. nov. and Neosartorya laciniosa sp. nov.
(1987), as modified by Smedsgaard (1997). In this study, strains previously identified as N. glabra based
on ascospore characteristics were divided into five groups.
Phylogenetic analyses. Genomic DNA was extracted according to The group N. glabra sensu stricto included the ex-type strain,
the procedure described by Lee & Taylor (1990). A fragment of the CBS 111.55T, and clustered with Neosartorya aurata, Neosar-
59 portion of the b-tubulin gene was amplified using the primers bt2a torya stramenia, Neosartorya hiratsukae and Neosartorya
and bt2b (Glass & Donaldson, 1995). The segment of the calmodulin fennelliae, but was placed far away from N. spinosa. Strains
gene was amplified using the primers cmd5 (59-CCGAGTACAAGG- CBS 112.55 and CBS 841.96 showed low sequence similarity
AGGCCTTC-39) and cmd6 (59-CCGATAGAGGTCATAACGTGG-39)
which were constructed in this study based on the complete Asper-
with N. glabra sensu stricto and were not clustered within
gillus oryzae sequence (GenBank accession number D44468). The the species. N. laciniosa, which has ascospores with a finely
amplified DNA fragments were purified using a QIAquick PCR puri- rough convex surface and more or less straight crests, was
fication kit (Qiagene). DNA sequences were determined using a located near to N. spinosa. N. laciniosa differed in three (two
BigDye Terminator v3.1 cycle sequencing kit (ABI) and an ABI 3100 continuous base deletions at positions 327 and 328 after
DNA sequencer. Both strands of each fragment were sequenced. primer bt2a) of 466 bases in the partial b-tubulin gene
DNA sequences were edited with the DNASTAR computer package and sequence compared with N. spinosa and the bootstrap of the
an alignment of the sequences was performed using the CLUSTAL W grouping was only 62 %. N. coreana, which has ascospores
program (Thompson et al., 1994). The neighbour-joining (NJ) method with more bent equatorial crests, was also located near N.
was used for the phylogenetic analysis. For NJ analysis, the data were spinosa, but separated by a 99 % bootstrap value.
first analysed using the Tamura–Nei parameter distance calculation
model with gamma-distributed substitution rates, which were then The primers cmd5 and cmd6 amplified about 580 bp of the
used to construct the NJ tree with MEGA version 2.1 (Kumar et al., calmodulin gene which contains introns 2, 3 and 4 and exons
2001). To determine the support for each clade, a bootstrap analysis
was performed with 1000 replications. 2, 3, 4 and partial exon 5. The phylogenetic relationships of
the calmodulin gene sequences for the 46 strains were
inferred from NJ analyses (see Supplementary Fig. S1 in
IJSEM Online). In the cladogram based on the calmodulin
RESULTS gene sequences, N. laciniosa was well separated from N.
The primers bt2a and bt2b amplified about 550 bp of the 59 spinosa with a bootstrap value of 98 %. However, there were
portion of the b-tubulin gene which contains introns 3, 4 two subgroups in this taxon. Apart from these differences,
and 5 and exons 3, 4, 5 and partial exon 6. The phylogenetic the topology of the calmodulin gene sequence phylogenetic
relationships of the b-tubulin gene sequences for the 46 tree was similar to that of the b-tubulin gene sequence tree.

http://ijs.sgmjournals.org 479
S.-B. Hong and others

Fig. 1. Phylogenetic tree inferred from NJ

analysis of partial b-tubulin gene sequences.
Bootstrap values (1000 replications) are
shown above or below the nodes. s. st.,
sensu stricto. Bar, 0?01 nucleotide changes
between taxa.

On the basis of Aspergillus anamorph morphology, N. projections and two straight equatorial crests. Strains of
spinosa, N. botucatensis, N. paulistensis, N. glabra, N. laci- N. laciniosa have microtuberculate ascospores with two
niosa and N. coreana are very similar. The vesicle diameter of distinct straight crests. In the furrow, two distinct equatorial
the six species ranges from 7?5 to 20 mm and conidia are rings with small projections are present (see Supplementary
(sub)globose to ellipsoid, ranging from 2?5 to 4?0 mm. The Fig. S2 in IJSEM Online). N. coreana has a rugose to weak
cleistothecia of N. glabra are yellow–white to pale yellow reticulate ornamentation of convex ascospores with two
(2-3A2-3; Kornerup & Wanscher, 1978), while the other distinct, and often bent, crests.
three species are pale yellow to light yellow (2-3A3-4). Based
on light microscopy, the ascospores of N. spinosa are rough, Our studies of extrolites showed that N. spinosa, N.
those of N. laciniosa and N. coreana are finely roughened paulistensis and N. botucatensis produced aszonalenins,
(Figs 2 and 3) and N. glabra has nearly smooth-walled 2-pyrovoylaminobenzamide and pseurotin (Table 2). In
ascospores. SEM observations showed that the ascospores of N. coreana and N. laciniosa, aszonalenins were also present,
N. spinosa CBS 483.65T have convex surfaces with distinct but the other compounds were absent. All strains of N.
spines. The ascospores of N. botucatensis and N. paulistensis laciniosa also typically produced a large number of trypto-
also have convex surfaces with spines (see Supplementary quivalins and tryptoquivalons. Aszonalenins were also
Fig. S2 in IJSEM Online). N. glabra ascospores have nearly found in strains of Neosartorya fischeri, but this species
smooth, convex surfaces with small microtuberculate produced a different extrolite profile consisting of fischerin,

Fig. 2. Neosartorya laciniosa sp. nov. CBS 117074T. (a) Colonies on MEA after 7 days at 25 6C; (b) ascomata; (c) asci; (d,
e) conidial heads; (f) ascospores under a light microscope; (g) ascospores as seen by SEM. Bars, 100 mm (b) and 10 mm
(c–g).

480 International Journal of Systematic and Evolutionary Microbiology 56

 Two novel Neosartorya species

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S.-B. Hong and others

482 International Journal of Systematic and Evolutionary Microbiology 56

 Two novel Neosartorya species

Table 2. Extrolites from Neosartorya species based on HPLC analysis of ex-type strains
The data were backed up by HPLC analyses of several further isolates of N. glabra, N. spinosa, N.
laciniosa, N. fischeri and N. pseudofischeri.

Species Extrolites

N. hiratsukae Avenaciolide
N. glabra Asperpentyn, avenaciolide, wortmannin-like compound
N. spinosa Aszonalenins, chrysogine, 2-pyrovoylaminobenzamide, pseurotin
N. paulistensis Aszonalenins, 2-pyrovoylaminobenzamide, pseurotin
N. botucatensis Aszonalenins, 2-pyrovoylaminobenzamide, pseurotin
N. laciniosa Aszonalenins, tryptoquivalins, tryptoquivalons
N. coreana Aszonalenins
N. fischeri Aszonalenins, fischerin, neosartorin, fiscalins, helvolic acid,
fumitremorgins
N. pseudofischeri Asperfuran, cytochalasin-like compound, fiscalin-like compound,
pyripyropens, gliotoxin

neosartorin, fiscalins, helvolic acid and fumitremorgins. N. have convex surfaces irregularly roughened by verruculose
glabra differed from the other taxa by the production of and small triangular projections up to 1 mm long. However,
asperpentyn, avenaciolide and wortmannin. in our study, we have not observed that these two taxa differ
significantly from N. spinosa with regard to their ascospore
morphology and growth characteristics. Together with their
DISCUSSION phylogenetic similarity, we consider N. botucatensis and N.
paulistensis to represent synonyms of N. spinosa.
Raper & Fennell (1965) described Neosartorya glabra (Raper
& Fennell) Kozakiewicz as having a smooth (or nearly so), Raper & Fennell (1965) described N. spinosa (as Aspergillus
convex ascospore surface and N. spinosa (Raper & Fennell) fischeri var. spinosus) with echinulate ascospores with pro-
Kozakiewicz as having ascospores with spine-like projec- jections ranging from <0?5 mm to as much as 2?0 mm. Our
tions ranging from <0?5 mm to as much as 2?0 mm. observations showed that N. spinosa has echinulate asco-
Peterson (1992) described the ascospores of Neosartorya spores with spines ranging from <0?5 mm to 5 (7) mm long
pseudofischeri S.W. Peterson as having triangular flaps. or with verruculose and small triangular, sometimes circu-
Among the six species, N. pseudofischeri was clearly separ- larly arranged, projections.
ated by the typical ascospore ornamentation, DNA–DNA
hybridization (Peterson, 1992) and b-tubulin gene sequence Neosartorya takakii Horie et al. was described with asco-
(Geiser et al., 1998; Varga et al., 2000b). Our b-tubulin and spores bearing roughly circularly arranged projections
calmodulin gene sequences confirmed that N. pseudofischeri (Horie et al., 2001), a feature which has been also observed
is a well defined taxon. In addition, N. pseudofischeri pro- in other N. spinosa strains. This indicates that this species is
duces a typical extrolite profile consisting of asperfuran, a further synonym of N. spinosa. However, the ex-type strain
pyripyropens, gliotoxin and compounds which are similar of N. takakii could not be obtained in spite of numerous
to cytochalasins and fiscalins. attempts. In order to establish the exact taxonomic position
of N. takakii, a more detailed molecular and extrolite study
Six strains of N. spinosa (including CBS 483.65T), CBS of the type strain is required.
114221 (ex-type strain of N. botucatensis) and CBS 114216
(ex-type strain of N. paulistensis) have almost the same b- N. glabra strains were divided into five groups based on
tubulin and calmodulin gene sequences. Strain CBS 117058 partial b-tubulin and calmodulin gene sequences. Of the five
differed in only two of 466 bases of the b-tubulin gene groups, N. glabra sensu stricto, CBS 112.55 and CBS 841.96
sequence when compared with the other strains. Strain clustered with N. fennelliae, N. hiratsukae, N. stramenia and
KACC 41647 differed in only one of 522 bases of the N. aurata (Fig. 1 and Supplementary Fig. S1). N. glabra
calmodulin gene sequence when compared with the others. sensu stricto has smaller and white cleistothecia, relatively
Horie et al. (1995) reported that the ascospores of N. botu- straight equatorial crests and smoother walled convex sur-
catensis Horie et al. have convex surfaces with long spines up faces compared with N. spinosa, N. laciniosa and N. coreana.
to 5 (maximum 7) mm long and N. paulistensis Horie et al. N. glabra grows somewhat slower (mean diameter of

Fig. 3. Neosartorya coreana sp. nov. CBS 117059T. (a) Colonies on MEA after 7 days at 25 6C; (b) ascomata; (c) asci;
(d, e) conidial heads; (f) ascospores under a light microscope; (g) ascospores as seen by SEM. Bars, 100 mm (b) and 10 mm
(c–g).

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S.-B. Hong and others

42–51 mm within 7 days at 25 uC) than the other species microtuberculate projections and thereby differing from
(mean diameter 41–68 mm within 7 days at 25 uC). How- N. spinosa and N. laciniosa.
ever, at 10 uC, strains of N. glabra grow (6 mm diameter in
7 days), but the other taxa do not show growth. Latin diagnosis of Neosartorya laciniosa S. B.
Hong, Frisvad et Samson sp. nov.
CBS 112.55 isolated from garden soil in Australia was
identified as N. glabra by Raper & Fennell (1965), but with Anamorph: Aspergillus laciniosus S. B. Hong, Frisvad et
somewhat smaller ascospores and narrower, more widely Samson sp. nov.
separated crests. The strain produced secondary metabolites
that were different from the ex-type strain of N. glabra, CBS Coloniae in agaro malti 53–55 mm diam. post 7 dies 25 uC,
111.55 T (Samson et al., 1990). Our study shows that CBS albae, planae, hyphis aeriis paucis superatae, margo irregu-
112.55 has more than 5 % dissimilarity based on partial laris, ascomata copiosa in zona marginali granulari. Species
b-tubulin and calmodulin gene sequences and did not group homothallica. Cleistothecia superficialia, alba vel dilute lutea,
with N. glabra sensu stricto (Fig. 1 and Supplementary globosa vel subglobosa, 300–400 mm diam., post 10 hebdo-
Fig. S1). However, the isolate did not differ from N. glabra mades matura, hyphis hyalinis vel flavidis, 2–4 mm latis laxe
in phenotypic characters, except that it grew somewhat circumdata. Asci 8-spori, globosi vel subglobosi vel ovoidei,
slower on CYA, MEA and CZA at 25 uC (mean diameter 11–13 mm diam., maturi evanescentes. Ascosporae late lenti-
25–29 mm in 7 days). From these results, the strain could culares, 4–5 mm, duabus crestis aequatorialibus, distantibus,
represent a novel species, but more isolates should be ad 2 mm latis circumdatae, valvae leves vel asperulatae.
available to determine its taxonomic position. Also CBS Conidiophora ex hyphis aeriis oriunda, stipites 3–4 mm lati;
841.96 had a special position in the cladograms based on vesiculae hemisphaericae vel lageniformes, 10–14 mm diam.
b-tubulin and calmodulin gene sequences (Fig. 1 and Aspergilli uniseriati, phialides 7–862–3 mm; conidia (sub)-
Supplementary Fig. S1), but it resembled N. glabra based globosa, nonnumquam late ellipsoidea, levia, 2?5–3?5 mm
on phenotypic characteristics. As is the case for CBS 112.55, diam. Massa conidiorum columnaris. Metabolita aszonaleni-
more isolates of CBS 841.96 will have to be studied to num, tryptoquivalina, tryptoquivalona.
warrant its classification as a novel species. Typus: CBS 117721T (=NRRL 35589T=KACC 41657T).
Based on the b-tubulin and calmodulin gene sequence
phylogeny, N. laciniosa was located distantly from N. glabra, Description of Neosartorya laciniosa S. B.
but closer to N. spinosa. Its separation from N. spinosa was Hong, Frisvad & Samson sp. nov.
not well supported, with a bootstrap value of 62 % for the Anamorph: Aspergillus laciniosus S. B. Hong, Frisvad &
b-tubulin gene sequence, but it was well supported on the Samson sp. nov.
basis of the calmodulin gene sequence, with a bootstrap
value of 98 %. N. laciniosa has microtuberculate ascospores Neosartorya laciniosa (la.cin.i.os9us. N.L. adj. laciniosus -a
with two distinct straight crests and two distinct equatorial -um laciniate, divided into deep, narrow, irregular segments).
rings with small projections, differing from N. spinosa which
has ascospores with long spines or roughly circularly arranged Rapidly growing colonies on MEA, 53–55 mm diameter in
projections on the convex walls. Based on the calmodulin 7 days at 25 uC, white, plane, loosely overgrown by aerial
gene sequences, two subgroups can be distinguished; one hyphae, margin irregular, cleistothecia abundantly produced
consists of strains KACC 41652, KACC 41657T, CBS 206.92 in marginal areas with granular appearance. Conidia heads
and CBS 315.89, and the second consists of strains IBT 6660, few in number; reverse greyish orange (5B6) to yellowish
KACC 41644, KACC 41645, KACC 41646, CBS 585.90 and orange (4B6). Colonies on CYA spread broadly, attaining
KACC 41648. However, we do not consider them to repre- a diameter of 56–58 mm in 7 days at 25 uC or more than
sent two separate species according to the Genealogical Con- 70 mm in 7 days at 37 uC, more or less sulcate, white to pale
cordance Phylogenetic Species Recognition (Taylor et al., yellow (2A3), granular due to the abundant production of
2000), because CBS 206.92 cross-located in the other sub- cleistothecia, loosely overgrown by aerial hyphae; conidial
group in the tree based on the b-tubulin gene sequence. heads few in number, scattered; reverse light orange (6A45).
Furthermore, no phenotypic characteristics support the Homothallic, cleistothecia superficial, white to light yellow,
separation, apart from the observation that strain IBT 6660 globose to subglobose, 300–400 mm in diameter, surrounded
produced asperfuran, in contrast to the strains of the first by a loose covering of hyaline to yellowish white, 2–4 mm
subgroup. wide hyphae. Cleistothecial peridium hyaline to light yellow,
consisting of angular, 4–15 mm diameter cells. Asci 8-spored,
N. coreana was located near, but clearly separated from, globose or subglobose to ovoid, 11–13 mm diameter, evanes-
N. spinosa based on the b-tubulin and calmodulin gene cent at maturity. Ascospores broadly lenticular, spore body
sequence trees (Fig. 1 and Supplementary Fig. S1). This 4–5 mm diameter, provided with two distinct equatorial
species had similar phenotypic characteristics to N. spinosa crests which are up to 2 mm wide with two distinct equato-
and N. laciniosa but, when observed by SEM, the species rial rings of small projections in the ascospore furrow;
had rugose to weak reticulate ornamentation of convex convex surfaces finely rough or smooth in light microscopy,
ascospore walls without the two equatorial rings of appearing rugose to tuberculate in SEM (Fig. 2). Conidial

484 International Journal of Systematic and Evolutionary Microbiology 56

 Two novel Neosartorya species

heads columnar. Conidiophores arising from aerial hyphae, Cleistothecia superficial, white to light yellow (34A45) in
3–4 mm wide at the middle; vesicles subclavate, 10–14 mm 3 weeks, globose to subglobose, 200–300 mm in diameter,
in diameter. Aspergilli uniseriate, phialides 7–862–3 mm, surrounded by a loose covering of hyaline to yellowish
covering the upper half of vesicle. Conidia, globose to sub- white, 2–3 mm wide hyphae. Cleistothecial peridium hyaline
globose, sometimes broadly elliptical, smooth, 2?5–3?5 mm to light yellow, membranaceous, consisting of angular, 4–
diameter. Extrolites: aszonalenins and several tryptoquiva- 18 mm diameter cells. Asci 8-spored, globose or subglobose
lins, tryptoquivalons; strain IBT 6660 also produces to ovoid, 10–13 mm diameter, evanescent at maturity. Asco-
asperfuran. spores, spore body 4–5 mm diameter, provided with two
well-separated, but often bent, equatorial crests which are
The type strain, CBS 117721T (=NRRL 35589T=KACC up to 2 mm wide and sometimes irregularly shaped; convex
41657T) (dried culture), was isolated from soil from a surfaces finely roughened in light microscopy appearing
tomato field in Buyeo, Korea. rugose to weakly reticulate in SEM (Fig. 3). Conidial heads
columnar. Conidiophores arising from aerial hyphae,
Latin diagnosis of Neosartorya coreana S. B. smooth, more or less sinuous, 3–4 mm wide at the middle;
Hong, Frisvad et Samson sp. nov. vesicles subclavate, 8–13 (15) mm diameter. Aspergilli uni-
seriate; phialides, 6–962–3 mm, covering the upper half of
Anamorph: Aspergillus coreanus S. B. Hong, Frisvad et
vesicle. Conidia subglobose to broadly elliptical, smooth,
Samson sp. nov.
2?5–3?5 mm diam. Extrolites: aszonalenins, pseurotin in
Coloniae in agaro malti 62–66 mm diam. post 7 dies 25 uC, CBS 117057 and several further metabolites for which the
albae vel luteo-albae, in medio elevatae, hyphis aeriis laxis structure has not yet been elucidated.
superatae, margo irregularis, ascomata copiosa dispersa,
The type strain, CBS 117059T (=NRRL 35590T=KACC
aspectu granulari. Species homothallica. Cleistothecia super-
41659T) (dried culture), was isolated from soil from a
ficialia, alba vel dilute lutea, post 3 hebdomades matura,
tomato field in Buyeo, Korea.
globosa vel subglobosa, 200–300 mm diam., hyphis hyalinis vel
flavidis, 2–3 mm latis laxe circumdata. Asci 8-spori, globosi vel
subglobosi vel ovoidei, 10–13 mm diam., maturi evanescentes.
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