Table Of ContentJournal of Applied Bacteriology 1993,7 5, 595-603
Taxonomic studies on some leuconostoc-like organisms from
fermented sausages: description of a new genus Weissella for
the Leuconostoc paramesenteroides group of species
M.D. Collins, J. Samelisl, J. Metaxopoulos’ and S. Wallbanks
AFRC Institute of Food Research, Department of Microbiology, Reading Laboratory, Reading, UK and
’Agricultural University of Athens, Department of Food Science and Technology, Athens, Greece
4509/03/93:a ccepted 3 July 1993
M.D. COLLINS, J. SAMELIS, J. METAXOPOULOS AND s. WALLBANKS. 1993. Taxonomic studies
were performed on some unknown Leuconostoc-like organisms from fermented Greek sausage.
Comparative 16s rRNA sequence analysis showed the unidentified organisms represent a new
line within the Leuconostoc pararnesenteroides group of species. On the basis of the results of
this and earlier phylogenetic investigations, it is proposed that Leuconostoc paramesenteroides
and related species be reclassified in a new genus Weissella. In addition a new species,
Weissella hellenica, is proposed for the isolates from fermented sausage.
INTRODUCTION acid but differed from currently-recognized species in a
number of biochemical tests. In this paper we describe the
Leuconstocs are a diverse group of Gram-positive catalase-
phenotypic characteristics of these isolates and the results
negative cocci which share many characteristics with the
of a comparative 16s rRNA phylogenetic analysis.
genus Lactobacillus and other ‘lactic acid bacteria’. Apart
from their typically irregular coccoid morphology, leuco-
MATERIALS AND METHODS
nostocs are distinguished from gas-forming hetero-
fermentative lactobacilli primarily by their inability to
Bacterial strains
produce ammonia from arginine and the formation of only
D( -)-lactate from glucose (Garvie 1986). Some atypical Strains were isolated from dry, naturally-fermented Greek
species of heterofermentative lactobacilli, however, e.g. sausage on MRS agar (De Man et al. 1960) incubated at
Lactobacillus viridescens and Lact. fructosus do not deami- 30°C for 3 d under anaerobic conditions. Two representa-
nate arginine and form predominantly D( -)-lactate. The tive strains have been deposited in the National Collection
reliable differentiation of leuconostocs from such hetero- of Food Bacteria (NCFB 2973 = LV346; NCFB
fermentative lactobacilli by phenotypic criteria is currently 2972 = LV338).
very difficult. Furthermore, recent molecular systematic
investigations have revealed these taxa are phylogenetically Biochemical tests
intermixed, In particular, 16s rRNA sequencing studies
Strains were tested for Gram reaction and catalase pro-
show leuconostocs comprise three distinct genetic lines : the duction (Harrigan and McCance 1976). Growth at different
genus Leuconostoc sensu stricto, the Leuconostoc paramesen- temperatures was observed in MRS broth after incubation
teroides group (which includes the atypical lactobacilli Lact. at 15°C for 5 d, 37°C and 45°C for 5 d, and at 4°C and
confusus, Lact. minor, Lact. kandleri, Lact. halotolerans and 10°C for 10 d. Growth in the presence of 8% and 10%
Lact. viridescens) and the species Leuc. oenos (Yang and NaCl was observed in MRS broth at 30°C for 5 d. The
Woese 1989; Martinez-Murcia and Collins 1990, 1991 ; ability to grow at pH 3.9 was tested in MRS broth adjusted
Collins et al. 1991). In the course of a survey of the lactic with HCl to pH 3.9. Strains were tested for the ability to
acid flora of dry naturally-fermented Greek sausage, we iso-
grow on acetate agar (Rogosa et al. 1951) adjusted to pH
lated a group of Leuconostoc-like organisms. The sausage 5.6 as described by Shaw and Harding (1984). Production
isolates resembled leuconostocs in producing D( -)-lactic of hydrogen peroxide was tested on manganese dioxide agar
(MRS agar supplemented with 0.75% manganese dioxide
and with 0.5% xanthan gum) (Whittenbury 1964). Gas
Correspondence to: Dr M.D. Collms, AFRC Institute of Food Research,
(CO,) production from glucose was determined in MRS
Microbiology Department, Reading Laboratory, Earley Gate, Whiteknights
Road, Reading RG6 ZEF, UK. broth with diammonium citrate replaced by ammonium
596 M.D. COLLINS ET AL.
sulphate (Hitchener et al. 1982). The fermentation of car- tinguish the genus Weisseila from leuconostocs was synthe-
bohydrates was tested according to Sharpe (1979) by the sized by the phosphoramidite method on a model 391 DNA
miniplate method described by Jayne-Williams (1976). synthesizer (Applied Biosystems, Warrington, UK). The
Sugar fermentation was also tested with the API 50 CHL probe was labelled with fluorescein with the ECL 3’ end
system. The configuration of lactic acid formed from labelling and detection kit (Amersham International, UK)
glucose was tested enzymatically by D-laCtate and L-lactate according to the manufacturer’s instructions.
dehydrogenase (Boehringer Mannheim). Production of rRNA gene products for hybridizations were generated
acetoin from glucose was tested by Voges-Proskauer test with two primers pA (sequence 5’ GAGAGTTTGA-
(Barritt’s modification; Harrigan and McCance 1976). TCCTGGCTCAGGA) and pE* (sequence 5’ TT-
Hydrolysis of arginine was tested in MRS broth without CGAATTAAACCACATGC). Amplified rDNA products
meat extract but containing 0.05% glucose and 0.3% argin- (ca 1 pg) were denatured by boiling for 5 rnin and applied
ine, and 0.2% sodium citrate replacing ammonium citrate to Hybond N + nylon membrane (Amersham International)
(Shaw and Harding 1984). Production of dextran (slime) with the Bio-Rad slot-blot apparatus (Bio-Rad, Hemel
from sucrose was observed on MRS agar in which glucose Hempstead, UK) as described by the manufacturer. DNA
was replaced by 5% sucrose (Hitchener et al. 1982). was fixed to the membrane by placing it on 3 mm paper
(Whatman Laboratory Division, Maidstone, UK) soaked in
DNA base composition 0.4 mol I-’ NaOH, followed by 2 x 15 rnin washes in
2 x SSC (standard saline citrate). Prehybridization,
Chromosomal DNA was isolated as described by Pitcher et
hybridization, post-hybridization washing and immunologi-
al. (1989). DNA base composition was estimated by
cal detection of the probe were performed according to the
thermal denaturation in standard saline citrate as described
ECL kit manufacturer’s instructions (Amersham
by Garvie (1978) with DNA from Escherichia coli NCDO
International). Hybridizations were performed for 2 h at
+
1984 (K12) (51.5 mol% G C) as standard.
42°C and probes were used at a concentration of 20
ng m l ~ P ost-hybridization stringency washes (2 x 15
16s rRNA gene sequence determination and analysis min) were performed in 1 x SSC containing 0.1% SDS at
47°C. After incubation with detection reagents, membranes
Approximately 2.5 pg of chromosomal DNA was subjected
were exposed to autoradiography film (Fuji, Genetic
to PCR amplification in a total volume of 100 p1 containing
Research Instrumentation Ltd, Dunmow, UK) for 1-3
1 unit of Tag polymerase (Amersham International). The
min. Stripping of membranes for re-probing with 16s
reaction involved 36 cycles of denaturation at 92°C for 2
rRNA universal probe (sequence 5’ TTACCGCGGCTG-
min, primer annealing at 55°C for 1 min and primer exten-
CTGGCACGT 3’) was achieved by boiling in 0.5% SDS
sion at 72°C for 1.5 min. DNA was extracted with chloro-
for 8 min followed by washing in 2 x SSC for a further 5
form and purified with the Geneclean I1 kit (BIO 101,
min.
Inc.) according to the manufacturer’s instructions. Qualit-
ative analysis of the DNA fragments was performed by
agarose gel electrophoresis. Sequencing of the amplified RESULTS AND DISCUSSION
product was performed with LY-~~dAST P and SEQUE-
All 11 sausage isolates were found to be phenotypically
NASE version 2.0 sequencing kit (USB) as described pre-
homogeneous and formed large coccoid (but sometimes
viously by Hutson et al. (1993). Reaction products were
lenticular) cells occurring in pairs or chains. The strains
separated on 55 cm wedge shaped (0.2-0.6 mm) 6%
were Gram-positive, catalase-negative, facultative anaer-
acrylamide-7 mol I-’ urea gels at 55°C with an LKB
obes. All grew slowly in MRS broth but failed to grow on
Macrophor 2010 sequencing unit operated at 50 W per gel.
acetate agar (pH 5.6) or in 10% NaCl. They were all het-
Sequences were aligned and similarity values were deter-
erofermentative although gas production was relatively
mined with the Wisconsin Molecular Biology package
poor. The strains were similar to leuconostocs in producing
(Devereux et al. 1984). Nucleotide substitution rates (K,,,,
D( -)-lactate.
values) were calculated, and an unrooted phylogenetic tree
The partial 16s rRNA gene sequences of representative
was produced by using the algorithm of Fitch and Margol-
strains of the sausage bacterium were investigated to deter-
iash (1967) contained in a program written by Felsenstein
mine their relationship with known leuconostocs and other
(1982).
lactic acid bacteria. The 165 rRNA gene sequence of strain
LV346 determined by direct sequencing of amplified pro-
Oligonucleotide probe hybridizations
ducts is shown in Fig. 1. The sequence consisted of a con-
Oligonucleotide probe Wgp (sequence 5’ CCTACCTC- tinuous stretch of 1525 nucleotides representing
TTAG(C/T)(A/G)GGGGATAA 3’) designed to dis- approximately 98% of the total 16s rRNA gene. Two frag-
MOLECULAR SYSTEMATICS OF LEUCONOSTOCS 597
CCUGGCUCAGGAUGAACGCUGGCGGCGUGCCAAUACAUGC~GUCGAAUGGGACCGCUUUGUGCUU~UUGAUAUGACGAGCUUGCUCUGAUUUGAUUUUUUGAUU
UCAAAGAGUGGCGAACGGGUGAGUAACACGUGGGUAACCUACCUCUUAGCAGGGGAUAACAUUUGG~CAAGUGCUAAUACCGUAUAAUACCAACAACC
GCAUGGUUGUUGGUUGAAAGAUGGUUCUGCUAUCACUAAGAGAUGGACCCGCGGUGCAUUAGCUAGUUGGU~GGUAAUGGCUUACCAAGGCAAUGAUGC
AUAGCCGAGUUGAGAGACUGAUCGGCCACAAUGGGACUGGGACUGAGACACGGCCCAUACUCCUACGGGAGGCAGCAGUAGGG~UCUUCCACAAUGGGCGCAAGC
CUGAUGGAGCAACGCCGCGUGUGUGAUGAAGGGGUUUCGGCUCGU~CACUGUUAUAAGAGAAGAACGGCACUGAGAGU~CUGUUCAGUGUGUGACGG
UAUCUUACCAGAAAGGAACGGCUAAAUACGUGCCAGCAGCCGCGGU~UACGUAUGUUCC~GCGUUAUCCGGAUUUA~GGGCGU~GCGAGCGCAGA
C G G U U A U U U A A G U C U G A A G U G A A A G C C C U C A G C C C U G C U C C A U G U G
UAGCGGUGAAAUGCGUAGAUAUAUGGAAGAACACCAGUGGCGAAGGCGGCUUUCUGGACUGUAACUGACGUUGAGGCUCG~GUGUGGGUAGC~CAG
GAUUAGAUACCCUGGUAGUCCACACCGUAAACGAUGAGUGCUAGAUGUUCGAGGGUUUCCGCCCUUGAGUGUCGCAGCU~CGCAUUAAGCACUCCGCCU
GGGGAGUACGACCGCAAGGUUGAAAGCCCUCACUCAAAGGAAUUGACGGGGACCCGCAC~GCGGUGGAGCAUGUGGUUU~UUCG~GCAACGCGAAGAACCUUAC
CAGGUCUUGACAUCCCUUGACAACGCUAGAAAUAGCGCGUUCCCUUCGGGGACAAGGUGACAGGUGGUGCAUGGUUGUCGUCAGCUCGUGUCGUGAGAUG
UUGGGUUAAGUCCCGCAACGAGCGCAACCCCUUAUUAUUAGUUGCCAGCAUUCAGUUGGGCACUCUAGUGAGACUGCCGGUGAU~CCGGAGGAAGGUGG
GGAUGACGUCRAAUCAUCAUGCCCCUUAUGACCUGGGCUACACACGUGCUACAUGGCAUAUACAACGAGUCGCUAACCCGCGAGGGUACGCUAAUCUCU
UAAAGUAUGUCUCAGUUCGGAUUGUAGGCUGCAACUCGCCUACAUG~GUCGGAAUCGCUAGUAAUCGCGGAUCAGAACGCCGCGGUG~UACGUUCCCG
GGUCUUGUACACACCGCCCGUCACACCAUGAGAGUUUGUACACCCAAAGCCGGUGGGGUAACCUUUUAGGAGCCAGCCGUCUAAGGUGGGACAGAUGAU
UAGGGUGAAGUCGUAACAAGUAGCCGU
Fig. 1 Nucleotide sequence of derived 16s rRNA of Weissellu heNenica (NCFB 2973)
ments (approx. positions 50 to 200 and 900 to 1100, which ences (18 mismatches, four unmatched for a comparison of
includes variable regions V1 and V6, nomenclature of Neefs 15 18 bases) between these two taxa, however, is signifi-
et al. 1990) of the 16s rRNA genes of two other strains cantly greater than that observed between strains of the
(LV338, LV315) were sequenced and found to be identical same species. The presence of characteristic ‘signatures’
to that of LV346, thereby demonstrating genetic homo- within the 16s rRNA of the unknown organism provides
geneity of the unknown isolates. Similarity values for a further evidence of its genospecific separatedness. Most
1330 nucleotide region (ranging from positions 107 to 1410 notable of these is diagnostic sequences in helix 1007/1022
of the Escherichia coli numbering system) of the 16s rRNA of variable region V6 which readily distinguishes the
sequence from strain LV346 and homologous sequences of unknown bacterium from all known species of the Leuc.
over 100 reference strains from the major phylogenetic lines paramesenteroides group (Table 2). Earlier 16s rRNA
within the lactic acid bacteria were determined. Approx- sequencing studies (Yang and Woese 1989; Martinez-
imately 100 nucleotides proximal to the 5’ end of the rRNA Murcia and Collins 1990, 1991) demonstrated that Leuc.
gene were omitted from the similarity calculations in order paramesenteroides and related species (viz. Lact. confusus,
to remove alignment problems arising from considerable Lact. kandleri, Lact. minor, Lact. halotolerans, Lact.
variation in the length of the V1 region (see Neefs et al. 1990 virzdescens) form a natural phylogenetic group, separate
for nomenclature) between species. A matrix of representa- from Leuconostoc sensu stricto and other lactic acid bacteria.
tive sequence similarities is shown in Table 1. Strain A recent comparative study of 23s rRNA gene sequences
LV346 exhibited very high sequence relatedness (98.6%) to (Martinez-Murcia et al. 1993) confirmed the phylogenetic
Leuc. paramesenteroides. Relatively high sequence simi- distinctiveness of the Leuc. paramesenteroides cluster and it
larities were also shown with other species of the Leuc. is now evident that this grouping merits separate generic
paramesenteroides group of organisms (ca 94-97 %), status. Therefore, based upon the results of the present and
although significantly lower values were displayed with earlier phylogenetic investigations, we propose Leuc. para-
members of Leuconostoc sensu stricto (ca 89-91 %). Evolu- mesenteroides and related species be reclassified in a new
tionary distance (KnUcv)a lues between the sausage isolates, genus for which we propose the name Weissella. We further
leuconostocs and other representative lactic acid bacteria propose the unknown bacterium from fermented sausage be
were calculated, and a distance matrix tree constructed recognized as a new species with the name Weissella hel-
.
from these data is shown in Fig. 2. The branching pattern lenica
of the tree reinforced the high phylogenetic affinity
between the unknown strain and the Leuc. paramesen-
Description of Weisseiia gen. nov.
teroides group of species.
In the present study we have characterized a new group (Weissellu M. L. dim. fem. n. named after Norbert Weiss, a
of Leuconostoc-like organisms from dry fermented Greek German microbiologist, known for his many contributions
sausage. It was evident from both 16s rRNA similarity cal- to the lactic acid bacteria). Cells are generally short rods
culations and results of the distance matrix analysis that the with rounded to tapered ends or coccoid in shape occurring
closest phylogenetic relative of the unknown bacterium is singly, in pairs or in short chains. Cells are Gram-positive
Leuc. paramesenteroides. The number of nucleotide differ- and non-motile. Endospores are not formed. Catalase and
I S
.r
0
-t
r-,
N
MOLECULAR SYSTEMATICS OF LEUCONOSTOCS 599
Fig. 2 Unrooted tree showing the Leu. mnas
phylogenetic interrelationshipso f members
of the genus Weissella and some other W bolotolerans
lactic acid bacteria. The tree is based on a
comparison of ca 1330 nucleotides. A, W wridescens
W. Xandferi
Aerococcus ;C , Carnobacterium; E , Lb. fermentum
\
Enterococcus ;L b, Lactobacillus ;L c, Leu follax
Lactococcus ;L eu, Leuconostoc; P, Leu cornosum
Pediococcus ;S , Streptococcus ; V,
Vagococcus; W, Weissella Leu pseudomesenteroides
Leu lochs
Lb ocidaphtlus
Lb bomsteri
Lb delbrubckii
\
LC IOCtlS
Lc gorvteoe
cytochromes are not produced. Chemo-organotrophs, Description of Welssella h8lotolerans (Kandier,
having complex nutritional requirements. Hetero- Schillinger and Welss) comb. nov.
fermentative. With the exception of Weissella paramesen-
A detailed species description may be found in Kandler et
teroides and W. hetlenica (which produce D( -)-lactic acid)
al. (1983) and Kandler and Weiss (1986).
species of the genus generally produce DL lactate from
glucose. Acidoduric. Growth occurs at 15°C; growth does
not occur at 45°C (with the exception of some strains of W.
confusa). Strains of some species hydrolyse arginine. The Description Of weissell8 k8ndleri (HolzaPfel and van
cell wall peptidoglycan is based upon lysine; the inter- WYk) comb- nov-
peptide bridge contains alanine, or serine and alanine, as
A detailed species description may be found in Holzapfel
typical constituents. The guanine plus cytosine content of
and van Wyk (1982) and Kandler and Weiss (1986).
DNA is 37-47 mol%. The type species is W. vzrzdescens.
Description of Weissella confus8 (Holzapfel and Description of Weissell8 mlnor (Kandler, Schllllnger
Kandier) comb. nov. and Weiss) comb. nov.
A detailed species description may be found in Holzapfel A detailed species description may be found in Kandler et
and Kandler (1969) and Kandler and Weiss (1986). al. (1983) and Kandler and Weiss (1 986).
Table 2 Diagnostic sequences of Weitsella
spp. in helix 1007/1022 of variable region W.p aramesenteroides UUGCUAAUCCUAGAAAUAGGACGGUUCC
W. hellenica sp. nov. UUGACAACGCUAGAAAUAGCGCG.UUCC
V6 of 16s rRNA
W. confusa UUGACNACUCCAGAGAGGAG.CG.UUC
W. halotolerans UUGACCACCUCAGAGAUGAGGC.UUUCC
W. kandleri UUGACCACUCCAG AGAUGG AGC.UUUCC
W. minor UUGACCACUUCAGA G AUGAAGC.UUUC
W. viridescens UUGACCACUUCAGAGAUGAAGC.UUUC
Paired nucleotides are shown underlined.
L,
W. hellenica nov. sp. D L ys-Ala-Ser Large spherical or lenticular cells d is -); D( D
ci
a
W. paramesenteroides d (4 + + + d NT + + d D Lys-Ala, ; Lys-Ser-Ala, Spherical or lenticular cells % more the lactic of or
0
9
W. confusa + + DL L ys-Ala Short rods thickened at one end d reaction; D,
e
y
a
or del
W. halotolerans + NT DL Ly s-Ala-Ser Irregular short coccoid rods ns positive; ( ),
ai
r
st
W. minor + DL Lys-Ser-Ala, Irregular short coccoid rods with rounded to tapered ends 1 I-89% the of
d,
Differential characteristics of species of the genus Weissella Table 3 W. Characteristic W. kandleri viridescens Acid produced from: L-Ara binose Cellobiose Galactose Maltose Melibiose Rafinose Ribose Sucrose Trehalose Xylose + - NH, from arginine + NT Dextran formation Lactic acid configuration DL DL L Murein type Lys- Ala-Gly- Ala, Ala-Ser s- y Cell morphology Irregular Small irregular rods rods ~ ~ ~ f, -, 90% more of strains positive; more strains negative; 90% or or NT, 25% more than of total lactic acid is not tested. +); L(
MOLECULAR SYSTEMATICS OF LEUCONOSTOCS 601
Table 4 Differential characteristics of species of the genus Leuconostoc
Leuc.
mesenteroades Leuc.
Leuc. Leuc. Leuc. Leuc. subsp. mesenteraides Leuc. Leuc.
Characteristic carnosum citreum gelidum lactis mesenteroides subsp. cremoris oenos pseudomesenteroides
Acid produced from:
+ + + +
L- Arabinose d
+ + +
Cellobiose d d
+ +
Galactose d - d
+ + +
Maltose d -
+ + +
Melibiose d d
+ +
Rafinose - d -
+
Ribose - d d d
+ + + +
Sucrose -
+ + + + +
Trehalose
+ +
Xylose d d d
NH, from arginine - - - - - - - -
+
Dextran information d NT d - - - NT
Lactic acid D D D D D D D D
configuration
Murein type NT NT NT Lys-Ala, Lys-Ser-Ala, Lys-Ser-Ala, Lys-Ser, ; Lys-Ser-Ala,
Lys-Ala-Ser
+, 90% or more strains positive; -, 90% or more strains negative; d, 11-89% of strains positive; D, 90% or more of the lactic acid is
D( -); NT, not tested.
Description of Weissella paramesenteroldes (Garvle) bacillus broth below pH 44-50. Slime is not produced
comb. nov. from sucrose. Voges-Proskauer test negative. Hydrogen
peroxide is produced. Acid is produced from L-arabinose,
A detailed species description may be found in Garvie
glucose, fructose, mannose, a-methyl-D-glucoside, N-
(1967, 1986).
acetyl-glucosamine, maltose, sucrose, trehalose, D-arabitol
and gluconate. Acid is not produced from glycerol, erythri-
Description of Weissella viridescens (Niven and tol, D-arabinose, ribose, xylose, adonitol, P-methyl-xyloside,
Evans) comb. nov. L-sorbose, rhamnose, dulcitol, inositol, mannitol, sorbitol,
a-methyl-D-mannoside, amygdalin, arbutin, salicin, cello-
A detailed species description may be found in Niven and
Evans (1957) and Kandler and Weiss (1986). biose, lactose, inulin, melezitose, D-raffinose, amidon, gly-
cogen, xylitol, fl-gentiobiose, D-lyxose, D-tagatose, fucose,
L-arabitol, 2-ceto-gluconate and 5-ceto-gluconate. A very
Description of Weissella hellenica sp. nov.
weak and delayed reaction with galactose, D-tUranOSe and
(Gr. adj. Hellenikos, Greek; N.L. fern. adj. hellenica, melibiose may be observed. The cell wall murein is type
Greece, from where the bacterium was first isolated). Lys-L-Ala-L-Ser(L-Ala). The DNA base compositions of
+
Gram-positive, non-motile, non-sporeforming, spherical strains LV346 and LV338 were 39-4 and 40.0 mol% G C
but sometimes lenticular cells usually occurring in pairs or (T,) respectively. The type strain is NCFB 2973
short chains, with a tendency to form clusters. Growth in ( = LV346). Isolated from fermented sausages.
MRS broth is slow with cells tending to precipitate. Colo-
nies are small, often pin-point, smooth, round and greyish Differentiation of the genus Weissella from the other
white. No growth occurs at 37°C. All strains grow at 10°C lactic acid taxa
and 4°C (delayed). Heterofermentative but gas production Members of the genus Weissella may be distinguished
is relatively poor. All strains produce more than 98% readily from homofermentative lactobacilli, pediococci,
D( -)-lactate. Catalase-negative. Arginine is not hydrolysed. enterococci, lactococci and streptococci by the formation of
Growth occurs in 8% NaCI, but not in 10% NaCl or on gas from carbohydrates. A cell wall murein based upon
acetate agar (pH 5.6). All strains fail to acidify Lacto- lysine with an interpeptide bridge containing alanine, or
602 M.D. COLLINS ET AL.
(0) CCTACCTCTTAG(C/T)(A/G)GGGGATA3A’) directed
I 2 3 4 5 6
against PCR-amplified 16s rDNA also provides an
unequivocal means of distinguishing species of the genus
Weissella from Leuconostoc sensu stricto (see Fig. 3).
Differentlation of Welssella hellenlca from other
Welssella species
Weissella hellenica can be distinguished from other species
of the genus Weissella by the characteristics shown in Table
3.
ib)
I 2 3 4 5 6
ACKNOWLEDGEMENTS
M.D. Collins and S. Wallbanks gratefully acknowledge the
support of the Ministry of Agriculture and Fisheries and
Food. We are grateful to Professor T.O. MacAdoo
(Virginia Polytechnic Institute and State University, USA)
for deriving the species name and to Dr N. Weiss
(Deutsche Sammlung von Mikroorganismen, Germany) for
Fig. 3 Autoradiograph of slot-blot hybridizations to amplified providing murein composition.
16s rRNA gene fragments of row A: 1, Werssella
paramesenterordes (NCDO 803); 2, W. confusa (NCDO 1586); 3,
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