This study assessed the taxonomic hierarchy of the phylum
Firmicutes as well as elucidated the isolation and classification
states of novel Firmicutes species isolated from Korean
territory. The hierarchical classification system of the phylum
Firmicutes has been developed since 1872 when the genus
Bacillus was first reported and has been generally adopted
since 2001. However, this taxonomic hierarchy is still being
modified. Until Feb. 2017, the phylum Firmicutes consisted
of seven classes (Bacilli, Clostridia, Erysipelotrichia, Limnochordia,
Negativicutes, Thermolithobacteria, and Tissierellia),
13 orders, 45 families, and 421 genera. Firmicutes species isolated
from various environments in Korea have been reported
from 2000, and 187 species have been approved as of Feb.
2017. All Firmicutes species were affiliated with three classes
(Bacilli, Clostridia, and Erysipelotrichia), four orders (Bacillales,
Lactobacillales, Clostridiales, and Erysipelotrichales), 17
families, and 54 genera. A total of 173 species belong to the
class Bacilli, of which 151 species were affiliated with the order
Bacillales and the remaining 22 species with the order
Lactobacillales. Twelve species belonging to the class Clostridia
were affiliated within only one order, Clostridiales. The
most abundant family was Bacillaceae (67 species), followed
by the family Paenibacillaceae (56 species). Thirteen novel
genera were created using isolates from the Korean environment.
A number of Firmicutes species were isolated from
natural environments in Korean territory. In addition, a considerable
number of species were isolated from artificial resources
such as fermented foods. Most Firmicutes species,
belonging to the families Bacillaceae, Planococcaceae, and Staphylococcaceae, isolated from Korean fermented foods and
solar salterns were halophilic or halotolerant. Firmicutes species
were isolated from the whole territory of Korea, especially
large numbers from Provinces Gyeonggi, Chungnam,
and Daejeon.
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Protein glycosylation, the most universal and diverse posttranslational
modification, can affect protein secretion, stability,
and immunogenicity. The structures of glycans attached
to proteins are quite diverse among different organisms and
even within yeast species. In yeast, protein glycosylation plays
key roles in the quality control of secretory proteins, and particularly
in maintaining cell wall integrity. Moreover, in pathogenic
yeasts, glycans assembled on cell-surface glycoproteins
can mediate their interactions with host cells. Thus, a
comprehensive understanding of protein glycosylation in various
yeast species and defining glycan structure characteristics
can provide useful information for their biotechnological
and clinical implications. Yeast-specific glycans are a target
for glyco-engineering; implementing human-type glycosylation
pathways in yeast can aid the production of recombinant
glycoproteins with therapeutic potential. The virulenceassociated
glycans of pathogenic yeasts could be exploited
as novel targets for antifungal agents. Nowadays, several glycomics
techniques facilitate the generation of species- and
strain-specific glycome profiles and the delineation of modified
glycan structures in mutant and engineered yeast cells.
Here, we present the protocols employed in our laboratory
to investigate the N- and O-glycan chains released from purified
glycoproteins or cell wall mannoproteins in several
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showed that the isolate was most closely related to
Conexibacter woesei DSM 14684T, Solirubrobacter pauli ATCC
BAA-492T, Patulibacter minatonensis JCM 12834T, with 93.8%,
92.4%, and 91.5% sequence similarity, respectively; each genus
represented a family in the order Solirubrobacterales. Strain
BR7-21T was Gram-reaction positive, non-spore forming,
aerobic, non-motile, and short rod-shaped. It grew well on
half-strength R2A medium. The G + C content of the genomic
DNA was 73.9%. It contained meso-diaminopimelic acid in
the cell wall and the major menaquinones were MK-7(H4)
and MK-8(H4). The major fatty acids were summarized as
(C16:1 ω7c/iso-C15:0 2-OH), iso-C16:0, and C17:0 cyclo. On the
basis of polyphasic evidence, it was proposed that strain BR7-
21T should be placed in a new genus and species, for which
the name Baekduia soli gen. nov., sp. nov. was proposed with
the type strain BR7-21T (= KCTC 22257T = LMG 24797T). The
family Baekduiaceae fam. nov. is proposed to encompass
the genus Baekduia gen. nov.
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The taxonomic position of bacterial strain, designated 15J16-
1T3AT, recovered from a soil sample was established using
a polyphasic approach. Phylogenic analysis based on the
16S rRNA gene sequence showed that strain 15J16-1T3AT
belonged to the family Cytophagaceae, phylum Bacteroidetes,
and was most closely related to ‘Larkinella harenae’ 15J9-9
(95.9% similarity), Larkinella ripae 15J11-11T (95.6%), Larkinella
bovis M2TB15T (94.7%), Larkinella arboricola Z0532T
(93.9%), and Larkinella insperata LMG 22510T (93.5%). Cells
were rod-shaped, Gram-stain-negative, aerobic, and nonmotile.
The isolate grew on NA, R2A, TSA, but not on LB
agar. The strain was able to grow at temperature range from
10°C to 30°C with an optimum at 25°C and pH 6–8. Menaquinone
MK-7 was the predominant respiratory quinone.
The major cellular fatty acids comprised C16:1 ω5c (48.6%)
and C15:0 iso (24.1%). Phosphatidylethanolamine, phosphatidylserine,
and an unidentified lipid were the major polar
lipids. The G + C content of the genomic DNA was 49.5
mol%. Strain 15J16-1T3AT could be distinguished from its
closest phylogenetic neighbors based on its phenotypic, genotypic,
and chemotaxonomic features. Therefore, the isolate
is considered to represent a novel species in the genus
Larkinella, for which the name Larkinella roseus sp. nov. is
proposed. The type strain is 15J16-1T3AT (= KCTC 52004T
= JCM 31991T).
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Gram-staining-positive, motile, rod-shaped bacteria, designated
as H31022T and H31024 was isolated from rumen contents
of a Holstein cow. Optimum growth occurred at 25°C
and pH 7.0 on R2A agar medium. Oxidase and catalase activities
are positive. The 16S rRNA gene sequence (1,452 bp)
of the new isolates revealed they belong to the genus Kurthia
of the phylum Firmicutes. Highest gene sequence similarities
were assessed to be with Kurthia massiliensis JC30T (98.4%),
Kurthia senegalensis JC8ET (97.5%), and Kurthia populi 10y-
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9758T (96.7%). DNA G + C content of strains H31022T and
H31024 were 34.4% and 39.7%. Strains H31022T and H31024
has the following chemotaxonomic characteristics; the major
fatty acids are iso-C15:0, iso-C14:0 and anteiso-C15; polar
lipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine
(PE), unknown aminophospholipids
(APL), unknown glycolipids (GL), unknown phospholipids
(PL), and unknown polar lipids (L); the major quinone is MK-7.
Based on polyphasic taxonomic analysis, strains H31022T
(= KCTC 33923T = JCM 19640T) and H31024 (= KCTC 33924T
= JCM 19641T) identified a novel species in the genus Kurthia
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All orchids maintain an obligate relationship with mycorrhizal
symbionts during seed germination. In most cases,
germination-enhancing fungi have been isolated from roots
of mature plants for conservation and cultivation purposes.
To understand the germination biology of Dendrobium devonianum,
an over-collected medicinal orchid, the seeds of
D. devonianum were inoculated with a fungal strain (FDd1)
isolated from naturally occurring protocorms of D. devonianum
and two other germination-enhancing fungal strains
(FDaI7 and FCb4) from D. aphyllum and Cymbidium mannii,
respectively. The fungal strain was isolated from five
protocorms of D. devonianum and identified as a species of
the genus Epulorhiza. In germination trials, treatments with
all of the three fungal strains showed a significant promoting
effect on seed germination and protocorm formation, compared
with the control treatment (no inoculation). However,
FDd1 fungal strain showed the greatest effectiveness followed
by FDaI7 and FCb4. For all inoculation and control treatments,
seeds developed to protocorms regardless of the presence
of illumination, whereas protocorms did not develop
to seedlings unless illumination was provided. The results
of our manipulative experiments confirmed the hypothesis
that mycorrhizae associated with orchid seedlings are highly
host-specific, and the degree of specificity may be life stagespecific
under in vitro conditions. The specific mycorrhizal
symbionts from protocorms can enhance restoration efforts
and the conservation of orchids such as D. devonianum.
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Myung Soo Park , John A. Eimes , Sang Hoon Oh , Hwa Jung Suh , Seung-Yoon Oh , Seobihn Lee , Ki Hyeong Park , Hyuk Joon Kwon , Soo-Young Kim , Young Woon Lim
J. Microbiol. 2018;56(1):49-55. Published online January 4, 2018
While symbiotic fungi play a key role in the growth of endangered
Calanthe orchid species, the relationship between
fungal diversity and Calanthe species remains unclear. Here,
we surveyed root associated fungal diversity of six Calanthe
orchid species by sequencing the internal transcribed spacer
(ITS) region using 454 pyrosequencing. Our results revealed
that Paraboeremia and Coprinopsis are dominant fungal genera
among Calanthe species. In terms of overall relative abundance,
Paraboeremia was the most common fungal genus associated
with Calanthe roots, followed by Coprinopsis. Overall
fungal diversity showed a significant degree of variation depending
on both location and Calanthe species. In terms of
number of different fungal genera detected within Calanthe
species, C. discolor had the most diverse fungal community,
with 10 fungal genera detected. This study will contribute toward
a better understanding of those fungi that are required
for successful cultivation and conservation of Korean Calanthe
species.
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grown in freshwater medium until late exponential phase and
subsequently subjected to high-salinity condition that induces
excessive lipid accumulation. Our initial experiment revealed
that the concentrated culture of Chlorella sorokiniana HS1
exhibited the intense fluorescence of Nile red at the NaCl
concentration of 60 g/L along with 1 g/L of supplemental bicarbonate
after 48 h of induction period without significantly
compromising cultural integrity. These conditions were further
verified with the algal culture grown for 7 days in a 1 L
bottle reactor that reached late exponential phase; a 12% increment
in the lipid content of harvested biomass was observed
upon inducing high lipid accumulation in the concentrated
algal culture at the density of 5.0 g DW/L. Although
an increase in the sum of carbohydrate and lipid contents of
harvested biomass indicated that the external carbon source
supplemented during the induction period increased overall
carbon assimilation, a decrease in carbohydrate content suggested
the potential reallocation of cellular carbon that promoted
lipid droplet formation under high-salinity stress. These results thus emphasize that the two-phase process can be successfully
implemented to enhance algal lipid productivity by incorporating high-salinity stress conditions into the pre-concentrated
sedimentation ponds of industrial algal production
system.
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Zinc is an important transition metal in all living organisms
and is required for numerous biological processes. However,
excess zinc can also be toxic to cells and cause cellular stress.
In the model fungus Saccharomyces cerevisiae, a vacuolar zinc
transporter, Zrc1, plays important roles in the storage and
detoxification of excess intracellular zinc to protect the cell.
In this study, we identified an ortholog of the S. cerevisiae
ZRC1 gene in the human fungal pathogen Cryptococcus neoformans.
Zrc1 was localized in the vacuolar membrane in C.
neoformans, and a mutant lacking ZRC1 showed significant
growth defects under high-zinc conditions. These results suggested
a role for Zrc1 in zinc detoxification. However, contrary
to our expectation, the expression of Zrc1 was induced
in cells grown in zinc-limited conditions and decreased upon
the addition of zinc. These expression patterns were similar
to those of Zip1, the high-affinity zinc transporter in the plasma
membrane of C. neoformans. Furthermore, we used the
zrc1 mutant in a murine model of cryptococcosis to examine
whether a mammalian host could inhibit the survival of C.
neoformans using zinc toxicity. We found that the mutant
showed no difference in virulence compared with the wildtype
strain. This result suggests that Zrc1-mediated zinc detoxification
is not required for the virulence of C. neoformans,
and imply that zinc toxicity may not be an important aspect
of the host immune response to the fungus.
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Understanding the characteristics and regulation mechanisms
of cell wall integrity (CWI) in yeast is important not
only for basic research but also in biotechnological applications.
We found significantly different CWIs in two representative
strains of the thermotolerant methylotrophic yeast
Hansenula polymorpha. Compared to the A16 strain (classified
as Ogataea polymorpha), the DL1-L strain (classified as
Ogataea parapolymorpha) has a thinner cell wall that was
found to be more fragile following long-term cultivation and
more sensitive to zymolyase. To gain a deeper insight into this
difference, we compared the characteristics of the Mpk1pmediated
CWI signaling pathway in the two strains. While
a DL1-L mutant deficient in Mpk1p (mpk1Δ) showed severe
growth retardation at both normal and high growth temperatures
and in the presence of cell-wall disrupting agents, the
A16 mpk1Δ mutant displayed only a mild defect in cell growth.
Sorbitol effect on rescuing growth retardation was different
in the two mpk1Δ strains, which could partly be ascribed to
subtle differences in the activation of HOG pathway. Among
the cell wall disruptors evaluated, only caffeine clearly increased
phosphorylation of Mpk1p in DL1-L, but not in A16.
A transcriptome analysis of the DL1-L strain revealed that
caffeine significantly increased the expression of a subset of
cell-wall related genes in an Mpk1p-dependent manner, but
not the expected Rlm1-target genes. Taken together, our data
support an essential role for Mpk1p in maintaining CWI in
H. polymorpha, although the requirement for Mpk1p and
its regulation under diverse stress conditions varies depending
on the strain background.
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