Several coprinoid fungi have been identified as promotors of Cremastra appendiculata seed germination, while others appear ineffective. This study aimed to discern which genera within the Psathyrellaceae family exhibit this capability and to identify the most effective coprinoid fungi for the cultivation of C.
appendiculata. We collected 21 coprinoid fungi from diverse sources and symbiotically cultured them with C. appendiculata seeds. 9 fungi were found to induce seed germination and support seed development, specifically within the genera Coprinellus, Tulosesus, and Candolleomyces. In contrast, fungi that failed to promote germination predominantly belonged to the genera Coprinopsis and Parasola. Notably, four fungi-Coprinellus xanthothrix, Coprinellus pseudodisseminatus, Psathyrella singeri, and Psathyrella candolleana-were documented for the first time as capable of enhancing C. appendiculata seed germination. Strain 218LXJ-10, identified as Coprinellus radians, demonstrated the most significant effect and has been implemented in large-scale production, underscoring its considerable practical value. These findings contribute vital scientific insights for the conservation and sustainable use of C. appendiculata resources.
Human papillomaviruses (HPVs) can increase the proliferation of infected cells during HPV-driven abnormalities, such as
cervical cancer or benign warts. To date, more than 200 HPV genotypes have been identified, most of which are classified
into three major genera: Alphapapillomavirus, Betapapillomavirus, and Gammapapillomavirus. HPV genomes commonly
encode two structural (L1 and L2) and seven functional (E1, E2, E4–E7, and E8) proteins. L2, the minor structural protein
of HPVs, not only serves as a viral capsid component but also interacts with various human proteins during viral infection. A
recent report revealed that L2 of HPV16 recruits polo-like kinase 1 (Plk1), a master regulator of eukaryotic mitosis and cell
cycle progression, for the delivery of viral DNA to mitotic chromatin during HPV16 infection. In this study, we verified the
direct and potent interactions between the polo-box domain (PBD) of Plk1 and PBD-binding motif (S–S–pT–P)-containing
phosphopeptides derived from L2 of HPV16/HPV18 (high-risk alphapapillomaviruses), HPV5b (low-risk betapapillomavirus),
and HPV4 (low-risk gammapapillomavirus). Subsequent structural determination of the Plk1 PBD bound to the
HPV18 or HPV4 L2-derived phosphopeptide demonstrated that they interact with each other in a canonical manner, in
which electrostatic interactions and hydrogen bonds play key roles in sustaining the complex. Therefore, our structural and
biochemical data imply that Plk1 is a broad binding target of L2 of various HPV genotypes belonging to the Alpha-, Beta-,
and Gammapapillomavirus genera.
Prions are infectious proteins that mostly replicate in self-propagating amyloid conformations (filamentous protein polymers)
and consist of structurally altered normal soluble proteins. Prions can arise spontaneously in the cell without any
clear reason and are generally considered fatal disease-causing agents that are only present in mammals. However, after the
seminal discovery of two prions, [PSI+] and [URE3], in the eukaryotic model microorganism Saccharomyces cerevisiae,
at least ten more prions have been discovered, and their biological and pathological effects on the host, molecular structure,
and the relationship between prions and cellular components have been studied. In a filamentous fungus model, Podospora
anserina, a vegetative incomparability-related [Het-s] prion that directly triggers cell death during anastomosis (hyphal
fusion) was discovered. These prions in eukaryotic microbes have extended our understanding to overcome most fatal human
prion/amyloid diseases. A prokaryotic microorganism (Clostridium botulinum) was reported to have a prion analog. The
transcriptional regulators of C. botulinum-Rho can be converted into the self-replicating prion form ([RHO-X-C+]), which
may affect global transcription. Here, we outline the major issues with prions in microbes and the lessons learned from the
relatively uncovered microbial prion world.
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Tubulysins are bioactive secondary metabolites produced by myxobacteria that promote microtubule disassembly. Microtubules
are required for protozoa such as Tetrahymena to form cilia and flagella. To study the role of tubulysins in myxobacteria,
we co-cultured myxobacteria and Tetrahymena. When 4000 Tetrahymena thermophila and 5.0 × 108
myxobacteria were
added to 1 ml of CYSE medium and co-cultured for 48 h, the population of T. thermophila increased to more than 75,000.
However, co-culturing tubulysin-producing myxobacteria, including Archangium gephyra KYC5002, with T. thermophila
caused the population of T. thermophila to decrease from 4000 to less than 83 within 48 h. Almost no dead bodies of T.
thermophila were observed in the culture medium. Co-culturing of T. thermophila and the A. gephyra KYC5002 strain with
inactivation of the tubulysin biosynthesis gene led to the population of T. thermophila increasing to 46,667. These results
show that in nature, most myxobacteria are preyed upon by T. thermophila, but some myxobacteria prey on and kill T. thermophila
using tubulysins. Adding purified tubulysin A to T. thermophila changed the cell shape from ovoid to spherical and
caused cell surface cilia to disappear.
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This study examined the changes in soil enzymatic activity, microbial carbon source metabolic diversity, and straw decomposition
rates in paddy fields treated with 1, 2, or 3 years of straw returning (SR1–SR3). The soil’s ability to decompose straw
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the numbers of straw returning years. Cellulolytic bacteria and inorganic phosphate bacteria were significantly positively
correlated with the decomposition rate (r = 0.783 and r = 0.375, P < 0.05). Based on 16S sequencing results, straw returning
improved the microbial diversity of paddy soils by increasing unclassified bacteria and keeping dominant soil microorganism
populations unchanged. The relative importance of individual microbial taxa was compared using random forest models.
Proteobacteria, ammoniating bacteria, and potassium dissolving bacteria contributed to peroxidase activity. The significant
contributors to phosphate monoesterase were Acidobacteriota, Desulfobacterota, ammoniating bacteria, cellulolytic bacteria,
and potassium-dissolving bacteria. Proteobacteria, ammoniating bacteria, cellulolytic bacteria, and potassium-dissolving bacteria
contributed to urease activity. Desulfobacterota, ammoniating bacteria, cellulolytic bacteria, and potassium-dissolving
bacteria contributed to the neutral invertase activity. In conclusion, soil microbial community structure and function were
affected within 2 years of straw returning, which was driven by the combined effects of soil organic carbon, available nitrogen,
available potassium, and pH. With elapsing straw returning years, soil properties interacted with soil microbial communities,
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methanogens in the nearshore sediments than in the offshore
sediments. The Mann–Whitney U test demonstrated that the
relative abundance of Methanococcoides was significantly
higher in the offshore sediments, while the relative abundances
of Methanogenium, Methanosarcina, Methanosaeta,
Methanolinea, and Methanomassiliicoccus were significantly
higher in the nearshore sediments (P < 0.05). The abundance
of the mcrA gene in the nearshore sediments was significantly
higher than that in the offshore sediments. Furthermore, a
similar vertical distribution of the methanogen and sulfatereducing
bacteria (SRB) abundances was observed in the SYS
sediments, implying there is potential cooperation between
these two functional microbes in this environment. Finally,
total organic carbon (TOC) was significantly correlated with
methanogen community composition.
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Helicobacter pylori infection causes chronic inflammation
in the stomach, which is linked to the development of gastric
cancer. The anti-inflammatory and anticancer effects of a
glycolysis inhibitor 2-deoxyglucose (2DG) and an antidiabetic
medication metformin (Met) have gotten attention. Using a
Mongolian gerbil animal model, we investigated H. pylorimediated
gastric pathogenesis and how this pathogenesis is
influenced by 2DG and Met. Five-week-old male gerbils were
infected with H. pylori strain 7.13. After 2 weeks of infection,
gerbils were fed 2DG-containing food (0.03% w/w), Met-containing
water (0.5% w/v), or both (Combi) for 2 (short-term)
or 10 weeks (long-term). Gastric pathogenesis and host response
to H. pylori infection were examined by macroscopic
and histopathologic analysis of gerbils’ stomach. As a result,
indicators of gastric pathogenesis by H. pylori infection including
infiltration of polymorphonuclear neutrophils and
lymphocytes, intestinal metaplasia, atrophy, and proliferation
of gastric epithelial cells were attenuated by short-term administration
of 2DG, Met, or Combi. When the infection was
sustained for long-term, gastric pathogenesis in drug-treated
gerbils was equivalent to that in untreated gerbils, with the
exception that the infiltration of neutrophil was reduced by
2DG. Colonization of H. pylori in stomach was unaffected
by both short- and long-term treatments. Our findings demonstrate
that the progression of gastric pathogenesis induced
by H. pylori infection can be attenuated by the shortterm
individual or combinational treatment of 2DG and
Met, implying that 2DG or Met could be considered as a
treatment option for gastric diseases in the early stages of
infection.
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We evaluated the Cre-lox and CRISPR-Cas9 systems as markerrecycling
tools in Saccharomyces cerevisiae recombinants containing
multiple-integrated expression cassettes. As an initial
trial, we constructed rDNA-nontranscribed spacer- or Ty4-
based multiple integration vectors containing the URA3 marker
flanked by the loxP sequence. Integrants harboring multiple
copies of tHMG1 and NNV-CP expression cassettes were obtained
and subsequently transformed with the Cre plasmid.
However, the simultaneous pop-out of the expression cassettes
along with the URA3 marker hampered the use of Cre-lox as
a marker-recycling tool in multiple integrants. As an alternative,
we constructed a set of CRISPR-Cas9-gRNA vectors containing
gRNA targeted to auxotrophic marker genes. Transformation
of multiple integrants of tHMG1 and NNV-CP
cassettes by the Cas9-gRNA vector in the presence of the URA3
(stop) donor DNA fragments generated the Ura- transformants
retaining multiple copies of the expression cassettes.
CRISPR-Cas9-based inactivation led to the recycling of the
other markers, HIS3, LEU2, and TRP1, without loss of expression
cassettes in the recombinants containing multiple
copies of tHMG1, NNV-CP, and SfBGL1 cassettes, respectively.
Reuse of the same selection marker in marker-inactivated
S. cerevisiae was validated by multiple integrations of the
TrEGL2 cassette into the S. cerevisiae strain expressing SfBGL1.
These results demonstrate that introducing stop codons into
selection marker genes using the CRISPR-Cas9 system with
donor DNA fragments is an efficient strategy for markerrecycling
in multiple integrants. In particular, the continual
reuse of auxotrophic markers would facilitate the construction
of a yeast cell factory containing multiple copies of expression
cassettes without antibiotic resistance genes.
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Extraintestinal pathogenic Escherichia coli (ExPEC) is an important
zoonotic pathogen that places severe burdens on public
health and animal husbandry. There are many pathogenic
factors in E. coli. The type VI secretion system (T6SS) is a
nano-microbial weapon that can assemble quickly and inject
toxic effectors into recipient cells when danger is encountered.
T6SSs are encoded in the genomes of approximately
25% of sequenced Gram-negative bacteria. When these bacteria
come into contact with eukaryotic cells or prokaryotic
microbes, the T6SS assembles and secretes associated effectors.
In the porcine ExPEC strain PCN033, we identified four
classic rearrangement hotspot (Rhs) genes. We determined
the functions of the four Rhs proteins through mutant construction
and protein expression. Animal infection experiments
showed that the Δrhs-1CT, Δrhs-2CT, Δrhs-3CT, and
Δrhs-4CT caused a significant decrease in the multiplication
ability of PCN033 in vivo. Cell infection experiments showed
that the Rhs protein is involved in anti-phagocytosis activities
and bacterial adhesion and invasion abilities. The results
of this study demonstrated that rhs1, rhs3, and rh4 plays an
important role in the interaction between PCN033 and host
cell. Rhs2 has contribution to cell and mice infection. This
study helps to elucidate the pathogenic mechanism governing
PCN033 and may help to establish a foundation for further
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Aquatic microorganisms in the sediment and water column
are closely related; however, their distribution patterns between
these two habitats still remain largely unknown. In this
study, we compared sediment and water microeukaryotic and
bacterial microorganisms in aquaculture ponds from different
areas in China, and analyzed the influencing environmental
factors as well as the inter-taxa relationships. We found that
bacteria were significantly more abundant than fungi in both
sediment and water, and the bacterial richness and diversity
in sediment were higher than in water in all the sampling
areas, but no significant differences were found between the
two habitats for microeukaryotes. Bacterial taxa could be
clearly separated through cluster analysis between the sediment
and water, while eukaryotic taxa at all classification
levels could not. Spirochaetea, Deltaproteobacteria, Nitrospirae,
Ignavibacteriae, Firmicutes, Chloroflexi, and Lentimicrobiaceae
were more abundantly distributed in sediment,
while Betaproteobacteria, Alphaproteobacter, Cyanobacteria,
Roseiflexaceae, Dinghuibacter, Cryomorphaceae, and Actinobacteria
were more abundant in water samples. For eukaryotes,
only Cryptomonadales were found to be distributed
differently between the two habitats. Microorganisms in sediment
were mainly correlated with enzymes related to organic
matter decomposition, while water temperature, pH, dissolved
oxygen, and nutrient levels all showed significant correlation
with the microbial communities in pond water. Intensive interspecific
relationships were also found among eukaryotes
and bacteria. Together, our results indicated that eukaryotic
microorganisms are distributed less differently between sediment
and water in aquaculture ponds compared to bacteria.
This study provides valuable data for evaluating microbial
distributions in aquatic environments, which may also be of
practical use in aquaculture pond management.
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Adaptation to changing environmental conditions is crucial
for the survival of microorganisms. Bacteria have evolved
various mechanisms to cope with osmotic stress. Here, we
report the identification and functional characterization of
the osmotic stress response operon, betIBA, in Acinetobacter
nosocomialis. The betIBA operon encodes enzymes that are
important for the conversion of choline to the osmoprotectant,
glycine betaine. The betIBA operon is polycistronic
and is under the regulation of the first gene, betI, of the same
operon. A bioinformatics analysis revealed the presence of
a BetI-binding motif upstream of the betIBA operon, and
electrophoretic mobility shift assays confirmed the specific
binding of BetI. An mRNA expression analysis revealed that
expression of betI, betB, and betA genes is elevated in a betIeletion
mutant compared with the wild type, confirming that
the autorepressor BetI represses the betIBA operon in A.
nosocomialis. We further found that the betIBA operon is
under the transcriptional control of the quorum-sensing (QS)
regulator, AnoR in, A. nosocomialis. A subsequent analysis
of the impact of BetI on expression of the QS genes, anoR
and anoI, demonstrated that BetI acts as a repressor of anoR
and anoI. In addition, it was noticed that the osmotic stress
response regulator, OmpR might play an important role in
controlling the expression of betIBA operon in A. nosocomialis.
Collectively, these data demonstrate that QS and osmotic
stress-response systems are correlated in A. nosocomialis
and that the expression of genes in both systems is
finely tuned by various feedback loops depending on osmolarity
conditions.
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Salterns are hypersaline extreme environments with unique
physicochemical properties such as a salinity gradient. Although
the investigation of microbiota in salterns has focused
on archaea and bacteria, diverse fungi also thrive in the brine
and soil of salterns. Fungi isolated from salterns are represented
by black yeasts (Hortaea werneckii, Phaeotheca triangularis,
Aureobasidium pullulans, and Trimmatostroma salinum),
Cladosporium, Aspergillus, and Penicillium species. Most
studies on saltern-derived fungi gave attention to black yeasts
and their physiological characteristics, including growth under
various culture conditions. Since then, biochemical and
molecular tools have been employed to explore adaptation of
these fungi to salt stress. Genome databases of several fungi
in salterns are now publicly available and being used to elucidate
salt tolerance mechanisms and discover the target genes
for agricultural and industrial applications. Notably, the number
of enzymes and novel metabolites known to be produced
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subjects to study fungal biodiversity and adaptive
mechanisms in extreme environments, but also valuable bioresources
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To screen for Aspergillus activity against Xanthomonas oryzae
pv. oryzae and analyse the antimicrobial components
involved, 60 Aspergillus spp. were isolated and purified from
fruits, soil and other habitats. As-75, an Aspergillus strain that
can antagonize Xanthomonas oryzae pv. oryzae, was identified
based on the zone of inhibition formed during co-culture.
According to morphological, ITS rDNA gene sequencing
and phylogenetic tree results, the strain showed close
homology to Aspergillus sclerotiorum. The biochemical characterization
tests showed that the fermentation broth of strain
As-75 exhibited a high capacity for environmental adaptation.
The results of the antimicrobial spectrum experiments demonstrated
that As-75 exhibited fairly strong antagonistic activity
against five plant pathogenic fungi and six plant pathogenic
bacteria in vitro. The fermentation broth of strain As-75
displayed maximum stability under fluorescent illumination
at temperatures below 60°C at pH 6.5. A substance with antagonistic
activity was obtained from strain As-75 via fractional
extraction, silica gel column chromatography and thinlayer
chromatography. Through mass spectrometry, nuclear
magnetic resonance and electrospray ionization mass spectrometry
(ESI-MS) analyses, the target compound was identified
as (2Z)-2-butenedioic acid-2-(1-methylethenyl)-4-methyl
ester; its molecular weight of 170.06 daltons and formula
of C8H10O4 identify it as a novel compound. Trials of
the preventative and curative effects demonstrated that compound
S1 exhibited a better control efficiency than the control
against rice bacterial blight. Additionally, the M1 processing method was better, and the efficiency of compound
S1 in preventing rice bacterial blight in six rice varieties,
TN1, IR24, ZF802, Zhonghua 11, Wuyunjing 21, and Nipponbare,
was 78.3%, 77.5%, 74.2%, 75.3%, 70.9%, and 72.1%,
respectively.
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Mucor circinelloides is a pathogenic fungus and etiologic agent
of mucormycosis. In 2013, cases of gastrointestinal illness
after yogurt consumption were reported to the US FDA, and
the producer found that its products were contaminated with
Mucor. A previous study found that the Mucor strain isolated
from an open contaminated yogurt exhibited virulence
in a murine systemic infection model and showed that this
strain is capable of surviving passage through the gastrointestinal
tract of mice. In this study, we isolated another Mucor
strain from an unopened yogurt that is closely related but
distinct from the first Mucor strain and subsequently examined
if Mucor alters the gut microbiota in a murine host
model. DNA extracted from a ten-day course of stool samples
was used to analyze the microbiota in the gastrointestinal
tracts of mice exposed via ingestion of Mucor spores. The
bacterial 16S rRNA gene and fungal ITS1 sequences obtained
were used to identify taxa of each kingdom. Linear regressions
revealed that there are changes in bacterial and fungal abundance
in the gastrointestinal tracts of mice which ingested
Mucor. Furthermore, we found an increased abundance of
the bacterial genus Bacteroides and a decreased abundance
of the bacteria Akkermansia muciniphila in the gastrointestinal
tracts of exposed mice. Measurements of abundances
show shifts in relative levels of multiple bacterial and fungal
taxa between mouse groups. These findings suggest that exposure
of the gastrointestinal tract to Mucor can alter the microbiota
and, more importantly, illustrate an interaction between
the intestinal mycobiota and bacteriota. In addition, Mucor was able to induce increased permeability in epithelial
cell monolayers in vitro, which might be indicative of unstable
intestinal barriers. Understanding how the gut microbiota is
shaped is important to understand the basis of potential methods
of treatment for gastrointestinal illness. How the gut
microbiota changes in response to exposure, even by pathogens
not considered to be causative agents of food-borne illness,
may be important to how commercial food producers
prevent and respond to contamination of products aimed at
the public. This study provides evidence that the fungal microbiota,
though understudied, may play an important role
in diseases of the human gut.
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Interspecific mycelial interactions between white rot fungi
are always accompanied by an increased production of laccase.
In this study, the potential of the white rot fungus Dichomitus
squalens to enhance laccase production during interactions
with two other white rot fungi, Trametes versicolor or Pleurotus
ostreatus, was assessed. To probe the mechanism of laccase
induction and the role that laccase plays during combative
interaction, we analyzed the differential gene expression profile
of the laccase induction response to stressful conditions
during fungal interaction. We further confirmed the expression
patterns of 16 selected genes by qRT-PCR analysis. We
noted that many differentially expressed genes (DEGs) encoded
proteins that were involved in xenobiotic detoxification
and reactive oxygen species (ROS) generation or reduction,
including aldo/keto reductase, glutathione S-transferases,
cytochrome P450 enzymes, alcohol oxidases and dehydrogenase,
manganese peroxidase and laccase. Furthermore, many
DEG-encoded proteins were involved in antagonistic mechanisms
of nutrient acquisition and antifungal properties, including
glycoside hydrolase, glucanase, chitinase and terpenoid
synthases. DEG analyses effectively revealed that laccase
induction was likely caused by protective responses to
oxidative stress and nutrient competition during interspecific
fungal interactions.
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