bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Ноя. 2, 2023
Abstract
Motile
cilia
are
complex
organelles
comprised
of
>800
structural
proteins.
Variants
in
58
these
genes
cause
primary
ciliary
dyskinesia
(PCD)
humans.
We
used
a
second-site
non-complementation
approach
diploid
Chlamydomonas
reinhardtii
strains
to
investigate
whether
reduced
dosage
different
combinations
motile
proteins
affects
growth
/
regeneration.
Temperature-sensitive
mutants
intraflagellar
transport
fail
complement
heterozygous
diploids
at
the
restrictive
temperature,
which
may
be
due
poisonous
interactions
or
reduction
protein
levels.
Diploid
for
211
double
mutant
21
phenotypically
recessive
when
unperturbed
and
do
not
show
SSNC.
Consequently,
we
developed
sensitized
screen.
When
synthesis
is
inhibited,
cells
utilize
existing
cytoplasmic
pool
proteins,
insufficient
build
full-length
cilia.
Six
dynein
arm
preassembly
factors
(DNAAFs)
pf13,
pf23,
wdr92,
oda8
regrow
that
shorter
than
wild-type
diploids,
suggests
heterozygosity
limits
assembled
arms
further.
isolated
new
null
allele
(
pf23-4)
shows
more
severe
loss
ODAs
IDAs
previously
studied
pf23-1
hypomorphic
allele.
also
identified
role
PF23
modification
IC138,
I1/
f
inner
arm.
In
our
SSNC
assay,
pf23-4
exhibits
phenotype
.
Whole
cell
extract
immunoblots
one-half
levels
both
single
genotypes.
The
pf13
with
two
outer
ODA6
,
ODA9.
suggest
multiple
needed
assembly
Our
data
support
as
scaffolding
hub
involved
assembly.
Author
Summary
essential
movement
fluids.
humans,
defects
(PCD),
rare
disease
characterized
by
recurring
respiratory
infections,
left-right
asymmetry
defects,
ear
infertility.
Many
function
use
study
effects
changes
results
help
fold
assemble
motors
cytoplasm
sensitive
factors.
Reductions
steps
pathway
prevents
efficient
regeneration
under
stressful
cellular
conditions.
The Plant Journal,
Год журнала:
2025,
Номер
122(2)
Опубликована: Апрель 1, 2025
SUMMARY
Centromeres
in
eukaryotes
are
defined
by
the
presence
of
histone
H3
variant
CENP‐A/CENH3.
Chlamydomonas
encodes
two
predicted
CENH3
paralogs,
CENH3.1
and
CENH3.2,
that
have
not
been
previously
characterized.
We
generated
peptide
antibodies
to
unique
N‐terminal
epitopes
for
each
paralogs
as
well
an
antibody
against
a
shared
epitope.
All
three
recognized
proteins
expected
size
on
immunoblots
had
punctate
nuclear
immunofluorescence
staining
patterns.
These
results
consistent
with
both
being
expressed
localized
centromeres.
CRISPR‐Cas9‐mediated
insertional
mutagenesis
was
used
generate
null
mutations
either
or
CENH3.2
.
Single
mutants
were
viable
but
cenh3.1
cenh3.2
double
recovered,
confirming
function
is
essential.
sequenced
assembled
chromosome‐scale
genomes
from
strains
CC‐400
UL‐1690
(a
derivative
CC‐1690)
complete
centromere
sequences
17/17
14/17
chromosomes
respectively,
enabling
us
compare
evolution
across
four
isolates
near
assemblies.
data
revealed
significant
changes
between
homologous
centromeres
including
mobility
degeneration
ZeppL‐LINE1
(ZeppL)
transposons
comprise
major
repeat
sequence
Chlamydomonas.
cleavage
under
targets
tagmentation
(CUT&Tag)
purify
map
CENH3‐bound
genomic
found
enrichment
CENH3‐binding
almost
exclusively
at
regions.
An
interesting
exception
chromosome
2
UL‐1690,
which
its
genetically
mapped
region
second,
distal
location,
centered
around
single
recently
acquired
ZeppL
insertion.
The
regions
17
ranged
63.5
kb
(average
lower
estimate)
175
upper
estimate).
relatively
small
suggests
may
be
useful
organism
testing
deploying
artificial
technologies.
Abstract
Background
Common
bean
is
a
significant
grain
legume
in
human
diets.
However,
the
lack
of
complete
reference
genome
for
common
beans
has
hindered
efforts
to
improve
agronomic
cultivars.
Findings
Herein,
we
present
first
telomere-to-telomere
(T2T)
assembly
(Phaseolus
vulgaris
L.,
YP4)
using
PacBio
High-Fidelity
reads,
ONT
ultra-long
sequencing,
and
Hi-C
technologies.
The
resulted
size
560.30
Mb
with
an
N50
55.11
Mb,
exhibiting
high
completeness
accuracy
(BUSCO
score:
99.5%,
quality
value
(QV):
54.86).
sequences
were
anchored
into
11
chromosomes,
20
22
telomeres
identified,
leading
formation
9
T2T
pseudomolecules.
Furthermore,
identified
repetitive
elements
accounting
61.20%
predicted
29,925
protein-coding
genes.
Phylogenetic
analysis
suggested
estimated
divergence
time
approximately
11.6
million
years
ago
between
P.
Vigna
angularis.
Comparative
revealed
expanded
gene
families
variations
YP4
G19833
associated
defense
response.
Conclusions
genomic
insights
presented
here
are
crucial
future
genetic
studies
not
only
but
also
other
legumes.
Journal of Phycology,
Год журнала:
2023,
Номер
59(4), С. 644 - 652
Опубликована: Июль 7, 2023
Abstract
Elucidating
biological
processes
has
relied
on
the
establishment
of
model
organisms,
many
which
offer
advantageous
features
such
as
rapid
axenic
growth,
extensive
knowledge
their
physiological
and
gene
content,
ease
with
they
can
be
genetically
manipulated.
The
unicellular
green
alga
Chlamydomonas
reinhardtii
been
an
exemplary
that
enabled
scientific
breakthroughs
over
decades,
especially
in
fields
photosynthesis,
cilia
function
biogenesis,
acclimation
photosynthetic
organisms
to
environment.
Here,
we
discuss
recent
molecular/technological
advances
have
applied
C.
how
further
fostered
its
development
a
“flagship”
algal
system.
We
also
explore
future
promise
this
leveraging
genomics,
proteomics,
imaging,
synthetic
biology
for
addressing
critical
issues.
PLoS ONE,
Год журнала:
2024,
Номер
19(2), С. e0297014 - e0297014
Опубликована: Фев. 8, 2024
Recent
advances
in
long-read
sequencing
technologies
have
enabled
the
complete
assembly
of
eukaryotic
genomes
from
telomere
to
by
allowing
repeated
regions
be
fully
sequenced
and
assembled,
thus
filling
gaps
left
previous
short-read
methods.
Furthermore,
can
also
help
characterizing
structural
variants,
with
applications
fields
genome
evolution
or
cancer
genomics.
For
many
organisms,
main
bottleneck
sequence
long
reads
remains
lack
robust
methods
obtain
high-molecular-weight
(HMW)
DNA.
this
purpose,
we
developed
an
optimized
protocol
extract
DNA
suitable
for
unicellular
green
alga
Chlamydomonas
reinhardtii
,
based
on
CTAB/phenol
extraction
followed
a
size
selection
step
molecules.
We
provide
validation
results
protocol,
as
well
statistics
obtained
Oxford
Nanopore
Technologies
sequencing.
Centromeres
play
a
crucial
and
conserved
role
in
cell
division,
although
their
composition
evolutionary
history
green
algae,
the
ancestors
of
land
plants,
remains
largely
unknown.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Сен. 3, 2024
Abstract
Latency
is
a
common
strategy
in
wide
range
of
viral
lineages,
but
its
prevalence
giant
viruses
remains
unknown.
Here
we
describe
the
activity
and
production
from
617
kbp
integrated
element
model
green
alga
Chlamydomonas
reinhardtii
.
We
resolve
region
using
long-read
sequencing
show
that
particles
are
produced
released
otherwise
healthy
cultures.
A
diverse
array
viral-encoded
selfish
genetic
elements
expressed
during
GEVE
reactivation
produce
proteins
packaged
virions.
In
addition,
field
isolates
sp.
harbor
latent
related
to
C.
exhibit
similar
infection
dynamics,
demonstrating
virus
latency
prevalent
natural
host
communities.
Our
work
reports
largest
temperate
documented
date
first
active
identified
unicellular
eukaryote,
substantially
expanding
known
limits
latency.
Genome Biology and Evolution,
Год журнала:
2024,
Номер
16(7)
Опубликована: Июнь 27, 2024
Abstract
Polar
regions
harbor
a
diversity
of
cold-adapted
(cryophilic)
algae,
which
can
be
categorized
into
psychrophilic
(obligate
cryophilic)
and
cryotrophic
(nonobligate
snow
algae.
Both
accumulate
significant
biomasses
on
glacier
habitats
play
major
roles
in
global
climate
dynamics.
Despite
their
significance,
genomic
studies
these
organisms
remain
scarce,
hindering
our
understanding
evolutionary
history
adaptive
mechanisms
the
face
change.
Here,
we
present
draft
genome
assembly
annotation
algal
strain
CCCryo
101-99
(cf.
Sphaerocystis
sp.).
The
haploid
is
122.5
Mb
length
represented
by
664
contigs
with
an
N50
0.86
Mb,
Benchmarking
Universal
Single-Copy
Orthologs
(BUSCO)
completeness
92.9%
(n
=
1,519),
maximum
contig
5.3
guanine-cystosine
(GC)
content
53.1%.
In
total,
28.98%
(35.5
Mb)
contains
repetitive
elements.
We
identified
417
noncoding
RNAs
annotated
chloroplast
genome.
predicted
proteome
comprises
14,805
genes
BUSCO
97.8%.
Our
preliminary
analyses
reveal
higher
repeat
compared
mesophilic
chlorophyte
relatives,
alongside
enrichment
gene
families
associated
photosynthesis
flagella
functions.
current
data
will
facilitate
future
comparative
studies,
improving
likely
response
polar
algae
to
warming
as
well
trajectories
permanently
cold
environments.
Genome Research,
Год журнала:
2023,
Номер
33(9), С. 1582 - 1598
Опубликована: Авг. 14, 2023
Telomeres
and
subtelomeres,
the
genomic
regions
located
at
chromosome
extremities,
are
essential
for
genome
stability
in
eukaryotes.
In
absence
of
canonical
maintenance
mechanism
provided
by
telomerase,
telomere
shortening
induces
instability.
The
landscape
ensuing
rearrangements
is
not
accessible
short-read
sequencing.
Here,
we
leverage
Oxford
Nanopore
Technologies
long-read
sequencing
to
survey
extensive
repertoire
telomerase
mutants
model
green
microalga
Research Square (Research Square),
Год журнала:
2023,
Номер
unknown
Опубликована: Сен. 8, 2023
Abstract
Background
Dunaliella
algae,
such
as
salina
rich
in
β-carotene
and
bardawil
is
lutein
α-carotene,
have
been
used
aquaculture,
supplements,
cosmetics
feed
industries.
The
genus
diverse
secretive,
so
characterization
of
novel
strains
breeding
new
varieties
through
mutagenesis
technology
will
promote
natural
carotenoid
bioproduction.
Results
Salt
stress
test
demonstrated
microalgae
strain
ZP-1
was
a
halotolerant
strain.
Morphology
observation
phylogeny
analysis
indicated
this
unicellular
green
algae
member
the
.
Biomass
RAM
medium
up
to
2.45
g/L,
showing
an
advantage
over
common
terms
yield.
Furthermore,
screen
on
ethyl
methanesulfonate
mutant
library
generated
from
high-biomass
conducted
improve
productivity.
A
selected
morphology
combined
with
quantification
by
HPLC
nominated
turn
yellow
dunaliella
4
(
tyd4
).
displayed
increased
productivity
28.55%
zeaxanthin
22.19%.
promoted
17.40%
continuous
culture
under
red
light.
Application
exogenous
1.0
µM
melatonin
lead
cell
density
improved
biomass.
Conclusions
results
together
support
that
EMS
effective
approach
for
improvement
sp.
ZP-1,
which
exhibiting
potential
some
extent
overcome
bottleneck
low
biomass
current
commercial
strains.
had
higher
contents
both
zeaxanthin,
whose
yield
could
be
further
elevated
light
at
lab-scale
culture.
Our
study
provided
resources
scientific
research
technical
reference
bioproduction
carotenoids.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Фев. 29, 2024
Background:
Centromeres
play
a
crucial
and
conserved
role
in
cell
division,
although
their
composition
evolutionary
history
green
algae,
the
ancestors
of
land
plants,
remains
largely
unknown.
Results:
We
constructed
near
telomere-to-telomere
(T2T)
assemblies
for
two
Trebouxiophyceae
species,
Chlorella
sorokiniana
NS4-2
pyrenoidosa
DBH,
with
chromosome
numbers
12
13,
genome
sizes
58.11
Mb
53.41
Mb,
respectively.
identified
validated
centromere
sequences
using
CENH3
ChIP-seq
found
that,
similar
to
humans
higher
centromeric
signals
algae
display
pattern
hypomethylation.
Interestingly,
centromeres
both
species
comprised
transposable
elements,
they
differed
significantly
composition.
Species
within
genus
more
diverse
composition,
major
constituents
including
members
LTR/Copia,
LINE/L1,
LINE/RTEX
families.
This
is
contrast
Chlamydomonas
reinhardtii,
Coccomyxa
subellipsoidea,
Chromochloris
zofingiensis,
which
instead
has
pronounced
single-element
Moreover,
we
observed
significant
differences
structure
among
chromosomes
strong
collinearity
genus,
suggesting
that
sequence
evolves
rapidly
than
non-centromeric
regions.
Conclusions:
study
not
only
provides
high-quality
data
comparative
genomics
but
gives
insight
into
early
laying
an
important
foundation
further
research
on
evolution.