Abstract
The
seeds
of
oilseed
rape
(
Brassica
napus
L.)
are
used
in
large-scale
production
one
the
most
health-promoting
plant
oils
food
industry,
as
well
for
animal
feed
and
biofuel
production.
Thus,
increasing
yield
this
crop
is
crucial
economic
ecological
importance.
However,
conventional
breeding
programs
slow,
laborious
time-consuming.
Hence,
along
with
discovery
possibility
to
apply
CRISPR/Cas
technology
edit
genomes
accelerate
process,
much
effort
has
been
put
into
applying
study
specific
genes
biosynthetic
pathways,
especially
species
many
gene
copies
such
B.
.
Here,
recent
improvements
generating
CRISPR/Cas-induced
mutations
genome,
delivering
reagents
cells,
fast-checking
efficiency
targeted
mutagenesis
reagents,
transformation
regeneration
procedures
described.
Finally,
new
applications
tools
precision
discussed,
focusing
mainly
on
verified
field.
International Journal of Molecular Sciences,
Год журнала:
2022,
Номер
23(18), С. 10202 - 10202
Опубликована: Сен. 6, 2022
CRISPR/Cas-based
genome
editing
technologies,
which
allow
the
precise
manipulation
of
plant
genomes,
have
revolutionized
science
and
enabled
creation
germplasms
with
beneficial
traits.
In
order
to
apply
these
CRISPR/Cas
reagents
must
be
delivered
into
cells;
however,
this
is
limited
by
tissue
culture
challenges.
Recently,
viral
vectors
been
used
deliver
cells.
Virus-induced
(VIGE)
has
emerged
as
a
powerful
method
several
advantages,
including
high
efficiency
simplified
process
for
generating
gene-edited
DNA-free
plants.
Here,
we
briefly
describe
editing.
We
then
focus
on
VIGE
systems
types
viruses
currently
CRISPR/Cas9
cassette
delivery
also
highlight
recent
applications
advances
in
Finally,
discuss
challenges
potential
Plants,
Год журнала:
2023,
Номер
12(17), С. 3106 - 3106
Опубликована: Авг. 29, 2023
Embryo
rescue
(ER)
techniques
are
among
the
oldest
and
most
successful
in
vitro
tissue
culture
protocols
used
with
plant
species.
ER
refers
to
a
series
of
methods
that
promote
development
an
immature
or
lethal
embryo
into
viable
plant.
Intraspecific,
interspecific,
intergeneric
crosses
allow
introgression
important
alleles
agricultural
interest
from
wild
species,
such
as
resistance
tolerance
abiotic
biotic
stresses
morphological
traits
crops.
However,
pre-zygotic
post-zygotic
reproductive
barriers
often
present
challenges
achieving
hybridization.
Pre-zygotic
manifest
incompatibility
reactions
hinder
pollen
germination,
tube
growth,
penetration
ovule
occurring
various
tissues,
stigma,
style,
ovary.
To
overcome
these
barriers,
several
strategies
employed,
including
cut-style
graft-on-style
techniques,
utilization
mixed
distinct
placenta
pollination,
pollination.
On
other
hand,
act
at
different
tissues
stages
ranging
early
subsequent
growth
reproduction
offspring.
Many
genera
result
abortion
due
failure
endosperm
development.
In
cases,
needed
hybrids.
holds
great
promise
for
not
only
facilitating
but
also
obtaining
haploids,
doubled
manipulating
ploidy
levels
chromosome
engineering
by
monosomic
disomic
addition
well
substitution
lines.
Furthermore,
can
be
shorten
cycle
propagation
rare
plants.
Additionally,
it
has
been
repeatedly
study
embryonic
development,
especially
embryo-lethal
mutants.
The
widely
procedure
is
embryos
taken
placed
directly
on
media.
certain
ovule,
ovaries
placentas
enables
young
zygote
stage
maturity.
Horticultural Plant Journal,
Год журнала:
2024,
Номер
10(4), С. 983 - 994
Опубликована: Июнь 28, 2024
Protoplast-based
transient
gene
expression
system
has
been
widely
used
in
plant
genome
editing
because
of
its
simple
operation
and
less
time-consuming.
In
order
to
establish
a
universal
protoplast-based
transfection
for
verifying
activities
vectors
containing
targets
Brassica,
we
systematically
optimized
factors
affecting
protoplast
isolation
expression.
We
established
an
efficient
(PTGE)
Chinese
cabbage,
achieving
high
yield
4.9
×
105
·
g−1
FW,
viability
over
95%,
efficiency
76%.
showed
the
first
time
that
pretreatment
protoplasts
with
hypotonic
MMG
could
significantly
enhance
efficiency.
Furthermore,
incubated
at
37
°C
6
min
improved
86%.
also
demonstrated
PTGE
worked
well
(more
than
50%
efficiency)
multiple
Brassica
species
including
Pak
Choi,
kale,
turnip.
Finally,
was
validating
CRISPR/Cas9
pak
choi,
demonstrating
broad
applicability
crops.
Cotton
has
tremendous
economic
value
worldwide;
however,
its
allopolyploid
nature
and
time-consuming
transformation
methods
have
hampered
the
development
of
cotton
functional
genomics.
The
protoplast
system
proven
to
be
an
important
versatile
tool
for
genomics,
tissue-specific
marker
gene
identification,
tracking
developmental
trajectories,
genome
editing
in
plants.
Nevertheless,
isolation
abundant
viable
protoplasts
suitable
single-cell
RNA
sequencing
(scRNA-seq)
remains
a
challenge
cotton.We
established
efficient
transient
expression
using
isolated
from
taproots.
enables
large
numbers
uses
optimized
PEG-mediated
transfection
protocol.
highest
yield
(3.55
×
105/g)
viability
(93.3%)
were
obtained
roots
grown
hydroponics
72
h.
scRNA-seq.
efficiency
(80%)
was
achieved
when
as
described
above
transfected
with
20
μg
plasmid
min
solution
containing
200
mM
Ca2+.
Our
protoplast-based
is
various
applications,
including
validation
CRISPR
vectors,
protein
subcellular
localization
analysis,
protein-protein
interaction
studies.The
protocol
developed
this
study
stable,
versatile,
time-saving.
It
will
accelerate
genomics
molecular
breeding
cotton.
Horticulture Research,
Год журнала:
2025,
Номер
12(5)
Опубликована: Март 12, 2025
Lobed
leaves
are
advantageous
for
gas
exchange,
canopy
architecture,
and
high-density
planting;
however,
the
genetic
mechanisms
of
leaf
lobe
formation
in
Brassica
crops
remains
poorly
understood.
Here,
lob10.1,
our
previously
identified
major
QTL
controlling
presence/absence
lobes
B.
rapa
(AA),
was
fine
mapped
to
a
confidence
interval
69.8
kb.
REDUCED
COMPLEXITY
ORGAN
(BrRCO,
BraA10g032440.3c),
homeodomain
leucine
zipper
class
I
(HD
ZIP
I)
transcription
factor,
predicted
be
most
likely
candidate
gene
underlying
lob10.1.
Null
mutations
BrRCO
by
CRISPR/Cas9
lobed-leaf
parent
RcBr
over-expression
counter-part
near
isogenic
lines
(NILRcBr)
lead
entire
lobed
leaves,
respectively.
Analysis
evolution
revealed
that
A10.
RCO
functions
as
core
generally
negatively
selected
rapa.
Moreover,
function
negative
regulator
directly
binding
promoters
BrACP5
repressing
its
expression.
The
ACID
PHOSPHATASE
TYPE
5
(BrACP5)
subsequently
confirmed
VIGS-BrACP5
produced
RcBr.
This
study
involved
development
elucidated
new
pathway
BrRCO-BrACP5
module.
These
findings
provide
theoretical
basis
crops.
Agronomy,
Год журнала:
2023,
Номер
13(3), С. 755 - 755
Опубликована: Март 5, 2023
Cucurbits
(Cucurbitaceae)
include
major
horticultural
crops
with
high
nutritional
and
economic
value
that
also
serve
as
model
plants
for
studying
plant
development
crop
improvement.
Conventional
breeding
methods
have
made
important
contributions
to
the
production
of
cucurbit
but
led
a
bottleneck
because
narrow
genetic
bases
low
variation
rates
these
crops.
With
molecular
techniques,
innovations
in
germplasm
through
transgenesis
gene
editing
breakthroughs
breeding.
Although
transformation
techniques
has
lagged
behind
other
crops,
great
progress
been
recent
years.
Here,
we
summarize
advances
improving
efficiency
including
screening
application
physical
treatments,
morphogenic
genes,
selection
markers.
In
addition,
review
technology
CRISPR
(clustered
regularly
interspaced
short
palindromic
repeat)/Cas9
(CRISPR-associated
nuclease
9)-mediated
knockout
base
editing.
This
work
provides
reference
