Transgene‐free genome editing in poplar
New Phytologist,
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 22, 2025
Summary
Precise
gene‐editing
methods
are
valuable
tools
to
enhance
genetic
traits.
Gene
editing
is
commonly
achieved
via
stable
integration
of
a
cassette
in
the
plant's
genome.
However,
this
technique
unfavorable
for
field
applications,
especially
vegetatively
propagated
plants,
such
as
many
commercial
tree
species,
where
cannot
be
segregated
away
without
breaking
constitution
elite
variety.
Here,
we
describe
an
efficient
method
generating
gene‐edited
Populus
tremula
×
P.
alba
(poplar)
trees
incorporating
foreign
DNA
into
its
Using
Agrobacterium
tumefaciens
,
expressed
base‐editing
construct
targeting
CCoAOMT1
along
with
ALS
genes
positive
selection
on
chlorsulfuron‐containing
medium.
About
50%
regenerated
shoots
were
derived
from
transient
transformation
and
free
T‐DNA.
Overall,
7%
chlorsulfuron‐resistant
T‐DNA
free,
edited
gene
nonchimeric.
Long‐read
whole‐genome
sequencing
confirmed
absence
any
tested
lines.
Additionally,
evaluated
CodA
negative
marker
eliminate
lines
that
stably
incorporated
their
Although
latter
not
essential
selecting
transgene‐free,
shoots,
it
may
prove
other
genotypes
or
varieties.
Язык: Английский
An efficient mRNA delivery system for genome editing in plants
Plant Biotechnology Journal,
Год журнала:
2025,
Номер
unknown
Опубликована: Фев. 10, 2025
Summary
Transgene‐free
genome
editing
is
important
for
crop
improvement
as
it
reduces
unanticipated
genomic
changes.
While
mRNA
delivery
systems
offer
a
powerful
method
achieving
transgene‐free
editing,
they
remain
inefficient
and
challenging
in
plants.
Here
we
describe
an
efficient
system
plants
with
substantially
improved
efficiency.
By
optimizing
the
5′
untranslated
regions
(5′UTRs)
poly(A)
tails
of
vitro
‐transcribed
(IVT)
mRNAs
coating
protamine
during
particle
bombardment,
have
developed
optimized
termed
v2_TMV/DEN2.
This
enhanced
efficiencies
knock‐out,
A‐to‐G
C‐to‐T
base
by
average
4.7‐,
3.4‐
2.5‐fold
at
various
endogenous
sites
compared
plasmid‐based
transient
via
bombardment
rice
suspension
cells
wheat
immature
embryos
48
h
post‐transformation.
Furthermore,
obtained
edited
5.0–180.8%
26.1–26.2%
using
v2_TMV/DEN2
wheat,
respectively,
0.0–43.2%
4.7–10.4%
plasmids.
Our
study
provides
convenient
Язык: Английский
Advancements in delivery strategies and non-tissue culture regeneration systems for plant genetic transformation
Advanced Biotechnology,
Год журнала:
2024,
Номер
2(4)
Опубликована: Сен. 26, 2024
Язык: Английский
Transgenic poplar for resistance against pest and pathogen attack in forests: an overview
Frontiers in Forests and Global Change,
Год журнала:
2024,
Номер
7
Опубликована: Дек. 18, 2024
Forests
are
potential
habitats
for
immense
terrestrial
ecosystems
and
aquatic
biodiversity,
performing
an
essential
role
in
ecological
preservation
regulation
of
climate.
The
anthropogenic
pressures
on
the
forests
lead
to
forest
loss,
fragmentation
degradation.
Requirements
sustainable
methodologies
protection
utmost
priority
under
climate
change
regime.
Among
trees,
poplar
trees
(
Populus
L.)
have
attracted
attention
global
forestry
as
a
promising
material
improving
quality
quantity
urban
landscapes.
These
plants
provide
wood,
which
can
be
utilized
raw
resources
paper
industry
source
biofuel.
However,
several
biotic
stresses,
such
attacks
by
pests
pathogens,
severely
affect
production
productivity.
improvement
through
conventional
tree
breeding
methods
is
restricted
due
their
long-life
cycles
lack
suitable
donors
with
resistance
genes.
has
been
model
plant
studying
gene
functions
its
highly
efficient
genetic
transformation
capabilities.
present
review
will
comprehensive
overview
pest
pathogen
poplar,
focusing
infection
mechanisms,
transmission
routes,
control
strategies.
Additionally,
it
examine
most
widely
used
(gene
gun-mediated,
Agrobacterium
tumefaciens
-mediated,
protoplast
transformation,
micro-RNA
mediated
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)-associated
(CRISPR-Cas)
systems
RNA
interference)
tolerance
against
pathogens
attack.
Furthermore,
delve
into
prospects,
challenges,
recent
advances
molecular
biology
tools
safe
application
improve
insect
trees.
Finally,
regeneration
transgenic
enhanced
resistance,
developed
various
engineering
techniques,
discussed.
Язык: Английский
Enhanced antifungal activity of sugarcane cv. NCo310 expressing chimeric chitinase 42
Physiological and Molecular Plant Pathology,
Год журнала:
2024,
Номер
133, С. 102341 - 102341
Опубликована: Июнь 17, 2024
Язык: Английский
Applications of CRISPR Technologies in Forestry and Molecular Wood Biotechnology
International Journal of Molecular Sciences,
Год журнала:
2024,
Номер
25(21), С. 11792 - 11792
Опубликована: Ноя. 2, 2024
Forests
worldwide
are
under
increasing
pressure
from
climate
change
and
emerging
diseases,
threatening
their
vital
ecological
economic
roles.
Traditional
breeding
approaches,
while
valuable,
inherently
slow
limited
by
the
long
generation
times
existing
genetic
variation
of
trees.
CRISPR
technologies
offer
a
transformative
solution,
enabling
precise
efficient
genome
editing
to
accelerate
development
climate-resilient
productive
forests.
This
review
provides
comprehensive
overview
applications
in
forestry,
exploring
its
potential
for
enhancing
disease
resistance,
improving
abiotic
stress
tolerance,
modifying
wood
properties,
accelerating
growth.
We
discuss
mechanisms
various
systems,
including
base
editing,
prime
multiplexing
strategies.
Additionally,
we
highlight
recent
advances
overcoming
key
challenges
such
as
reagent
delivery
plant
regeneration,
which
crucial
successful
implementation
also
delve
into
ethical
considerations
using
gene
drive
population-level
alterations,
well
importance
containment
strategies
mitigating
risks.
emphasizes
need
continued
research,
technological
advancements,
extensive
long-term
field
trials,
public
engagement,
responsible
innovation
fully
harness
power
shaping
sustainable
future
Язык: Английский
CRISPR/Cas基因组编辑在木本植物性状改良中的应用
Xuening Yuan,
Fengge Yao,
Yi An
и другие.
Chinese Science Bulletin (Chinese Version),
Год журнала:
2023,
Номер
unknown
Опубликована: Дек. 1, 2023
Genome editing in cells of apple cultivar ‘Fuji’ using Ggeminivirus-derived replicons for transient expression of CRISPR/Cas9 components.
Plant Biotechnology,
Год журнала:
2024,
Номер
41(4), С. 425 - 436
Опубликована: Ноя. 21, 2024
The
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)/CRISPR-associated
protein
9
(Cas9)
system
has
been
used
for
genome
editing
in
various
fruit
trees,
including
apple
(Malus
×
domestica).
In
previous
studies,
transfer
DNA
(T-DNA)
expressing
tools,
Streptococcus
pyogenes
Cas9
(SpCas9)
and
single
guide
RNA
(sgRNA),
was
stably
integrated
into
the
via
Agrobacterium-mediated
transformation.
However,
due
to
self-incompatibility,
long
generation
period,
high
heterozygosity
of
apple,
removing
only
T-DNA
from
by
crossbreeding
while
maintaining
introduced
varietal
trait
is
difficult.
Therefore,
an
efficient
SpCas9-sgRNA
delivery
without
transgene
insertion
required
apple.
this
study,
we
geminivirus-derived
replicons
(GVRs)
transient
expression
tools.
Small
vectors
were
deconstructed
splitting
elements
necessary
production
GVRs
bean
yellow
dwarf
virus
two
vectors.
Production
using
these
demonstrated
Arabidopsis
cells.
Genome
improved
GVR-producing
with
tools
protoplasts.
use
SpCas9
sgRNA
leaves
levels
sgRNA,
enabling
detection
targeted
mutations
endogenous
genome.
These
findings
demonstrate
utility
gene
It
can
be
expected
that
our
GVR-based
technology
potential
transgene-free
Язык: Английский