29
Tomato
(
Lycopersicon
esculentum,
Mill
.
)
plant
regeneration
in
absence
of
exogenous
growth
regulators
Plana,
D
.
,
Alvarez,
M
.
,
Florido,
M
.
,
Lara,
R
.
M
.
,
Moya,
C
.
INCA,
Genetic
and
Plant
Breeding
Dept
.
,
San
José
de
Las
Lajas,
La
Habana,
Cuba
.
Email:
dagmara@inca
.
edu
.
cu
Development
of
an
in
vitro
regeneration
system
without
exogenous
growth
regulators
is
important
for
genetic
manipulations
of
particular
plant
species
(Herrera
-
Estrella,
1999)
.
The
objective
of
the
present
work
was
to
obtain
tomato
regenerated
plants
independently
of
exogenous
growth
regulators
in
the
culture
medium
.
Dried
mature
seeds
of
tomato
(
Lycopersicon
esculentum,
Mill
.
)
from
five
cultivars
were
used
as
the
source
of
explants
.
After
sterilization,
the
seeds
were
sown
on
filter
paper
that
had
been
wetted
with
sterilized
water
and
precultured
for
zero
to
three
days
in
the
light
at
25
o
C
.
Precultured
seeds
were
cut
into
two
parts
(Ezura
1993);
the
portion
consisting
of
the
proximal
part
of
the
hypocotyls
of
the
embryo
was
cultured
on
Murashigue
and
Skoog
(MS)
medium
(1962),
with
30
g
.
L
-
1
commercial
sugar,
2g
.
L
-
1
gel
rite,
mio
-
inositol
100
mg
.
L
-
1
,
thiamine
4
mg
.
L
-
1
and
pH
adjusted
to
5,7
before
sterilizing
by
autoclave
at
121
0
C
.
The
number
of
regenerated
adventitious
shoots
were
counted
for
three
weeks
(Table
1)
.
The
elongated
shoots
were
excised
individually
from
the
explants
and
subcultured
on
MS
medium
for
rooting
.
The
plantlets
were
transplanted
into
pots
with
a
mixture
of
litonite
and
soil
(1:1)
for
growing
under
greenhouse
conditions
.
All
experiments
were
repeated
twice
.
The
percentage
of
explants
with
adventitious
shoots
ranged
from
15
to
69%
at
the
third
week
(Table
1)
.
At
the
same
time
the
adventitious
shoots
were
excised
for
rooting
and
the
calluses
subcultivated
on
fresh
medium
to
obtain
more
adventitious
shoots
.
The
Amalia
cv
.
shoots
scored
for
up
to
2
.
8
two
weeks
after
the
first
subculture
.
Morphological
characteristics
of
regenerated
and
nonregenerated
plants
and
fruits
were
similar;
on
the
other
hand,
forty
regenerated
plants
showed
no
variations
in
chromosome
number
.
The
advantages
of
this
regeneration
method
are:
it
does
not
employ
exogenous
growth
regulators,
it
has
feasible
handling,
and
also
regenerated
plants
are
obtained
in
a
short
time
.
Literature
cited:
Ezura,
H
.
;
Nishimiya,
S
.
;
Kasumi,
M
.
(1993)
.
Efficient
regeneration
of
plant
independent
of
exogeneous
growth
regulators
in
bell
pepper
(
Capsicum
annumm,
L
.
)
.
Plant
Cell
Reports
12:676
-
680
.
Herrera
-
Estrella,
L
.
(1999)
.
Plant
genetic
transformation:
technology
and
applications
.
In:
Métodos
en
Biología
Molécular
Vegetal
.
First
International
INIA
-
Carrillanca
Workshop
and
Course
on
Plant
Biothecnology
.
58
-
65p
.
Murashige,
T
.
and
Skoog,
F
.
(1962)
.
A
revised
medium
for
rapid
growth
and
biassays
with
the
tobacco
tissues
cultures
.
Physiol
.
Plantarum
.
15:
473
-
497
.
Table
1,
next
page