othman, yasmin rofina husin nurain khalid centre for research...
TRANSCRIPT
Nur
ain
Hus
in, R
ofin
aYa
smin
Oth
man
, * N
orzu
laan
iKha
lid
Cen
tre
for R
esea
rch
in B
iote
chno
logy
for A
gric
ultu
re
(CEB
AR
) U
nive
rsity
Mal
aya
Mal
aysi
a
®In
trod
uctio
n &
Pro
blem
sta
tem
ent
®Pr
otoc
ol m
odifi
catio
n°
Cal
lus
indu
ctio
n°
Mai
ntai
nanc
e of
em
bryo
geni
c ce
ll su
spen
sion
cu
lture
s°
Dev
elop
men
t pro
cess
°Pl
ant r
egen
erat
ion
®Fi
eld
asse
ssm
ent
®C
oncl
usio
n®
Ack
now
ledg
emen
t
®Ba
nana
is o
ne o
f the
pri
ority
frui
t cro
p in
Mal
aysi
a®
Mus
a ac
umin
ata
cv b
eran
gan
is o
ne o
f the
ban
anas
gr
own
com
mer
cial
ly®
Mai
nly
as d
esse
rt, n
ow d
evel
oped
into
food
pro
duct
s®
Susc
etib
le to
bio
tic a
nd a
biot
ic s
tres
s®
Nee
d fo
r bio
tech
nolo
gies
for c
rop
impr
ovem
ent
®Em
bryo
geni
c ce
ll su
spen
sion
is a
n id
eal t
arge
t sys
tem
fo
r m
igro
prop
agat
ion
and
adop
ting
biot
echn
olog
ical
m
etho
d (tr
ansg
enic
s)
Mic
ropr
opag
atio
nvi
a sh
oot m
eris
tem
cultu
res
: ®
not e
ffici
ent f
or p
lant
pro
duct
ion
®la
bour
inte
nsiv
e®
not i
deal
targ
et ti
ssue
for p
lant
tran
sfor
mat
ion
Embr
yoge
nic
cell
susp
ensi
on :
®10
X m
ore
effic
ient
for p
lant
pro
duct
ion
®C
an b
e au
tom
ated
in b
iore
acto
rs®
Sing
le c
ell t
rans
form
atio
n
®C
urre
nt p
roto
cols
for b
anan
a so
mat
ic
embr
yoge
nesi
s ar
e lim
ited
by lo
w e
mbr
yo
germ
inat
ion,
low
pla
nt re
gene
ratio
n ra
tes
and
long
cul
ture
per
iod
®O
ptim
isat
ion
of m
ultip
licat
ion,
dev
elop
men
t an
d re
gene
ratio
n to
impr
ove
the
num
ber o
f pl
antle
ts re
cove
ry fr
om s
omat
ic e
mbr
yos.
®D
iffer
ent c
once
ntra
tions
of t
wo
amin
o ac
ids
(L-
glut
amin
e an
d L
-pro
line)
in d
evel
opm
enta
l m
edia
(liq
uid
and
solid
).®
Diff
eren
t pla
ting
dens
ity o
f em
bryo
s fo
r ge
rmin
atio
n
Expl
ants
pre
-trea
ted
on s
olid
med
ia w
ith
23µM
of 2
, 4-D
and
sub
sequ
ently
tran
sfer
red
to m
edia
with
10
µM 2
, 4-D
for 3
-4 m
onth
s
Embr
yoge
nic
callu
s in
duct
ion
Prop
agat
ion
med
ia
Mur
ashi
gean
d Sk
oog
(MS)
bas
al m
edia
1.
1 m
g/L
2,4-
D0.
25 m
g/L
zeat
in10
mg/
L as
corb
ic a
cid
20 g
/L s
ucro
se
Prop
agat
ion
of e
mbr
yoge
nic
cell
susp
ensi
on
®D
iffer
ent d
evel
opm
ent m
edia
con
sist
ing
of
vary
ing
conc
entr
atio
n of
pro
line
and
glyc
ine
in
liqui
d or
sol
id m
edia
wer
e te
sted
. ®
Prol
ine
and
glyc
ine
have
pro
mot
ive
effe
cts
for
embr
yo d
evel
opm
ent
®H
igh
conc
entr
atio
n of
pro
line
in li
quid
med
ia
how
ever
pro
ve to
indu
ce a
bnor
mal
ity in
the
embr
yo d
iffer
entia
tion
stag
e ®
Abn
orm
aliti
es o
bser
ved
incl
ude
form
atio
n of
ps
eudo
-rad
icle
stru
ctur
e, e
xten
sive
bro
wni
ng a
nd
stun
ted
grow
th w
hich
eve
ntua
lly c
ause
d th
e em
bryo
s to
be
necr
otic
.
Embr
yo D
evel
opm
ent M
edia
DE
GIJ
Abn
orm
al e
mbr
yos
in p
rolin
e-su
pple
men
ted
liqui
d M
3 m
edia
. Em
bryo
s w
ith
long
(A-B
) and
sho
rt (C
-D) p
seud
o-ra
dicl
e st
ruct
ure.
E) r
etar
ded
embr
yos
5 m
onth
s af
ter c
ultu
re in
dev
elop
men
t med
ia. F
) Em
bryo
s w
ith e
xten
sive
bro
wni
ng
and
brow
ning
cou
ple
with
root
-like
form
atio
n (G
). (H
) A d
etac
hed
root
-like
st
ruct
ure
and
(I) fu
sed
root
-like
str
uctu
re.
Con
trol
®D
evel
opin
g so
mat
ic e
mbr
yos
on s
olid
ified
M3
med
ia w
ere
cultu
red
with
diff
eren
t em
bryo
pla
ting
dens
ity®
200
µl o
f sus
pend
ed em
bryo
geni
ccel
l sus
pens
ion
wer
e di
sper
sed
on M
4GS
med
ia(M
S ba
sal
med
ium
, 400
mg/
l L-g
luta
min
e, 3
0 g/
l suc
rose
, 2
g/l P
hyta
gelTM
)in
thre
e cu
lture
den
sitie
s®
Cel
ls w
ere
eith
er s
prea
d ev
enly
thro
ugho
ut th
e pe
trid
ish
(low
den
sity
), or
par
tially
spr
ead
on 2
/3
(mod
erat
e de
nsity
) or 1
/3 o
f the
pet
ridi
sh (h
igh
dens
ity).
Com
pari
son
betw
een
diffe
rent
em
bryo
cul
ture
den
sity
on
solid
med
ia a
nd
deve
lopm
enta
l sta
ges
of m
atur
atio
n af
ter 3
mon
ths
in c
ultu
re
low
den
sity
mod
erat
ede
nsity
high
den
sity
Cul
ture
den
sity
stu
dies
sho
wed
that
hig
hly
dens
e em
bryo
inoc
ulat
ion
deve
lope
d po
orer
in
term
of d
urat
ion
take
n to
mat
ure
and
time
take
n to
sub
cultu
re a
s co
mpa
red
to le
ss d
ense
in
ocul
atio
n
®Th
e m
inim
um d
urat
ion
need
ed to
obt
ain
mat
ure
embr
yos
in li
quid
med
ia w
as s
how
n to
be
ear
lier (
1.5
mon
th) t
han
that
nee
ded
in s
olid
m
edia
(2.5
mon
ths)
. ®
In a
ll m
edia
test
ed, r
egen
erat
ed p
lant
lets
ob
tain
ed in
liqu
id M
3 m
edia
was
hig
her t
han
that
obt
aine
d in
sol
id c
ount
erpa
rts.
Com
pari
son
in th
e nu
mbe
r of p
lant
lets
obt
aine
d in
di
ffere
nt d
evel
opm
ent m
edia
Mor
pho-
hist
odiff
eren
tiatio
nst
ages
of
bana
na (c
vbe
rang
an) s
omat
ic e
mbr
yos.
Ro
oted
ber
anga
npl
antle
. Acc
limat
ized
be
rang
anpl
antle
ts re
ady
to b
e pl
ante
d in
the
field
.
Tiss
ue c
ultu
re d
eriv
ed p
lant
s in
the
field
6 m
onth
s ; o
il pa
lm
Ope
n fie
ld ;
2 m
onth
s
-1001020304050
56
78
910
percentage of plants (%)
no. o
f han
ds p
er b
unch
trea
tmen
t
cont
rol
-1001020304050
56
78
910
percentage of plants (%)
no o
f han
ds p
er b
unch
trea
tmen
t
cont
rol
Ope
n fie
ld
IN B
ETW
EEN
OIL
PA
LM
IN B
ETW
EEN
OIL
PA
LM
TREE
STR
EES
OPE
N F
IELD
-10010203040506070
23
45
percentage of plants (%)
heig
ht (m
eter
)
trea
tmen
t
cont
rol
-20
-1001020304050607080
23
45
percentage of plants (%)
heig
ht (m
eter
)
trea
tmen
t
cont
rol
IN B
ETW
EEN
OIL
PA
LM
IN B
ETW
EEN
OIL
PA
LM
TREE
STR
EES
OPE
N F
IELD
01020304050607080
9-11
cm
12-1
4 cm
15 c
m
percentage of plants (%)
pseu
dost
em d
iam
eter
(cm
)
trea
tmen
t
cont
rol
-1001020304050607080
9-11
cm
12-1
4 cm
15-1
7 cm
percentage of plants (%)
pseu
dost
em d
iam
eter
(cm
)
trea
tmen
t
cont
rol
IN B
ETW
EEN
OIL
PA
LM
IN B
ETW
EEN
OIL
PA
LM
TREE
STR
EES
OPE
N F
IELD
-1001020304050
percentage of plants (%)
bunc
h w
eigh
t (kg
)
trea
tmen
t
cont
rol
-50510152025303540
12-1
4 kg
15-1
7 kg
18-2
0 kg
21-2
3 kg
24-2
6 kg
27-2
9 kg
30-3
2 kg
percentage of plants (%)
bunc
h w
eigh
t (kg
)
trea
tmen
t
cont
rol
IN B
ETW
EEN
OIL
PA
LM
IN B
ETW
EEN
OIL
PA
LM
TREE
STR
EES
OPE
N F
IELD
-20
-100102030405060
56
78
9
percentage of plants (%)
flow
erin
g tim
e (m
onth
)
trea
tmen
t
cont
rol
-20
-100102030405060
56
78
9
percentage of plants (%)
flow
erin
g tim
e (m
onth
)
trea
tmen
t
cont
rol
IN B
ETW
EEN
OIL
PA
LM
IN B
ETW
EEN
OIL
PA
LM
TREE
STR
EES
OPE
N F
IELD
-1001020304050607080
89
1011
percentage of plants (%)
harv
estin
g tim
e (m
onth
)
trea
tmen
t
cont
rol
-10010203040506070
89
1011
percentage of plants (%)
harv
estin
g tim
e (m
onth
)
trea
tmen
t
cont
rol
¸2,
4d p
ulse
impo
rtan
t for
em
bryo
geni
cca
llus
indu
ctio
n¸
Liqu
id m
edia
for d
evel
opm
ent c
an e
nhan
ce e
mbr
yo
prod
uctio
n¸
Low
to m
oder
ate
embr
yo c
ultu
re d
ensi
ty c
an
impr
ove
mat
urat
ion
peri
od¸
Fiel
d as
sess
men
t-ba
sed
on a
nnov
aof
the
aver
age
resu
lts s
how
ed in
sign
ifica
nt d
iffer
ence
bet
wee
n m
eris
tem
and
cell
susp
ensi
on d
eriv
ed p
lant
s fo
r p-
valu
e m
ore
than
0.0
5 (a
t 95%
con
fiden
ce le
vel)
®Pr
of. D
r. Ro
fina
Yasm
in O
thm
an®
Nur
ain
Hus
in-p
ost g
rad
stud
ent
®M
r.Tan
pr
ovid
ing
farm
s®
Nur
Saf
ina
tech
nica
l ass
ista
nce
®U
niv
Mal
aya
and
Min
istr
y of
Sc
ienc
e, T
echn
olog
y an
d In
nova
tion
fund
ing
and
faci
litie
s
Nor
zula
aniK
halid
Uni
vers
ity M
alay
ala
ni@
um.e
du.m
y
Nor
zula
aniK
halid
Uni
vers
ity M
alay
ala
ni@
um.e
du.m
y
