Effect of salinity stress on some date palm cultivars during
proliferation stage in vitro
(Received: 15.05.2008; Accepted: 20.05.2008)
S. F. El-Sharabasy;** W. H. Wanas * and A. Y. Al-Kerdany **
*Faculty of Agriculture,Ain Shams University
** Central Laboratory of Date Palm Research and Development, ARC
ABSTRACT
This study was carried out at the the Central Laboratory of Date Palm Research and
Development ,Agricultural Research Center ,Ministry of Agriculture ,Giza ,Egypt during 2005 and
2006 to evaluate an in vitro technique for salt tolerance of three date palm cultivars Samani, Sewy
and Bartamuda cultured on MS medium with 0.0 ,4000,8000and 12000 p.p.m NaCl after 12 weeks
for three subculturs. The effect of salinity on shoot length showed insignificant differences under
0.0 p.p.m, but it increased significantly for the three cultivars especially Bartamuda cv. to 4000
p.p.m then decreased significantly at 8000 and 12000 p.p.m NaCl. Number of shoots showed the
high significant value at 4000 p.p.m for the Bartamuda cv., however, at 8000 and 12000 p.p.m it
decreased and the differences between Sewy and Bartamda cvs. Were insignificant follwed by
Samani cv. The number of burned leaves was the highest at 12000 p.p.m NaCl for the Sewy and
Samani cvs. as compared with Bartmuda cv. Significant differences was recorded between
Bartamuda and the other two cultivars. The growth vigor of Bartmuda was still the highest even at
12000 p.p.m. NaCl.
Key words: Date palm, in vitro proliferation, salinity strees.
INTRODUCTION
S
alinity is one of the major environmental
stresses that challenge plant growth and
crop productivity worldwide (Sen and
Mohammed, 1994). Producing sustainable and
profitable crops under these conditions needs
technological and biological approaches,
including selection of new, more salt tolerant
cultivars of plants using conventional breeding
programs or tissue culture techniques (Al
Mansoori et al., 2006). Tissue Culture
technique is an excellent tool for elucidating
the correlation between organizational level
and salt tolerance because of the possibility it
offers for studying the physiology of intact
plantlets together with that of organs and
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
single cells using homogeneous plant material
under uniform environmental conditionse.
(Mills., 1989).Water availability is one of the
principle limitations of crop production
particularly in the arid and semi arid regions
where date palm is predominantly grown. The
use of in vitro cultures to study stress
responses is based on the fact that in vitro
cultured cells behave similarly to cell of intact
plants subjected to water deficit and salinity
conditions (Attree et al.,1991). Therefore, in
the present study proliferation stage in vitro of
Samani, Sewy and Bartamuda date palm cvs
were evaluated for salinity tolerance
represented by NaCl levels in culture medium.
274
S. F. El-Sharabasy and W. H .Wanas
MATERIALS AND METHODS
This study was carried out at the Tissue
Culture Laboratory of the Central Laboratory of
Date Palm Research and Development
,Agricultural Research Center ,Ministry of
Agriculture ,Giza ,Egypt during 2005 and 2006
to evaluate in vitro technique for salt tolerance
in the three date palm cultivars (Samani, Sewy
and Bartamuda) A small cluster consisted of 46 developed shoots from date palm cvs Samani
(SM), Sewy (SE) and Bartamuda (BA) was
cultured on MS basal nutrient medium
supplemented with 0.0, 4000, 8000 and 12000
p.p.m NaCl, 30gl-1 sucrose and 3gl-1 phytagel.
Media were dispensed into the culture vessels
in aliquots of 35ml per small jars (150 ml) and
capped with polypropylene closures. Media
were then autoclaved for 20 min. at 121°C and
1.2 Kg/cm2. All culture vessels were incubated
in the growth room for 6 weeks under (27±2°C)
and 16 hr illumination of 3000 Lux (Mater,
1986).
The following data were recorded after 12
weeks:
1- Average shoot length (mm) / cluster.
2- Average number of shoots /cluster.
3- Average number of burned leaves / cluster.
4- Growth vigor [G.V was calculated visually
as scores according to the method described by
(Pottino, 1981)].
a- Negative growth
=1 (The lowest
growth)
b- Below average growth = 2
c- Average growth
=3
d- Above average growth = 4
e- Good growth
=5 (The highest growth).
All of the experiment treatments and
cultivars were arranged in a factorial expe-
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
riment in a completely randomized design (3
cultivars X 4 treatments X 3 replicates) with
one shoot for each replicate .Duncan multiple
range test at 5% level was used to verify the
differences between means of the treatments
(Snedecor and Cochran, 1982) .
RESULTS AND DISCUSSION
I. Effect of NaCl treatments on shoot length
(cm) of some date palm cvs. in vitro during
proliferation stage
The data presented in Fig. (1) and Table
(1) showed the effect of salinity treatments
NaCl (4000, 8000 and 12000 p.p.m) on shoot
length for some date palm cvs after 12 weeks.
The data on the effect of salinity indicated that
NaCl treatment at 4000 p.p.m gave the highest
mean value (5.13) followed by control (3.43)
then 8000 (3.76) and 12000p.p.m (3.11). The
highest mean shoot length of cultivars was
obtained for Bartamuda cv (4.38) followed by
Samani and Sewy cvs (3.61 and 3.58,
respectively) and the difference between
Bartamuda and the other cvs was significant.
The interactions between different cultivars and
salinity concentrations showed insignificant
differences in the average shoot length under
0.0 p.p.m but it increased significantly for the
three cvs especially BA cv to 4000 p.p.m then
decreased significantly at 8000 and 12000
p.p.m .The highest value was obtained at 4000
p.p.m for the BA cv (5.69cm) with a significant
difference from SM cv .While the lowest value
was recorded at 12.000 p.p.m (2.78cm) for SE
cv .In general, results revealed that, low level
of salinity (4000 p.p.m) in culture medium
enhanced shoot length of the three cultivars
significantly; BA was significantly superior in
this respect.
Salinity stress for date palm in vitro
275
Table (1): Effect of NaCl salt treatments on shoot length (cm) of some date palm cvs after 12
weeks in vitro during proliferation stage .
NaCl Salt concentration (p.p.m)
Cultivar
SAMANI
SEWY
BARTAMUDA
Means of treatments
CONTROL
4000
8000
12000
Means of cvs.
3.58c
4.60b
3.29cd
2.97cd
3.61B
3.20cd
3.51c
3.43BC-
5.09ab
5.69a
5.13A-
3.23cd
4.77b
3.76B-
2.78d
3.57c
3.11C-
3.58B
4.38A
Samani
Sewy
Bartamuda
6
5.5
5
Av g. shoot
le ngth
4.5
4
3.5
Bartamuda
Sewy
3
Co
ntr
ol
40
80
00 p
00 p
.p.
m
.p
Samani
12
00
.m
0 p.
p.m
Salinity tre atme nts
Fig. (1): Effect of NaCl salt treatments on shoot length (cm) of some date palm cvs after 12 weeks
in vitro during proliferation stage.
Effect of NaCl salt treatments on number of
shoots of some date palm cvs in vitro during
proliferation stage.
The data presented in Table (2) and Fig
(2) showed the effect of salinity treatments on
number of shoots of some date palm cvs. after
12 weeks. The data on the effect of salinity
indicated that NaCl treatment 4000 p.p.m gave
the highest mean value (9.50) then decreased
at 8000 p.p.m (7.83) and 12000 p.p.m (5.50)
with a significant differences between them and
control (5.41), without significant difference
between them. The highest mean of cultivars
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
was obtained for BA cv (8.25) followed by SE
cv (7.0) and SM cv (5.93) cvs and the
difference between BA and the other cvs was
significant. The interactions between cultivars
and salinity concentrations were significant for
the number of shoots. The highest significant
value was obtained at 4000 p.p.m by the BA cv
(11.00) .However at 8000 and 12000 p.p.m the
differences between SE and BA cvs were
insignificant. The lowest value was recorded at
12.000 p.p.m (4.500) for SM cv without a
significant difference between control 0.0 p.p.m
and this concentration.
276
S. F. El-Sharabasy and W. H .Wanas
Table (2): Effect of NaCl salt treatments on number of shoots of some date palm cvs after 12
weeks in vitro during proliferation stage.
NaCl Salt concentration (p.p.m)
4000
8000
12000
8.25b
6.25cd
4.50e
9.25b
8.50b
5.50de
11.00a
8.75b
6.50cd
9.50A7.83B5.50C-
Cultivar
SAMANI
SEWY
BARTAMUDA
Means of treatments
CONTROL
4.75e
4.75e
6.75c
5.41C-
Samani
Sewy
Means of cvs.
5.93C
7.00B
8.25A
Bartamuda
11
10
9
8
Number of shoots
7
6
5
Bartamuda
Sewy
4
Co
ntr
ol
40
00
80
p.p
00
p.p
.m
.m
Samani
12
00
0p.
p.m
Salinity treatment
Fig. (2): Effect of NaCl salt treatments on number of shoots of some date palm cvs. after 12 weeks
in vitro during proliferation stage.
Effect of NaCl salt treatments on number of
burned leaves for some date palm cvs in vitro
during proliferation stage.
The data presented in Table (3) and Fig
(3) showed the effect of salinity treatments on
number of burned leaves of some date palm cvs
after 12 weeks in vitro. The data on the effect
of salinity indicated that NaCl treatment 12000
p.p.m gave the highest significant mean value
(1.91) followed by 8000 p.p.m (1.00) with a
significant differences among them, then 4000
p.p.m (1.00) and control (1.00), without
significant difference between them. The
highest mean number of burned leaves of
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
cultivars was obtained for SE (1.25) and SM
(1.25) cvs then BA cv (1.18), without
significant differences among the cultivars. The
interactions between cultivars and salinity
concentrations were significant for the number
of burned leaves The highest number of burned
leaves was obtained at 12000 p.p.m for the SE
and SM cvs (2.00) as compared with BA cv
(1.75) with a significant difference between BA
and the other two cvs .The rest of values under
all concentrations recorded (1.00) as a number
of burned leaves for the three cvs without any
significant differences among them.
Salinity stress for date palm in vitro
277
Table (3): Effect of NaCl salt treatments on number of burned leaves for some date palm vs. after
12 weeks in vitro during proliferation stage.
Cultivar
SAMANI
SEWY
BARTAMUDA
Means of treatments
NaCl Salt concentration (p.p.m)
4000
8000
12000
1.00c
1.00c
2.00a
1.00c
1.00c
2.00a
1.00c
1.00c
1.75b
1.00B1.00B1.91A-
CONTROL
1.00c
1.00c
1.00c
1.00B-
Samani
Sewy
Means of cvs.
1.25A
1.25A
1.18A
Bartamuda
2
1.8
1.6
Number of
burned leaves
1.4
1.2
1
Bartamuda
Sewy
0.8
Co
nt r
ol
40
00
80
p.p
00
p.p
.m
.m
Samani
12
00
0p
.p.
m
salinity treatments
Fig. (3): Effect of NaCl salt treatments on number of burned leaves for some date palm cvs. after
12 weeks in vitro during proliferation stage.
Effect of NaCl salt treatments on growth
vigor of some date palm cvs in vitro during
proliferation stage
The data presented in Table (4) and Fig
(4) showed the effect of salinity treatments on
growth vigor of some date palm cvs after 12
weeks in vitro. The effect of salinity indicated
that growth vigor increased significantly at
4000 p.p.m (4.58) as compared to all other
treatments. Afterwards, a significant gradual
decrease in growth vigor took place at 8000 and
12000 p.p.m (3.58 and 2.75) where the latest
value was insignificantly different from the
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
control .The highest mean growth vigor of
cultivars was obtained by BA cv (3.87)
followed by SE (3.31) then SM (2.93) cv and
the differences among them were significant.
The interactions between cultivars and salinity
concentrations showed that the highest
significant value of growth vigor was obtained
at 4000 p.p.m for the BA cv (5.00) followed by
SE cv (4.75) without a significant difference
between them. The lowest value of growth
vigor was recorded at 0.0 p.p.m (2.00) for SM
cv .The growth vigor of BA still the highest
even at 12000 p.p.m
278
S. F. El-Sharabasy and W. H .Wanas
Table (4): Effect of NaCl salt treatments on growth vigor of some date palm cvs after 12 weeks in
vitro during proliferation stage.
NaCl Salt concentration (p.p.m)
Cultivar
CONTROL
4000
8000
12000
Means of cvs.
SAMANI
2.00d
4.00b
3.25c
2.50d
2.93C
SEWY
2.50d
4.75a
3.50bc
2.50d
3.31B
BARTAMUDA
3.25c
5.00a
4.00b
3.25c
3.87A
Means of treatments
2.58C-
4.58A-
3.58B-
2.75C-
Samani
Sewy
Bartamuda
5
4.5
4
Growth vigor
3.5
3
2.5
Bartamuda
Sewy
2
Co
ntr
ol
40
00 p
80
00 p
.p.
m
.p.
m
Samani
12
00
0 p.
p.m
Salinity treatments
Fig. (4): Effect of NaCl salt treatments on growth vigor for some date palm cvs in vitro during
proliferation stage.
Results revealed that low level of salinity
(4000 p.p.m) in culture medium significantly
enhanced shoot length for the three date palm
cultivars and BA cv was significantly superior
in this respect. High salinity levels of 8000 and
12000 p.p.m NaCl caused a decrease in shoot
length and number of shoots and also the
growth vigor. Al-Naber et al., (1998) stated that
growth behavior of Atriplex nummularia
plantlets under 3 concentrations of NaCl (0.1,
0.2 and 0.3 M) showed
no significant
differences for number or length of shoots,
however, number of leaves per plantlet
decreased with increasing levels of salinity and
drought stresses. Also, Shilbi and Al-Juboory,
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
(2002) mentioned that salinity and water deficit
significantly reduced shoot growth and rooting
at treatments higher than 75 mM of NaCl. The
same were published by Sidky, (2004) and ElAdawe, (2005) who reported that high salinity
levels of 4000, 8000 and 12000 p.p.m
decreased shoot length, number of roots and
root length of plantlets.
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El-Adawe, A.S. (2005). Using of in vitro
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Mater, A.A. (1986). In vitro propagation of
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Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.
280
S. F. El-Sharabasy and W. H .Wanas
Arab J. Biotech., Vol. 11, No. (2) July (2008): 273-280.