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RESEARCH PAPER
Abstract
In the present investigation, three tomato cultivars Pusa Ruby, Vaibhav and Arkamegali (Solanum lycopersicum
L.) were selected for multiple shoot induction. The explants cotyledon, hypocotyl and epicotyl of all the cultivars
of tomato were cultured on MS medium fortified with different concentrations of BAP (1.0 to 4.0 mg/l), TDZ (1.0
to 4.0 mg/l), Zeatin (0.5 to 1.0 mg/l), AS (5.0 to 20 mg/l) and 0.1 mg/l IAA . Adventitious shoot buds were
induced at the cut ends of the explants after three weeks of culture. The shoots were induced from cotyledon,
hypocotyl and epicotyl explants on different concentrations of growth regulators (RM 1 to RM 10). Among ten
media combinations tested, RM 9 was the best combination for induction of shoots from cotyledon, hypocotyl
and epicotyl explants compared to RM 2. The highest number of shoots was induced on 0.5 mg/l Zn + 10 mg/l AS
+ 0.1 mg/l IAA (RM 9) followed by 2.0 mg/l BAP + 10 mg/l AS + 0.1mg/l IAA (RM 2). Among the three genotypes
tested Pusa ruby showed highest number of shoots per explant followed by Vaibhav and Arkamegali. Highest
number of adventitious shoots was induced from cotyledonory compared to hypocotyl and epicotyl explants in
Pusa ruby and Arka meghali, where as Vaibhav induced highest shoots from epicotyl explants. Rooting was
achieved on MS medium supplemented with 0.1 mg/l IAA in all the genotypes. Hence the plant regeneration was
found to be influenced by the genotype, different explants with hormonal combinations in the media.
* Corresponding
Introduction
consuming.
with
Hence,
breeding
associated
1992),
and
somatic
embryogenesis
(Kaprakis
MS-medium
(Murashige
and
Skoog,
1962)
Explant selection
factors
influencing
high
efficient
and
rapid
days.
growth
parameters
like
regeneration
percentage,
shoot
hormones
in
the
morphogenetic
Table 1. Combination of various growth hormones used in association with MS basal medium for
standardization of regeneration in tomato cultivars.
Media code
Nutrient medium
BAP (mg/L)
TDZ (mg/L)
ZR (mg/L)
IAA (mg/L)
RM 1
MS
1.0
0.1
RM 2
MS
2.0
10
0.1
RM 3
MS
3.0
15
0.1
RM 4
MS
4.0
20
RM 5
MS
1.0
0.1
RM 6
MS
2.0
10
0.1
RM 7
MS
3.0
15
0.1
RM 8
MS
4.0
20
0.1
RM 9
MS
10
0.5
0.1
RM 10
MS
15
1.0
0.1
0.1
Table 2. Percentage of regeneration in three cultivars of tomato in MS medium supplemented with growth
regulators.
Media
Arka Meghali
Hypocotyl
RM 1
2.880.03
Vaibhav
Epicotyl
Cotyledon
2.620.03 41.550.05
Mean
15.68
Hypocotyl
d
Epicotyl
Pusa Ruby
Cotyledon
5.070.03 32.850.05
7.140.04
Mean
15.02
Hypocotyl
64.560.03
53.950.03
76.490.04
91.590.04
90.720.02
95.850.04
93.55
83.563.27
80.173.33
89.542.67
84.42
69.824.47
59.984.22
73.795.20
50.934.42
30.414.42
56.755.54
46.03
86.503.05
84.722.67
86.503.05
85.91
74.823.06
79.625.16
80.174.22
8.393.33
4.573.51
7.575.77
6.84
92.700.05
92.850.05
96.950.03
96.95
88.790.04
85.140.04
94.960.02
83.68
RM 3
76.59
57.23
RM 4
30.414.42 50.123.33
RM 5
39.96
4.573.33
3.373.27 16.985.33
8.31
RM 6
81.854.47
71.785.33 84.722.67
79.45
RM 7
RM 8
3.373.51
3.373.51
3.80
66.24
73.72
7.084.71
4.61
RM 9
93.23
RM 10
1.842.66
2.57
87.73
90.72
3.373.51
62.30
2.493.33
1.842.94
5.976.29
39.43
3.614.47
82.13
Mean
56.33
51.16
63.84
SEm
0.01
0.01
0.02
0.01
0.02
0.02
0.02
0.01
0.02
CD
0.07
0.05
0.09
0.06
0.09
0.08
0.10
0.07
0.11
0.04
0.04
0.05
0.03
0.05
0.04
0.05
0.04
0.07
57.1
44.40
56.93
47.87
Mean
37.765.16
Cotyledon
RM 2
Mean
Epicotyl
49.73
71.32
65.88
75.96
65.00
31.90
89.47
67.86
78.20
f
j
72.74
49.09
19.38
77.20
d
f
71.41
4.67
92.49
87.49
89.63
71.05
(1%)
CV
Data - Mean SE of 10 replications. The data were taken 3 weeks after culture. The values followed by same
letters are not significantly different @ P= 0.05.
cent
sodium
hypochlorite
hypochlorite
for
20
min
has
increased
and
obtained
the
100%
Table 3. Shoot primordia per regenerating explants in three cultivars of tomato in MS medium supplemented
with growth regulators.
Media
Arka Meghali
Vaibhav
Hypocotyl
Epicotyl
Cotyledon
Mean
RM1
2.440.04
1.860.03
5.680.03
3.33
RM 2
11.420.03
8.780.03
12.710.05
RM 3
7.160.24
6.640.30
11.120.44
8.31
RM 4
3.820.93
3.600.84
9.270.26
5.56
RM 5
2.400.81
1.780.53
5.190.94
3.123
RM 6
7.760.24
7.170.20
10.980.21
8.64
RM 7
5.031.44
5.800.34
7.930.33
6.26
RM 8
1.640.42
1.600.37
1.910.35
RM 9
11.850.05
10.080.03 17.220.03
RM 10
10.000.05 8.040.04
12.950.01
10.97
e
g
h
Pusa Ruby
Hypocotyl Epicotyl
Cotyledon
Mean
2.040.03 5.040.03
2.390.04
3.16
8.590.04 11.840.04
9.840.03
5.540.26 7.560.26
6.860.23
6.65
3.280.55 6.540.30
3.260.57
4.36
1.350.47
2.400.81
2.381.27
Mean
Hypocotyl
Epicotyl
Cotyledon
Mean
7.770.03
7.590.03
9.230.03
8.20
17.880.03
12.86
15.670.33
13.320.49
19.770.32
16.25
10.40
13.670.30
10.060.31
17.480.26
13.74
8.070.23
4.780.86
10.870.27
7.91
d
b
10.09
2.04
10.350.33
17.370.33
13.36
5.690.70 5.931.39
4.691.26
11.080.23
7.670.21
15.350.42
11.37
1.200.21
1.340.26
4.801.30
2.721.07
4.551.88
1.72
8.790.04 15.920.03
11.160.02
5.820.03 11.760.04
7.040.02
13.05
10.33
5.44
1.39
10.96
0.01
0.01
0.01
CD
0.05
0.05
0.10
0.08
0.06
0.07
0.06
0.07
0.05
0.28
0.36
0.37
0.18
0.17
0.23
0.23
0.36
0.32
5.85
0.01
7.93
2.38
8.21
19.280.03 14.34c
0.01
4.82
7.69
j
15.16
11.850.03
0.02
7.01
11.900.05
0.02
9.34
4.022
10.55
11.96
0.01
5.46
0.01
6.23
4.36
12.360.33
1.640.42
7.88
6.20
12.31
9.82
15.76
10.360.30 9.64c
SEm
6.860.23 11.690.53
Mean
4.89
12.6
(1%)
CV
Data - Mean SE of 10 replications. The data were taken 3 weeks after culture. The values followed by same
letters are not significantly different @ P= 0.05.
Effects of different hormonal combinations in tomato
shoot differentiation
shoot length.
was
the
most
efficient
medium
for
tomato
Arka Meghali
Vaibhav
Pusa Ruby
Hypocotyl
Epicotyl
Cotyledon
Mean
Hypocotyl
Epicotyl
Cotyledon
Mean
RM 1
3.120.01
3.050.03
4.290.10
3.49
2.97
RM 2
5.630.05
4.390.01
6.160.03
5.75
RM 3
4.140.24
3.370.16
5.140.24
4.22
RM 4
2.520.22
2.630.21
3.730.24
RM 5
3.050.32
1.580.21
4.000.31
Epicotyl
Cotyledon
Mean
5.010.06
3.870.06
6.840.01
5.24
9.660.07
7.390.02
11.730.47
9.59
8.680.21
7.690.15
10.880.23
9.08
7.060.23
5.460.22
8.970.25
4.720.29
3.540.22
6.150.24
2.87
RM 6
5.050.23
3.680.29
6.450.26
5.06
4.81
6.860.23
5.810.34
8.850.31
7.18
RM 7
4.230.26
3.170.30
5.950.25
4.45
RM 8
1.320.16
1.320.16
1.580.21
6.39
4.88
1.41
RM 9
4.470.04
3.330.01
7.220.01
4.47
RM 10
3.980.02
4.380.06
5.600.03
2.070.01
4.770.08
2.070.02
4.630.04
5.840.02
5.060.05
5.11
3.370.16
4.550.22
3.730.24
3.88
3.460.16
3.700.29
2.900.25
2.050.24
3.340.34
2.630.21
2.67
3.020.23
5.650.26
4.330.26
4.33
2.810.23
4.740.24
3.840.23
3.80
1.230.15
1.580.21
1.280.22
1.37
5.920.06
7.750.08
5.860.04
6.51
5.390.02
5.630.08
4.160.02
2.96
4.84
Mean
Hypocotyl
3.35
4.26
10.990.12
8.660.04
12.820.06
10.82
7.620.03
6.680.02
10.960.14
5.13
3.17
4.72
3.52
6.87
5.54
8.97
0.02
0.09
0.02
0.08
0.02
0.10
0.04
0.19
0.03
0.13
0.02
0.09
0.02
0.09
2.70
0.99
0.55
0.83
0.94
1.36
1.00
0.87
1.06
3.70
5.52
5.320.30
4.80
3.45
8.460.26
0.04
0.19
3.100.24
2.77
3.80
8.42
7.12
4.49
4.550.22
0.13
0.66
5.73
4.350.22
3.54
6.81
6.140.29
SEm
CD
(1%)
CV
3.90
Mean
7.16
d
f
2.34
a
7.26
6.13
Mean number of shoots per regenerating explants, presented as mean SE of 10 replications. Data was recorded
at 5 weeks of culture. The values followed by same letters are not significantly different @ P= 0.05.
Table 5. Shoot length per regenerating explants in three cultivars of tomato in MS medium supplemented with
growth regulators.
Media
Arka Meghali
Vaibhav
Pusa Ruby
Hypocotyl
Epicotyl
Cotyledon
Mean
(cm)
Hypocotyl
Epicotyl
Cotyledon
Mean
(cm)
RM 1
5.060.01
5.250.01
3.570.01
4.63
5.950.01
7.800.00
7.85O.01
7.87
RM 2
4.510.04
5.510.02
6.350.07
4.820.00
9.810.02
5.770.00
RM 3
5.540.10
2.880.03
2.600.12
3.640.10
7.310.08
5.270.08
RM 4
2.450.11
3.350.10
0.630.03
2.14
2.230.11
4.760.09
3.150.09
3.38
RM 5
2.920.10
3.42 0.13
2.700.12
3.01
6.390.19
8.800.10
7.860.07
8.02
RM 6
2.060.12
2.67 0.09
1.740.12
2.16
4.500.08
8.200.11
5.980.08
6.23
RM 7
1.270.10
1.880.11
0.750.06
1.30
4.510.09
6.820.17
5.690.10
5.67
RM 8
0.690.03
1.06 0.06
0.550.03
3.760.07
5.630.10
4.280.04
RM 9
5.570.02
4.550.03
7.710.03
7.530.01
9.860.01
8.790.02
RM 10
4.510.01
5.880.04
3.040.03
6.800.02
9.430.05
8.660.01
Mean
SEm
CD
(1%)
CV
3.30
0.01
0.03
4.31
0.01
0.07
2.23
0.01
0.03
0.16
0.32
0.22
4.30
5.16
0.77
5.61
4.48
3.28
4.97
0.01
0.05
7.84
0.02
0.10
6.30
0.01
0.04
0.65
0.89
0.32
Mean
Hypocotyl
Epicotyl
Cotyledon
Mean
( cm)
4.84
4.830.03
5.710.00
3.970.03
3.260.04
4.860.02
5.720.01
3.360.10
4.750.07
3.250.07
1.610.07
3.230.07
1.150.07
1.99
2.880.11
4.120.13
2.730.07
3.91
2.810.12
4.350.10
2.510.12
3.22
2.540.11
3.240.09
1.640.09
2.47
1.590.07
2.450.11
1.190.07
5.520.03
5.500.01
6.700.03
2.870.03
5.090.01
5.320.03
6.13
5.41
4.56
8.46
8.30
6.37a
3.32
0.02
0.08
4.67
0.01
0.07
2.56
0.01
0.07
0.93
0.61
0.52
5.78
4.61
3.40
3.79
d
f
4.79
2.51
4.98
3.87
3.15
1.74
2.36
6.66
5.80
5.91
4.64
3.52b
Mean number of shoot length per regenerating explants, presented as mean SE of 10 replications. Data was
recorded at 5 weeks of culture. The values followed by same letters are not significantly different @ P= 0.05.
Shoot primordia per regenerating explant
grown under
number
of
days
required
for
complete
plant
413418.
121.
Bhatnagar-Mathur P, Vadez _K V, Sharma K.
References
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esculentum
to
callogenesis
and
1138-1140.
223-231.
242.
regeneration
in
tomato
(Lycopersicon
25, 36-71.
Selvi
DT,
Khader
MA.
1993.
in-vitro
optimization
esculentum
Mill.)
var.
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