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ABSTRACT
Lawsonia inermis L.
IAA, IBA, 2, 4-D, KN
Callus
Different concentration of phytohormones affected Callus formation of Lawsonia inermis L. Explant of Lawsonia
inermis was cultured in MS medium supplemented with different concentration of IAA, IBA, 2,4-D, NAA,
KN, BAP. Medium supplemented with various phytohormones for organogenesis. Cultures were kept on
252 temperature and 16 hr photoperiod while callus was observed on different concentration of auxin or
cytokinin alone or in combination. Most suitable medium for callus formation from shoot tips was that 2, 4
D (2.5 mgL-1) + IAA (0.5 mgL-1).
Received on :
21.08.2010
Accepted on :
27.12.2010
*Corresponding
author
INTRODUCTION
Lawsonia inermis L. (Lythraceae) commonly has known as
Henna, Mehandi an important medicinal plant. It is a glabrous
much branched deciduous shrub (Muhammad and Mustafa,
1994). This plant is worldwide known cosmetic agent used to
colour hair, skin and nails (Hanna et al., 1998). However it is
not only relevant to cosmetic Henna was also reported to
have tuber culostatic activity. The leaves are also used as
prophylactic against skin disease. They are used extremely in
the form of paste or decoction against boils, burns, bruises
and skin inflammation. A decoction is used as a gargle against
sore throat (Rout et al., 2001). The roots of this plant are useful
in burning sensation, leprosy, strangury and premature graying
of hair. The major phytochemical constituents of Henna,
Lowsone is found to posses significant anti-inflammatory,
analgestic and antiphyretic activities (Ali et al., 1995)
Table 1: Effects of various concentrations of auxin incorporated in MS Medium on callus induction from shoot tip culture of Lawsonia inermis
L. after 4 weeks
Conc.
% culture showing callus formation
of Phytohormones2, 4 D
IAA
concentration in A
B
C
A
MgL-1
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
9.5
10
9
21
35
71
96
81
64
58
55
50
38
21
18
15
12
9
8
4
2
NR
9
17
33
77
98
83
63
55
56
46
34
25
19
17
11
10
6
3
1.5
NR
8
16
36
76
91
81
65
57
54
45
37
27
21
20
10
9
8
4
1.5
2
10
18
24
40
60
83
80
79
71
60
45
32
25
20
19
10
8
7
5
2
IBA
A
NAA
A
8
22
29
49
70
80
78
72
69
58
47
35
28
19
17
10
8
6
7
1
9
16
35
56
69
84
76
71
64
52
45
37
23
16
21
7
9
7
6
1
5
10
25
32
38
34
48
46
42
40
38
35
30
23
20
16
12
9
5
3
6
7
21
31
34
40
40
44
40
36
31
34
29
21
20
18
12
10
6
2
8
17
22
29
31
41
46
45
41
37
34
30
25
20
19
14
10
5
5
3
NR
10
16
21
38
45
63
75
82
87
86
77
68
56
44
31
25
25
16
10
NR
9
21
34
48
57
62
76
81
86
84
78
63
59
46
35
35
28
14
9
NR
10
20
34
46
55
61
78
83
86
82
77
61
54
46
32
32
26
11
6
Culture media
Semisolid MS media (Murashige and Skoog, 1962) containing
3% sucrose with varying concentration of phytohormones
was used for callus formation and roots and shoot
regeneration. Combination of auxin (IAA, IBA, NAA, 2, 4-D)
was also used for root and shoot regeneration. Coconut milk
(CW 20% v/v) was also used in MS basal medium for
regeneration system. The pH of media was adjusted to 5.8
before gelling with Agar Agar (0.8 w/v) by adding 0.1%
NaOH or 0.1% HCl and autoclaved for 15 to 20 minutes at
15PSI and 1210 surface sterilized leaves were inoculated into
the culture medium in culture room.
Effect of Cytokinins
When MS medium was supplemented with 5mgL -1
concentration of cytokinins (KN, BAP) callus formation was
induced. The callus was not fast growing one and did not
grow further sub-culture in medium even similar composition.
Root differentiation was never observed in any such cultures.
Culture condition
Culture was inoculated at 252C under cool fluorescent
light (1500 2000 Lux) with a 16 hr/8 hr light dark cycle. Each
treatment consisted of minimum 15 explants and all
experiments were carried out under sterile condition. All
inoculation tools like forceps, needle, scissors, blade, scalpel
etc. were thoroughly autoclaved. Culture area, glass wares
and all the tools used were properly sterilized with UV light
and floor surface were swabble with 95% ethyl alcohol.
Inoculation tools were flamed time to time.
Figure 1 and 2: (1) 14 days old callus from shoot tip culture on MS
medium containing 2, 4-D; (2) 3 weeks old callus from shoot tip on
MS medium containing NAA
Table 2: Effects of various concentrations of cytokinin amd combination of cytokinin and auxin on cellus induction and shoot differentiation
from shoot tip culture of Lawsonia inermis L.
MS Medium supplemented with
-1
KN (1mgL )
KN (5mgL-1)
BAP (1mgL-1)
BAP (1.5mgL-1)
BAP (2mgL-1)
BAP (5mgL-1)
Group
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
3rd week
4th week
10
9
8
7
8
9
13
15
14
16
15
19
14
15
17
7
8
6
41
42
43
21
24
25
19
20
18
42
40
41
13
15
14
6
7
4
19
18
20
20
21
22
34
33
35
42
45
44
35
34
33
18
19
17
72
75
74
53
55
54
40
41
42
73
74
75
36
35
37
19
20
18
22
23
24
28
29
27
37
36
38
59
60
59
58
60
59
21
22
20
84
86
87
63
64
62
55
53
54
94
93
95
51
52
50
24
25
23
12
13
15
15
16
17
22
23
21
30
31
29
30
29
31
14
15
13
59
61
59
45
46
44
27
28
26
60
62
61
28
29
27
13
14
16
41
REFERENCES
Ali, B. H., Bashir and Tanira, M. O. 1995. Anti inflammatory,
antipyretic and analgesic effect of Lawsonia inermis L. (henna) in rats.
Pharmacology. 51: 356 - 363.
42