Khảo sát khả năng ứng dụng kỹ thuật nuôi cấy mô sẹo, tạo dịch huyền phù cây Drosera burmanni Vahl trong thu nhận hợp chất anthraquinone
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Drosera burmanni Vahl
N
2003-2007
U
9/2007
C
**************************
Drosera burmanni Vahl
PGS.TS.
TS.
ThS.
T
9/2007
iii
, .
:
PGS.
,
.
,
,
,
, . HCM.
.
,
.
29
.
,
, ,
,
,
.
9/2007
iv
, Drosera burmanni
h
,
,
,
:
- M
D. burmanni Vahl
B5
: NAA (0,2 mg/l)/2,4-D (0,2
mg/l),
20 g/l, PVP (1,25 mg/l) 5,8,
.
- ,
D. burmanni in vitro.
Drosera burmanni Vahl.
-
:
3,03% so
,
(2,78%); 2,8%
0,78%;
0,02% (
), trong khi
.
v
SUMMARY
The thesis
Drosera burmanni was carried out
at Plant Biotechnology lab, University of Natural Sciences; and Chromatography
room, Research Institute for Biotechnology and Enviromental Technology,
University of Agriculture and Forestry, HCM city from March to August, 2007.
Results:
- Callus of D. burmanni Vahl was cultured successfully by using
Gamb
2,4-D (0,2 mg/l), succrose (20g/l), PVP (1,25 mg/l), pH was adjusted to
5,8 before autoclaving.
- Column Chromatography and Thin Layer Chromatography were used to
purify anthraquinone compound in D. burmanni. This compound was
used as authentic sample for HPLC technique.
- Roots of D. burmanni contained 3,01% dry weight (DW) of anthraquinone,
larger than 2,78% in leaves.
- Anthraquinone content of red-leaf D. burmanni Vahl took 0,78% of DW,
compared with 2,78% DW in green-leaf trees.
- Biomass of D. burmannis suspension contained 0,02% of fresh weight of
anthraquinone, while no quantifiable amounts of anthraquinone has been
found in spent medium.
Ho Chi Minh city, September, 2007.
Hoang Thi Thu
vi
..................................................................................................................... .i
................................................................................................................. iii
..................................................................................................................... iv
Summary ..................................................................................................................... v
..................................................................................................................... vi
.......................................................................................... ix
..................................................................................................... x
................................................................................ xii
.................................................................................................. xiii
.................................................................................................. 1
1.1. ....................................................................................................... 1
1.2. .......................................................................................................... 2
1.3. ............................................................................................................ 2
1.4. .................................................................................................. 2
...................................................................... 3
2.1. Drosera burmanni Vahl .......................................................... 3
2.1.1. ................................................................................................ 3
2.1.2. ........................................................................................................... 3
2.1.3. ............................................................................................... 4
2.1.4. ............................................................................................ 6
2.1.5. .......................................................................................... 7
2.1.6. ....................................................................................... 8
2.1.7. .............................................................................. 11
2.1.8. ............................................................................................ 12
2.1.8.1. ...................................................................................... 12
2.1.8.2. ................................................. 12
2.1.8.3. ........................................................................................... 12
2.1.8.4. ........................................................................ 14
2.1.8.5. ................................................................................................ 15
2.1.9. ............................................................ 16
h ........................................................................................................ 17
2.2.1. ......................................................................... 17
2.2.2. ........................................................................................... 18
2.2.2.1. ......................................................................................... 18
2.2.2.2. .................................................................... 19
2.2.3. .............................................................................. 19
2.2.3.1. ......................................................................... 19
2.2.3.2. ............................................................................ 19
vii
2.2.3.3. ............................... 20
2.2.3.4. ......................................... 20
2.2.3.5.
.................................................................................................. 22
2.2.4.
........................................................................................................... 22
2.2.4.1. ..................................................................... 22
2.2.4.2. ......................................... 22
2.2.4.3. ......................................................................................... 22
2.2.4.4. o ............................................................................................. 23
2.2.4.5. ............................................................................ 23
2.2.4.6. .......................................................................... 23
2.3. .................................................... 24
2.3.1. .................................................. 24
2.3.1.1. .................................................................................................... 24
2.3.1.2. ..................... 26
2.3.1.3. ........................................................................... 27
2.3.2. ..... 28
.......................................................... 31
................................................................................ 31
3.1. ......................................................... 31
3.1.1. ........................................................................................................ 31
3.1.2. ................................................................................................ 33
3.1.2.1.
.......................................... 33
3.1.2.2.
........................................................................................... 34
3.1.2.3. 3:
2,4-
............................................................. 35
3.1.2.4.
........................................................... 36
3.1.2.5.
................................................................................ 37
3.2.
D. burmanni ............ 38
3.2.1. ........................................................................................................ 38
3.2.2. ................................................................................................ 39
in vitro .............. 40
........................................................................................................ 40
............................................................................................... 40
......................................... 40
............................................................................... 41
viii
ong
........................................................................... 41
in vitro .................................................................... 43
............................................... 44
............................................................ 45
............................................................................................. 45
- .................................................................. 46
4.1. ........................................................... 46
.......................................... 46
........................................................................................... 48
4.1.3. 3:
2,4-
............................................................. 49
........................................................... 54
4.1.5. 5:
................................................................................ 56
4.2.
D. burmanni ............ 58
4.3.
............................. 59
............................................................................... 59
................................................................................... 61
in vitro .................................................................... 62
...................................................... 64
............................................................. 66
................................................................... 68
5.1. ....................................................................................................... 68
5.1.1. .......................................................... 68
5.1.2. ......... 68
5.2. ........................................................................................................ 69
..................................................................................... 70
ix
2,4-D : 2,4-dichlorophenoxy acetic acid
BAP : 6-benzylaminopurine
CRD : Completely Randomized Design ()
HPLC : High Performane Liquid Chromatography (
)
IAA : 3-indolyl-acetic acid
IBA : 3-indolebutyric acid
MS : Murashige & Skoog, 1962
NAA : naphthalene acetic acid
PVP : poly vinyl pyrrolidone
TCL : Thin Cell Layer (
)
TLC : Thin Layer Chromatography (
)
x
TRANG
Drosera burmanni .................. 4
nh Drosera burmanni Vahl .................................................... 4
D. burmanni ........................................................................................ 5
nh 2.4. D. burmanni Vahl ........................................................... 7
............ 10
2.6.
Drosera ......................... 15
................... 21
........................................... 21
2.9.
............................................................................................... 24
2.10.
............................................................. 26
................................................................................... 27
2.12.
...................................... 29
D. burmanni in vitro ................................................ 31
4.1.
D.burmanni
5
(0,2 mg/l) 2,4-
i ..................................................................................................... 52
4.2.
D. burmanni
5
(0,2 mg/l) 2,4-D
............................................................................................... 53
.............................................. 55
........................................... 55
.............................................. 55
4.4.
D. burmanni ........................................................................................ 57
pm ................... 60
................... 60
xi
................... 60
................... 60
70 ppm ................ 60
D. burmanni
............................................................................................................ 61
D. burmanni
.............................................................................................................. 61
D. burmanni Vahl ........................... 63
4.13.
D. burmanni Vahl .................... 63
D. burmanni .......... 65
D. burmanni
............................................................................................................ 65
...................................................................................................... 66
4.17. ................... 66
xii
TRANG
2.1.
........................ 18
3.1. Quy t .......................... 39
3.2.
............................................................................ 42
3.3.
.......................................................... 45
3.4.
(
) ........... 58
4.1
................................................................................... 46
.... 54
..................................................................... 59
xiii
TRANG
Drosera
7-methyljuglone ..................................................................................................... 8
Drosera ................... 9
Drosera ............................................................................................... .13
v
....................................... 33
............................................................... 34
ng 3.3.
2,4-D
..................................... 35
................................................................................ 36
................................................................... 37
in vitro ............................................................... 43
c ................................................................ 44
................... 46
................................................................................................ 47
............................................................................................................. 48
....................................................................................... 48
xiv
....................................................................................................... 49
......................................................... 54
.... 56
............. 59
in vitro ............................................................................................ 62
4.10.
................................................................................................ 64
4.11.
....... 66
1
1
1.1.
,
. o
m .
in vitro
Drosera Drosera
burmanni
in vitro
2
,
Drosera burmanni Vahl TRONG
1.2.
Drosera burmanni Vahl.
Drosera burmanni Vahl.
1.3.
D. burmanni Vahl.
D. burmanni
HPLC.
1.4.
D. burmanni
.
D.
burmanni
3
2
2.
2.1. TDrosera burmanni Vahl
2.1.1.
[14]:
.
.
.
.
.
.
Drosera.
: Drosera burmanni Vahl.
burmese sundew.
cburman.
2.1.2.
Drosera burmanni
Nam . . . [33], [34].
Drosera burmanni Thanh
-
1
4
h
2.1: Khu v Drosera burmanni gii [30].
2.1.3.
Drosera burmanni cao 5 - 30 cm,
hay
[6], [15], [33], [35].
2.2: Drosera burmanni Vahl [33], [35].
h].
Drosera burmanni Vahl
Drosera burmanni
Vahl
A
F
E D
C
B
5
L
.
Drosera
burmanii ].
2.3: D. burmanni [42].
; .
C, D: C.
Hoa (Hnh 2.2 D, E)
-
A
D
C
B
6
-
9], [14].
N
2.1.4.
: S
: C
t keo
25], [34], [35].
7
2.1.5.
2.4: D. burmanni [3], [40].
[37].
Drosera
- 80%)
5 - 35
O
- 25
O
5
O
, t -
8
2.1.6.
h
Drosera in vitro.
Naphthoquinone
Drosera h plumbagin
-methyljuglone (2.1) [21].
(B
2.2 29].
2.1: Drosera -methyljuglone [21]
Plumbagin 7 Methyljuglone
D. andersoniana
D. auriculata
D. binata
D. bulbosa
D. capensis
D. capillaris
D. cistiflora
D. dichotoma
D. erythrorhiza
D. intermedia
D. longifolia
D. lunata
D. macrophylla
D. madagascariensis
D. microphylla
D. modesta
D. natalensis
D. peltata
D. adelae
D. aliciae
D. aliciae x capensis
D. anglica
D. auriculata
D. burkeana
D. burmanii
D. capensis
D. cistiflora
D. cuneifolia
D. dielsiana
D. filiformis
D. hamiltonii
D. hilaris
D. indica
D. intermedia
D. longifolia
D. longifolia x rotundifolia
9
D. platypoda
D. prolifera
D. pygmaea
D. regia
D. rotundifolia
D. slackii
D. stolonifera, spp. Rupicola
D. venusta
D. villosa
D. madagascariensis
D. ramentacea
D. regia
D. rotundifolia
D. spathulata
D. stolonifera, spp. Stolonifera
D. tracii
D. trinervia
D. villosa
2.2: Drosera [21]
Naphthoquinone
Droserone
Hydroxydroserone
Biramentaceone
a
Droserone-5-glucoside
Rossoliside
Hydroxydroserone-5-glucoside
2-Methylnaphtazarin-5-O-
glucoside
b
D. whittakeri
D. rotundifolia
D. whittakeri
D. rotundifolia
D. ramentaceae
D. rotundifolia
D. rotundifolia
D.spatulata
D.rotundifolia
D.rotundifolia
D.spatulata
In vivo
In vivo
In vivo
In vivo
In vivo
In vivo
In vivo
In vitro
In vitro
In vivo
In vitro
In vitro
10
hhthoquinone-
Drosera [21].
R
1
R
2
R
3
R
4
R
5
[1]
CH
3
H H H OH
7-Methyljuglone
[2]
H H CH
3
H OH
Plumbagin
[3]
H H CH
3
OH OH
Droserone
[4]
OH H CH
3
OH OH
Hydroxydroserone
[5]
H H CH
3
OH OGlc
Droserone-5-glucoside
[6]
OH H CH
3
OH OGlc
Hydroxydroserone-5-gluside
R
6
R
7
[7]
CH
3
H
Rossoliside
[8]
H CH
3
2-Methyl-hydrojuglone-4-glucoside
Flavonoid
Drosera ch flavonoid gl
isoquercetin, kaempferol, astragalin, hyper].
Quercetin Kaempferol
Myricetin Hyperin
2.5: C29].
R
4
O
O
R
2
R
3
R
5
R
1
OH
OGlc
R
7
OH
R
6
11
2.1.7. P
N[3]
T
C
T
V
in vitro
Drosera
Drosera
in vitro Drosera
