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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. burmannis 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. TDrosera burmanni Vahl
2.1.1. 
 [14]:
.
.
.
.
.
.
Drosera.
: Drosera burmanni Vahl.
burmese sundew.
cburman.

2.1.2. 
Drosera burmanni 
            
Nam . . . [33], [34].
Drosera burmanni Thanh
 - 
1 

4


h














2.1: Khu v Drosera burmanni  gii [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 (Hnh 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


hhthoquinone-
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: C29].

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 

×