




 

 


Dendrobium T






H   








H   




 2007
 
TS.  

CN. 






iii














:
- 
- TS. 
- 










 , 







, 
Nai  



.


  , , 








.
 








iv






: TS. 

, TS. , CN. 



.






: 









 , 





,  , 
 . , 














  Dendrobium, 


, 



, 








, 











 .




, 







 10 

Dendrobium 


 . 6 

 : OPAC10, OPAH13, 0PB01, OPB08, S1384,
U693 











 




 . 



, x

 



SUMMARY

Genetic diversity analysis of wild Dendrobium stem at Ho Chi Minh city
and Nam Cat Tien forest by RAPD
Supervisor: Tran Thi Dung
1
, PhD; Vo Thai Dan
2
, PhD; Luu Phuc Loi
1
, BSc
Student: Truong Minh Dung
1
1
Department of Biotechnology, Nong Lam university, Ho Chi Minh city
2
Faculty of Agriculture, Nong Lam university, Ho Chi Minh city
To analysis the genetic diversity of wild Dendrobium orchid, six decamers were
used for 10 wild Dendrobium species, collected from Ho Chi Minh city and Nam
Cat Tien forest (Dong Nai province). Collecting 10 species from Ho Chi Minh city
and Nam Cat Tien forest (Dong Nai province), optimizing of protocols for DNA
extraction and purification, constructing of protocol for RAPD, analyzing the
genetic of 10 wild Dendrobium species by RAPD and constructing the dendrogram
by NTSYSpc 2.1 software, Winboot program are the objective of the study.
Methods
- To identify 10 wild Dendrobium species with genotype.
- Extraction of total DNA from fresh tissue of 10 wild Dendrobium species.
- Six random primer: OPAC10, OPAH13, OPB01, OPB08, S1384, U693 were
used to analysis of genetic diversity.
- Following RAPD reactions, dendrogram were constructed by NTSYSpc 2.1

 ..................................................................... ix






 ............................................................................ x








........................................................................ x






 .............................................................................. xi

1. 



 ..................................................................................................... 1

 .................................................................................. 18
 .............................. 18
 ............................................................. 18
 ............................................................. 19
  ...................................................... 19
2.5. 









Winboot .. 26
2.5.1. NTSYSpc version 2.1 ............................................................................ 26
2.5.2. Winboot ................................................................................................. 28
2.6 ................................................... 29
2.6.1.  .......................................................... 29
2.6.2. nh h .......................................................... 31
3. 

























 .................. 45

viii
4.2.  


















10 

Dendrobium
d  ................................................................................ 54
4.5. 







10 

Dendrobium
b 2.1 Winboot ........................................ 60


-  .............................................................................. 69
5.1. 

 ..................................................................................................... 69
5.2. 






AFLP: Amplified Fragment Length Polymorphism
bp: base pair
CTAB: Cetyl Trimethyl Ammonium Bromide
DNA: Deoxyribonucleic acid
dNTP: Deoxyribonucleotide triphosphate
EB: extraction buffer
EDTA: Ethylene Diamine Tetra acetic Acid
mM: milimolar (milimol/lite)
NCT: 


OD: Optical density
PCR: Polymerase Chain Reaction
pmol: Picomol
RAPD: Random Amplified Polymorphic DNA
RFLP: Restriction Fragment Length Polymorphism
RNA: Ribonucleic acid
Rnase: Ribonuclease
SSR: Simple Sequence Repeat
Ta : Annealing t
TAE: Tris  Acetate  EDTA
TE: Tris  EDTA

TP. HCM: 




Dendrobium .. 46
4.2. 












Dendrobium ..................................................................................................... 47
B





 ...... 50
4.4. 

(ng/)  .............................. 51
 ................................. 53
 ................................................................................. 53
 ............... 55




 Trang
Dendrobium ..................................................... 5
 .......................................................... 13
 ....................................................... 21
 ....................................................... 23
 ....................................................... 25
.  ......................................... 26
2.7. 

2.10m ............................................................. 27
2.8. 



Winboot ..................................................... 29
Dendrobium  ....................... 33
4.1. 











2. 
Dendrobium 

 

. ................ 58
3. 
Dendrobium 

. ................................. 59
4. 
Dendrobium . ................ 59
5. 
 Dendrobium 

. ................................. 60
6. 
 Dendrobium  



. ................ 60
7. 
g Dendrobium 

. ................................. 61
8. 


n Dendrobium 

Dendrobium . ............................................................................. 65
1







1.1. 
   

 






 
 


  

Dendrobium.


 ,  ,
m 

 

 

 Dendrobium 










.  ,              
lan 















 : 




ng di truyn ca ging lan rng Dendrobium thu th

  H




 







- 







 Dendrobium 
























 .
- 











 Dendrobium 


primer RAPD.
- 










.
-  RAPD , 



.
- Dendrobium 
.
- M Dendrobium 


Winboot.
3



2.1.  Dendrobium
Dendrobium   Epidendreae   Epidendroideae  
Orchidaceae.
Lan Dendrobium 
Dendrobium 
.
 Dendrobium             


,  

 : h

 g

    


                 
  
             


Hoa: h        


nh




nh
 q- Q

 

- 
5

- - - 
 15cm - 
 
 7
D. densiflorum ( )cm -  
 - 4, thon, 
 
              
b
D. farmeri 
cao 20cm -  - 

                

D. fimbriatum ng
       cm -       
cm - 
 inh
, Kon Tu 
D. heterocarpum 
20cm - cm - 13cm
 
 
.
D. primulinum (Long Tu) ;
cm - 10
 v



          
m Tris  HCl 1M (pH 7.5), NaCl 5M, EDTA 0.5M, sodium dodecyl



9

protein.
        

   polysaccharide      
           


 
            

                




Mc 
         , m     



260nm




280nm



 

260nm
/OD
280nm
- 2.0.



 


 
 11
Log u = Log u
0
 K

, DN

DNA in vitro.

primer (primer 
 12
 s i tip
nhau, mm 3 :
: - denaturation

- 

0
C - 95
0

 - hybridization
 

0
C - 70
0


 - elongation
      
0