Tuyển non dòng vô tính cao su kháng bệnh rụng lá corynespora

BỘ GIÁO DỤC VÀ ĐÀO TẠO
TRƢỜNG ĐẠI HỌC NÔNG LÂM TP.HCM
BỘ MÔN CÔNG NGHỆ SINH HỌC






KHÓA LUẬN TỐT NGHIỆP




TUYỂN NON DÕNG VÔ TÍNH CAO SU (Hevea brasiliensis
Muell. Arg.) KHÁNG BỆNH RỤNG LÁ CORYNESPORA
(Corynespora Leaf Fall Disease)







Ngành: CÔNG NGHỆ SINH HỌC
Niên khóa: 2003 – 2007
Sinh viên thực hiện: NGUYỄN NGỌC THANH TRANG


2007




BỘ GIÁO DỤC VÀ ĐÀO TẠO
TRƢỜNG ĐẠI HỌC NÔNG LÂM TP.HCM
BỘ MÔN CÔNG NGHỆ SINH HỌC






KHÓA LUẬN TỐT NGHIỆP




TUYỂN NON DÕNG VÔ TÍNH CAO SU (Hevea brasiliensis
Muell. Arg.) KHÁNG BỆNH RỤNG LÁ CORYNESPORA
(Corynespora Leaf Fall Disease)

Giáo viên hƣớng dẫn: Sinh viên thực hiện:
TS. TRẦN VĂN CẢNH NGUYỄN NGỌC THANH TRANG
ThS. PHAN THÀNH DŨNG
TS. PHAN PHƢỚC HIỀN







2007
iii
LỜI CẢM ƠN



 

 

 





.  ThS. 
 .

  
 





Con xin   anh
 



NGUYỄN NGỌC THANH TRANG
iv
TÓM TẮT

.
TUYỂN NON DÕNG VÔ TÍNH CAO SU (Hevea brasiliensis Muell. Arg.)
KHÁNG BỆNH RỤNG LÁ CORYNESPORA (Corynespora Leaf Fall
Disease)”. 03/03/2007  ,
            
Su  VNCCSVN.
   TS. Trần Văn Cảnh, ThS. Phan Thành Dũng 
TS. Phan Phƣớc Hiền.
 



                
; 



+ 
+ 100% dv  Corynespora      

 .
    D       ;
T 
; 

 D
Dvt ca


v
SUMMARY
NGUYEN NGOC THANH TRANG, Nong Lam University. “SCREENING
RUBBER CLONES (Hevea brasiliensis Muell. Arg.) RESISTANT TO
CORYNESPORA LEAF FALL DISEASE”. The research was carried out from
03/03/2007 to 30/07/2007 at the laboratoy of Crop Protection Division, Molecular
biology laboratory,   Rubber Technology Center  Rubber
Research Institute of Vietnam (RRIV).
Supervisors: PhD. Tran Van Canh, Msc. Phan Thanh Dung and
PhD. Phan Phuoc Hien.
Materials and Methods:
+ Corynespora disease rubber leaflets (10  12 days old) used for
estimation of POD activity.

+ Free  diseased leaflets (10  12 days old) used for artificial inoculation
with Corynespora cassiicola and estimation of the number of stomas.
+ 60 rubber clones were screened for resistance to Corynespora leaf fall
disease using detached leaflets technique and droplets of spore suspension.
Results:
+ Screening rubber clones resistant to CLFD using detached leaflets
technique is a reliable method.
+ 100% clones investigated were infected to CLFD at various susceptible
levels. Among 60 clones, 15 clones are very severely infected; 12 clones
are severely infected; 22 clones are moderately infected and 11 clones are
lightly infected.
+ POD activities and the number of stoma are different from various
clones. Generally, the more susceptible clones are, the more the number of
stoma they have. The more POD activities the clones have, the less they are
susceptible to the disease. But there is no linear correllation found between
POD activity or the number of stoma and the disease severity.
vi
MỤC LỤC

 TRANG
 ............................................................................................................ iii
 .................................................................................................................. iv
SUMMARY ................................................................................................................ v
 .................................................................................................................. vi
 ................................................................................. x
 ................................................................................................ xi
................................................................................................. xii
 ......................................................................................... xiii
Chƣơng 1: MỞ ĐẦU ................................................................................................. 1
1.1.  ...................................................................................................... 1

1.2.  ...................................................................................... 2
1.2.1. M .................................................................................................. 2
1.2.2.  .................................................................................................... 2
1.3.  ............................................................................................... 3
Chƣơng 2: TỔNG QUAN TÀI LIỆU ...................................................................... 4
2.1.  .................................................................................... 4
2.1.1.  ...................................................... 4
2.1.2.  .......................................................... 5
2.2. C. cassiicola ........................................................................ 6
2.2.1.  ............................................................................................ 6
2.2.2.  ................................................................................... 7
2.2.3.  ...................................................................................... 7
2.2.4.  ....................................................................................... 7
2.2.5.  .................................................................................... 8
2.2.6. 
C. cassiicola ................................................................................. 8
vii
2.3. Corynespora  ............................... 9
2.3.1.  ..................................................................................... 9
2.3.2. Corynespora  .............................. 10
2.3.2.1.  .............................................................................. 10
2.3.2.2.  ....................................................................... 11
2.3.3.  ................................................................ 12
2.3.4.  ......... 13
2.4.  ...................................................................................... 13
2.4.1.  ............................................................................................. 13
2.4.2.  ......................................................................................... 13
2.4.3.  ..................................................................................... 14
2.4.4. yme ................................ 14
2.4.5.  ............................ 14

2.4.6.  .............. 16
2.4.7.  ....................... 17
2.4.8. Peroxidase .............................................................................................. 17
2.4.8.1.  .................................................................... 17
2.4.8.2.  ..................................... 18
2.5.  ................................................................................... 18
2.5.1.  ................................................................................................... 18
2.5.2.  ............................................................................................. 19
2.5.3. Vai  .................................................. 20
2.5.4.  ............................................ 20
Chƣơng 3: VẬT LIỆU VÀ PHƢƠNG PHÁP TIẾN HÀNH ............................... 21
3.1.  .................................................................. 21
3.1.1.  ................................................................................................ 21
3.1.2.  ................................................................................................. 21
3.1.3.  ............................................................................ 21
3.2.  ................................................................................ 21
viii
3.2.1.   ........................................................................ 21
3.2.2. C. cassiicola .............................................. 22
3.2.2.1.  ....................................................................... 22
3.2.2.2.  .......................................................................... 22
3.2.2.3.  ..................................................................... 22
3.2.3. Corynespora 
 ................................................................. 23
3.2.3.1.  ........................................................................ 23
3.2.3.2.  .................................................................................. 23
3.2.3.3.  .................................................... 25
3.2.4.  ....... 25
3.2.4.1.  ........................................................................................ 25
3.2.4.2.  ......................................................... 25

3.2.4.3.  .................................................................................. 25
3.2.4.4.  .................................. 27
3.2.5.  ... 27
3.2.5.1.  ........................................................................................ 27
3.2.5.2.  ....................................................................... 27
3.2.5.3.  .................................................................................. 27
3.2.5.4.  .................................. 28
Chƣơng 4: KẾT QUẢ VÀ THẢO LUẬN ............................................................. 29
4.1. Corynespora  ........................................ 29
4.2.  .......................................................................................... 31
4.3. Corynespora 
 ................................................................................. 32
4.4. 
 ............................................................. 36
4.4.1.  .. 36
4.4.2.  ................................... 38
ix
4.5.  h 
 ..................................................... 40
Chƣơng 5: KẾT LUẬN VÀ ĐỀ NGHỊ .................................................................. 43
5.1.  ........................................................................................................ 43
5.2.  ........................................................................................................ 43
 ......................................................................................... 45
 .................................................................................................................. 50

x
DANH SÁCH CHỮ VIẾT TẮT

BVTV 
C. cassiicola Corynespora cassiicola
CLFD Corynespora Leaf Fall Disease
CRD Completely Randomized Design
Dvt 
H. brasiliensis Hevea brasiliensis
NT 
PDA Potato Dextrose Agar
POD Peroxidase
PR proteins Pathogenesis Related Proteins
PSA Potato Sucrose Agar
UV Ultra Violet

VNCCSVN S

xi
DANH SÁCH BẢNG

 TRANG
Bng 3.1: Bp b ................... 24
Bng 4.1: Kt qu u tra CLFD  i  ................ 31
Bng m nhim CLFD ca 60 dvt cao su ................................. 33
B ph h n m mn cm ca con lai di truyn t
m .............................................................................................................................. 34
B ph h n m mn cm ca con lai di truyn t
b ............................................................................................................................... 34
Bng 4.5: Bng kt qu u bi chng .......... 37
Bng 4.6: Bng kt qu nh ................................................... 38
Bng 4.7: Bng kt qu m s ng ..................................................... 41

xii
DANH SÁCH HÌNH

 TRANG
: Cu tng ..................................................................................... 19
u to m ng........................................................... 20
p b ........................................... 24
: Mt s triu ch





Corynespora  ...................... 30
: Mt s triu ch





Corynespora  ..................... 30
Khun lc C. cassiicola ng PSA ....... 32
: Khun lc C. cassiicola ng PSA ....... 32
C. cassiicola  ................................. 32
 Dvt LH 94/612 sau 5 o CLFD.......... Error!
Bookmark not defined.
 .............................................................. 41

xiii
DANH SÁCH BIỂU ĐỒ

 TRANG
Bi 4.1: Hoi chu bnh ca 12 dvt cao su
... 38
Bi 4.2: Ma hoc nhim bnh CLFD ........ 39
Bi 4.3: Mi quan h gia s n cm vi CLFD ca
12 dvt cao su .............................................................................................................. 42

1

Chƣơng 1
MỞ ĐẦU
1.1. Đặt vấn đề
(Hevea brasiliensis Muell. Arg.)  


  ,   
            
   Corynespora    Corynespora cassiicola (Berk. and
Curt.) Wei.               
L 

 

     (dvt)          
  

C. cassiicola 

m 
   .
Tr
       25%. 
Report/Report2000.htm#Corynespora).   CLFD     
1999, C 


, CLFD 
... P
2

         
 
 








 Vi
(VNCCSVN) 
dvt  




.
P


               ,
             
 , t “Tuyển non dòng
vô tính cao su (Hevea brasiliensis Muell. Arg.) kháng bệnh rụng lá
Corynespora (Corynespora Leaf Fall Disease)”.
1.2. Mục đích và yêu cầu
1.2.1. Mục đích
Corynespora 
60 dvt ca
Kperoxidase 

K

1.2.2. Yêu cầu

C. cassicola 
C. cassiicola 
             


3


 POD 



1.3. Giới hạn đề tài
 
 m 


  b

 

4


Chƣơng 2
TỔNG QUAN TÀI LIỆU
2.1. Sơ lƣợc về cây cao su
2.1.1. Cây cao su và tầm quan trọng của nó
Cao su (Hevea brasiliensis Muell. Arg.)  
Euphorbiaceae)  
             khi


 
  , Char

 1996).
 
 Nnr
, 
  

 

     ,         
         
 1996).
 . Vo
 n
      
                
5

. 
          

    http://210.245.64.232/BaocaoNH/2007/T7/T7_Caosu.pdf).
  0.000  





5
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8
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 



...


2.1.2. Tình hình bệnh hại trên cây cao su
   Trong
 http://www
.daff.gov.au/__data/assets/pdf_file/0009/146925/management_of_plant_pathogen_collec
tions_vietnamese.pdf).
r, 
n


Theo Chee (1976), 
g (1997), 
, 2004).   
 bao
6

 hr Corynespora,
b.
,  
 

Colletotrichum gloeosporioides 
   Odium hevea          
Corynespora do Corynespora cassiicola (Berk. and Curt.) Wei.   


  
20  .
2.2. Sơ lƣợc về nấm C. cassiicola
2.2.1. Phân loại học

               
(Eucaryotae 


  Corynespora   


 Ascomycota
Dothideomycetes
Pleosporomycetidae
Pleosporales
Corynesporascaceae
 Corynespora
Corynespora cassiicola

7


2.2.2. Đặc điểm hình thái
Chee (1988), 
(H. brasiliensis (23,42  132,6 x 2,60  
               
 P. T. Dung (1995; 2004), 

              
 
 

Theo   s (1969),  
pC. cassiicola 

 .
2.2.3. Đặc điểm sinh lý
C. cassiicola 
  ctv  
 C. cassiicola    
P. T. Dung (1995), C. cassiicola 
               
  
 Chee (1988) C. cassiicola 
cao su (H. brasiliensis

 C. cassiicola 
 Mushrif, 2006).
2.2.4. Khả năng tồn tại
, C. cassiicola     
 
 . 
8


su, s


  

  C. cassiicola 
 

(, 2006).
 

 2004).
P. T. Dung (1995), C. cassiicola  

2.2.5. Khả năng phát tán
C. cassiicola 
 
chu k 
 
     
 sau khi 
 



a nhau (Mushrif, 2006).
2.2.6. Con đƣờng xâm nhập, phạm vi phân bố, phổ kí chủ và khả năng gây
bệnh của nấm C. cassiicola


9





 Magnaporthe grisea hay Botrytis cinerea 

(N
C. cassiicola 
cellulase





  2000).
2.3. Giới thiệu về bệnh rụng lá Corynespora trên cây cao su
2.3.1. Lịch sử phát hiện
   Corynespora         Corynespora
cassiicola (Berk. and Curt.) Wei., x

   r         
, 
7 (Jinji , 2007).
 
 
N. T. 
lo 

 
             

10


    Corynespora     dvt     
               

      

2.3.2. Triệu chứng của bệnh Corynespora trên cây cao su

2.3.2.1. Trên vƣờn ƣơm
T

  
   
  (Jacob, 2006).
               
  
              
Kothandaraman, 1996).
 
 
              
   .    

Jayasinghe, 2000).
 
 
 



h
11



uy

 



        
 
hay         do n       
,   
 



        
   n
xa 
.
         

     do  
(Jacob, 2006).
2.3.2.2. Trên vƣờn khai thác
Corynespora  


 (stag  horn)
 

12


 


 



  
 .
 . Khi
 


 .
  .

.  
 


2.3.3. Khả năng hình thành nòi mới
Ndvt 
            
2000) 

 
dvt cao su.


, 