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BỘ NÔNG NGHIỆP VÀ PHÁT TRIỂN NÔNG THÔN






GIÁO TRÌNH MÔ ĐUN

CHUẨN BỊ HOÁ CHẤT VÀ MÔI
TRƢỜNG VI NHÂN GIỐNG
MÃ SỐ: 03

NGHỀ: VI NHÂN GIỐNG CÂY LÂM NGHIỆP

Trình độ: Sơ cấp nghề

Bắc Giang, tháng 10 năm 2008











2



TUYÊN BỐ BẢN QUYỀN






MÃ TÀI LIỆU:


































3

LỜI GIỚI THIỆU

Vi nhân giống cây lâm nghiệp là nghề sản xuất giống cây lâm nghiệp
chất lượng cao đáp ứng nhu cầu trồng rừng kinh doanh ở Việt Nam, trong
chương trình đào tạo nghề ngắn hạn cho lao động nông thôn từ nay đến năm
2020, nhằm trang bị cho học viên một số kiến thức và kỹ năng cơ bản để thực
hiện các bước công việc nhân giống cây lâm nghiệp bằng vi nhân giống.
Giáo trình Vi nhân giống cây lâm nghiệp được xây dựng và phát triển
theo các bước: phân tích nghề, phân tích công việc và xây dựng chương trình,
giáo trình dạy nghề theo mô đun.
Giáo trình mô đun: Chuẩn bị hóa chất và môi trường vi nhân giống là
mô đun thứ 3 trong 6 mô đun của chương trình dạy nghề: Vi nhân giống cây
lâm nghiệp nhằm trang bị cho học viên những kiến thức và kỹ năng xác định,
pha chế, bảo quản hóa chất và môi trường dùng trong nghề Vi nhân giống cây
lâm nghiệp.
Giáo trình mô đun gồm 3 bài: Bài 1: Xác định các loại hóa chất dùng

trong vi nhân giống; Bài 2: Pha chế và bảo quản dung dịch mẹ; Bài 3: Pha chế
và bảo quản môi trường vi nhân giống
Để hoàn thành giáo trình chúng tôi nhận được sự giúp đỡ của các nhà
khoa học ở các viện nghiên cứu, các cán bộ kỹ thuật ở các cơ sở sản xuất, các
giảng viên ở các trường đại học, cao đẳng, dạy nghề và khoa Lâm nghiệp
Trường Đại học Nông - Lâm Bắc Giang. Nhân dịp này cho phép chúng tôi gửi
lời cảm ơn đến lãnh đạo Bộ Nông nghiệp & PTNT, các viện nghiên cứu, các
trường, các nhà khoa học, các cán bộ kỹ thuật, các thày cô giáo đã tham gia
chương trình và đóng góp nhiều ý kiến quý báu, tạo điều kiện thuận lợi để
chúng tôi hoàn thành giáo trình này.
Trong quá trình biên soạn giáo trình mô đun chắc chắn không tránh khỏi
những thiếu sót. Chúng tôi rất mong nhận được những ý kiến đóng góp của quý
báu của các nhà khoa học, các nhà quản lý và các bạn đọc để hiệu chỉnh và
hoàn thiện giáo trình phục vụ sự nghiệp đào tạo nghề ngắn hạn cho lao động
nông thôn ở nước ta.







4

MC LC

TIấU TRANG
1
2
4

5
5
BI 1:
5
5
5
1. Vai trò, đặc tính lý, hoá học và sinh học của
các hoá chất dùng trong vi nhân giống 5
5
1.2. 10
1.3. Cỏc vitamin 15
1.4. 17
23
23
23
25
28
30
30
30
30

trờn bao bỡ 30

30
32
BI 2: P 33
I. 33
33
33

33
34
36
36
37

5

 38
  39
 40
 40
 40
 46
BÀI 3:  47
 47
 47
 47
 48
 53
 54
 55
 56
 56
 56
 56
 57
Bài 1:  57
 59
Bài 2:  59

 61
Bài 3:  61
 63
 64
 64
 64
 64
 65
 65
 65
 66
 66
5.1. Bài 1 66
5.2. Bài 2 66
5.3. Bài 3 67
 68





6


CÁC THUẬT NGỮ CHUYÊN MÔN, CHỮ VIẾT TẮT































7

Mễ UN CHUN B HO CHT
V MễI TRNG VI NHN GING
Mó s mụ un: M 03


Gii thiu mụ un:





8

Bi 1:
XC NH CC LOI HểA CHT DNG TRONG VI NHN GING

Mc tiờu:
Sau khi hc xong bi ny hc viờn cú kh nng:
-

-
-

A. Ni dung:
1. Vai trò, đặc tính lý, hoá học và sinh học của các
hoá chất dùng trong vi nhân giống





3

+

v NO
3
-

1.1. Cỏc nguyờn t a lng
Ni t (N):

8

nitrat (NO
3
-

4
+

(KNO
3
), nitrat amon (NH
4
NO
3
) và canxi nitrat (Ca(NO
3
)
2
.4H
2





 



3
)
2
.4H
2
O, KNO
3
, NaNO
3


4
NO
3

4
)
2
SO
4

4
NO
3

.
 
NO
3
+
và NH
4
+







Phốt pho: (P)

  


hay dihyd

2
PO
4
-






         




4
2-


-O-

 bào.

9
Kali (K):
K
+


+


+

t
+


3

,
KCl. 6H
2
O, KH
2
PO
4


+



  200 mM.



+

 8 là pH thích


+


+
là ion trung hoà quan

+




+






+

+



+

Nó trung ho


+

ion NO
3
-
           

+



+


+

bào r
Canxi (Ca):

10


+
và Mg
+
vì Ca
2+





2+


2+

               
cytokinin.
Ca
2+

         
  Ca
2+
    

Ion Ca
2+


2+


2+

  
2+
         
(Ca
2+


2+


            


2+




 



2+

 

               
         
polygalacturonaza     
     
           
  
2+



11
Ion Ca
2+

Ca
2+



2+
. Ion Ca

2+
  
           

 


2+


           
2+


2+

Magiê (Mg):
 
              
photphate. Ion Mg
+


+


  
4
 
+

duy

2+


Vai trò Mg
2+

2+

  


2

2+

- 20% ion Mg
2+

Mg
2+
và K
+
    
-  nào


Vai trò Mg
2+


2+

-
   
2+
   

2+


2+


 
2+


12

2+


2+
thì n

Ngoài ra Mg
2+
          


Mg
2+

2+

2
và Vmax.
Vai trò Mg
2+



uá trình này, Mg
2+

Mg
2+


2+




Lƣu huỳnh (S):
  
4
2-
        
 


4

4
. 7H
2
O
1.2 Các nguyên tố vi lượng (Fe, B, Cl, Co, Cu, Mn, Mo, Zn )
 , 


  . 





 : iron
(Fe), manganese (Mn), zinc (Zn), boron (B), copper (Cu), 
(Mo).






 , 





 . 
 . 








 (EDTA) - chelate Fe thay cho citrate Fe , 












. Tuy nhiên, 
 .











 (Co), iodine (I)
 (Na), 





 














 .






13
 


           

 
       







Sắt (Fe):
           
2+
,
Fe
3+

-10

2+


3+

2+


3+
).

       -    
3+
- citrate hay
- 

 > Fe(III) + e
-


 


   
2
(SO
4
)
3
và FeCl
3
   


-

2
ETDA và FeSO
4
. 7H
2
0
Bo (B):





14








bào v
- 











H
3
BO
3
Đồng (Cu):



Cu
2+
   /l. Các ion Cu
2+
     

2

CuNO
3.

2+
hay
        
2+
    



- 
         
[(Cu
2+
)/(Cu
+


           
              


15


qua


       


Mangan (Mn):

n



2+
không liên k


cây.
             




Coban (Co):


     






Coban 
             
 

               
               
         -


16

-
2,3- 


Molybden (Mo):

4
2-



4
2-


sau:
MoO
4
2-
> HMoO
4
-
>H
2
MoO
4


2

    
2
MoO

4
.2H
2
         
-molybden.
-

Kẽm (Zn):



 

        
-


2+


2+




               
tryptophan.
Clo (Cl):
Ion Cl
-

 


-


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


17
    
          
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  



   
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     






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


trình trao 
1.3 Các vitamin




 


            

    


(B1), acid nicotinic (PP), pyridoxine (B6) và myo-   
vitami
-10 mg/l.
        

-
-10 mg/l.
Myo-

-inositol
             
phosphoinositide và phosphatidylinositol có vai trò quan 

18
-
-5.000 mg/l.

         
acid, vitamin E (tocopherol), riboflavin và p-
             
           



           
(B1), pyridoxine HCl (B6), nicotinic axít, tr     


g môi
  


     ông

nicotinic axit (vitamin B
3
), canxi pantothenate (vitamin B
5

              


a. Vitamin B1 (Thiamine.HCl, Aneurin)
 . 













 itamin B 1 






 1, 

















 1. 
 
b. Vitamin B2 (Riboflavin, Lactoflavin)

 
  u vitamin này không hình
 










 0,01 ppm, 


10 - 50 ppm.
c. Vitamin B6 (Pyridoxine, Adermin)
 - phosphate-









 . 












19
Vitamin B6 là coenzin c


d. Myo-Inositol (Bios I)
 , 
polygalacturon

 . 








 . 












 100 ppm. 





















 .
e. Nicotinic acid va
̀
pyridoxine
































 . 
 , ascorbic acid, vitamin E (tocophenol), -aminobenzoic
aci











 , 

















 .  , 


0,1-10,0 ppm.
1.4 Các chất điều hoà sinh trưởng
B



               


a. Nhóm các auxin
Các au

-











  : 1H-
indole-3-acetic acid (IAA), 1-naphtaleneacetic acid (NAA), 1H-indole-3-
butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D) 
acid (NOA). 









 ;  , IBA,
2,4-

  , 

































 (auxin-
oxydase) 






.
            






20
- 
- 
- 
- 


            bào. Các



loãng (Razdan 1994)


     -indolebutiric axid),
IAA (3-indole acetic axid), NAA (Napthaleneaxetic axid), 2,4-D (2,4-D-
Dichlorophenoxyaxetic axid) và 2,4,5-T (Trichlorophenoxyacetic axid). Trong


-D và 2,4,5-





















4-chlorophenoxyacetic acid (4-CPA) 

-chlorophenoxyacetic acid (PCPA),
2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 2-methyl-4-chlorophenoxyacetic
acid (MCPA), 4-amino-3,5,6-trichloropicolinic acid (picloram),  3,6-
dichloro-2-methoxybenzoic acid (dicamba).
Vai trò của các chất thuộc nhóm auxin:

- 


 




- 

-  



21
- 
- 
- -D)
b. Nhóm các cytokinin
 , 




 , 











 .  6-
benzylaminopurine (BAP) 

 6-benzyladenin (BA), 6---dimethyl-
aminopurine (2-iP), N-(2-furfurylamino )-1-H-purine-6-amine (kinetin),  6-
(4-hydroxy-3-methyl-trans-2-butanylamino ) purine (zeatin). 

 2-


 ,   . 
 .






















 -diphenylurea (DPU), thidiaziron, N-2-chloro-4-
puridyl-N-phenyl urea (CPPU)  .
































 .
 /






(morphogenesis) 



 . 
(embryogenesis),  /cytokinin cao, trong khi
 

. 
 .





 2,4-







 5,0 ppm (mg/l) 
 Agrostis 





 0,1 








 2 








 /

 1. 















 
(transfer RNA).  , 











 .















khi k





 . 







,  ,






























 (





)  .












 , 









 , 



 hân
 . 












 (zea: ngô).






 , 










 .




 













22











 , 


















 , 




:
6-Benzylaminopurine (BAP):  




















 .


























 ,  
.


hát sinh

Chức năng chủ yếu của các cytokinin:
- 
- 
- 
- 


- 
- 
- 
- 
- 
- 
c. Gibberellin










 1930. 
 1895 









 . 
1926,  Gibberella fujikuroi gây ra. 



 30, 









 , 






gibberellin.
 1950, 





















 . T, 








60  . 



3
.
















  : 











 , 







,  .

23

















 . 

































.
            



Gibberellin có các chức năng cơ bản sau:
- 



- 
- ,

- 
- -

- 

- 
- 

- 
e. Abscisic axít (ABA)


















 ,  , 





 .


































 , 
 .

 

- 


- 


24
-   
cành.
- 
f. Ethylene

- Gây già hoá 

- 
- 

- 
-     

Bảng 1. Phân loại các chất điều hoà sinh trƣởng thực vật
Chất điều hoà sinh trƣởng tự nhiên
(phytohocmon)
Chất điều hoà sinh trƣởng nhân tạo
ulator)
Auxin (AIA, IAN, APA)
Gibberellin (A
1
, A
2
, A
3
, A
54
)
Cytokinin (zeatin, zeatin ribozit,
diphenyl ure )
   
2,4D; 2,4,5T;
    
PBA)

AAB, các   
jasnomic

Retardant (MH, CCC, ATIB
C  Ethylene
ACEP

AIA: axit  - indol axetic
IAN: axit  - indolyl axetonitril
APA: axit phenyl axetic
 - ANA: axit  - naphtil axetic
2,4D: axit 2,4 - diclo phenoxiaxetic

BA: benzyl adenin
PBA: tetrahidro piranyl benzyl adenin
MH: malein hidrazit
CCC: clo colin clorit
ATIB: axit 2,3,5 - triiot benzoic
ACEP: axit 2 - 

25
1.5 Những chất thường dùng khác:
* Đường: 
 

                
 - 5%.
* Agar: 
o
C agar

o





* Than hoạt tính: 
- 
hoá.
-           

- 
* Các chất kháng sinh nhân

* Nước dừa: -


- 20%.
* Dịch chiết nấm men
- 1%.

2. Phân loại hóa chất trong vi nhân giống



2.1. Nhóm hóa chất khử trùng
* Cồn:




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