Ảnh hưởng của kinetin và sự kết hợp giữa kinetin và 2,4d đến quá trình hình thành protocorm like bodies ở cây lan kim tuyến (anoectochilus setaceus blume)

ẢNH HƯỞNG CỦA KINETIN VÀ SỰ KẾT HỢP GIỮA KINETIN
VÀ 2, 4-D ĐẾN QUÁ TRÌNH HÌNH THÀNH PROTOCORM-LIKE
BODIES Ở CÂY LAN KIM TUYẾN (Anoectochilus setaceus Blume)
Trần Thị Hồng Thúy
1
, La Việt Hồng
2
, Phạm Bích Ngọc
3
, Chu Hoàng Hà
4

Tóm tắt:     Anoectochilus setaceus    phong lan
(Orchidaceae), Chi Anoectochilus,  Anoectochilus setaceus
     protocorm-  
 

.L
-1

2, 4-
 ml.L
-1
 0,5 mg.L
-1
  
86,25%, PLBs si-
-1


-1


         
nghi

Từ khóa: protocorm-like bodies, in vitro, kinetin, lan kim tuyến, vi nhân giống

1. ĐẶT VẤN ĐỀ
 Anoectochilus setaceus Blume hay Anoectochilus roxburghii Wall. Ex
Lindl)   Phong lan (Orchidaceae
cao [1 -
                 

k 2   
1]. 

-like bo
                
(Cymbidium
in vitro
protocorm-like bodies (PLBs) [5]. Theo Ishiiet al (1998) [13], Huan et al 

1
Học viên Cao học, K16, Trường ĐHSP Hà Nội 2
2
Th, Trường ĐHSP Hà Nội 2
3
TS, Viện Công nghệ Sinh học, Viện Hàn lâm KH&CN Việt Nam
4
PGS.TS, Viện Công nghệ Sinh học, Viện Hàn lâm KH&CN Việt Nam



 
 
 

Phalaenopsis

Oncidium
(Jheng et al. 2006) [14]. PLBs 

 

in vitro  in
vitro.
 

 in vitro 
2. NỘI DUNG NGHIÊN CỨU
2.1. Vật liệu và phương pháp nghiên cứu
2.1.1. Vật liệu nghiên cứu
in vitro 
             -D (2, 4-
Dichlorophenoxy acetic acid).
2.1.2. Phương pháp nghiên cứu
Bố trí thí nghiệm
   
--like bodies.

+ 100 ml.L
-1


mg.L
-1
 + 100 ml.L
-1


mg.L
-1
 + 100 ml.L
-1


3: 1,5 mg.L
-1
 + 100 ml.L
-1


 + 100 ml.L
-1

 X         4-    
    100 ml.L
-1
  

1,5 mg.L
-1
kinetin + 0,5 mg.L

-1
2, 4-D 
 + 100 ml.L
-1

1,0 mg.L
-1
kinetin + 0,5 mg.L
-1
2, 4-D 
 + 100 ml.L
-1

 mg.L
-1
kinetin + 0,5 mg.L
-1
2, 4-D 
 + 100 ml.L
-1
.
in vitro 
 

0
C,

Phương pháp xác định chỉ tiêu nghiên cứu
C; 
      ; 

; , 
2.2. Kết quả và thảo luận
2.2.1. Ảnh hưởng của kinetin đến sự hình thành protocorm-like bodies từ chồi của
lan kim tuyến
Protocorm-

 
   
-
    
protocorm-

Bảng 1: Ảnh hưởng của kinetin đến sự hình thành protocorm-like bodies từ chồi của lan
kim tuyến






LBs




PLBs
CT ĐC
-
0
0

-
CT 1

12
86,25
0,23 ± 0,004
Cụm PLB
s
khỏe, nhiều
PLB
s
nhỏ/cụm PLB
s
, màu
trắng xanh
CT 2
12
83,20
0,18 ± 0,040
Số PLB
s
/Cụm PLB
s
ít,
màu trắng xanh
CT 3
10
48,34
0,21 ± 0,060
Nhiều cụm PLB

s
bị teo
nhỏ, màu thâm nâu

                 

- 
 
-1

1 (0,5 mg.L
-1

-1


-1



-1




-1



-1




-1




Hình 1: Quá trình hình thành protocorm-like bodies của cây lan kim tuyến từ chồi trên
môi trường bổ sung kinetin 0,5 mg.L
-1
. A: mẫu chồi, B, C, D, E, F, G, H: hình thành cụm
PLBs trong 30 ngày, Thanh kẻ ngang màu trắng: tương ứng 1 mm
2.2.2. Ảnh hưởng của 2, 4-D kết hợp kinetin đến sự hình thành protocorm-like
bodies từ chồi của lan kim tuyến

-


Bảng 2: Ảnh hưởng của kinetin kết hợp 2, 4-D đến sự hình thành protocorm-like bodies từ
chồi của lan kim tuyến












PLBs
CT ĐC
-
0
0
-
CT 1
7
73,20
0,15 ± 0,004
Cụm PLB
s
phồng to, màu
trắng
CT 2
7
77,11
0,15 ± 0,010
Cụm PLB
s
phồng to, màu
trắng
CT 3
7
84,25
0,16 ± 0,008
Cụm PLB
s

phồng to, màu
trắng


-1
2, 4-
    
 

  
-
-1

-
-
Onchidium [7], [15].

Hình 2: Protocorm-like bodies của cây lan kim tuyếntừ chồi trên môi trường
bổ sung kinetin kết hợp 2, 4-D sau 30 ngày nuôi cấy
3. KẾT LUẬN VÀ KIẾN NGHỊ
in vitro 
ch  -like bodies. M  
   
-l
 
ml.L
-1
au

-1

-D 0,5 mg.L
-1
-like bodies

TÀI LIỆU THAM KHẢO
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nghip, H,. 2005.
2. B Khoa h, Sách đỏ Việt Nam (Phn II. Thc vt - 320 tr), Nxb Khoa
hc T , H., 2007.
3.  c CHXHCN Vit Nam, Ngh nh s -CP, 2006.
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dc, H., 2010.
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512. 2008.
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and orientation on direct somatic embryogenesis from leafexplants of Phalaenopsis
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EFFECT OF KINETIN AND KINETIN COMBINED 2, 4-D ON PROTOCORM-LIKE
BODIES FORMATION OF Anoectochilus setaceus Blume
Tran Thi Hong Thuy, La Viet Hong, Pham Bich Ngoc, Chu Hoang Ha
Abstract
Anoectochilus setaceus Blume belongs to Ochid genus and is one of rare medicinal
plants in Vietnam. The generation of protocorm-like bodies (PLBs) plays an important role in
micropropagating orchid process. In this study, shoots of Anoectochilus setaceus Blume were
used explants to induced PLBs formation on liquid Knudson C media which was added 100
ml.L
-1
coconut water and kinetin or kinetin combinated 2, 4-D. The results shown that PLBs
were the highest induced in liquid cultured Knudson C contains 100 ml.L
-1
coconut water,
1% saccharose and 0,5 mg.L
-1-
kinetin, frequency of explants regenerated PLBs was 86.25%,
PLBs were strongly growth. The combination of 0.5 mg.l
-1
2, 4-D with some different
concentrations of kinetin has not affected PLBs formation of Anoectochilus setaceus Blume.
This is the first report on inducing PLBs from shoots of Anoectochilus setaceus Blume. These
results provides great potential for research and production of Anoectochilus setaceus Blume
in industrial scale via PLBs formation.
Keywords: Anoectochilus setaceus Blume, protocorm-like bodies, in vitro, kinetin,
micropropagation