K THUT NUI CY M CHUI

1. Chun b vt liu
a) Chun b mi trng nui cy:
Chun b mi trng khi lm mu cy l c mi trng sn sng.
Mi trng MS
TT

Tn ho cht

1
I
1
2
3
4
5
II
1
2
3
4
5
6
7
III
1
2
IV

2
a lng
NH4NO3
KNO3
MnSO4. 7H2O
KH2PO4
CaCl2. 2H2O
Vi lng
H3BO3
MnSO4. 4H2O
ZnSO4. 7H2O
KI
CuSO4. 5H2O
MoO4Na2. 2H2O
CoCl2. 6H2O
St
FeSO4. 7H2O
Na2EDTA
Vitamin
Axit Nicotinic
(B5)
Thiamin HCl
Pyrodoxin
Inositol
ng Saccaroza
Agar
PH mi trng

1
2
3
V
VI
VII
VIII

Lng pha 1 lt
dung dch
m
3
30,0gr
38,0gr
7,4gr
3,4gr
8,8gr
620,0mg
2300,0mg
860,0mg
83,0mg
2,5mg
25,0mg
2,5mg
5,56gr
7,46gr
100,0mg
20,0mg
100,0mg
100,0mg
20,0gr
6,5gr
5,0 - 5,5

Lng ly cho
1 lt mi trng nui cy
4

Ghi ch
5

Ly 50ml/lt

Pha chung

Ly 50ml/lt

Pha ring

Ly 10ml/lt

Pha chung

Ly 5ml/lt

Pha chung

Ly 2ml/lt

Pha chung

100gr/lt
20gr/lt
6,5gr/lt

Pha chung

Mi trng ti sinh chi: MS + 4 mg/l BAP + 0,5 mg/l IAA + 30g ng + 6g agar
Mi trng nhn nhanh: MS +5mg/l BAP +0,1 mg/l IAA + 30g ng + 6 agar

Mi trng to cy hon chnh: MS BAP 2 mg/l + 20g/l ng + 0,1% than hot

tnh + 6g agar.
b) Chun b chi phi m bo cc yu cu sau:
- Cao t mt t n nh (l bp): 70-80 cm.
- ng knh gc thn gi: 8-10 cm.
- Thn gi c hnh bp mng
- C t 3-4 l hnh li mc.
- Chi tch t cy m c kim dch cn trnh cc bnh hi chnh.
Chi ly v cn ct b phn thn gi, li phn thn ngm c nh sinh trng.
Tch b nhng lp thn gi v phn r c bn ngoi, kh trng bng dung dch
HgCl2 v cn 70%.
2. Nui cy
Sau khi kh trng, ct mu cy thnh tng mu c kch thc 1,5 x 1,5 x 1,0 cm,
a vo mi trng ti sinh chi. Sau 4-5 tun, s c khong 10-12 chi c ti
sinh.
Tch mu cy ti sinh ra tng cm nh, cy sang mi trng nhn nhanh cm
chi, cy chuyn sau 4-5 tun/ ln.
Ti sinh cy hon chnh: sau khi ti sinh s lng cy, tch chi cy sang mi
trng ra r. Chi s vn di v ra r sau 3-4 tun nui cy.
3. Ra ngi cy trong nh li
Cy ti sinh trong bnh cy s c chuyn ra nh li gi trong iu kin mt sau
2-3 tun trc khi ra ngi. Cy ra ngi cao trn 5 cm s c trng trc tip trong
bu.
- Gi th lm bu phi m bo ti xp, thong kh, c m t 60-70% v
dinh dng cho b r v cy pht trin kho.
- Ti bu c mu en, chc chn, c bn m bo cho qu trnh vn chuyn
- Kch thc ti bu: rng 15cm x cao 18cm

4. Tiu chun cy ging nui cy m

- m bo ng u
- Cy ging khng c bin d dip lc phin l.
- Cy ra r trong ng nghim phi c ln ng u, thn gi to kho cao t 5-8
cm, ng knh thn 5-7 mm, c t 3 r tr ln, r trng, di t 2-4 cm, c 3-4 l
tht
- Cy ging nui cy m trong bu t khi xut vn phi c ln ng u, thn
gi to kho cao t 25-30 cm, ng knh thn 10-15 mm, c 5-7 l tht.