|
| |
Tissue culture of Hemerocallis has been explained by Kyte
and by Hartmann et. al.
We shall follow the process suggested by Kyte. The Kyte approach was based upon
the work of Heuser and Harker.
The following Table describes the process.
|
Process |
Comments |
|
Explant |
2 mm sections of
young inflorescence scapes. Flower petals and sepals from 1 mm flower
buds. |
|
Treatment |
Remove and discard
bracts from buds. Wash inflorescence with 0.1 % Tween 20. Rinse. Mix 10%
bleach with Tween for 20-30 minutes. Rinse three times in sterile water.
In hood, moisten sterile toweling with sterile antioxidant. Slice scape
into 2 mm sections. Place sections down upside down i medium in test
tube. |
|
Media |
Modified MS with
KH2PO4 casein hydrolysate, malt extract, adenine sulfate. Hormones NAA,
kinetin for callus formation with NAA lowered for plantlet formation. |
|
Light |
Continuous dark for
4-8 weeks for callus followed by 300-1,000 f.c. continuous for plantlet. |
|
Temperature |
26 C |
|
General |
Daylily meristems
are not used because they are hard to clean |
The following Table depicts the media for callus and
plantlets that is recommended.
|
Compound |
Callus |
Plantlet |
|
NH4NO3 |
1650 |
1650 |
|
KNO3 |
1900 |
1900 |
|
CaCl2,
2H2O |
440 |
440 |
|
KH2PO4 |
300 |
300 |
|
MgSO4,
7H2O |
370 |
370 |
|
MS minor salts |
32.8 |
32.8 |
|
FeSO4,
7H2O |
27.8 |
27.8 |
|
Na2EDTA |
37.8 |
37.8 |
|
Inositol |
100 |
100 |
|
Thiamine HCl |
0.4 |
0.4 |
|
Kinetin |
0.1 |
0.1 |
|
AdSO4 |
160 |
160 |
|
NAA |
10 |
0.5 |
|
Malt Extract |
500 |
500 |
|
Casein
hydrolysate |
500 |
500 |
|
Sucrose |
60000 |
30000 |
|
Agar |
6000 |
6000 |
|
pH 5.5 |
|
|
|
Telmarc 
