Slow Growth Storage
Slow Growth Storage
In situ Ex situ
In vitro collections
Slow growth
storage
(°C , O2 , H2O Cryopreservation
, medium ~) (-196°C)
✓ Slow growth storage (=conservation at above freezing temperatures)
1st...................4th..................8th......................12th
Several months
?
Conservation at low temperature
Conservation of shoot culture at low temperature (=slow growth storage)
determining the
Initial material
32.4
62.4
3 3
2,13
2.01 2
2
1
1 0,27
0,32
0
0
standard culture after 16 mon post-storage standard culture after 16 mon post-storage
Inizio
conditions 16
thsmesi
at 4°C Rec.
regrowth Inizio
conditions 16 mesi
ths at 4°C Rec.
regrowth
Sucrose 60 g/l
S60
Sucrose 30 g/l + Mannitol 15S30+M15
g/l
5
5 4,83 4,31 4,43
3,81 4
4
3
3
2 2
1 1
0,29 0,19
0
0
standard culture after 16 mon post-storage
Inizio
conditions
16 mesi
ths at 4°C
Rec.
regrowth
standard culture
Inizio
after 16 mon
16 mesi
post-storage
Rec.
conditions ths at 4°C regrowth
Gisela 5® 12 months: morphological evaluation of survival and
quality (% of jars with material still conservable)
100
+/- ++ +++ +
90 100 100 100
80
70
60 66,7
50
%
40
30
20
10
0
15
12,65
12
8,85
9
6 5,52
3,81
3
0
S30 S45 S60 S30+M15
WM (Workable Material) = (no jars still in storage/total n° of initial jars) x (average gr of WM per jar)
Gisela 5® After 20 months
- Gas permeability +
Slow growth storage
Storage conditions:
✓ 4°C
✓ darkness
✓ different containers
✓ 6 months
Glass jars,
500 cc
Plastic tubes,
80 cc
Standard
1.7 Proliferation Rate:
2.0
‘Domenica’ Conservation at 4°C in the dark: 4 months
Standard
2.0 Proliferation Rate:
3.7
Conservation at 4°C in the dark: 6 months
‘Rosa Sant’Antonio
di Padova’
‘Domenica’
2-3 subcultures
Stop complaining!
I told you! We don’t have
money to buy a cold cham-
ber for the laboratory!