tissue culturing

1. Micropropogation of saccharum
(sugarcane)
By: Maira sultan

2. Introduction
• Grass family (Poaceae)
• Main product = sucrose fig. 1: sugarcane feilds
• World's largest crop by production quantity
• Brazil =largest producer
• Pakistan =Punjab = 60 - 65 %

3. Tissue culture needs
• Rapid, large-scale production
• Disease-free
• Newly developed varieties
• Speed up the breeding
• Commercialization process

4. Tissue culture Protocols
1. Explant source
2. Medium and condition
3. Callus induction
4. Shoot regeneration
5. Rooting medium
6. Environmental condition fig.2: invitro cultures
7. Acclimatization

5. Explant source
• Healthy young meristems
• Removing leaf sheath
• Cut into 1.0 -1.5 cm length
• Washed for 20-30 minutes
• Alcohol (70 %) for 30 seconds
fig.3: young sugarcane plants

6. medium and conditions
• Semisolid basal MS medium
• Concentrations and combinations of different
PGRs

7. Callus induction
• range of concentration of auxins
• 2,4-D (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
• IBA (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
• NAA (0.5,1.0,2.0,2.5,3.0 and 4.0 mg/l)
Fig.4: callus formation in sugarcane

8. Shoot regeneration
• White friable calli
• Cultured on MS medium
• Combinations and concentrations of:
BAP,
Kinetin (0.5- 4.0mg/l)
IBA, NAA (0.1-1.0mg/l)
• Multiple shoot regeneration individually or in
combination

9. Rooting
• Half-strength (1/2 MS)
• IBA
• NAA
• IAA (0.5-3.0mg/l)
fig.5: roots of sugarcane

10. Environmental conditions
• pH = 5.8 before gelling
• Photoperiod = 16h
• Temperature = 25 ± 3 C
• Relative humidity = 70-80%

11. Acclimatization
• Culture medium
• plastic trays
• 1/8 MS basal salt
solution
• After 30 days transplanted in to
field condition

12. Results
• Callus Induction:
• Regeneration of Micro shoots:

13. Thank you