Micropropogation of saccharum(sugarcane)

By: Maira sultan

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 %

Tissue culture needs

• Rapid, large-scale production

• Disease-free

• Newly developed varieties

• Speed up the breeding

• Commercialization process

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

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

medium and conditions

• Semisolid basal MS medium

• Concentrations and combinations of different PGRs

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

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

Rooting

• Half-strength (1/2 MS)

• IBA

• NAA

• IAA (0.5-3.0mg/l)

fig.5: roots of sugarcane

Environmental conditions

• pH = 5.8 before gelling

• Photoperiod = 16h

• Temperature = 25 ± 3 C

• Relative humidity = 70-80%

Acclimatization

• Culture medium

• plastic trays

• 1/8 MS basal saltsolution

• After 30 days transplanted in to field condition

Results

• Callus Induction:

• Regeneration of Micro shoots:

Thank you