Micropropagation is a technique used to propagate selected sugarcane varieties using in vitro culture. It allows for rapid multiplication of plants while eliminating diseases. The process involves culturing small explants on nutrient media to induce shoot formation, multiplying the shoots, rooting the plantlets, and hardening them for field planting. Specific media compositions and plant growth regulators are used at different stages to optimize growth. Micropropagation provides benefits for sugarcane improvement by allowing widespread distribution of new varieties and selection of plants with desired traits.

2. MICROPROPAGATION TECHNIQUE &
PROPAGATION TECHNIQUE OF
SUGARCANE
DEPARTMENT OF PLANT BIOTECHNOLOGY
COLLEGE OF AGRICULTURE , GWALIOR
GUIDED BY : DR. ASHOK AHUJA
SUBMITTED BY : PRIYA TIWARI
141A48
B.Sc.(Ag.) 4th year ELP

3. WHAT IS MICROPROPAGATION ?
Micropropagation is the true to type
propagation of selected genotype or
varieties using in vitro culture technique.
OR
The production of whole plants from small
sections of a plant, called an “explant”

4. ADVANTAGES OF MICROPROPAGATION
 Rapid rate of multiplication ensuring spread of new
varieties.
 Disease elimination .
 Plants having small seeds can be grown reliably.
 High fecundity rates.
 Process is independent of seasons.
 In dioecious species, plants of desired sex can be
selected and multiplied .

5. SUGARCANE – AN INTRODUCTION
 Sugarcane is an important cash crop of India.
 Sugarcane ( Saccharum spp.) ,a C4 perennial
grass, is a member of the Gramineae family.
 It constitutes a major source of edible sugars.
 Sugarcane is an oldest major crop known to man, a
major crop of tropical and subtropical regions
worldwide.
 India is the second largest producer of sugarcane in
world.

6. TYPES OF MICROPROPAGATION TECHNIQUE IN
SUGARCANE
 Shoot tip culture.
 Meristem culture .
 Callus culture.

7. TOP PRODUCERS OF SUGARCANE IN WORLD
2013-14
COUNTRY PRODUCTION
( THOUSAND METRIC TONS)
Brazil 734 000
India 342 382
China 115 124
Thailand 95 950
Pakistan 55 309
Mexico 49 735
Phillippines 34000

8. STAGES OF MICROPROPAGATION
 Stage 1 : Establishment of shoot cultures
 Stage 2 : Multiplication of shoots
 Stage 3 : Rooting and hardening
 Stage 4 : Field planting

9. CULTURE MEDIA COMPOSITION
 For shoot initiation and rooting medium ,White’s medium is
used. For shoot multiplication medium, modified MS medium is
used.
 In shoot initiation medium (White’s medium) , PGRs used are
following :
 Kinetin (1.07 mg/l)
 Gibberellic acid (0.5 mg/l)
 IBA (1 mg/l)
 In shoot multiplication medium , PGRs used are following:
 Kinetin (1.07 mg/l)
 Gibberellic acid (0.5 mg/l)
 NAA ( 0.5 mg/l)
 6-BAP (0.25 mg/l)
 In root medium , PGRs used are following:
 NAA (1 mg/l )

10. STAGE 1 : ESTABLISHMENT OF SHOOT
CULTURE : STERILIZATION
 Collect actively growing shoots from 3-4 month old field
grown healthy plants.
 Remove outer sheath by wiping sheath with rectified spirit.
 Explant prepared by cutting growing apices 7-8 cm long
with few whorls of leaves.
 Wash it with surf for 5 min. with several change of water.
 Treat it with 70% ethyl alcohol for one min. and rinse it with
water for 4-5 times.
 Treat with 10% sodium hypochlorite solution & cover with
aluminium foil and shake vigorously for 20 min.

11. MATERIAL COLLECTION AND EXPLANT
PREPRATION

12. B ) INOCULATION
 In Laminar air flow, the material is washed thoroughly 3-
4 times with sterile distilled water till the chlorine smell is
removed.
 The apical dome along with two leaf primordial is
excised with help of a sterile sharp blade and
immediately placed on a filter paper support and
immersed in the shoot apex medium .
 The culture are incubated at 26° ± 1° C and under 16
hr light (2500 lux).
 Transfer of the meristem to fresh media is essential for
a few days.
 After 15 days elongated shoot apices are generally
transferred to bigger container for further growth and
elongation.

13. Inoculated shoot tip

14. Elongated shoot tip
Axillary shoot development

15.  After 45 to 60 days depending on the growth of the
shoot apices, the material is transferred to
multiplication medium .
 After 45 days new shoots will arise from the axils of
the developing shoots.
 These are separated in small groups in fresh
medium.
STAGE 2 : MULTIPLICATION OF SHOOTS

16. Multiplication of Shoots

17. STAGE 3 : ROOTING AND HARDENING
 The stage involves the induction of roots in vitro by
transferring the well grown plants into rooting
medium .
 A group of 4-5 plants can be kept in culture tubes
containing the medium.
 Within 15-30 days roots will be formed.
 Plantlets with well developed roots are planted in
polybag and hardened.
 The mixture for planting is prepared by mixing
sieved sand, silt presumed in 1:1:1 ratio.

18. Rooted plants

19. HARDENING OF PLANTS
Pre-hardening
Hardened plants

20. STAGE 4 : FIELD PLANTING
 Established plants after 45-60 day are transferred
to field in normal crop season and survival rate is
85-90 per cent.
 A spacing of 45 cm between plants and 90 cm
between rows is recommended. Canes are ready
for harvest at 8 to 10 months for seed purposes.

21. MEDIA COMPOSITION
Ingredients Shoot apex
medium (mg/l)
(whites basal)
Multiplication
medium (mg/l)
(modified MS)
Rooting
medium
(mg/l)
(whites
medium)
Ammonium
nitrate
_ 1640 _
Potassium
nitrate
_ 1900 _
Calcium chloride _ 440 _
Magnesium
sulphate
720 _ 720
Sodium sulphate 200 _ 200
Potassium
chloride
65 _ 65

22. Ingredients Shoot apex
medium (mg/l)
(whites basal)
Multiplication
medium (mg/l)
(modified MS)
Rooting
medium
(mg/l)
(whites
medium)
Potassium nitrite 80 _ 80
Calcium nitrite 300 _ 300
Sodium
dihydrogen
orthophosphate
16.5 16.5
Potassium
dihydrogen
orthophosphate
_ 170 _
Boric acid 1.5 6.2 1.5
Cobalt chloride _ 0.025 _
Sodium
molybdate
_ 0.25 _
Cupric sulphate _ 0.25 _

23. Ingredients Shoot apex
medium (mg/l)
(whites basal)
Multiplication
medium (mg/l)
(modified MS)
Rooting
medium
(mg/l)
(whites
medium)
Potassium iodide 0.75 _ 0.75
Zinc sulphate 3 _ 3
Manganese
sulphate
7 22.5 7
Ferric EDTA 36.7 36.7 36.7
Meso inositol 100 100 _
Kinetin 1.07 1.07 _
Gibberlic acid 0.5 0.5 _
Indole Butyric
acid
1 _ _
Glycine 2 2 3

24. Ingredients Shoot apex
medium (mg/l)
(whites basal)
Multiplication
medium (mg/l)
(modified MS)
Rooting
medium
(mg/l)
(whites
medium)
Nicotinic acid _ 0.5 0.5
Pyridoxine _ 0.5 0.1
Thiamine _ 0.1 0.1
Naphthalene
acetic acid
_ 0.5 1
6 Benzyl amino
purine
_ 0.25 _
Calcium
pantithenate
_ _ 1
Riboflavin _ _ 1
Coconut water 100 ml 100 ml _
Sucrose 20 g 20 g 20 g
pH 5.8 5.8 7.0

25. REFERENCES:
 Jalaja, N.C. 2001. a practical manual for the sugarcane
micropropagation. A Publication
from Sugarcane breeding Institute, Coimbatore.
 Sreenivasan, T.V. and N.C. Jalaja. 1992.
Micropropagation of sugarcane varieties for
increasing cane yield. Sugar Journal, South
India Sugarcane and Sugar Technologists
Association, 19(4): 61-64.
 Sundara, B. and Jalaja, N.C. 1944. Effect of intra – row
spacing on mericlone transplants
of Sugarcane (Saccharum officinarum). Indian Journal
of Agricultural Sciences. 64(8): 546-549.
 Sundara, B. 1955. Economics of using sugarcane
mericlones for commercial planting.
Co-operative sugar, 26(6): 459-461.