This document outlines a plan for a sugarcane tissue culture project. It begins with an introduction on sugarcane production and importance of quality seed. The objectives are then stated as producing more cane and sugar through tissue culture with disease-free varieties in less time and at lower cost compared to conventional methods. The methodology involves shoot tip culture, meristem culture, and callus culture. A 4-year plan is provided with the first year focusing on setup and small-scale production, expanding to large-scale in years 3-4. Required facilities, estimated quarterly budget, and anticipated outcome of more profitable sugarcane plantation establishment through higher propagation rates are described.

1. Production of sugarcane by
tissues culture

3. Contents
1. Title
2. Origin and definition of the problem
3. Objectives
4. Scientific utility of the project
5. Review of research conducted/ being conducted on the subject in India and abroad
6. Methodology and experimental technique
7. Yearwise plan of work
8. Facility available and required
9. Budget estimation
10. Outcome of the project.

4. INTRODUCTION
•Sugarcane , Saccharum spp.belongs to family Poaceae. It is the main sugar production crop that contributes
more than 75% of the total sugar pool at the global level. Sugarcane is cultivated as a commercial crop in nearly
60 countries spread over the world. India is the largest producer of sugarcane in the world.
•Quality seed plays an immense value to keep phase in productivity and sugar recovery. Due to limitation of time
taking process to mass multiplication of elite popular varieties there is an importance of substitutes methodology
to fasten the multiplication rate and to evolve desirable clones to the changed climatic condition.
•Tissue culture provide an alternative method for the crop improvement. Plant regeneration from tissue culture of
sugarcane has been successfully applied to breeding programs for rapid screening of clones for disease
resistance, salt tolerance, herbicide resistance and early maturity and high sugar.

5. USES AND IMPORTANCE OF
SUGARCANE
•Sugarcane is mainly an industrial crop as the cane is supplied to sugar industries.
•Sugarcane’s products like sugar and fermented products are very important in making and
preserving various kinds of medicines like syrups, liquids, capsules etc.
•Sugarcane provide a juice, which is used for making white sugar, and jaggery and by-products
like bagasse and molasses.
•Green top of cane are a good source of fodder for cattle. Its remains are good manure in
alkaline and saline soils.

6. PROBLEM
Seed multiplication of newly released varieties of sugarcane is one of the major constraints.
Once a desired clone is identified, it usually takes 6-7 years to produce sufficient quality of
improved seed material. This long duration causes a major bottleneck in breeding
programmes.

7. OBJECTIVE
1. Better rate of propagation (micro propagation offers a practical and fast method for mass propagation )
2. More cane and sugar yield than the conventional seed
3. Disease free variety
4. Save time
5. money .

8. SCIENTIFIC UTILITY OF
PROJECT
 Use as a nutritional drink
 comprises significant amount of minerals, vitamins, and hydrophilic compounds
 The presence of pharmacological activities is proven in sugarcane juice and its unripened products such as
brown sugar, molasses, and jaggery are considered as richest sources of phenolic compounds, such as phenolic
acids, flavonoids, and different glycosides.
 The lipophilic compounds including various policosanols and phytosterols are the important components of
sugarcane wax present in sugarcane leaves and shoots are observed with several pharmacological effects such
as sympathomimetic, antihypercholesterolemic, and antithrombotic activities.

9. • Shenk and Hildebrandt (1972) have reported requirement of high Concentration of auxin for rooting in
sugarcane.
• Barba et al. (1977) reported that root development requires higher sugar levels in nutrition media.
• Barba and Nickell (1969) reported that Sugarcane tissues subcultured for Over 4 years had lost the
capability to differentiate shoots.
• Micropropagation is an in vitro method for clonal multiplication of plants using meristematic or non-
meristematic cells/tissues as the explant. Plants can be regenerated directly (adventitiously) from the
explant (Geijskes et al., 2003) or indirectly (de novo) through the callus derived from the explant (Heinz and
Mee, 1969).
• In sugarcane, plants have been produced by direct regeneration from both apical and axillary meristems
(Taylor and Dukie, 1993) and from immature leaf tissues (Lakshmanan et al., 2002; Geijskes et al., 2003).
As with other plant species,
REVIEW OF RESEARCH CONDUCTED

10. METHODOLOGY AND
TECHNIQUE
 Types of micropropogation technique in sugarcane
1. Shoot tip culture
2. Meristem culture
3. Callus culture

11. 1. Shoot tips were collected from juvenile sugar-cane plants (3-4 months age) and were used as explants.
2. Sterilization of explants was carried out using 0.1% HgCl2 after washing thoroughly under tap water for
7-10 min.
3. Subsequently the explants were washed gently with sterile DDH2O (double distilled water) in aseptic
condition under laminar flow hood.
4. Shoot tips of 2-4 mm were excised and placed on MS (medium supplemented with different
combinations of auxin and cytokinin to identify the appropriate media combinations for regeneration of
sugar-cane through shoot tip culture.
• Media were consisted of 3% sucrose, 0.6% agar, pH was adjusted to 5.7 before addition of agar and
autoclaved at 120°C for 15 min.
5. Explants were incubated at 25±2°C under 16 h photoperiod regime.
SHOOT TIP CULTURE

12. 6. The regenerated shoots were multiplied manifolds when they were sub-cultured in the same medium within
three weeks.
7. The regenerated shoots were devoid of roots. So, for root induction the shoots were excised separately and
placed on rooting media.
8. The proper stage of root development was another criterion for selecting plantlets to be transferred to the
soil.
9. In vitro regenerated plantlets were transferred to small pots containing mixture of soil and sand for future
establishment.

14. Year wise plan
1ST YEAR- In first year there will be hardly any profit as, all the money will be
spend on set up. And the set up will be on the small scale.
2ND YEAR- profit will increase as compare to the previous year , as set up will
be already done and we will focus on selling our product and in making a secure
place in market for our product .
3rd and 4th year –this will be the year of expanding to large scale, only if
when our profit will be more than 50% in small scale. We will buy more land and will
increase our production and supply.

15. Facility available and required
1. Fund
2.Tissue culture lab
3. Reagents & chemicals
4. Electricity
5 .Water supply
6.Land
7.labour

16. Budget estimation
Quarter 1 Quarter 2 Quarter 3 Quarter 4
Funding 50,00,000
land 21,00,000
electricity 15,000 15,000 20,000 25,000
water 10,000 10,000 13,000 15,000
labor 150,000 150,000 170,000 175,000
chemical 100,000 100,000 30,000
Tissue culture
lab
5,00,000
equipment 2,00,000 15,000
bonus 20,000
medical 10,000 5,000 5,000
vehicle 4,00,000
petrol 10,000 10,000 25,000 25,000
maintenance 5,000 7,000

17. OUTCOME OF THE PROJECT
The use of tissue cultured plant source is by far more profitable than using the conventional plant
source in term of the rate of propagation. Thus in the multitude challenges of sugarcane
plantation establishment we get more profit .
Now, we have large number of sugarcane plant in short period of time .

18. THANK YOU