This document summarizes recent research on sugarcane tissue culture. It discusses that sugarcane is an important crop grown for sugar and ethanol production. Tissue culture techniques like micropropagation through shoot tip culture and callus culture are used for rapid multiplication of sugarcane varieties. The document provides details of explants used, sterilization processes, growth media, and hormones found most effective for callus induction and shooting. It also discusses using tissue culture for somaclonal variation to develop salt tolerant varieties and mentions challenges in obtaining genetically stable plants.

1. Case study report on Sugarcane Submitted by : Arun N ID Number: 008078 Module: D2D001 School of Bioscience, Nottingham University Recent advances in research on plant tissue culture of Sugarcane 1

2. Sugarcane is one of the most important crops in the world. Brazil is the largest sugarcane grower, they grow it for: Sugar – Production of food & alcoholic beverages Ethanol – Gasoline + ethanol blend as fuel In sugarcane, micropropagation is important for rapid multiplication of elite genotypes/clones and for the spreading of new varieties. Research attention on tissue culture of sugarcane was intensified due to its economic importance as a cash crop. 2 Sugarcane tissue culture

3. In vitro plant regeneration using shoot tip culture. In vitro micropropagation through callus culture. Somaclonal variation – screening for salt tolerance. 3 Recent research study on

4. Explants used are: Young leaves Shoot tips Meristem cells Leaf sheath 4 Laboratory details Common surface sterilization techniques are followed: Steps highlighted in green color are carried out inside the flow chamber

6. In vitro shoots are inoculated on half-strength MS medium for better rooting.

7. 2,4-D conc. 2.5-3.0mg/L is best for callus induction.

8. NAA 3.0mg/L along with ½ strength MS medium is good for rooting.5

9. In vitro plant regeneration using shoot tip culture 6 For shoot regeneration, cytokinin BAP was more effective than Kn and IBA. Auxin is essential for root initiation. NAA is best for rooting. Whereas, IAA or IBA produce poor quality roots. Rate of multiplication is high in adventitious shoots.

10. Plants regenerated from meristem (shoot tip) are very similar, both Phenotypically Genotypically, to the parent plant. Shoot tip culture is better than leaf roll culture for plant production. For shoot regeneration the combination of auxin and cytokinin is essential. Regeneration potential of callus was specific and genotype dependent and also parallel with hormonal conc. and combination. 7 In vitro propagation of Sugarcane

11. 8 In vitro callus regeneration and plant establishment (1) Callus regeneration in MS+2.5mg/l 2,4-D (2) Callus regeneration in MS+2.5mg/l NAA (3&4) Multiple shoot emergence from callus tissue in MS+2.0 mg/l BAP+0.5mg/l IBA (5) Micro shoots rooted in 1/2MS+NAA(2.5 mg/l). (6) Hardening of rooted plant lets in plastic trays.

12. Somaclonal variation The term somaclonal variation was first introduced by Scow Croft and Larkin (1981) In Saccharum Sp somaclonal variation is termed as sub-clonal variation. Salt tolerant plants were produced by tissue culture techniques Khan et al., (2004) Explant – young leaf Regeneration medium – Fe-EDTA 1mg/L, Casein Hydrolyzate 500mg/L, Cystine free base 30mg/L Salt tolerant soma-clones performed better in the above mentioned characteristics 9

13. Over the past 10 years tissue culture technology of sugarcane has been developed, but not without some challenges. Work on Tissue culture should integrate hand in hand with molecular techniques, it will be helpful in obtaining genetically stable plants. Mass propagation of Sugarcane through Tissue culture approach will result in availability of disease free germplasm, propagation of newer germplasm with improved agronomic characters for the ultimate benefit of the farmers. Sugarcane tissue culture work can be directed towards germplasm conservation also. 10 Conclusion

14. In order to ascertain the true to type character of the Tissue culture raised plantlets through shoot proliferation, it is essential to check at the genomic level, it is felt that further experimentation is needed towards achieving the result. 11 Further studies

15. Alam, M. H. (1995). Highly frequency in vitro regeneration in sugarcane. Sugarcane, 6:20-21. Baksha, R. A. (2002). In vitro shoot tip culture of sugarcane (Saccharum) variety Isd-28. Biotechnology, 1(2-4):67-72. Begum, S. H. (1995). Efficient regeneration of plants from leaf base callus in sugarcane. Plant tissue culture, 5:1-5. Behera, K. K. (2009). Rapid in vitro micro propagation of sugarcane (Saccharum officinarum L. Cv-Nayana) through callus culture. Nature and Science, 7(4). Biradar, S. B. (2009). In vitro plant regeneration using shoot tip culture in commercial cultivar of sugarcane. Karnataka J. Agric. Sci., 22(1):21-24. Heinz, D. a. (1969). Plant differentiation from callus tissue of Saccharum species. Crop Science, 9:346-348. Karim, M. A. (2002). Micropropagation of two sugarcane (Saccharum officinarum) varieties from callus culture. Online Journal of Biological Science, 2(10): 682-685. Khan, S. K. (2004). Somaclonal variation in sugarcane through tissue culture are subsequent screeing of salt tolerance. Asian Journal of Plant Science, 3(3): 330-334. Liu, M. (1983). In vitro methods applied Sugarcane improvement. In: Thope T A (ed) Plant tissue cultures: methods and applications in agriculture, pp 299-323. Maretzki, A. (1987). Tissue culture: Its prospects and problems In: Sugarcane Improvement through breeding. (Ed.) D.J.Heinz. Elsevier Science publisher B.V., 343-384. 12 References

16. 13 Thank you 