Optimization of Temperature Conditions for Screening Thermotolerance in Sugarcane through Temperature Induction Response (TIR) Technique

Author(s): R. Gomathi, S. Shiyamala, S. Vasantha, A. Suganya

An experiment was conducted with the objective of standardizing temperature conditions for screening thermotolerance in sugarcane variety Co 86032 under in vivo and in vitro conditions. Lethal temperature condition for settlings (in vivo) and callus (in vitro) were identified by subjecting the 30 days old settling and callus 7 temperatures condition (39, 41, 43, 45, 47, 48 and 49 ºC) with four time durations (5, 10, 15 and 20 hrs). Temperature conditions at 48ºC with 20 h and 48 ºC with 15 h of heat stress treatment were identified as critical temperature condition for settlings and callus respectively. To determine the optimum induction temperature conditions (sub lethal temperature) for settlings, the variety Co 86032 was exposed to gradual increase in temperature from 38, 40, 42 to 44 ºC with two time durations (10 and 15 h), followed by lethal temperature (48 ºC with 10 h). The optimum induction temperature condition for developing thermo tolerance in settlings was worked out to be 40 ºC with 10 h stress treatment. Similarly, the optimum induction temperature condition for calli for develop thermo tolerance was worked out as 42ºC with 10 h. Adaptive response of Co 86032 by heat acclimation was investigated under in-vivo and in-vitro conditions through temperature induction response technique. It was found that induced settlings and calli for thermotolerance recorded higher soluble protein, proline, glycine betaine, total phenols, POX activity and SOD activity than non-induced.

Sugarcane, temperature induction response (TIR) technique, in vivo, in vitro, thermotolerance, ROS scavenging enzymes, osmolytes

1. Ashraf, M., & Foolad, M.R. (2007). Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environmental and Experimental Botany 59: 206–216
2. Ashraf, M., & Hafeez, M. (2004). Thermotolerance of pearl millet and maize at early growth stages: growth and nutrient relations. Biol. Plant. 48:81–86
3. Bates, I.S., Waldren, R.P.,& Teare, I.D. (1973). Rapid determination of free proline for water stress studies. Plant and Soil 39: 205-207
4. Beauchamp, C., & Fridovich, I., (1971). Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal. Biochem .44(1): 276–287
5. Burke, J.J. (1998). Characterization of acquired thermotolerance in soyabean seedlings. Plant Physiol. Biochem. 36: 601 – 7
6. Chen, T.H.H., Shen, Z.Y. & Lee, P.H. (1982). Adaptability of crop plants to high temperature stress. Crop Science. 22: 719–725
7. Cushman, J.C. & Bohnert, H.J. (2000). Genomic approaches to plant stress tolerance. Current Opinion in Plant Biol. 3:117–124
8. Gomathi, R., Vasantha, S., Hemaprabha, G., Alarmelu, S., & Shanthi, R.M. (2011). Evaluation of elite sugarcane clones for drought tolerance. Jl. Sugar Res.1: 55–62
9. Gong, M., Chen, S.N., Song, Y.Q., & Li, Z.G. (1997). Effect of calcium and calmodulin on intrinsic heat tolerance in relation to antioxidant systems in maize seedlings. Aust. Jl Pl. Physiol. 24: 371–379
10. Gong, M., Knight, M.R., & Trewavas, A.J. (1998). Heat-shock induced changes of intracellular Ca2? level in tobacco seedlings in relation to thermotolerance. Plant Physiol. 116: 429–437
11. Gopalakrishna, R. (2001). Cloning and characterization of moisture stress responsive genes from stress-tolerant groundnut (Arachis hypogaea L.). PhD Dissertation submitted in University of Agricultural Sciences, Bangalore.
12. Grieve, C.M., & Grattan, S.R. (1983). Rapid assay for determination of water soluble quaternary ammonium compounds. Plant and Soil. 70: 303–307
13. Hahn, G.M., & Li, G.C. (1990). Thermo-tolerance, thermo-resistance and thermo-sensitization. In: Morimoto R.I., Tissieres A. and Georgopoulos, C. (edits). Stress proteins in biology and medicine. p. 79–100, Cold Spring Harbor, New York
14. Hatice, G., & Atilla, E. (2003). Some physiological changes in strawberry plants under heat stress. Jl. Horti. Sci. and Biotech. 78: 894–898
15. Heath, R.L., & Packer, L. (1968). Photoperoxidation in isolated chloroplasts. Arch. Biochem. Biophy.125: 189–198.
16. Hong, S.W., Lee, U., & Vierling, E. (2003). Arabidopsis hot mutants define multiple functions required for acclimation to high temperatures. Plant Physiol. 132:757–67
17. Howarth, C.J., Pollock, C.J., & Peacock, J.M. (1997). Development of laboratory – based methods for assessing seedling thermotolerance in pearl millet. New Phytol. 137: 129 – 39
18. Jain, M., Mathur, G., Koul, S., & Sarin, N.B. (2001). Ameliorative effects of proline on salt stress induced lipid peroxidation in cell lines of groundnut (Arachis hypogea L). Plant Cell Rep.20: 463-468
19. Jiang, Y.W., & Huang, B.G. (2001). Effects of calcium on antioxidant activities and water relations associated with heat tolerance in two cool-season grasses. J. Exp. Bot. 52:341–349
20. Kumar, G., Krishnaprasad, B.T., Savitha, M., Gopalakrishna, R., Mukhopadhyay, K. Ramamohan, G., & Udaya Kumar, M. (1999). Enhanced expression of heat shock proteins in thermotolerant lines of sunflower and their progenies selected on the basis of temperature induction response (TIR). Theoretical and Applied Genetics. 99: 359-367
21. Laemmli, U.K. (1970). Cleavage of structure proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685
22. Larkindale, J., & Huang, B. (2004). Thermotolerance and antioxidant systems in Agrostis stolonifera: involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. Jl of Plant Physiol.161: 405–413
23. Larkindale, J., Hall, J.D., Knight, M.R., & Vierling, E. (2005a). Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiol. 138:882–97
24. Larkindale, J., Mishkind, M,. & Vierling, E. (2005b). Plant responses to high temperature, In: Jenks M. A. and Hasegawa P. M. (edits). Plant abiotic stress. p:71-95. Oxford: Blackwell Scientific Publications. London, UK
25. Leopold, A.C., Musgrave, M.E., & Williams, K.M. (1981). Solute leakage resulting from leaf desiccation. Plant Physiol. 68:1222–1225
26. Levitt, J. (1980). Responses of plants to environmental stresses. Chilling, Freezing and High Temperature Stresses. Vol. 1. Academic Press.. pg. 1-.449
27. Liang, Y., Sun, W., Zhu, Y.G., & Christie, P. (2007). Mechanisms of silicon-mediated alleviation of abiotic stresses in higher plants: A review. Environ. Pollut. 147: 422–428
28. Lindquist, S.L., & Craig, E. (1988). The heat shock proteins. Annu. Rev. Genet. 22: 631–637
29. Lowry, O.H., Rosebrough, N.J., Farr, A.L., & Randall, R.J. (1951). J. Biol. Chem.193- 265
30. Malik, C.P., & Singh, M.B.(1980). Plant Enzymology and Histo-enzymology. Kalyani Publishers , New Delhi
31. Moyer, R.A., Hummer, K.E., Finn, C.E., Frei, B.,& Wrolstad, R.E. (2002). Anthocyanins, phenolics, and antioxidant capacity in diverse small fruits: vaccinium, Rubus, and Ribes. J. Agri. Food Chem. 50: 519–525
32. Nadonly, L., & Sequeiria, L. (1980). Increase in peroxidase activities are not directly involved in induced resistance in tobacco. Physiol. Plant Pathol.16: 1-8
33. Nelson, V., Adger, J., & Brown. (2008). Adaptation to Environmental Change: Contributions of a Resilience Framework. Valerie Nelson, Richard Lamboll and Adele Arendse (Edits). In. Climate Change Adaptation, Adaptive Capacity and Development Discussion Paper, UK
34. Pareek, A., Singla, S.L., & Grover, A. (1997). Short term salinity and high temperature stress associated ultra- structural alterations in young leaf cell of Oryza sativa L. Ann. Bot. 80:629–39
35. Sairam, R.K., & Tyagi, A. (2004). Physiology and molecular biology of salinity stress tolerance in plants. Curr. Sci. 86: 407-421
36. Sakamoto, A. & Murata, N. (2002). The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant Cell Environ. 25: 163–171
37. Sangwan, V., & Dhindsa, R.S. (2002). In vivo and in vitro activation of temperature-responsive plant map kinases. FEBS Letters. 531: 561–564
38. Sanjam MJ, Tucic B., & Matic, G. (2010). Differential expression of Heat Shock proteins Hsp70 and Hsp90 in vegetative and reproductive tissues of Iris pumila. Acta Physiol. Planta. 33: 233–240
39. Scandalios, J, G., Acevedo, A., & Ruzsa, S. (2000). Catalase gene expression in response to chronic high temperature stress in maize. Plant Sci. 156: 103–110
40. Scharf, K.D., Heider, H., Hohfeld, I., Lyck, R., Schmidt, E, & Nover, L. (1998). The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules. Mol. Cellular Biol.18: 2240–2251
41. Senthil-Kumar, M., Srikanthbabu, V., Mohan Raju, B., Ganeshkumar,N., Shivaprakash, N., & Udayakumar, M. (2003). Screening of inbred lines to develop a thermotolerant sunflower hybrid using the temperature induction response (TIR) technique: a novel approach by exploiting residual variability. J. Exp. Bot. 54(392): 2569-2578
42. Srikanthbabu, V., Ganesh Kumar, Krishnaprasad, B.T., Gopalakrishna, R., Savitha, M., & Udaya Kumar. M. (2002). Identification of pea genotypes with enhanced thermo tolerance using temperature induction response (TIR) technique. J.Pl. Physiol. 59:535–545
43. Srikanthbabu, V. (2004). Identification of thermotolerant lines from the mutated population of groundnut (Arachis hypogea L.) based on the temperature induction response and expression of HSFs and HSPs. Ph.D Dissertation submitted to the University of Agricultural Sciences, Bangalore – 65
44. Steel, R.G.D., Torrie, J.H., & Dickey, D.A. (1996). Principles and procedures of statistics: a biometrical approach, 3rd ed. NewYork: McGraw Hill
45. Sung, D.Y., Kaplan, F., Lee, K.J., & Guy, C.L. (2003). Acquired tolerance to temperature extremes. Trends Plant Sci. 8: 179–87
46. Taiz, L., & E. Zeiger (2006). Plant Physiology. Massachusetts: Sinauer Associates Inc. Publishers.1-410
47. Tripathy, J.N., Zhang, J., Robin, S., Nguyen, T.T., & Nguyen, H.T. (2000). QTLs for cell membrane stability mapped in rice (Oryza sativa L.) under drought stress. Theoretical and Applied Genetics.100: 1197–1202
48. Uma, S., Prasad, T.G., & Udayakumar, M. (1995). Genetic variability in recovery growth and synthesis of stress proteins in response to polyethylene glycol and salt stress in Finger millet. Ann. Bot. 76:43–9
49. Vierling, E. (1991). The role of heat shock proteins in plants. Ann. Rev. Pl. Physiol. and Pl. Mol. Biol. 42:579–620.
50. Wahid, A., & Ghazanfar, A. (2006). Possible involvement of some secondary metabolites in salt tolerance of sugarcane. J. Pl. Physiol. 163: 723–730
51. Wahid, A., & Shabbir, A. (2005). Induction of heat stress tolerance in Barely seedlings by pre sowing seed treatment with glycine betaine. Pl. Growth Regul. 46: 133–141
52. Wahid, A., Gelani, S., Ashraf, M., & Foolad, M.R. (2007). Heat tolerance in plants: An overview. Environ. Exp. Bot. 61. 199–223
53. Wahid, A., & Close, T.J. (2007). Expression of dehydrins under heat stress and their relationship with water relations of sugarcane leaves. Biol. Plant. 51: 104–109
54. Wang, W., Vinocur, B., & Altman, A. (2003). Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta. 218:1–14
55. Wu, C.H., Caspar, T., Browse, J., Lindquist, S., & Somerville, C. (1988). Characterization of an hsp70 cognate gene family in Arabidopsis. Plant Physiol. 88: 731–740
56. Xu, S., Li, J., Zhang, X., Wei, H., & Cui, L. (2006). Effects of heat acclimation pretreatment on changes of membrane lipid peroxidation, antioxidant metabolites, and ultrastructure of chloroplasts in two cool-season turf grass species under heat stress. Environ. Exp. Bot. 56: 274–285
57. Ye, L.A., Gao, H.Y., & Zou, Q. (2000). Responses of antioxidant system and xanthophylls cycle in Phaseolus vulgaris to combined stress of high irradiance and high temperature. Photosynthesis. 38: 205–210
58. Yin, H., Chen, Q.M., & Yi, M.F. (2008). Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum. Pl. Growth Regul. 54: 45–54
59. Yoshida, S., Forno, D.A., Cock, J.H., & Gomaz, K.U. (1976). Laboratory Manual for Physiological Studies of Rice. IRRI, Los Baños
60. Yuan, Y., Qian, H., Yu, Y., Lian, F., & Tang, D. (2011). Thermotolerance and antioxidant response induced by heat acclimation in Freesia seedlings. Acta Physiol. Plant. 33: 1001–1009
61. Zhaolong, W. & Bingru, H. (2004). Physiological recovery of Kentucky bluegrass from simulation drought and leaf stress. Crop Sci.44: 1729-1736