Assessment of Polyamines and Trehalose in Wheat Microspores Culture for Embryogenesis and Green Regenerated Plants


Most aspects of microspore culture protocol have the capacity to cause stress to microspores, hence, less stressful treatments might be required to avoid deleterious effects. In stressed plants, polyamines and trehalose can act as compatible solutes or osmoprotectants by stabilizing proteins and biological membranes. To improve green plant regeneration in wheat microspore culture, this study assessed the effects of polyamines (putrecine, spermidine, spermine) and trehalose on androgenic response namely embryogenesis, green plant regeneration and ploidy of green plants regenerated in three spring wheat genotypes. Microspores of the genotypes produced significant numbers of embryos and green plants among polyamine treatments but trehalose had no effect (P ≤ 0.05). Polyamine treatments for 30 min generally produced more green plants per 100 microspores than the 60 min treatments in all three genotypes. At least three out of twelve polyamine treatments in each genotype improved the production of double haploid plants and seed setting in regenerants. Wheat genotype, concentration and duration of polyamine treatment had significant impact on embryogenesis and regeneration of green plants in this study. The study also showed that polyamines could be used to accelerate cultivar development in wheat breeding.

Share and Cite:

A. Redha and P. Suleman, "Assessment of Polyamines and Trehalose in Wheat Microspores Culture for Embryogenesis and Green Regenerated Plants," American Journal of Plant Sciences, Vol. 4 No. 11, 2013, pp. 2218-2226. doi: 10.4236/ajps.2013.411275.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Y. P. S. Bajaj, “In-Vitro Production of Haploids and Their Use in Cell Genetics and Plant Breeding,” In: Y. P. S. Bajaj, Ed., Biotechnology in Agriculture and Forestry. Haploids in Crop Improvement, Springer-Verlag, Berlin, Heidelberg, 1990, pp. 3-44.
[2] S. Teparkum and R. E. Veilleux, “Indifference of Potato Anther Culture to Colchicine and Genetic Similarity among Anther-Derived Monoploid Regenerants Determined by RAPD Analysis,” Plant Cell Tissue and Organ Culture, Vol. 53, No. 1, 1998, pp. 49-58.
[3] B. P. Forster, E. Heberle-Bors, K. J. Kasha and A. Touravev, “The Resurgence of Haploids in Higher Plants,” Trends in Plant Science, Vol. 12, No. 8, 2007, pp. 368-375.
[4] O. Slama-Ayed, J. De Buyser, E. Picard, Y. Trifa and H. S. Amara, “Effect of Pretreatment on Isolated Microspore Culture Ability in Durum Wheat (Triticum turgidum subsp. durum Desf.),” Journal of Plant Breeding and Crop Science, Vol. 2, No. 2, 2010, pp. 30-38.
[5] P. S. Baeziger, W. K. Russell, G. L. Graef and B. T. Campbell, “Improving Lives: 50 Years of Crop Breeding, Genetics and Cytology (C-1),” Crop Science, Vol. 46, No. 5, 2006, pp. 2230-2244.
[6] A. Touraev, B. P. Foster and S. M. Jain, “Advances in Haploid Production in Higher Plants,” Sringer Science + Business Media B.V., Dordrecht, 2009, pp. 1-208.
[7] J. Guzy-Wróbelska and I. Szarejko, “Molecular and Agronomic Evaluation of Wheat Double Haploid Lines Obtained through Maize Pollination and Anther Culture Methods,” Plant Breeding, Vol. 122, No. 4, 2003, pp. 305-313.
[8] S. K. Basu, M. Datta, M. Sharma and A. Kumar, “Haploid production technology in wheat and some selected higher plants,” Australian Journal of Crop Science, Vol. 5, No. 9, 2011, pp. 1087-1093.
[9] M. Y. Zheng, “Microspore Culture in Wheat (Triticum aestivum)-Doubled Haploid Production via Induced Embryogenesis,” Plant Cell Tissue and Organ Culture, Vol. 73, No. 3, 2003, pp. 213-230.
[10] A. Redha and P. Suleman, “Effect of Exogenous Application of Polyamines on Wheat Anther Cultures,” Plant Cell Tissue and Organ Culture, Vol. 105, No. 3, 2010, pp. 345-353.
[11] H. P. Bais and G. A. Ravishankar, “Role of Polyamines in the Ontogeny of Plants and Their Biotechnological Applications,” Plant Cell Tissue and Organ Culture, Vol. 69, No. 1, 2002, pp. 1-34.
[12] A. Bouchereau, A. Azis, F. Larher and J. Martin-Tanguy, “Polyamines and Environmental Challenges: Recent Development,” Plant Science, Vol. 140, No. 2, 1999, pp. 103-125.
[13] J. Martin-Tanguy, “Metabolism and Function of Polyamines in Plants: Recent Development (New Approaches),” Plant Growth Regulation, Vol. 34, No. 1, 2001, pp. 135-148.
[14] V. Kuznetsov, N. L. Radyukina and N. I. Shevyakova, “Polyamines and Stress: Biological Role, Metabolism, and Regulation,” Russian Journal of Plant Physiology, Vol. 53, No. 5, 2006, pp. 583-604.
[15] A. Altman, R. Kaur-Sawhney and A. W. Galston, “Stabilization of Leaf Protoplast through Polyamine-Mediated Inhibition of Senescence,” Plant Physiology, Vol. 60, No. 4, 1977, pp. 570-574.
[16] R. T. Besford, C. Richardson, J. L. Campos and A. F. Tiburcio, “Effect of Polyamines on Stabilization of Molecular Complexes in Thylakoid Membranes of Osmotically-Stressed Oat Leaves,” Planta, Vol. 189, No. 2, 1993, pp. 201-206.
[17] A. F. Tiburcio, R. T. Besfor, T. Capell, A. Borrell, P. S. Testillano and M. C. Rsueno, “Mechanisms of Polyamine Action during Senescence Response Induced by Osmotic Stress,” Journal of Experimental Botany, Vol. 45, No. 12, 1994, pp. 1789-1800.
[18] K. Gupta, A. Dey and B. Gupta, “Plant Polyamines in Abiotic Stress Responses,” Acta Physiologiae Plantarum, Vol. 35, No. 7, 2013, pp. 2015-2036.
[19] C. Kevers, T. Gaspar and D. Jacques, “The Beneficial Role of Different Auxins and Polyamines at Successive Stages of Somatic Embryo Formation and Development of Panax ginseng in Vitro,” Plant Cell Tissue and Organ Culture, Vol. 70, No. 2, 2002, pp. 181-188.
[20] M. K. Rajesh, E. Radha, K. Anitha and V. A. Parthasarathy, “Plant Regeneration from Embryo-Derived Callus of Oil Palm—The Effect of Exogenous Polyamines,” Plant Cell Tissue and Organ Culture, Vol. 75, No. 1, 2003, pp. 41-47.
[21] B. P. Hema and H. N. Murthy, “Improvement of in Vitro Androgenesis in Niger Using Amino Acids and Polyamines,” Biologia Plantarum, Vol. 52, No. 1, 2008, pp. 121-125.
[22] N. Benaroudj, D. H. Lee and A. L. Goldberg, “Trehalose Accumulation during Cellular Stress Protects Cells and Cellular Proteins from Damage by Oxygen Radicals,” Journal of Biological Chemistry, Vol. 276, 2001, pp. 24261-24267.
[23] H. Schluepmann, A. van Dijken, M. Aghdasi, B. Wobbes, M. Paul and S. Smeekens, “Trehalose Mediated Growth Inhibition of Arabidopsis Seedlings Is Due to Trehalose-6-Phosphate Accumulation,” Plant Physiology, Vol. 135, No. 2, 2004, pp. 879-890.
[24] P. J. Eastmond, A. van Dijken, M. Spileman, A. Tissier, H. G. Dichinson, J. D. Jones, S. C. Smeekens and I. A. Graham, “Trehalose-6-Phosphate Synthase I, Which Catalalyses the First Step in Trehalose Synthesis, Is Essential for Arabidopsis Embryo Maturation,” The Plant Journal, Vol. 29, No. 2, 2002, pp. 225-235.
[25] J. E. Schmid, M. Winzeler, B. Keller, B. Bütter, P. Stamp and H. Winzeler, “Induction and Use of Doubled Haploids in Wheat and Spelt Breeding Programs,” In: New Methods in Cereal Breeding, Eucarpia Cerial Section, Prospectives of Cereal Breeding in Europe, Landquart, 1994, pp. 146-147.
[26] A. Touraev, A. Indriato, I. Wratschko, O. Vicente and E. Heberle-Bors, “Efficient Microspore Embryogenesis in Wheat (Triticum aestivum L.) Induced by Starvation at High Temperature,” Sexual Plant Reproduction, Vol. 9, No. 4, 1996, pp. 209-215.
[27] W. Liu, M. Y. Zheng, E. Polle and C. F. Konzak, “Highly Efficient Doubled-Haploid Production in Wheat (Triticum aestivum L.) via Induced Microspore Embryogenesis,” Crop Science, Vol. 42, No. 3, 2002, pp. 686-692.
[28] M. E. Shariatpanahi, K. Belogradova, L. Hessamvaziri, E. Heberle-Bors and A. Touraev, “Efficient Embryogenesis and Regeneration in Freshly Isolated and Cultured Wheat (Triticum aestivum L.) Microspores without Stress Pretreatment,” Plant Cell Reports, Vol. 25, No. 12, 2006, pp. 1294-1299.
[29] J. E. Schmid, “In Vitro Production of Haploids in Triticum spelta,” In: Y. P. S. Bajaj, Ed., Biotechnology in Agriculture and Forestry, Vol. 13: Wheat, Springer-Verlag, Berlin, Heidelberg, 1990, pp. 363-381.
[30] A. Redha and T. Attia, “Improvement of Green Plant Regeneration by Manipulation of Anther Culture Induction Medium of Hexaploid Wheat,” Plant Cell Tissue and Organ Culture, Vol. 92, No. 2, 2008, pp. 141-146.
[31] J. M. Widholm, “The Use of Fluorescein Diacetate and Phenosafranine for Determining Viability of Cultured Plant Cells,” Biotechnic & Histochemistry, Vol. 47, No. 4, 1972, pp. 189-194.
[32] R. Phillips, M. C. Press and A. Eason, “Polyamines in Relation to Cell Division and Xylogenesis in Cultures Explants of Heliantus tuberosus: Lack of Evidence for Growth-Regulatory Action,” Journal of Experimental Botany, Vol. 38, No. 1, 1987, pp. 164-172.
[33] I. El-Hadrami, M. P. Carron and J. D’Auzac, “Clonal variability of the embryogenic potential of Hevea brasiliensis: Relations with Polyamines ( PA ) and Peroxidases ( PO ) in Calli,” Comptes Rendus-Academie des Sciences Paris, Vol. 308, No. 111, 1989, pp. 299-305.
[34] M. B. P. Calheiros, L. G. E. Vieira and S. R. L. Fuentes, “Effect of Exogenous Polyamines on Direct Somatic Embryogenesis in Coffee,” Revista Brasileira de Fisiologia Vegetal, Vol. 6, No. 2, 1994, pp. 109-114.
[35] O. Faure, M. Mengoli, A. Nougarede and N. Bagni, “Polyamine Pattern and Biosynthesis in Zygotic and Somatic Embryo Stages of Vitis vinifera,” Journal of Plant Physiology, Vol. 138, No. 5, 1991, pp. 545-549.
[36] I. Couée, I. Hummel, C. Sulmon, G. Gouesbet and A. El Armani, “Involvement of Polyamines in Root Development,” Plant Cell Tissue and Organ Culture, Vol. 76, No. 1, 2004, pp. 1-10.
[37] I. S. Dewi and B. S. Purwoko, “Role of Polyamines in Inhibition of Ethylene Biosynthesis and Their Effects on Rice Anther Culture Development,” Indonesian Journal of Agricultural Science, Vol. 9, No. 2, 2008, pp. 60-67.
[38] A. W. Galston, R. Kaur-Sawhney, T. Atabella and A. F. Tiburcio, “Plant Polyamines in Reproductive Activity and Response to Abiotic Stress,” Botanica Acta, Vol. 110, No. 3, 1997, pp. 197-207.
[39] S. Hoekstra, M. H. van Zijderveld, F. Heidekamp and F. van der Mark, “Microspore Culture of Hordeum vulgare L.: The Influence of Density and Osmolality,” Plant Cell Reports, Vol. 12, No. 12, 1993, pp. 661-665.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.