Doubled Haploid Production from Spanish and Central European Spelt by Anther Culture

Authors
1 Department of Plant Genetics and Production, Experimental Station of Classroom Dei-CSIC, Spanish National Research Council, Avda Montañana 1005, 50.059 Zaragoza, Spain.
2 Departamento Genética y Producción Vegetal, Estación Experimental de Aula Dei-CSIC
3 Field Crops Program, IRTA (Institute for Food and Agricultural Research and Technology), Avda Alcalde Rovira 191, 25198 LLeida, Spain.
Abstract
In recent years, spelt (Triticum aestivum (L.) ssp. Spelta) has become an added-value alternative crop to modern wheat. Spanish spelt constitutes a unique separate gene pool from central European germplasm. The availability of spelt Doubled Haploid (DH) production protocol is a great advantage to speed up breeding programs. This is the first study evaluating the ability of DH plant production, by anther culture, of five Spanish spelt landraces and three F5 lines derived from Spanish spelt x bread wheat crosses. Two central European commercial varieties were also included in the analysis. DH plants were obtained from all material with the exception of one F5 line. The Spanish spelt landraces produced more embryos/100 anthers (73-166.3) than the two European varieties (8.6-22.2). The main bottleneck in the Spanish germplasm was the high number of albino plants regenerated, with percentage of green plants lower than 13% in three of the landraces. Nevertheless, up to 15.6 and 1.8 green plants/100 anthers were obtained from the Spanish and the central European germplasm, respectively. A great variation in the percentage of spontaneous chromosome doubling was obtained, with 4 lines producing around 80% and 2 lines less than 15%. The ovary genotype used for anther co-culture is a critical factor to increase the efficiency of the system. Bread wheat ‘Caramba’ ovaries increased almost 6 times the number of green plants as compared to spelt landrace ‘BG-1987’ ovaries. This study shows that DH plants can be produced efficiently from Spanish spelt to be used in breeding programs.

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1. Bohanec, B. 2009. Doubled Haploids via Gynogenesis. In: “Advances in Haploid Production in Higher Plants”, (Eds): Touraev, A., Forster, B. P. and Jain, S. M. Springer Science+Business Media BV, Netherlands, PP. 35-46.
2. Caballero, L., Martín, L. M. and Alvarez, J. B. 2001. Allelic Variation of the HMW Glutenin Subunits in Spanish Accessions of Spelt Wheat (Triticum aestivum ssp. spelta L. em Thell). Theor. Appl. Genet., 103: 124-128.
3. Campbell, K. G. 1997. Spelt: Agronomy, Genetics and Breeding. Plant Breed. Rew., 15: 187-213.
4. Castillo, A. M., Cistué, L., Vallés, M. P. and Soriano, M. 2009. Chromosome Doubling in Monocots. In: “Advances in Haploid Production in Higher Plants”, (Eds): Touraev, A., Forster, B. P. and Jain, S. M. Springer Science+Business Media BV, Netherlands, PP. 329-338.
5. Castillo, A. M., Sánchez-Díaz, R. A. and Vallés, M. P. 2015. Effect of Ovary Induction on Bread Wheat Anther Culture: Ovary Genotype and Developmental Stage, and Candidate Gene Association. Front. Plant Sci., 6: 402.
6. Devaux, P. 2003. The Hordeum bulbosum (L.) Method. In: “Doubled Haploid Production in Crop Plants”, (Eds.): Maluszynski, M., Kasha, K. J., Forster, B. P. and Szarejko, I. Kluwer Academic Publishers, Dordrecht, PP. 15-19.
7. Dwivedi, S. L., Britt, A. B., Tripathi, L., Sharma, S., Upadhyaya, H. D. and Ortiz, R. 2015. Haploids: Constraints and Opportunities in Plant Breeding. Biotech. Adv., 33: 812-829.
8. Echavarri, B., Soriano, M., Cistué, L., Valles, M. P. and Castillo, A. M. 2008. Zinc Sulfate Improved Microspore Embryogenesis in Barley. Plant Cell Tiss. Organ. Cult., 93: 295–301.
9. Elia, M. 2007. Estudios Previos a la Mejora Genética de la Escanda (Triticum spelta L.): Uso de la Colección Española. Doctoral Thesis, Univ Lleida, Lleida, Spain.
10. Elia, M., Moralejo, M., Rodriguez-Quijano, M. and Molina-Cano, J. L. 2004. Spanish Spelt. A Separate Gene Pool within the Spelt Germplasm. Plant Breed., 123: 297-299.
11. Escarnot, E., Jacquemin, J. M., Agneessens, R. and Paquot, M. 2012. Comparative Study of the Content and Profiles of Macronutrients in Spelt and Wheat: A Review. Biotechnol. Agron. Soc. Environ., 16(2): 243-256.
12. Escarnot, E., Tibaut, C. and Forgeois, P. 2014. Study of the Impact of Growth Substance Treatment and Maize (Zea mays L.) Variety in Spelt (Triticum spelta L.) Haplodiploidization. Biotechnol. Agron. Soc. Environ., 18(1): 32-36.
13. Guzmán, C., Caballero, L., Moral, A. and Alvarez, J. B. 2010. Genetic Variation for Waxy Proteins and Amylose Content in Spanish Spelt Wheat (Triticum spelta L). Genet. Resour. Crop Evol., 57: 721-725.
14. Holme, I. B., Olesen, A. and Andersen, S. B. 1999. Anther and Isolated Microspore Culture Response of Wheat Lines from Northwestern and Eastern Europe. Plant Breed., 118: 111–117.
15. Hu, H. and Kasha, K. J. 1997. Improvement of Isolated Microspore Culture of Wheat (Triticum aestivum L.) through Ovary Co-Culture. Plant Cell Rep., 16: 520-525.
16. Hunter, C. P. 1987. European Patent Application Nr0245898 A2. PP. 1-8
17. Jensen, C. J. 1977. Monoploid Production by Chromosome Elimination. In: “Applied and Fundamental Aspects of Plan Cell, Tissue and Organ Culture”, (Eds.): Reinert, J. and Bajaj, Y. P. S. Springer, Berlin, PP. 299-330.
18. Kim, M., Park, E. -J., An, D. and Lee, Y. 2013. High-Quality Embryo Production and Plant Regeneration Using a Two-Step Culture System in Isolated Microspore Cultures of Hot Pepper (Capsicum annuum L.). Plant Cell Tiss. Organ. Cul., 112: 191-201.
19. Konvalina, P., Stehno, Z., Capouchová, I., Zechnaer, E., Berger, S., Grausgruber, H., Janovská, D. and Moudry, J. 2014. Differences in Grain/Straw Ratio, Protein Content and Yield in Landraces and Modern Varieties of Different Wheat Species under Organic Farming. Euphytica, 199(1-2): 31-40.
20. Lantos, C., Weyen, J., Orsini, J.M., Gnad, H., Schlieter, B., Lein, V., Kontowski, S., Jacobi, A., MihÄly, R., Broughton, S. and Pauk, J. 2013. Efficient Application of In Vitro Anther Culture for Different European Winter Wheat (Triticum aestivum L.) Breeding Programmes. Plant Breed., 132: 149–154.
21. Lantos, C., Jenes, B., Bóna, L., Cserháti, M. and Pauk, J. 2017. High Frequency of Doubled Haploid Plant Production in Spelt Wheat. Acta Biologica Cracoviensia S Botanica, 58(2): 107-112.
22. Longin, C.F.H. and Würschum, T. 2014. Genetic Variability and Correlation among Agronomic and Disease Resistance Traits in a Diversity Panel and Elite Breeding Material of Spelt Wheat. Plant Breed., 133: 459-464.
23. Longin, C. F. H. and Würschum, T. 2016. Back to the Future: Tapping into Ancient Grains for Food Diversity. Trends Plant Sci., 21(9): 731-737.
24. Maluszynski, M., Kasha, K.J., Foster, B. P. and Szarejko, I. 2003. Doubled Haploid Production in Crop Plants: A Manual. Kluwer Academic Publishers, Dordrecht, PP.1-428.
25. Ruibal-Mendieta, N. L., Delacroix, D., Mignolet, E., Pycke, J. M., Marques, C., Rozenberg, R., Petitjean, G., Habib-Jiwan, J. L., Meurens, M., Quetin-Leclercq, J., Delzenne, N. M. and Larondelle, Y. 2005. Spelt (Triticum aestivum ssp Spelta) as a Source of Breadmaking Flours and Bran Naturally Enriched in Oleic Acid and Minerals but Not Phytic Acid. J. Agric. Food Chem., 53: 2751-2759.
26. Schmid, J. 1990. In Vitro Production of Haploids in Triticum spelta. In: “Biotechnology in Agriculture and Forestry- 13 Wheat”, (Ed.): Bajaj, Y. P. S. Springer-Verlag, Berlin, Heildelberg, PP. 363-381.
27. Soriano, M., Cistué, L. and Castillo, A. M. 2008. Enhanced Induction of Microspore Embryogenesis after n-Butanol Treatment in Wheat (Triticum aestivum L.) Anther Culture. Plant Cell Rep., 27: 805-811.
28. Soriano, M., Cistué, L., Vallés, M. P. and Castillo, A. M. 2007. Effects of Colchicine on Anther and Microspore Culture of Bread Wheat (Triticum aestivum L.). Plant Cell Tiss. Organ. Cul., 91: 225-234.
29. Tadesse, W., Inagaki, M., Tawkaz, S., Baum M. and van Ginkel, M. 2012. Recent Advances and Application of Doubled Haploids in Wheat Breeding. Afr. J. Biotech., 11(89): 15484-15492.
30. Wedzony, M., Forster, B. P., Zur, I., Golemiec, E., Szechynska-Hebda, M., Dubas, E. and Gotebiowska, G. 2009. Progress in Doubled Haploid Technology in Higher Plants. In: “Advances in Haploid Production in Higher Plants”, (Eds): Touraev, A., Forster, B. P. and Jain, S. M. Springer Science+Business Media BV, Netherlands, PP. 1-34.
31. Wojnarowiez, G., Jacquard, C., Devaux, P., Sangwan, R. S. and Clément, C. 2002. Influence of Copper Sulphate on Anther Culture in Barley (Hordeum vulgare L). Plant Sci., 162: 843-847.