Caracterización botánica de variedades de caña de azúcar mediante marcadores fenotípicos y moleculares
Keywords:
descriptores de la UPOV, SSR, TRAP, UPOV´s descriptorsAbstract
El Programa de Mejoramiento Genético de la Caña de Azúcar (PMGCA) de la Estación Experimental Agroindustrial Obispo Colombres (EEAOC) genera continuamente nuevas variedades de caña de azúcar, especialmente para la provincia de Tucumán, Argentina. Una precisa identificación varietal es esencial para proteger los derechos de propiedad intelectual de los nuevos cultivares. Para caracterizar tres cultivares de caña de azúcar recientemente liberados por el PMGCA, en este trabajo se utilizaron marcadores moleculares TRAP y SSR (de sus siglas en inglés de “Target Region Amplified Polymorphism” y “Simple Sequence Repeat”, respectivamente), en combinación con marcadores fenotípicos propuestos por la Unión Internacional para la Protección de las Obtenciones Vegetales (UPOV). Se utilizaron cinco combinaciones de cebadores TRAP y cinco pares de cebadores SSR. Los productos de amplificación se separaron mediante electroforesis en geles de poliacrilamida en condiciones desnaturalizantes. Para la caracterización fenotípica se utilizaron los nueve descriptores obligatorios de la UPOV. Las bandas y los datos morfológicos se transformaron en matrices binarias de 0 y 1, se calculó la similitud y se generaron los dendrogramas utilizando el análisis UPGMA (de sus siglas en inglés de “Unweighted Pair–Group Method using Arithmetic averages”). Ambos marcadores moleculares revelaron claramente la relación filogenética entre genotipos. La caracterización morfológica permitió diferenciar correctamente las variedades analizadas, revelando únicamente la semejanza externa entre los genotipos. La caracterización completa permitió diferenciar inequívocamente los cultivares recientemente liberados para registrarlos y proteger los derechos de los mejoradores.
ABSTRACT
Botanical characterization of new sugarcane cultivars through molecular and phenotypical markers
The Sugarcane Breeding Program of Agroindustrial Experimental Station Obispo Colombres continuously generates new varieties of sugarcane, especially for the province of Tucumán, Argentina. Accurate varietal identification is essential to protect the intellectual property rights of new varieties. In this work, Target Region Amplified Polymorphism (TRAP) and Simple Sequence Repeat (SSR) molecular markers were used, in combination with phenotypic markers proposed by the International Union for Plant Variety Protection (UPOV), to characterize three sugarcane varieties recently released by the local breeding program. Five combinations of TRAP primers and five pairs of SSR primers were used. The amplification products were separated by electrophoresis in polyacrylamide gels under denaturing conditions. For the phenotypic characterization, the nine mandatory UPOV descriptors were used. The bands and morphological data were transformed into binary matrixes of 0 and 1, the similarity was calculated and the dendrograms were generated by the Unweighted pair-group method using arithmetic averages (UPGMA) analysis. Both molecular markers clearly revealed the phylogenetic relationship among genotypes. The morphological characterization correctly differentiated the varieties analyzed; only revealing the external similarity between genotypes. The entire characterization allowed us to unambiguously differentiate the recently released cultivars to register and protect plant breeder’s rights.
References
Aljanabi, S.; L. Forget and A. Dookun. 1999. An improve and rapid protocol for the isolation of polysaccharide and polyphenol free sugarcane DNA. Plant Mol. Biol. Rep. 17: 281.
Alwala, S.; A. Suman; J. A. Arro; J. C. Veremis and C. A. Kimbeng. 2006. Target region amplification polymorphism (TRAP) for assessing genetic diversity in sugarcane germoplasm collections. Crop Sci. 46:448-455.
Cordeiro, G. M.; R. Casu; C. L. McIntyre; J. M. Manners and R. J. Henry. 2001. Microsatellite markers from sugarcane (Saccharum spp.) ESTs cross transferable to Erianthus and Sorghum. Plant Sci.160: 1115-1123.
Cordeiro, G. M.; G. O. Taylor and R. J. Henry. 2000. Characterisation of microsatellite markers from sugarcane (Saccharum sp.), a highly polyploid species. Plant Sci. 155: 161-168.
Costilla, D. D.; E. R. Chavanne; M. I. Cuenya.; M. B. García y M. Arias. 2011. Descripción y registro de variedades de caña de azúcar producidas y difundidas por la Estación Experimental Agroindustrial Obispo Colombres. Avance Agroind. 32 (1): 8-12.
D’Hont, A.; P. Seshagiri Rao; S. Alleyne; J. C. Glaszmann and P. Feldmann. 1997. Application of molecular markers in sugarcane breeding. Proceedings West Indies sugar technologists, pp. 77–79.
Di Rienzo, J.; F. Casanoves; M. Balzarini; L. Gonzalez; M. Tablada y C. Robledo. 2009. InfoStat. Grupo InfoStat. FCA, Universidad Nacional de Córdoba, Argentina.
Dutilleul, P.; J. D. Stockwell; D. Frigon and P. Legendre. 2000. The Mantel Test versus Pearson’s correlation analysis: assessment of the differences for biological and environmental studies. Journal of Agricultural, Biological, and Environmental Statistics 5 (2):131-150.
Gallacher, D. J. 1997. Optimised descriptors recommended for Australian sugarcane germplasm (Saccharum spp. Hybrid). Aust J Agric Res. 48 (6): 775–779.
García, M. G.; M. Ontivero; J. C. Díaz Ricci and A. P. Castagnaro. 2002. Morphological traits and high resolution RAPD markers for the identification of the main strawberry varieties cultivated in Argentina. Plant Breed. 121: 76-80.
Li, G. and C. F. Quiros. 2001. Sequence-related amplified polymorphism (SRAP), a new marker system based on a simple PCR reaction: its application to mapping and gene tagging in Brassica. TAG. 103:455-461.
Perera, M.; M. Arias; D. Costilla; A. C. Luque; M. B. García; M. I. Cuenya; J. Racedo; S. Ostengo; M. P. Filippone and A. P. Castagnaro. 2012. Genetic diversity assessment and genotype identification in sugarcane based on DNA markers and morphological traits. Euphytica. 185: 491-510. DOI: 10.1007/s10681-012-0661- 9. 185 (3):491–510.
Powell, W.; G. Machray and J. Provan. 1996. Polymorphism revealed by simple sequence repeats. Trends Plant Sci 1: 215–222.
Salem, K. F. M.; A. M. El-Zanaty and R. M. Esmail. 2008. Assessing wheat (Triticum aestivum L.) genetic diversity using morphological characters and microsatallite markers. World J. of Agric. Sci. 4: 538-544.
Sneath, P. H. and R. R. Sokal. 1973. Numerical taxonomy: the principles and practices of numerical taxonomy. Freeman, San Francisco.
Union for the Protection of New Varieties of Plants (UPOV). 2005. Directrices para la ejecución del examen de distinción, homogeneidad y estabilidad tg/186/1. www.Upov.Int/es/publications/tg-rom/tg186/tg_186_1.Pdf (accessed 1 April 2019).
Downloads
Published
How to Cite
License
Copyright (c) 2020 Perera et. all.
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.