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Evaluation of Rhizomania-Resistance Segregating Sequences and Overall Genetic Diversity Pattern Among Selected Accessions of Beta and Patellifolia

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Evaluation of Rhizomania-Resistance Segregating Sequences and Overall Genetic Diversity Pattern Among Selected Accessions of Beta and Patellifolia Euphytica (2016) 207:685–706 DOI 10.1007/s10681-015-1570-5 Evaluation of rhizomania-resistance segregating sequences and overall genetic diversity pattern among selected accessions of Beta and Patellifolia. Potential implications of breeding for genetic bottlenecks in terms of rhizomania resistance Anna Litwiniec . Maria Gos´ka . Beata Choin´ska . Kamilla Kuzdowicz_ . Aleksander Łukanowski . Barbara Skibowska Received: 21 April 2015 / Accepted: 27 September 2015 / Published online: 7 October 2015 Ó The Author(s) 2015. This article is published with open access at Springerlink.com Abstract Sugar beet is hypothesized to have a representatives of Beta and Patellifolia and sugar beet narrowed genetic base due to its origin as White cultivars, with a special focus on the complex Beta Silesian Beet and from numerous breeding selections vulgaris. Another purpose was to determine the and practices. High sugar quality, yield of recoverable potential usefulness and conformity of selected sugar, cytoplasmic-male sterility system, monoger- molecular markers in different groups of materials in mity, pests and disease resistance and bolting resis- the context of rhizomania resistance. To reach these tance constitute some of the adaptations that goals, molecular RAPD, ISSR techniques, literature- significantly influenced the existing genetic back- selected rhizomania resistance-segregating sequences ground of the crop. In this study we aimed to evaluate as well as mitochondrial markers were used. The the extent of genetic diversity existing in wild beet comparison of genetic diversity in wild and cultivated Beta forms shows that the population differentiation values and distance values are relatively high in & A. Litwiniec ( ) Á B. Choin´ska cultivars. Moreover, the diversity component seemed Plant Breeding and Acclimatization Institute–National Research Institute, Bydgoszcz Research Center, to be compromised rather on the level of population Department of Genetics and Breeding of Root Crops, (Hs) than in total (Ht) in cultivars. Our results shed a Laboratory of Biotechnology, Powstan´co´w new light on the expected genetic bottlenecks existing Wielkopolskich 10, 85-090 Bydgoszcz, Poland in cultivars and revealed features specific for individ- e-mail: [email protected]; [email protected] ual taxa (Patellifolia, Corollinae). Some degree of distinctiveness was suggested between genetic deter- M. Gos´ka Á K. Kuzdowicz_ Á B. Skibowska minants of rhizomania resistance in modern cultivars Plant Breeding and Acclimatization Institute–National in comparison with wild resistance sources. In addi- Research Institute, Bydgoszcz Research Center, Department of Genetics and Breeding of Root Crops, tion, we document here an internal heterogeneity Laboratory of Cytogenetics and Breeding Methods, existing in selected wild/weedy accessions at the level Powstan´co´w Wielkopolskich 10, 85-090 Bydgoszcz, of crucial sequences using high resolution melting. Poland A. Łukanowski Keywords Wild beets Á Sugar beet Á Genetic UTP University of Science and Technology, Faculty of diversity Á PCR Á Molecular markers Á Rhizomania Agriculture and Biotechnology, Department of Molecular resistance Phytopathology, Kordeckiego 20, 85-225 Bydgoszcz, Poland 123 686 Euphytica (2016) 207:685–706 Introduction stimulate more rational and flexible management of genetic resources in the future (Frese et al. 2001). Sugar beet (Beta vulgaris L. ssp. vulgaris) is a crop Additionally as a result of interfertile B. vulgaris with great economic importance, especially in tem- coexisting in seed production areas, spontaneous perate climate zones where it constitutes a valuable outcrossings between wild and cultivated beets promote sucrose source. It contributes about one fifth to the the origin of weed beets (Boudry et al. 1993). Despite its global sugar production, which for the 2007–2011 stochastic nature and basically unwanted effects in that period reached approximately 157 million metric tons particular case, the phenomenon itself may also have (Koo and Taylor 2012). Crop rotation is necessary in some positive implications, via increasing the variabil- beet production due to disease problems (Koo and ity level in the available gene pool. Taylor 2012), which may also be effectively elimi- One of the best examples for implementing desired nated by resistance breeding approaches. characters successfully in wild sea beet (Beta vulgaris Breeding contributed above all to the increase in ssp. maritima)duringbreedingisrhizomaniaresistance sugar yield. Some hybrids of fodder beets selected for genes. Early partially resistant sugar beet cultivars were relatively high sugar content, i.e. ‘‘White Silesian’’, developed based on Munerati’s pool of these accessions constituted the origin for all subsequently developed selected in the Po estuary for Cercospora leaf spot sugar beet varieties (Fischer 1989; Biancardi et al. resistance (Van Geyt et al. 1990). Presently it is 2010). At the same time other breeding practices and believed that all the resistance sources identified thus directions, for example cytoplasmic male sterility and far, designated from Rz1 to Rz5, originate most genetic monogermity systems, used throughout the probably from some wild sea beet ancestors (Biancardi history of the modern crop’s development, are sup- et al. 2002), although their identity and independence posed to impinge significantly on its genetic structure from each other has not been fully confirmed yet. and, as a consequence, to result in genetic bottlenecks, In this study we aimed to evaluate the extent of thus reducing available diversity and hampering in a genetic diversity existing in wild beet representatives way the progress in genetic combinations for new belonging to the genera Beta and Patellifolia as cultivars’ releases (Biancardi et al. 2010). compared to sugar beet cultivars with a special A considerably higher genetic diversity is expected emphasis on the complex Beta vulgaris using molec- to exist in wild relatives of sugar beet, serving thus as a ular RAPD, ISSR techniques and literature-selected reservoir of potentially useful traits and genes, such as putative rhizomania resistance-segregating markers as those determining tolerance to biotic and abiotic well as mitochondrial markers. stresses, yield parameters or other advantageous mor- Our results not only shed a new light on the phological and physiological features (Van Geyt et al. expected genetic bottlenecks existing in cultivars and 1990; Stevanato et al. 2013). Hybridization between B. revealed some features specific for individual taxa vulgaris and exotic germplasm belonging to Beta is (Patellifolia, Corollinae), but also suggest that some possible, often providing fertile progeny, fully compat- degree of distinctiveness may exist between genetic ible at the chromosomal level. This phenomenon has determinants of rhizomania resistance in modern been employed in sugar beet enhancement programs cultivars when compared with wild resistance sources. since the beginnings of the twentieth century, as growth in cultivated acreage and expanding distribution areas were inevitably related to increased pathogen pressure Materials and methods and disease spread (Panella and Lewellen 2007; Biancardi et al. 2010). From this time on, many directed Plant material and internationally coordinated initiatives have been undertaken by research and breeding communities, The materials selected for the study comprise repre- especially in France and the USA, aiming not only at the sentatives of five wild beet species, i.e. B. vulgaris ssp. introgression of selected valuable traits into the sugar maritima (4 accessions: B.m.01, B.m.27—resistant, beet crop, but also at broadening of its genetic base. B.m.40 and B.m.72), B. macrorhiza (B.macr.21), B. Such an integrated approach, although requiring further corolliflora (B.c.20), P. procumbens (P.p.25) and P. selection cycles, is at the same time expected to patellaris (P.pat.22), one weed beet accession (wbM), 123 Euphytica (2016) 207:685–706 687 and five sugar beet cultivars with contrasting pheno- standard, i.e. GeneRulerTM 100 bp Plus DNA Ladder types for rhizomania resistance (B.v.14, B.v.15 and (Thermo Scientific). All the sequences of primers B.v.16—diploid, tolerant, B.v.13 and B.v.17—tri- included in the study are presented in Table 2. Cyto- ploid, susceptible). Wild accessions were obtained plasmic diversity was examined using primers designed from international genebanks, and a weed beet pop- by Nishizawa et al. (2000) for mitochondrial minisatel- ulation was collected from a sugar beet field in lites according to the protocol described previously by Minikowo (Poland). Cultivated materials were Fe´nart et al. (2008). obtained from commercial sugar beet breeding com- panies (Strube, KHBC Ltd.). Ten genotypes per each HRM analysis accession were individually analyzed, unless other- wise indicated. Further single individuals of wild B. Experiments were performed on CFX96TM Real-Time vulgaris ssp. maritima/B. vulgaris ssp. vulgaris, System (BIO-RAD) using AmpliQ 59 HOT EvaGreenÒ representing the following accessions: B.m.28, HRM Mix (Novazym). Primer sequences were designed B.m.29 (resistant), B.m.30, B.m.02, B.v.33 and using Primer BLAST (NCBI). B.m.31 (susceptible) were included as well for addi- tional, more in-depth comparisons of the molecular Statistical analyses marker efficiencies on an expanded group of materials and some for HRM (high resolution melting) standards Dendrograms of genetic distance were
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