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      Genetic Characterization of Old Grapevines collected in Oases of the Atacama Desert Translated title: Caracterización Genética de Vides Antiguas colectadas en Oasis del Desierto de Atacama

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          Abstract

          Old grapevine (Vitis vinifera L.) accessions are a source of genes that could be rescued for use per se or in modern breeding programs. The first step in this rescuing is collecting and characterizing the germplasm from a particular region. This study presents the genetic characterization of 21 grapevine accessions collected from the Atacama Desert in the far North of Chile. Characterization was based on 12 microsatellites (Simple Sequence Repeats, or SSRs) supplemented with Amplified Fragment Length Polymorphic (AFLP) markers. Most of the collected accessions produced red berries and shared the genetic characteristics of the cv. País, an old genotype found throughout America. However, among those red-berried accessions, one showed a severe abortive phenotype (22S7), and another (6S4) differed from ’País’ in one allele. Both could be examples of somatic mutations, even though no variations in their AFLP patterns were found. On the other hand, the only accession with red berries that exhibited genetic characteristics different from those of ’País’ (5CN) corresponded to ’Gros Colman’, a supposedly Georgian genotype introduced to this region by the mid-20th century. Greater genetic diversity was detected among the white and pink accessions, which were classified into five clades based on their SSR allelic patterns. Of these genotypes, 11Si was identified as ’Emperatriz’ or ’Red Seedless’, an Argentinean variety; accessions 16H1 and 17H2 corresponded to a product of crossing ’País’ and ’Muscat of Alexandria’; and, finally, accession 20S5 was identified as ’Ahmeur bou Ahmeur’, an Algerian genotype harboring pink berries. Two seeded genotypes harboring small and large white berries were not identified as known varieties. The possible use of these accessions for breeding to enhance survival in the harsh environment of the Atacama Desert is discussed.

          Translated abstract

          Las accesiones de vid (Vitis vinifera L.) de antigua data son una fuente de variantes génicas que pueden ser rescatadas para su uso directo o su incorporación en programas de fitomejoramiento. La primera etapa de este rescate es la colecta y caracterización del germoplasma propio de una región particular. Este estudio describe la caracterización genética de 21 accesiones de vid colectadas en el desierto de Atacama, en el Norte de Chile. La caracterización se basó en 12 marcadores de microsatélites (SSR), complementado con marcadores de tipo fragmentos amplificados de largo variable (AFLP). La mayoría de las accesiones colectadas con bayas negras corresponden al genotipo ’País’, un cultivar antiguo que se encuentra distribuido en toda América. Sin embargo, entre las accesiones de baya negra, una de ellas mostró un fenotipo abortivo severo (22S7), y otro (6S4) difirió en un alelo con respecto a ’País’. Ambos podrían ser ejemplos de mutaciones somáticas, aun cuando no fue posible detectar variaciones en sus patrones de AFLP. Por otra parte, la única accesión con bayas negras que exhibió características genéticas diferentes a ’País’ correspondió a ’Gros Colman’ (5CN), un genotipo supuestamente proveniente de Georgia, Rusia, introducido en esta región a mediados del siglo XX. Una mayor diversidad genética fue detectada entre las accesiones de bayas blancas y rosadas, que se clasificaron en cinco clados en base a sus alelos de microsatélites. De estos genotipos, 11Si se identificó como ’Emperatriz’ o ’Red Seedless’, una variedad de origen argentino, mientras que las accesiones 16H1 y 17H2 correspondieron a un cruzamiento de ’Moscatel de Alejandría’ × ’País’. Por último, la accesión 20S5 se identificó como ’Ahmeur bou Ahmeur’, un genotipo argelino de bayas de color rosado. Dos genotipos semillados de bayas blancas, pequeñas o grandes, no se asimilaron con alguna variedad conocida. Se discute también la posibilidad de utilizar algunas de estas accesiones en fitomejoramiento, para aumentar la tolerancia a estreses propios del ambiente del desierto de Atacama.

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          Most cited references26

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          AFLP: a new technique for DNA fingerprinting

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            Development of a standard set of microsatellite reference alleles for identification of grape cultivars.

            In order to investigate the comparability of microsatellite profiles obtained in different laboratories, ten partners in seven countries analyzed 46 grape cultivars at six loci (VVMD5, VVMD7, VVMD27, VVS2, VrZAG62, and VrZAG79). No effort was made to standardize equipment or protocols. Although some partners obtained very similar results, in other cases different absolute allele sizes and, sometimes, different relative allele sizes were obtained. A strategy for data comparison by means of reference to the alleles detected in well-known cultivars was proposed. For each marker, each allele was designated by a code based on the name of the reference cultivar carrying that allele. Thirty-three cultivars, representing from 13 to 23 alleles per marker, were chosen as references. After the raw data obtained by the different partners were coded, more than 97% of the data were in agreement. Minor discrepancies were attributed to errors, suboptimal amplification and visualization, and misscoring of heterozygous versus homozygous allele pairs. We have shown that coded microsatellite data produced in different laboratories with different protocols and conditions can be compared, and that it is suitable for the identification and SSR allele characterization of cultivars. It is proposed that the six markers employed here, already widely used, be adopted as a minimal standard marker set for future grapevine cultivar analyses, and that additional cultivars be characterized by means of the coded reference alleles presented here. The complete database is available at http://www.genres.de/eccdb/vitis/ Cuttings of the 33 reference cultivars are available on request from the Institut National de la Recherche Agronomique Vassal collection (didier.vares@ensam.inra.fr).
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              The direction of microsatellite mutations is dependent upon allele length.

              Microsatellites, comprising tandemly repeated short nucleotide sequences, are ubiquitous in eukaryotic genomes. Mutations within microsatellites are frequent, altering their overall length by insertion or deletion of a small number of repeat units, with a rate as high as 10(-3) in humans. Despite their high mutability, stable allele frequency distributions are typically observed for microsatellites in humans as well as other primates, although the mechanism maintaining these stable distributions remains unclear. Previous studies have suggested that microsatellite mutations occur more frequently in longer alleles and favour expansion. Generalizing these results has been hindered because the sample sizes were small, only a small subset of alleles for any marker was studied and the direction of mutation (expansion or contraction) was not rigorously determined. Here we examine 236 mutations at 122 tetranucleotide repeat markers and find that the rate of contraction mutations increases exponentially with allele size, whereas the rate of expansion mutations is constant across the entire allele distribution. The overall rate of expansion mutations does not differ from that of contractions. Our findings offer an explanation for the stationary allele distribution of microsatellites.
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                Author and article information

                Journal
                chiljar
                Chilean journal of agricultural research
                Chil. j. agric. res.
                Instituto de Investigaciones Agropecuarias, INIA (Chillán, , Chile )
                0718-5839
                September 2011
                : 71
                : 3
                : 476-482
                Affiliations
                [02] Santiago orgnameInstituto de Investigaciones Agropecuarias orgdiv1Centro de Investigación La Platina Chile phinrichsen@ 123456inia.cl
                [03] Alcalá de Henares Madrid orgnameInstituto Madrileño de Investigació y Desarrollo Rural orgdiv1Agrario y Alimentario IMIDRA orgdiv2Laboratorio de Biotecnología España
                [01] Iquique orgnameUniversidad Arturo Prat orgdiv1Departamento Agricultura del Desierto y Biotecnología Chile
                Article
                S0718-58392011000300020 S0718-5839(11)07100300020
                f0045987-22e8-4dc4-97ce-e5c46ecd510b

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 04 January 2010
                : 03 May 2011
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 29, Pages: 7
                Product

                SciELO Chile

                Categories
                SCIENTIFIC NOTES

                SSR,AFLP,Atacama Desert,germplasm,’Listán Prieto’,Vitis vinifera,desierto de Atacama,germoplasma

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