Microsatellite DNA in peach (<i>Prunus persica</i> L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars

Abstract

We isolated and sequenced 26 microsatellites from two genomic libraries of peach cultivar 'Redhaven', enriched for AC/GT and AG/CT repeats, respectively. For 17 of these microsatellites, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was assayed in 50 peach and nectarine cultivars. Of the 1300 PCRs carried out, all but two produced amplified products of the expected size. All microsatellites were polymorphic, showing 2-8 alleles per locus. Heterozygosity ranged from 0.04-0.74 (mean 0.47); the discrimination power (PD) ranged from 0.04-0.84 (mean 0.60). Cultivar heterozygosity varied greatly, with one cultivar ('Independence') being homozygous at all loci. The set of microsatellites discriminated all cultivars investigated, except several sport mutations, i.e., 'Dixitime' vs. 'Springcrest', 'Compact Redhaven' vs. 'Redhaven', and two pairs of cultivars, 'Venus' vs. 'Orion' and 'Elegant Lady' vs. 'Rome Star', whose pedigrees are controversial. We were able to analyze the paternity of several cultivars. In most cases, the parenthood was confirmed. The comparison of three long-living 'Redhaven' accessions supplied by different repositories did not provide any evidence of somatic instability of microsatellites. Hence, microsatellites, ranked according to their information content, are recommended as markers of choice for peach fingerprinting and suggestions are provided for interpreting band profiles and the correct sizing of alleles.

Molecular Genetic Markers for Root-Knot Nematode Resistance in Some Peach Rootstocks

“…These results agreed with the findings of Wang et al (2002) who found that most SSRs are not specifically linked to gene loci of immediate interest and developing an SSR map is very time consuming and expensive. Either these results were in agreement with Testolin et al (2000) who gave some examples to the utility of SSRs for pedigree determination of several peach cultivars. Aranzana et al, (2003) reported that the available SSRs on the Prunus saturated map allowing them to develop a resource useful for map comparison or MAS in fruit crops.…”

“…Microsatellite or Simple Sequence Repeats (SSRs) are the best available choice of markers for peach genetics and breeding, they are generally codominant, highly polymorphic, and can be found in large numbers covering the whole genome of any species (Sosinski et al, 2000). Many SSRs have been developed in peach (Testolin et al, 2000;Aranzana et al, 2002;Dirlewanger et al, 2002). The objectives of this study were to:…”

Molecular and quantitative signatures of biparental inbreeding depression in the self-incompatible tree species Prunus avium

“…The DNA was quantified on a NanoDrop system (Thermo Fisher Scientific, Schwerte, Germany) and diluted to a standardized DNA concentration of 10 ng ml À1 . We amplified eight microsatellite loci (Testolin et al, 2000;Dirlewanger et al, 2002;Schueler et al, 2003), using the combinations and conditions described in , with the only difference being the labelling from primers UDP98_411 (FAM), UDP98_412 (FAM) and BPPCT_040 (HEX). Fragments were analysed on a Megabace genetic analyser (Amersham Biosciences, Buckinghamshire, UK) using the Megabace ET 400-R size standard (GE Healthcare, Munich, Germany).…”

Impact of negative frequency-dependent selection on mating pattern and genetic structure: a comparative analysis of the S-locus and nuclear SSR loci in Prunus lannesiana var. speciosa

“…This method could certainly visualize the variations of both two S-alleles, which respective individuals possess, implying that null alleles are unlikely to be detected at the S-locus. Genotypes at 11 nuclear SSR loci, BPPCT005, 014, 026, 028, 034, 037, 038 and 040 (Dirlewanger et al, 2002), UDP96-001 (Cipriani et al, 1999), UDP98-412 (Testolin et al, 2000) and pchcms5 (Sosinski et al, 2000), were also scored using the method described by Kato et al (2011). Part of the genotypes had been already determined in our previous studies (Kato and Mukai, 2004;Kato et al, 2007Kato et al, , 2011.…”