Microsatellite repeats in grapevine reveal DNA polymorphisms when analysed as sequence-tagged sites (STSs)

Abstract

Microsatellite repeat sequences were investigated as sequenced-tagged site (STS) DNA markers to determine the potential for genetic analysis of the grapevine genome. The PCR-generated markers detect codominant alleles at a single locus or site in the genome. The marker type is very informative detecting high heterozygosity (69%-88%) within individual grapevine cultivars and high genetic variation between cultivars, making it a useful marker type for plant genome mapping and genome typing. For five loci a screening of 26 V. vinifera cultivars found 13, 12, 8, 5, and 4 different length alleles respectively with some alleles more common than others. The genomic DNA sequences surrounding microsatellite sequences were conserved within the genus permitting STS primers to amplify STSs from other Vitis species. These Vitis species were found to have some unique alleles not present in V. vinifera.

Fine-scale genetic structure of grape phylloxera from the roots and leaves of Vitis

“…Root pieces were collected from vines sampled at both sites and DNA typed using five microsatellite loci: VVS2, VVMD6, VVMD7, VVMD28, VVMD31 (Thomas and Scott, 1993;Bowers et al, 1996Bowers et al, , 1999. The root pieces were frozen and transported in liquid nitrogen and stored at À801C prior to DNA extraction.…”

Detection of downy and powdery mildew resistance QTL in a ‘Regent’ × ‘RedGlobe’ population

“…Extracted DNA was quantified using a Nanodrop spectrophotometer and the concentrations were standardised to fall within the range 25-35 ng/ll. SSRs were selected from the literature and the NCBI database (Thomas and Scott 1993;Bowers et al 1996Bowers et al , 1999Scott et al 2000;Decroocq et al 2003;Adam-Blondon et al 2004;Di Gaspero et al 2005;Merdinoglu et al 2005;Doligez et al 2006;Cipriani et al 2008; http://www.ncbi.nlm.nih.gov/) with the aim to achieve comprehensive genome coverage and increased marker density for known minor and major mildew resistance QTL regions on chromosomes 5, 12, 15 and 18. A standard set of PCR conditions (1.8 mM MgCl 2 , 0.75 U Supertherm Taq, 5 mM dNTP and 0.3 pmol/ll of each primer) was used for all reactions.…”

Genealogy of wine grape cultivars: ‘Pinot’ is related to ‘Syrah’

“…Primer pairs for most of the VMC microsatellite markers are unpublished (except VMC7F2 in Pellerone et al, 2001) and belong to the Vitis Microsatellite Consortium (www.agrogene.com). Primer pairs for VVMD microsatellites were published in Bowers et al (1996) and Bowers et al (1999b); for VrZAG in Sefc et al (1999); for VVS in Thomas and Scott (1993) and Thomas et al (1998). All 60 markers were already mapped (Grando et al, 2003;Riaz et al, 2004) and they have been chosen in 18 out of the 19 linkage groups of the grape genome (Adam-Blondon et al, 2004), so that they are evenly distributed throughout the genome (average distance between the markers is 12 cM).…”

“…The advent of PCR-based microsatellite markers in the 1990s revolutionized grape cultivar identification and parentage analysis (Sefc et al, 2001). Thomas and Scott (1993) were the first to distinguish grape cultivars with microsatellite markers and to show their Mendelian inheritance by following the segregation of a single marker in recent deliberate crosses. Parentage analyses based on exclusion, using 30 polymorphic microsatellites allowed Bowers and Meredith (1997) to identify the parents of a traditional cultivar for the first time: 'Cabernet Sauvignon', the noble Bordeaux variety that gives some of the world's finest wines, was shown to be a progeny of two other Bordeaux cultivars, 'Cabernet Franc' and 'Sauvignon Blanc'.…”