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Titolo:
Conversion of AFLP bands into high-throughput DNA markers
Autore:
Meksem, K; Ruben, E; Hyten, D; Triwitayakorn, K; Lightfoot, DA;
Indirizzi:
So Illinois Univ, Ctr Excellence Soybean Res Teaching & Outreach, Dept Plant Soil & Gen Agr, Carbondale, IL 62901 USA So Illinois Univ Carbondale ILUSA 62901 en Agr, Carbondale, IL 62901 USA
Titolo Testata:
MOLECULAR GENETICS AND GENOMICS
fascicolo: 2, volume: 265, anno: 2001,
pagine: 207 - 214
SICI:
1617-4615(200104)265:2<207:COABIH>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
BACKCROSS QTL ANALYSIS; HIGH-RESOLUTION MAP; DEATH SYNDROME SDS; ARABIDOPSIS-THALIANA; FIELD-RESISTANCE; CYST-NEMATODE; GENOME; PCR; BARLEY; LOCUS;
Keywords:
amplified fragment length polymorphism (AFLP); soybean; positional cloning; disease resistance; soybean cyst nematode;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
46
Recensione:
Indirizzi per estratti:
Indirizzo: Meksem, K So Illinois Univ, Ctr Excellence Soybean Res Teaching & Outreach, Dept Plant Soil & Gen Agr, Room 176, Carbondale, IL 62901 USA So IllinoisUniv Room 176 Carbondale IL USA 62901 , IL 62901 USA
Citazione:
K. Meksem et al., "Conversion of AFLP bands into high-throughput DNA markers", MOL GENET G, 265(2), 2001, pp. 207-214

Abstract

The conversion of AFLP bands into polymorphic sequence-tagged-site (STS) markers is necessary for high-throughput genotype scoring. Technical hurdlesthat must be overcome arise from genome complexity (particularly sequence duplication), from the low-molecular-weight nature of the AFLP bands and from the location of the polymorphism within the AFLP band. We generated six STS markers from ten AFLP bands (four AFLPs were from co-dominant pairs of bands) in soybean (Glycine max). The markers were all linked to one of two loci, rhg1 on linkage group G and Rhg4 on linkage group A2, that confer resistance to the soybean cyst nematode (Heterodera glycines I.). When the polymorphic AFLP band sequence contained a duplicated sequence or could not beconverted to a locus-specific STS marker, direct sequencing of BAC clones anchored to a physical map generated locus-specific flanking sequences at the polymorphic locus. When the polymorphism was adjacent to the restrictionsite used in the AFLP analysis, single primer extension was performed to reconstruct the polymorphism. The six converted AFLP markers represented 996bp of sequence from alleles of each of two cultivars and identified eight insertions or deletions, two microsatellites and eight single-nucleotide polymorphisms (SNPs). The polymorphic sequences were used to design a non-electrophoretic, fluorometric assay (based on the TaqMan technology) and/or develop electrophoretic STS markers for high-throughput genotype determination during marker-assisted breeding for resistance to cyst nematode. We conclude that the converted AFLP markers contained polymorphism at a 10- to 20-fold higher frequency than expected for adapted soybean cultivars and that the efficiency of AFLP band conversion to STS can be improved using BAC libraries and physical maps. The method provides an efficient tool for SNP and STS discovery suitable for marker-assisted breeding and genomics.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 09:02:45