High-resolution melt analysis to identify and map sequence-tagged site anchor points onto linkage maps: a white lupin (Lupinus albus) map as an exemplar.
* The provision of sequence-tagged site (STS) anchor points allows meaningful comparisons between mapping studies but can be a time-consuming process for nonmodel species or orphan crops. * Here, the first use of high-resolution melt analysis (HRM) to generate STS markers for use in linkage mapping is described. This strategy is rapid and low-cost, and circumvents the need for labelled primers or amplicon fractionation. * Using white lupin (Lupinus albus, x = 25) as a case study, HRM analysis was applied to identify 91 polymorphic markers from expressed sequence tag (EST)-derived and genomic libraries. Of these, 77 generated STS anchor points in the first fully resolved linkage map of the species. The map also included 230 amplified fragment length polymorphisms (AFLP) loci, spanned 1916 cM (84.2% coverage) and divided into the expected 25 linkage groups. * Quantitative trait loci (QTL) analyses performed on the population revealed genomic regions associated with several traits, including the agronomically important time to flowering (tf), alkaloid synthesis and stem height (Ph). Use of HRM-STS markers also allowed us to make direct comparisons between our map and that of the related crop, Lupinus angustifolius, based on the conversion of RFLP, microsatellite and single nucleotide polymorphism (SNP) markers into HRM markers.
Croxford
Adam E
AE
Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, SY23 3DA, UK.
Rogers
Tom
T
KG Fruits Ltd, Tatlingbury Oast, Five Oak Green, Tonbridge, Kent TN12 6RG, UK.
Caligari
Peter D S
PDS
Instituto de Biología Vegetal y Biotecnología, Universidad de Talca, 2 Norte 685 Talca, Chile.
Wilkinson
Michael J
MJ
Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, SY23 3DA, UK.
D054458:Amplified Fragment Length Polymorphism Analysis; D001483:Base Sequence; D016009:Chi-Square Distribution; D002874:Chromosome Mapping / Q000379:methods*; D003433:Crosses, Genetic; D018745:Genome, Plant; D020180:Heteroduplex Analysis; D007364:Intercalating Agents; D031297:Lupinus / Q000235:genetics*; D018895:Microsatellite Repeats; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D040641:Quantitative Trait Loci; D016324:Sequence Tagged Sites
D007364:Intercalating Agents