Development and implementation of high-throughput SNP genotyping in barley.
BACKGROUND
High density genetic maps of plants have, nearly without exception, made use of marker datasets containing missing or questionable genotype calls derived from a variety of genic and non-genic or anonymous markers, and been presented as a single linear order of genetic loci for each linkage group. The consequences of missing or erroneous data include falsely separated markers, expansion of cM distances and incorrect marker order. These imperfections are amplified in consensus maps and problematic when fine resolution is critical including comparative genome analyses and map-based cloning. Here we provide a new paradigm, a high-density consensus genetic map of barley based only on complete and error-free datasets and genic markers, represented accurately by graphs and approximately by a best-fit linear order, and supported by a readily available SNP genotyping resource.


RESULTS
Approximately 22,000 SNPs were identified from barley ESTs and sequenced amplicons; 4,596 of them were tested for performance in three pilot phase Illumina GoldenGate assays. Data from three barley doubled haploid mapping populations supported the production of an initial consensus map. Over 200 germplasm selections, principally European and US breeding material, were used to estimate minor allele frequency (MAF) for each SNP. We selected 3,072 of these tested SNPs based on technical performance, map location, MAF and biological interest to fill two 1536-SNP "production" assays (BOPA1 and BOPA2), which were made available to the barley genetics community. Data were added using BOPA1 from a fourth mapping population to yield a consensus map containing 2,943 SNP loci in 975 marker bins covering a genetic distance of 1099 cM.


CONCLUSIONS
The unprecedented density of genic markers and marker bins enabled a high resolution comparison of the genomes of barley and rice. Low recombination in pericentric regions is evident from bins containing many more than the average number of markers, meaning that a large number of genes are recombinationally locked into the genetic centromeric regions of several barley chromosomes. Examination of US breeding germplasm illustrated the usefulness of BOPA1 and BOPA2 in that they provide excellent marker density and sensitivity for detection of minor alleles in this genetically narrow material.
Close
Timothy J
TJ
Department of Botany & Plant Sciences, University of California (UCR), Riverside, CA, 92521, USA. timothy.close@ucr.edu
Bhat
Prasanna R
PR
Lonardi
Stefano
S
Wu
Yonghui
Y
Rostoks
Nils
N
Ramsay
Luke
L
Druka
Arnis
A
Stein
Nils
N
Svensson
Jan T
JT
Wanamaker
Steve
S
Bozdag
Serdar
S
Roose
Mikeal L
ML
Moscou
Matthew J
MJ
Chao
Shiaoman
S
Varshney
Rajeev K
RK
Szucs
Péter
P
Sato
Kazuhiro
K
Hayes
Patrick M
PM
Matthews
David E
DE
Kleinhofs
Andris
A
Muehlbauer
Gary J
GJ
DeYoung
Joseph
J
Marshall
David F
DF
Madishetty
Kavitha
K
Fenton
Raymond D
RD
Condamine
Pascal
P
Graner
Andreas
A
Waugh
Robbie
R
D000483:Alleles; D008040:Genetic Linkage; D005819:Genetic Markers; D005821:Genetic Techniques; D005838:Genotype; D001467:Hordeum / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide
D005819:Genetic Markers