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A DArT platform for quantitative bulked segregant analysis

journal contribution
posted on 2023-05-16, 19:23 authored by Wenzl, P, Raman, H, Wang, Junping, Meixue ZhouMeixue Zhou, Huttner, E, Kilian, A
Background: Bulked segregant analysis (BSA) identifies molecular markers associated with a phenotype by screening two DNA pools of phenotypically distinct plants for markers with skewed allele frequencies. In contrast to gel-based markers, hybridization-based markers such as SFP, DArT or SNP generate quantitative allele-frequency estimates. Only DArT, however, combines this advantage with low development and assay costs and the ability to be deployed for any plant species irrespective of its ploidy level. Here we investigate the suitability of DArT for BSA applications using a barley array as an example. Results: In a first test experiment, we compared two bulks of 40 Steptoe/ Morex DH plants with contrasting pubescent leaves (mPub) alleles on chromosome 3H. At optimized levels of experimental replication and marker-selection threshold, the BSA scan identified 433 polymorphic markers. The relative hybridization contrast between bulks accurately reflected the between-bulk difference in the frequency of the mPub allele (r = 0.96). The 'platform noise' of DArT assays, estimated by comparing two identical aliquots of a DNA mixture, was significantly lower than the 'pooling noise' reflecting the binomial sampling variance of the bulking process. The allele-frequency difference on chromosome 3H increased in the vicinity of mPub and peaked at the marker with the smallest distance from mPub (4.6 cM). In a validation experiment with only 20 plants per bulk we identified an aluminum (Al) tolerance locus in a Dayton/Zhepi2 DH population on chromosome 4H with < 0.8 cM precision, the same Al-tolerance locus that had been mapped before in other barley populations. Conclusion: DArT-BSA identifies genetic loci that influence phenotypic characters in barley with at least 5 cM accuracy and should prove useful as a generic tool for high-throughput, quantitative BSA in plants irrespective of their ploidy level. © 2007 Wenzl et al; licensee BioMed Central Ltd.

Funding

Grains Research & Development Corporation

History

Publication title

BMC Genomics

Volume

8

Issue

28 June 2007

Pagination

196

ISSN

1471-2164

Department/School

Tasmanian Institute of Agriculture (TIA)

Publisher

BioMed Central Ltd

Place of publication

United Kingdom

Repository Status

  • Restricted

Socio-economic Objectives

Barley

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