Validation of the high-throughput marker technology DArT using the model plant Arabidopsis thaliana.
Diversity Arrays Technology (DArT) is a microarray-based DNA marker technique for genome-wide discovery and genotyping of genetic variation. DArT allows simultaneous scoring of hundreds of restriction site based polymorphisms between genotypes and does not require DNA sequence information or site-specific oligonucleotides. This paper demonstrates the potential of DArT for genetic mapping by validating the quality and molecular basis of the markers, using the model plant Arabidopsis thaliana. Restriction fragments from a genomic representation of the ecotype Landsberg erecta (Ler) were amplified by PCR, individualized by cloning and spotted onto glass slides. The arrays were then hybridized with labeled genomic representations of the ecotypes Columbia (Col) and Ler and of individuals from an F(2) population obtained from a Col x Ler cross. The scoring of markers with specialized software was highly reproducible and 107 markers could unambiguously be ordered on a genetic linkage map. The marker order on the genetic linkage map coincided with the order on the DNA sequence map. Sequencing of the Ler markers and alignment with the available Col genome sequence confirmed that the polymorphism in DArT markers is largely a result of restriction site polymorphisms.
Wittenberg
Alexander H J
AH
Department of Plant Sciences, Laboratory of Plant Breeding, The Graduate School Experimental Plant Sciences, Wageningen University and Research Centre, P.O. Box 386, 6700 AJ Wageningen, The Netherlands.
van der Lee
Theo
T
Cayla
Cyril
C
Kilian
Andrzej
A
Visser
Richard G F
RG
Schouten
Henk J
HJ
D017360:Arabidopsis / Q000235:genetics*; D002874:Chromosome Mapping; D032461:Chromosomes, Plant / Q000235:genetics; D003433:Crosses, Genetic; D008040:Genetic Linkage; D005819:Genetic Markers / Q000235:genetics*; D018745:Genome, Plant; D005838:Genotype; D046228:Microarray Analysis; D005810:Multigene Family / Q000235:genetics; D011110:Polymorphism, Genetic / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length
D005819:Genetic Markers