Abstract
A set of 398 simple sequence repeat markers (SSRs) have been developed and characterised for use with genetic studies of Brassica species. Small-insert (250–900 bp) genomic libraries from Brassica rapa, B. nigra, B. oleracea and B. napus, highly enriched for dinucleotide and trinucleotide SSR motifs, were constructed. Screening the clones with a mixture of oligonucleotide repeat probes revealed positive hybridisation to between 75% and 90% of the clones. Of these, 1,230 were sequenced. Primer pairs were designed for 398 SSR clones, and of these, 270 (67.8%) amplified a PCR product of the expected size in their focal and/or closely related species. A further screen of 138 primers pairs that produced a PCR product in B. napus germplasm found that 86 (62.3%) revealed length polymorphisms within at least one line of a test array representing the four Brassica species. The results of this screen were used to identify 56 SSRs and were combined with 41 SSRs that had previously shown polymorphism between the parents of a B. napus mapping population. These 97 SSR markers were mapped relative to a framework of RFLP markers and detected 136 loci over all 19 linkage groups of the oilseed rape genome.
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Acknowledgements
We would like to thank staff at IACR Long Ashton and the John Innes Centre who provided support and help during laboratory work. This work was primarily funded by a grant (ref D08078) from the Biotechnology and Biological Sciences Research Council. Supporting finance was provided by IACR Long Ashton, John Innes Centre, BBSRC and the Natural Environment Research Council.
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Lowe, A.J., Moule, C., Trick, M. et al. Efficient large-scale development of microsatellites for marker and mapping applications in Brassica crop species. Theor Appl Genet 108, 1103–1112 (2004). https://doi.org/10.1007/s00122-003-1522-7
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DOI: https://doi.org/10.1007/s00122-003-1522-7