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map was significantly high but the total length of the genetic map
was extremely longer than that constructed from non-GBS based
markers53; over 800 cM for each chromosome.9 It is reasonable to
assume that this is not 800 cM, but error data. In this study, the linkage map was longer in the F2 population than in the F6:7 RILs, although the effect of having different parents in the F2 population and
the F6:7 RILs had to be considered. This is because the F2 population
has more heterozygous loci than the F6:7 RILs (Supplementary Fig.
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a small amount of error data will not have a significant impact on
the results of the analysis. However, more accurate genotype information may be required to determine the order of scaffolds in the de
novo assembly of the genome. Since the cost of creating a library for
MIG-seq is low, it is possible to increase the number of reads to be
acquired and call genotypes or to sequence each strain multiple times
and use only the data that match multiple times.
In this study, a clear distortion of the segregation ratio on chromosome 5B was also observed in the linkage map of F6:7 RILs, which
was also observed in our previous study (Nishimura et al.49), where
the F5 population derived from cross between TN26 and TN28 was
genotyped using SSR markers (Supplementary Data S4). Therefore,
it is not considered a bias caused by the MIG-seq method.
In this study, we discovered that genome size is associated with
the number of bases that can be sequenced by MIG-seq, and as a result, a relatively large number of SNPs can be detected in wheat.
Genotyping data with over 3,000 markers and a low defective rate
could be obtained without precise normalization of DNA concentration between emmer wheat and durum wheat, and the possibility of
constructing a linkage map of more than 1,000 markers with many
tetraploid wheat combinations was shown. These results show that
MIG-seq can be used for high-throughput genotyping of wheat.
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