Fazlikhani Leila, Keilwagen Jens, Kopahnke Doris, Deising Holger, Ordon Frank, Perovic Dragan
Institute for Resistance Research and Stress Tolerance, Federal Research Centre for Cultivated Plants, Julius Kühn-Institute (JKI), Quedlinburg, Germany.
Department of Phytopathology and Plant Protection, Institute of Agricultural and Nutrition Sciences, Faculty of Natural Sciences III, Martin Luther University of Halle-Wittenberg, Halle, Germany.
Front Plant Sci. 2019 May 22;10:640. doi: 10.3389/fpls.2019.00640. eCollection 2019.
Isolation of disease resistance genes in barley was hampered by the large genome size, but has become easy due to the availability of the reference genome sequence. During the last years, many genomic resources, e.g., the Illumina 9K iSelect, the 50K Infinium arrays, the Barley Genome Zipper, POPSEQ, and genotyping by sequencing (GBS), were developed that enable enhanced gene isolation in combination with the barley genome sequence. In the present study, we developed a fine map of the barley leaf rust resistance gene . 537 segmental homozygous recombinant inbred lines (RILs) derived from 4775 F-plants were used to construct a high-resolution mapping population (HRMP). The Barley Genome Zipper, the 9K iSelect chip, the 50K Infinium chip and GBS were used to develop 56 molecular markers located in the target interval of 8 cM. This interval was narrowed down to about 0.07 cM corresponding to 0.44 Mb of the barley reference genome. Eleven low-confidence and 18 high-confidence genes were identified in this interval. Five of these are putative disease resistance genes and were subjected to allele-specific sequencing. In addition, comparison of the genetic map and the reference genome revealed an inversion of 1.34 Mb located distally to the resistance locus. In conclusion, the barley reference sequence and the respective gene annotation delivered detailed information about the physical size of the target interval, the genes located in the target interval and facilitated the efficient development of molecular markers for marker-assisted selection for
大麦抗病基因的分离曾因基因组庞大而受阻,但由于参考基因组序列的可得性,现在变得容易了。在过去几年里,开发了许多基因组资源,例如Illumina 9K iSelect、50K Infinium芯片、大麦基因组拉链、POPSEQ以及测序基因分型(GBS),这些资源与大麦基因组序列相结合,能够增强基因分离能力。在本研究中,我们绘制了大麦叶锈病抗性基因的精细图谱。利用从4775株F植株衍生而来的537个片段纯合重组自交系(RIL)构建了一个高分辨率定位群体(HRMP)。使用大麦基因组拉链、9K iSelect芯片、50K Infinium芯片和GBS开发了56个位于8 cM目标区间的分子标记。该区间被缩小到约0.07 cM,对应于大麦参考基因组的0.44 Mb。在这个区间内鉴定出11个低可信度基因和18个高可信度基因。其中5个是假定的抗病基因,并对其进行了等位基因特异性测序。此外,遗传图谱与参考基因组的比较揭示了一个位于抗性基因座远端的1.34 Mb倒位。总之,大麦参考序列和相应的基因注释提供了有关目标区间物理大小、位于目标区间的基因的详细信息,并促进了用于标记辅助选择的分子标记的高效开发。