Department of Breeding Research, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), OT Gatersleben, Corrensstrasse 3, 06466, Seeland, Germany.
Department of Plant Physiology, Institute of Biology, Martin-Luther University Halle-Wittenberg, Weinbergweg 10, 06120, Halle, Germany.
Plant Cell Rep. 2017 Dec;36(12):1871-1881. doi: 10.1007/s00299-017-2200-6. Epub 2017 Aug 30.
Polymorphic probes identified via a sequence-based approach are suitable to infer the genotypes of recombinant inbred lines from hybridisation intensities of GeneChip transcript profiling experiments. The sequences of the probes of the ATH1 GeneChip exactly match transcript sequences of the Arabidopsis thaliana reference genome Col-0, whereas nucleotide differences and/or insertions/deletions may be observed for transcripts of other A. thaliana accessions. Individual probes of the GeneChip that show sequence polymorphisms between different A. thaliana accessions may serve as single-feature polymorphism (SFP) markers, provided that the sequence changes cause differences in hybridisation intensity for the accessions of interest. A sequence-based approach identified features on the high-density oligonucleotide array that showed sequence polymorphisms between A. thaliana accessions Col-0 and C24. Hybridisation intensities of polymorphic probes were extracted from genome-wide transcript profiles of Col-0/C24 and C24/Col-0 recombinant inbred lines and assessed after standardisation via sliding window analyses to identify SFP markers. The genotypes of the recombinant inbred lines were determined with the SFP markers and the resulting data were integrated with information, which had been established previously with single nucleotide polymorphism and insertion/deletion markers, to enrich the linkage map of the Col-0/C24 and C24/Col-0 recombinant inbred populations. Congruence between the molecular marker map and the sequence maps of the A. thaliana Col-0 chromosomes proved the reliability of the genotype information which was deduced from the transcript profiles of the Col-0/C24 and C24/Col-0 recombinant inbred lines.
基于序列的方法识别的多态性探针适合从杂交基因芯片转录谱实验的杂交强度推断重组自交系的基因型。ATH1 GeneChip 的探针序列与拟南芥参考基因组 Col-0 的转录序列完全匹配,而其他拟南芥品系的转录本可能观察到核苷酸差异和/或插入/缺失。在不同拟南芥品系之间显示序列多态性的 GeneChip 个别探针可作为单特征多态性 (SFP) 标记,只要序列变化导致感兴趣品系的杂交强度差异。基于序列的方法鉴定了高密度寡核苷酸阵列上的特征,这些特征在 Col-0 和 C24 拟南芥品系之间显示出序列多态性。从 Col-0/C24 和 C24/Col-0 重组自交系的全基因组转录谱中提取多态性探针的杂交强度,并通过滑动窗口分析进行标准化后评估,以识别 SFP 标记。使用 SFP 标记确定重组自交系的基因型,然后将所得数据与先前使用单核苷酸多态性和插入/缺失标记建立的信息集成,以丰富 Col-0/C24 和 C24/Col-0 重组自交群体的连锁图谱。分子标记图谱与 Col-0 染色体的拟南芥序列图谱之间的一致性证明了从 Col-0/C24 和 C24/Col-0 重组自交系的转录谱中推断出的基因型信息的可靠性。