Genomics-assisted dissection of barley morphology and development (ERA-PG 046)

我们将利用Illumina Origo Posay(OPA)技术对480个鲍曼回交获得的近等基因突变系进行基因分型，这些突变系位于3072个定位的遗传座位上(6144个等位基因)。我们将与它们的轮回亲本Bowman进行简单的配对比较，以确定包含突变等位基因的片段的边界，并基于大麦、水稻和新兴的Brachypodium和玉米基因组序列，为每个座位编制详细的比较推测的基因含量图。虽然OPA基因分型将把每个突变基因的位置缩小到一个很小的遗传区间，但从所有潜在的候选基因中识别突变基因可能仍然需要一种正向遗传学方法。为了促进这一点，以及未来所有类似的研究，我们建议为建立大麦基因组遗传锚定的物理图谱的分阶段开发的国际战略做出贡献，通过末端测序BAC，这些BAC目前正在进行高信息含量指纹分析和与&gt；12,000个非基因的超额杂交。这将提供高达400 Mbp的大麦基因组DNA序列信息，并将有助于将物理图谱锚定到大麦基因图谱(~5-6000个定位基因)和水稻基因组序列。为了证明开发的信息、资源和方法的实用性，每个合作实验室将确定跨越形态/发育突变位点的最小BAC平铺路径，确定候选基因，并通过比较等位基因测序(WT与突变)和使用小组可用的功能基因组学工具(TILLING、转基因、VIGS)的组合来启动它们的功能表征。所有信息将通过现有的基于网络的信息学资源向社会公开，以供更广泛的利用。

We will use Illumina Oligo Pool Assay (OPA) technology to genotype 480 Bowman backcross derived nearly isogenic mutant lines at 3072 mapped genetic loci (6144 alleles). We will use simple pair-wise comparison with Bowman, their recurrent parent, to define the boundaries of the segments containing mutated alleles and compile a detailed comparative putative gene content map for each locus based on the barley, rice and emerging Brachypodium and maize genome sequences. While OPA genotyping will narrow the location of each mutant locus to a small genetic interval, identifying the mutated gene from all potential candidates will likely still require a forward genetics approach. To facilitate this, and all similar studies in the future, we propose to contribute to an established international strategy for the staged development of a genetically anchored physical map of the barley genome by end sequencing BACs that are currently undergoing High Information Content Fingerprint analysis and Overgo hybridisation to >12,000 unigenes. This will provide up to 400 Mbp of barley genomic DNA sequence information and will assist in anchoring the physical map to the barley gene map (~5-6000 mapped genes) and rice genome sequence. To demonstrate the utility of the developed information, resources and approach, each partner lab will identify a minimum BAC tiling path spanning a morphological / developmental mutant locus, identify candidate genes and initiate their functional characterisation by a combination of comparative allele sequencing (WT vs. Mutant) and the use of functional genomics tools available to the group (TILLING, transgenics, VIGS). All information will be made publicly available to the community for broader exploitation through existing web-based informatics resources.