Department of Plant Biology and Genome Center, University of California, Davis, Davis, CA 95616, USA.
Department of Plant Biology, Michigan State University, East Lansing, MI 48824, USA.
Genetics. 2021 Nov 5;219(3). doi: 10.1093/genetics/iyab137.
Large-scale structural variations, such as chromosomal translocations, can have profound effects on fitness and phenotype, but are difficult to identify and characterize. Here, we describe a simple and effective method aimed at identifying translocations using only the dosage of sequence reads mapped on the reference genome. We binned reads on genomic segments sized according to sequencing coverage and identified instances when copy number segregated in populations. For each dosage-polymorphic 1 Mb bin, we tested independence, effectively an apparent linkage disequilibrium (LD), with other variable bins. In nine potato (Solanum tuberosum) dihaploid families translocations affecting pericentromeric regions were common and in two cases were due to genomic misassembly. In two populations, we found evidence for translocation affecting euchromatic arms. In cv. PI 310467, a nonreciprocal translocation between chromosomes (chr.) 7 and 8 resulted in a 5-3 copy number change affecting several Mb at the respective chromosome tips. In cv. "Alca Tarma," the terminal arm of chr. 4 translocated to the tip of chr. 1. Using oligonucleotide-based fluorescent in situ hybridization painting probes (oligo-FISH), we tested and confirmed the predicted arrangement in PI 310467. In 192 natural accessions of Arabidopsis thaliana, dosage haplotypes tended to vary continuously and resulted in higher noise, while apparent LD between pericentromeric regions suggested the effect of repeats. This method, LD-CNV, should be useful in species where translocations are suspected because it tests linkage without the need for genotyping.
大片段结构变异,如染色体易位,会对适合度和表型产生深远影响,但难以识别和描述。在这里,我们描述了一种简单有效的方法,仅使用映射到参考基因组的测序reads 的剂量来识别易位。我们根据测序覆盖度大小将reads 划分到基因组片段上,并识别出种群中出现的拷贝数分离情况。对于每个剂量多态性 1 Mb 分箱,我们测试了与其他可变分箱的独立性,这实际上是一种明显的连锁不平衡(LD)。在九个马铃薯(Solanum tuberosum)二倍体家系中,影响着丝粒区的易位很常见,其中两个易位是由于基因组组装错误引起的。在两个群体中,我们发现了易位影响常染色质臂的证据。在 cv. PI 310467 中,7 号和 8 号染色体之间发生了非相互易位,导致相应染色体末端发生了 5-3 拷贝数变化,影响了几个 Mb 的区域。在 cv. "Alca Tarma"中,4 号染色体的末端臂易位到 1 号染色体的末端。使用基于寡核苷酸的荧光原位杂交(oligo-FISH)探测探针,我们对 PI 310467 中的预测排列进行了测试和验证。在 192 个拟南芥(Arabidopsis thaliana)自然群体中,剂量单倍型往往连续变化,导致噪声增加,而着丝粒区之间的明显 LD 表明重复的影响。这种方法,LD-CNV,在怀疑有易位的物种中应该是有用的,因为它无需进行基因分型即可测试连锁。