Akai Kotaro, Asano Kenji, Suzuki Chika, Shimosaka Etsuo, Tamiya Seiji, Suzuki Takako, Takeuchi Toru, Ohki Takehiro
Memuro Upland Farming Research Division, Hokkaido Agricultural Research Center, National Agricultural Research Organization, Memuro, Hokkaido 082-0081, Japan.
Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido 060-8589, Japan.
Breed Sci. 2023 Apr;73(2):168-179. doi: 10.1270/jsbbs.22078. Epub 2023 Apr 13.
The isolation of disease resistance genes introduced from wild or related cultivated species is essential for understanding their mechanisms, spectrum and risk of breakdown. To identify target genes not included in reference genomes, genomic sequences with the target locus must be reconstructed. However, assembly approaches of the entire genome, such as those used for constructing reference genomes, are complicated in higher plants. Moreover, in the autotetraploid potato, the heterozygous regions and repetitive structures located around disease resistance gene clusters fragment the genomes into short contigs, making it challenging to identify resistance genes. In this study, we report that a assembly approach of a target gene-specific homozygous dihaploid developed through haploid induction was suitable for gene isolation in potatoes using the potato virus Y resistance gene as a model. The assembled contig containing -linked markers was 3.3 Mb in length and could be joined with gene location information from the fine mapping analysis. was successfully identified in a repeated island located on the distal end of the long arm of chromosome 9 as a Toll/interleukin-1 receptor-nucleotide-binding site-leucine rich repeat (TIR-NBS-LRR) type resistance gene. This approach will be practical for other gene isolation projects in potatoes.
从野生或相关栽培物种中导入的抗病基因的分离对于了解其机制、作用谱和失效风险至关重要。为了鉴定参考基因组中未包含的目标基因,必须重建具有目标位点的基因组序列。然而,在高等植物中,用于构建参考基因组的全基因组组装方法很复杂。此外,在同源四倍体马铃薯中,抗病基因簇周围的杂合区域和重复结构将基因组片段化为短重叠群,这使得鉴定抗病基因具有挑战性。在本研究中,我们报告称,通过单倍体诱导开发的目标基因特异性纯合双单倍体的组装方法适用于以马铃薯Y病毒抗性基因为例的马铃薯基因分离。包含连锁标记的组装重叠群长度为3.3 Mb,并且可以与精细定位分析的基因定位信息相结合。该基因在9号染色体长臂远端的一个重复岛中成功鉴定为Toll/白细胞介素-1受体-核苷酸结合位点-富含亮氨酸重复序列(TIR-NBS-LRR)型抗性基因。这种方法对于马铃薯的其他基因分离项目将是实用的。