低温危害一直是小麦生产发展的重要限制因子，关于小麦冻害涉及的生理生化机理有一定研究，但在低温冻害遗传基础及其相关基因方面的研究较少。本项目以课题组发现鉴定的低温敏感突变体BN044371为材料，在前期对低温敏感基因wchs1（暂定名）定位的基础上，应用下一代转录组测序技术（NGS）和集群分离分析（BSA）相结合的BSA RNA-Seq 方法，开发与wchs1 紧密连锁的分子标记，构建高密度遗传连锁图谱并精细定位。结合粗山羊草参考基因组和普通小麦测序信息最新进展，应用图位克隆技术，结合gene capture技术和NGS，实现对wchs1的克隆。研究结果将有助于解析小麦苗期对低温敏感的遗传基础，对小麦抗低温育种理论和实践均具有重要的意义。研究方法和策略对普通小麦D基因组重要性状功能基因遗传图谱构建、精细定位和基因克隆有重要借鉴和参考价值。

Freezing temperature limits the geographical distribution of wheat (T. aestivum L.) and often causes severe losses in productivity. Therefore, understanding the genetic mechanism responding to low temperature is very important for wheat tolerance breeding. We characterized a wheat chilling-sensitive mutant BN044371, which carries a chilling-sensitive recessive gene temporarily named wchs1.This mutant grew normally at 3°C and above but showed yellowish and collapsed leaves phenotypes when shifted to temperatures below 0- 2°C after seven or more days at seedling stage. The wchs1 was mapped on the 1DS according to the SSR and SNP markers. In this proposal, next generation sequencing (NGS) and bulked segregant analysis (BSA) techniques were applied to develop polymorphic SNP markers tightly linked to wchs1, a high-density genetic linkage map of wchs1 will be constructed which is essential for fine mapping and map-based cloning of the gene. Based on the advanced genome sequencing of Aegilops tauschii and wheat, using map-based gene cloning technology combining with the gene capture array and the NGS, the wchs1 gene will be cloned. The proposed research will interpret the genetic mechanism responding to low temperature, which is necessary for wheat tolerance theory and breeding practice. The methodology applied in this research will demonstrate its potential power in developing high density genetic linkage maps of agronomical important traits, fine mapping and cloning genes in common wheat.