The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley
The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley
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DOI：
10.1007/s00122-008-0813-4
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发表时间：
2008-09-01
影响因子：
5.4
通讯作者：
McNamara, Raymond B.
中科院分区：
文献类型：
--
作者：
Lehmensiek, Anke;Sutherland, Mark W.;McNamara, Raymond B.
利用已建立的遗传图谱，将单基因调控覆盖黑穗病抗性Ruh.7H定位到亚历克西斯/单倍体大麦群体7HS染色体端粒区。距离Ruh.7H， abg704最近的标记为7.5 cM。利用水稻序列数据，应用比较基因组学方法鉴定了7HS远端13个可能含有单核苷酸多态性（snp）的位点。其中，一个位点产生不同大小的多态性共显性带，而另外两个位点含有snp，这些snp是使用最近开发的高分辨率熔化（HRM）技术鉴定的。其中两个标记位于Ruh.7H两侧，近端标记位于3.8 cM处，远端标记位于2.7 cM处。这是首次将HRM技术应用于植物中已知的cleaved amplified polymorphic sequence （CAPS）标记的SNP检测和快速得分。这种简单、精确的后pcr技术应该广泛应用于复杂谷物和其他植物基因组中感兴趣的遗传区域的精细定位。
Using an established genetic map, a single gene conditioning covered smut resistance, Ruh.7H, was mapped to the telomere region of chromosome 7HS in an Alexis/Sloop doubled haploid barley population. The closest marker to Ruh.7H, abg704 was 7.5 cM away. Thirteen loci on the distal end of 7HS with potential to contain single nucleotide polymorphisms (SNPs) were identified by applying a comparative genomics approach using rice sequence data. Of these, one locus produced polymorphic co-dominant bands of different size while two further loci contained SNPs that were identified using the recently developed high resolution melting (HRM) technique. Two of these markers flanked Ruh.7H with the proximal marker located 3.8 cM and the distal marker 2.7 cM away. This is the first report on the application of the HRM technique to SNP detection and to rapid scoring of known cleaved amplified polymorphic sequence (CAPS) markers in plants. This simple, precise post-PCR technique should find widespread use in the fine-mapping of genetic regions of interest in complex cereal and other plant genomes.