期刊
FUNCTIONAL & INTEGRATIVE GENOMICS
卷 14, 期 1, 页码 85-100出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10142-014-0360-9
关键词
Barley; Fr-H2; Physical mapping; 454 sequencing; CBF genes
资金
- Fondazione Cassa di Risparmio di Modena, Progetto di Ricerca Internazionale FROSTMAP Physical mapping of the barley Frost resistance-H2 (Fr-H2) locus
- GENOMORE project of the Fondazione Cassa di Risparmio di Carpi
Frost resistance-H2 (Fr-H2) is a major QTL affecting freezing tolerance in barley, yet its molecular basis is still not clearly understood. To gain a better insight into the structural characterization of the locus, a high-resolution linkage map developed from the Nure x Tremois cross was initially implemented to map 13 loci which divided the 0.602 cM total genetic distance into ten recombination segments. A PCR-based screening was then applied to identify positive bacterial artificial chromosome (BAC) clones from two genomic libraries of the reference genotype Morex. Twenty-six overlapping BACs from the integrated physical-genetic map were 454 sequenced. Reads assembled in contigs were subsequently ordered, aligned and manually curated in 42 scaffolds. In a total of 1.47 Mbp, 58 protein-coding sequences were identified, 33 of which classified according to similarity with sequences in public databases. As three complete barley C-repeat Binding Factors (HvCBF) genes were newly identified, the locus contained13 full-length HvCBFs, four Related to AP2 Triticeae (RAPT) genes, and at least five CBF pseudogenes. The final overall assembly of Fr-H2 includes more than 90 % of target region: all genes were identified along the locus, and a general survey of Repetitive Elements obtained. We believe that this gold-standard sequence for the Morex Fr-H2 will be a useful genomic tool for structural and evolutionary comparisons with Fr-H2 in winter-hardy cultivars along with Fr-2 of other Triticeae crops.
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