Rapid cloning of genes in hexaploid wheat using cultivar-specific long-range chromosome assembly

0482335 - ÚEB 2018 RIV US eng J - Journal Article
Thind, A. K. - Wicker, T. - Šimková, Hana - Fossati, D. - Moullet, O. - Brabant, C. - Vrána, Jan - Doležel, Jaroslav - Krattinger, S.G.
Rapid cloning of genes in hexaploid wheat using cultivar-specific long-range chromosome assembly.
Nature Biotechnology. Roč. 35, č. 8 (2017), s. 793-796. ISSN 1087-0156. E-ISSN 1546-1696
R&D Projects: GA MŠMT(CZ) LO1204
Institutional support: RVO:61389030
Keywords : adult-plant resistance * leaf rust * sequence capture * genome * arabidopsis * virulence * barley * canada * locus * lr22a
OECD category: Plant sciences, botany
Impact factor: 35.724, year: 2017

Cereal crops such as wheat and maize have large repeat-rich genomes that make cloning of individual genes challenging. Moreover, gene order and gene sequences often differ substantially between cultivars of the same crop species1-4. A major bottleneck for gene cloning in cereals is the generation of high-quality sequence information from a cultivar of interest. In order to accelerate gene cloning from any cropping line, we report ` targeted chromosome-based cloning via long-range assembly' (TACCA). TACCA combines lossless genome-complexity reduction via chromosome flow sorting with Chicago long-range linkage5 to assemble complex genomes. We applied TACCA to produce a high-quality (N50 of 9.76 Mb) de novo chromosome assembly of the wheat line CH Campala Lr22a in only 4 months. Using this assembly we cloned the broad-spectrum Lr22a leaf-rust resistance gene, using molecular marker information and ethyl methanesulfonate (EMS) mutants, and found that Lr22a encodes an intracellular immune receptor homologous to the Arabidopsis thaliana RPM1 protein.
Permanent Link: http://hdl.handle.net/11104/0277748