Fazlikhani, Leila

Ordon, Frank

Perovic, Dragan

English

JKI Open Journal Systems (Julius Kühn-Institut)

11th Young Scientists Meeting 2018, Braunschweig, Germany, November 14-16 Fazlikhani et al. Map-based cloning of RphMBR102 conferring resistance to barley leaf rust Leila Fazlikhani, Dragan Perovic and Frank Ordon Julius Kühn Institute, Institute for Resistance Research and Stress Tolerance, Quedlinburg E-mail of corresponding author: leila.fazlikhani@julius-kuehn.deLeaf rust of barley is a serious disease caused by the biotrophic fungus Puccinia hordei Otth., which under favorable con-ditions may cause yield losses up to 62%. So far, twenty-five race-specific leaf rust resistance genes (Rph1-Rph25) have been mapped, but no one have been yet isolated. However, some of these have been overcome by new pathotypes of P.hordei, indicating the need for intro-ducing new sources of resistance intobarley breeding as well as the need forisolating known ones towards decipher-ing the structure and function offeringthe possibility of developing functionalmarkers for breeding. In this respect, re-cent advances in the development of bar-ley genomic resources i.e. 9K and 50K iSe-lect arrays, genome zipper, POPSEQ, andGBS maps, as well as the barley referencesequence, enhance the possibility of nar-rowing down the target region harboringrespective resistance genes. The RphMBR1012 gene previously mapped inthe distal region of the short arm of bar-ley chromosome 1H is effective againstthe highly virulent barley leaf rust (Puc-cinia hordei) isolate I-80. In order to posi-tionaly clone the RphMBR1012 gene, a highresolution mapping population (HRMP)was constructed based on the cross“MBR1012 (resistant) x Scarlett (suscep-tible)”. 537 segmental homozygous re-combinant inbred lines (RILs) derivedfrom 4775 F2-plants corresponding to a resolution of 0.010% recombination were identified by analyzing the popula-tion with two co-dominant flanking SSR markers (QBS94 and QBS113) spanning an interval of 8.0 cM. To down size the target interval, initially 37 SSRs and SNP markers derived from the 9K chip and the genome zipper were mapped at the HRMP, resulting in shortening the target interval to 0.1 cM, flanked by QBS97 and QBS98. Further marker saturation was done by employment of 19 additional SNP markers derived from currently available barley genomic resources i.e. the 50K iSelect arrays and GBS. All se-lected markers from 50K iSelect and GBS were converted to KASPar markers. The target interval was downsized to 0.01 cM in the window between two KASP mark-ers QBS127 and QBS98. Using BlastN search to the barley genome reference sequence, markers were anchored to the reference barley sequence revealing a physical size of 0.44 Mb. 18 high-confi-dence and 11 low-confidence genes were detected of which five are related to dis-ease resistance. Allele specific re-se-quencing of all 29 candidate genes was conducted to reduce the number of pu-tative candidate genes.  Berichte aus dem Julius Kühn-Institut 200 43

Map-based cloning of RphMBR102 conferring resistance to barley leaf rust

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Map-based cloning of RphMBR102 conferring resistance to barley leaf rust

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