caa a22 4500 378901990 CHVBK 20180305123520.0 cr unu---uuuuu 161128e20040701xx s 000 0 eng 10.2298/hel0440001k doi (NATIONALLICENCE)gruyter-10.2298/hel0440001k MAP-BASED CLONING STRATEGY FOR ISOLATING THE RESTORER GENE Rf1 OF THE PET1 CYTOPLASM IN SUNFLOWER (Helianthus annuus L.) [Elektronische Daten] ESTRATEGIA DE CLONACIÓN PARA EL AISLAMIENTO DEL GEN RESTAURADOR Rf1 EN EL CITOPLASMA PET1 EN GIRASOL (Helianthus annuus L.) A BASE DE LOS MAPAS CONSTRUIDOS / STRATÉGIE DE CLONAGE POUR ISOLER LE GÈNE RESTAURATEUR Rf1 DANS LE CYTOPLASME PET1 DU TOURNESOL (Helianthus annuus L.) À LA BASE DES CARTES CONSTRUITES / [B. Kusterer, W. Friedt, E. Lazarescu, M. Prüfe, N. Özdemir, S. Tzigos, R. Horn] Map-based cloning of the restorer gene Rf1, responsible for fertility restoration of the PET1 cytoplasm in sunflower, requires tightly linked markers to the gene of interest. Screening 1200 decamer primers by bulked segregant analyses identified seven RAPD markers mapping on the same linkage group as the restorer gene Rf1. In the F2 population of the cross RHA325(cms) × HA342 (183 individuals) one of the RAPD markers, OPK13_454, mapped 0.9 cM from Rf1, followed by OPY10_740 with 2.2 cM. Bulked segregant analyses using 1024 AFLP primer combinations identified 282 polymorphisms in 203 primer combinations. Selected single plants from the bulks and recombinant plants were screened to further reduce the number of primer combinations to be mapped in the F2 population. The map for the linkage group carrying the restorer gene now consists of 43 markers (7 RAPD-, 1 SSR-, and 35 AFLPmarkers) and covers 191.9 cM. E32M36_155 and E44M70_275 were mapped 0.7 cM and 0.1 cM from the restorer gene, respectively. Two of the RAPD markers, OPK13_454 and OPY10_740, were successfully converted into SCAR markers, HRG01 and HRG02, respectively. Amplification of the markers OPK13_454/ HRG01 and OPY10_740/HRG02 was investigated for a set of 11 restorer and nine maintainer lines of PET1. For cloning the restorer gene Rf1, colony hybridizations against high-density BAC filters of our sunflower BAC library and 3D-PCR pooling strategies were used to identify positive BAC clones. BAC fingerprinting using different restriction enzymes in combination with hybridizations was performed to develop a contig around the restorer locus Rf1. This article is distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. BAC nationallicence CMS nationallicence fertility restoration nationallicence Helianthus nationallicence sunflower nationallicence Kusterer B. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Friedt W. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Lazarescu E. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Prüfe M. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Özdemir N. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Tzigos S. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut Horn R. Universität Rostock, Fachbereich Biowissenschaften, Institut für Molekulare Physiologie und Biotechnologie, Albert Einstein Str.3, 18051 Rostock, Germany, Phone: +49(0)381 498 6111/6170, Fax: +49(0)381 498 6112 aut HELIA De Gruyter 27/40(2004-07-01), 1-14 1018-1806 27:40<1 2004 27 helia https://doi.org/10.2298/hel0440001k text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.2298/hel0440001k text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kusterer B. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Friedt W. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Lazarescu E. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Prüfe M. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Özdemir N. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Tzigos S. Institut für Pflanzenbau und Pflanzenzüchtung I, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, Germany aut NATIONALLICENCE 700 1- Horn R. Universität Rostock, Fachbereich Biowissenschaften, Institut für Molekulare Physiologie und Biotechnologie, Albert Einstein Str.3, 18051 Rostock, Germany, Phone: +49(0)381 498 6111/6170, Fax: +49(0)381 498 6112 aut NATIONALLICENCE 773 0- HELIA De Gruyter 27/40(2004-07-01), 1-14 1018-1806 27:40<1 2004 27 helia CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter