Undesired fertility restoration in msm1 barley associates with two mTERF genes

  • Timm BernhardEmail author
  • Michael Koch
  • Rod J. Snowdon
  • Wolfgang Friedt
  • Benjamin Wittkop
Original Article


Key message

The novel Rfm3 locus causing undesired fertility restoration in the msm1 cytoplasm of winter barley is located on the short arm of chromosome 6H.


Undesired fertility restoration of cytoplasmic male sterile (CMS) mother lines in absence of the functional Rfm1 restorer gene is a significant problem for hybrid breeding in winter barley. Here, we describe that a novel restorer locus on the short arm of chromosome 6H, designated Rfm3, is closely linked to two mitochondrial transcription termination factor family (mTERF) protein coding genes. Genome-wide association studies in a multiparental mapping population revealed that two of the most significantly associated markers are located very close to these genes, with one marker lying directly within one mTERF gene sequence. Sequences of the candidate genes in the parental lines, segregating individuals and an independent set of breeding lines clearly revealed haplotypes discriminating completely sterile, partially fertile and Rfm1-restorer lines. The haplotypes segregate for several single nucleotide polymorphisms, a 6 bp insertion–deletion (InDel) polymorphism and another 2 bp InDel. CMS-unstable genotypes carrying haplotypes associated with undesired fertility restoration showed significantly higher grain setting on bagged spikes when plants were subjected to elevated temperatures during anthesis, indicating a temperature influence on pollen fertility. SNPs associated with desirable Rfm3 haplotypes can be implemented in marker-assisted selection of stable CMS mother lines.



Cytoplasmic male sterility


Fertility index




Mitochondrial transcription termination factor


Principal component analysis


Pentatricopeptide repeat


Standard deviation


Within sum of squares



This project was part of the collaborative projects “HybGPS” and “SpeedBarley” coordinated by the Federal Agency of Renewable Resources (FNR) and the Federal Office for Agriculture and Food, respectively, and funded by the Federal Ministry of Food and Agriculture. Thanks are due to the collaborative companies German Seed Alliance GmbH, Ackermann Saatzucht GmbH & Co. KG, Deutsche Saatveredelung AG (DSV), Nordsaat Saatzuchtgesellschaft mbH, Saaten-Union Recherche s.a.s. (SUR), W. von Borries-Eckendorf GmbH & Co. KG (WvB) and Saaten-Union Biotech GmbH (SUB) for their support and cooperation. We are particularly grateful to Dr. Jutta Ahlemeyer, Dr. Jens Vaupel and Astrid Hoffmann (DSV), Dr. Laszlo Cselenyi and Ulrike Avenhaus (WvB), Dr. Charles Snijders (SUR), Dr. Eberhard Laubach (Nordsaat) and Jutta Förster (SUB) for actively supporting this study and for valuable discussions. We also thank Annette Plank, Birgit Keiner, Stavros Tzigos, Lisa Unterberg, Petra Kretschmer and Nadine Biermann (DSV) for their valuable experimental assistance.

Author contribution statement

TB conducted the greenhouse and climate chamber trials, did the statistical analyses and the genome-wide association study, prepared the figures and tables and wrote the manuscript. MK and TB developed the sequencing primers and guided the reprocessing of the sequencing data for the candidate gene. TB, BW, WF and RJS conceived the research. BW, WF and RJS provided advice on trial conductance and on genome analysis and data interpretation. MK, BW, WF and RJS edited the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The authors declare that the experiments comply with the current laws of Germany.

Supplementary material

122_2019_3281_MOESM1_ESM.docx (26 kb)
Supplementary material 1 (DOCX 25 kb)


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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Plant Breeding, IFZ Research Centre for Biosystems, Land Use and NutritionJustus Liebig UniversityGiessenGermany
  2. 2.Deutsche Saatveredelung AGSalzkottenGermany

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