Improving Binding of Rf1 and Rf3 Proteins to Their RNA Targets for Hybrid Wheat Breeding

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Cytoplasmic male sterility (CMS) is a mitochondrial trait that is often used in hybrid breeding in autogamous plants to control self-pollination and produce sterile plants. To restore fertility in the F1 generation, Rf (Restorer-of-fertility) genes, which suppress the expression of the CMS trait, are applied. Rf proteins often belong to the pentatricopeptide repeat protein (PPR) family. CMS-Rf systems have been successfully applied in rice, maize and rye hybrid breeding and thus this approach is likely to be feasible for producing commercial hybrid wheat seed cheaply. Recently, orf279 has been determined as the genetic cause of CMS in wheat carrying Triticum timopheevii cytoplasm and the sequences of the Rf1 and Rf3 restorer genes have been identified. However, F1 fertility restoration conferred by both Rf genes and their level of expression were reported to vary in different genetic backgrounds. In addition, RNA Electrophoretic Mobility Shift Assay (REMSA) has shown weak binding of Rf1 and Rf3 proteins to their RNA targets. Here, we hypothesise that the poor RNA binding of Rf1 and Rf3 contributes to the inconsistent fertility restoration. My studies aim to improve the RNA binding affinity of Rf1 and Rf3. I will analyse the binding site and the protein sequences of Rf1 and Rf3, identify suboptimal amino acid combinations and change them by site-directed mutagenesis. Protein variants with improved binding affinity will be tested in vivo using transient transformation of wheat protoplasts. This study will be beneficial to hybrid wheat breeding programmes using CMS to control male fertility.
Original languageEnglish
Publication statusPublished - 11 Sep 2021
EventPerth Protein Group Annual General Meeting 2021 - Edith Cowan University, Bunbury Campus, Bunbury, Australia
Duration: 10 Sep 202112 Sep 2021


ConferencePerth Protein Group Annual General Meeting 2021
Abbreviated titlePPG2021
Internet address


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