Abstract
The combination of cytoplasmic male sterility (CMS) in one parent and a restorer gene (Rf) to restore fertility in another are indispensable for the development of hybrid varieties. We have found a rice Rf-1 gene that restores BT-type CMS by applying a positional cloning strategy. Using linkage analysis in combination with 6,104 BC1F3 progeny derived from a cross between two near-isogenic lines (NILs) differing only at the Rf-1 locus, we delimited the Rf-1 gene to a 22.4-kb region in the rice genome. Duplicate open reading frames (Rf-1A and Rf-1B) with a pentatricopeptide (PPR) motif were found in this region. Since several insertions and/or deletions were found in the regions corresponding to both the Rf-1A and Rf-1B genes in the maintainer’s allele, they may have lost their function. Rf-1A protein had a mitochondria-targeting signal, whereas Rf-1B did not. The Rf-1B gene encoded a shorter polypeptide that was determined by a premature stop codon. Based on the function of the Rf-1 gene, its product is expected to target mitochondria and may process the transcript from an atp6/orf79 region in the mitochondrial genome. Since the Rf-1A gene encodes a 791-amino acid protein with a signal targeting mitochondria and has 16 repeats of the PPR motif, we concluded that Rf-1A is the Rf-1 gene. Nine duplications of Rf-1A homologs were found around the Rf-1 locus in the Nipponbare genome. However, while some of them encoded proteins with the PPR motif, they do not restore BT-type CMS based on the lack of co-segregation with the restoration phenotype. These duplicates may have played diversified roles in RNA processing and/or recombination in mitochondria during the co-evolution of these genes and the mitochondrial genome.
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Acknowledgements
The authors thank Dr. Shinjyo for kindly providing the seeds of the near-isogenic lines of rice. This research was supported in part by a Grant-in-Aid for Scientific Research C (No. 12660008) from the Ministry of Education, Science and Culture, Japan.
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Communicated by M. Mackill
Sequence data from this article have been deposited with the EMBL/Genebank Data Libraries under accession nos. AB112808, AB112809, AB112810, AB112811.
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Akagi, H., Nakamura, A., Yokozeki-Misono, Y. et al. Positional cloning of the rice Rf-1 gene, a restorer of BT-type cytoplasmic male sterility that encodes a mitochondria-targeting PPR protein. Theor Appl Genet 108, 1449–1457 (2004). https://doi.org/10.1007/s00122-004-1591-2
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DOI: https://doi.org/10.1007/s00122-004-1591-2