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Identification and characterization of a semi-dominant restorer-of-fertility 1 allele in sugar beet (Beta vulgaris)

Theoretical and Applied Genetics volume 132, pages 227–240 (2019)Cite this article

Abstract

Key message

The sugar beet Rf1 locus has a number of molecular variants. We found that one of the molecular variants is a weak allele of a previously identified allele.

Abstract

Male sterility (MS) caused by nuclear-mitochondrial interaction is called cytoplasmic male sterility (CMS) in which MS-inducing mitochondria are suppressed by a nuclear gene, restorer-of-fertility. Rf and rf are the suppressing and non-suppressing alleles, respectively. This dichotomic view, however, seems somewhat unsatisfactory to explain the recently discovered molecular diversity of Rf loci. In the present study, we first identified sugar beet line NK-305 as a new source of Rf1. Our crossing experiment revealed that NK-305 Rf1 is likely a semi-dominant allele that restores partial fertility when heterozygous but full fertility when homozygous, whereas Rf1 from another sugar beet line appeared to be a dominant allele. Proper degeneration of anther tapetum is a prerequisite for pollen development; thus, we compared tapetal degeneration in the NK-305 Rf1 heterozygote and the homozygote. Degeneration occurred in both genotypes but to a lesser extent in the heterozygote, suggesting an association between NK-305 Rf1 dose and incompleteness of tapetal degeneration leading to partial fertility. Our protein analyses revealed a quantitative correlation between NK-305 Rf1 dose and a reduction in the accumulation of a 250 kDa mitochondrial protein complex consisting of a CMS-specific mitochondrial protein encoded by MS-inducing mitochondria. The abundance of Rf1 transcripts correlated with NK-305 Rf1 dose. The molecular organization of NK-305 Rf1 suggested that this allele evolved through intergenic recombination. We propose that the sugar beet Rf1 locus has a series of multiple alleles that differ in their ability to restore fertility and are reflective of the complexity of Rf evolution.

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Acknowledgements

This work was supported in part by JSPS KAKENHI Grant Number 18K05564 (TK) and NARO Bio-oriented Technology Research Advancement Institution (BRAIN) (Research program on development of innovative technology, Grant Number 30001A) (KT, KK and TK). TA is a recipient of a JSPS Research Fellowship for Young Scientists (16J01146).

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Author notes

  1. Takumi Arakawa and Sachiyo Ue have equally contributed to this study.

Affiliations

  1. Research Faculty of Agriculture, Hokkaido University, N-9, W-9, Kita-ku, Sapporo, 060-8589, Japan

    Takumi Arakawa, Sachiyo Ue, Chihiro Sano, Muneyuki Matsunaga, Hiroyo Kagami, Yu Yoshida, Kazuyoshi Kitazaki & Tomohiko Kubo

  2. Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization, Shinsei Minami 9-4, Memuro, 082-0081, Japan

    Yosuke Kuroda & Kazunori Taguchi

Authors
  1. Takumi Arakawa
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  2. Sachiyo Ue
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  3. Chihiro Sano
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  4. Muneyuki Matsunaga
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  9. Kazuyoshi Kitazaki
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Correspondence to Tomohiko Kubo.

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Additional information

Communicated by Mingliang Xu.

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Supplementary Fig.

 1. Agarose (2%) gel electrophoresis of PCR products amplified with primers for cytoplasmic DNA markers (PDF 76 kb)

Supplementary Fig.

 2. PCR products amplified with primers for DNA marker o7 (PDF 67 kb)

Supplementary Fig.

 3. DNA gel blot analysis of an NK-305 plant probed with an orf20-like 3′-UTR (PDF 58 kb)

Supplementary Fig.

 4. Alignment of nucleotide sequences of orf20NK-198 and orf20NK-305-1 coding and flanking regions (PDF 68 kb)

Supplementary Fig.

 5. Alignment of nucleotide sequences of orf20LS and orf20NK-305-2 coding and flanking regions (PDF 70 kb)

Supplementary Fig.

 6. Alignment of amino acid sequences of the protein products deduced from orf20NK-198 and orf20NK-305-1 nucleotide sequences (PDF 33 kb)

Supplementary Fig.

 7. Alignment of amino acid sequences of the protein products deduced from orf20LS and orf20NK-305-2 nucleotide sequences (PDF 33 kb)

Supplementary Fig.

 8. Alignment of nucleotide sequences of the upstream regions of orf19 and orf20NK-305-2 (PDF 46 kb)

Supplementary Fig.

 9. Alignment of nucleotide sequences of the upstream regions of orf20LS and orf20NK-305-1 (PDF 60 kb)

Supplementary Fig.

 10. Immunoblot analysis of proteins from transgenic suspension cells separated by SDS-PAGE (PDF 91 kb)

Supplementary Fig.

 11. Immunoblot analysis of proteins from transgenic suspension cells separated by BN-PAGE (PDF 196 kb)

Supplementary Table

 1. Nucleotide sequences of primers used in this study (PDF 35 kb)

Supplementary Table

 2. Segregation of male fertility and o7 marker types in an F2 population (PDF 36 kb)

Supplementary Table

 3. Segregation of male fertility and s17 marker types in an admixture population (PDF 37 kb)

Supplementary Table

 4. Segregation of male fertility and s17 type in a BC2F2 population (PDF 36 kb)

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Arakawa, T., Ue, S., Sano, C. et al. Identification and characterization of a semi-dominant restorer-of-fertility 1 allele in sugar beet (Beta vulgaris). Theor Appl Genet 132, 227–240 (2019). https://doi.org/10.1007/s00122-018-3211-6

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  • DOI: https://doi.org/10.1007/s00122-018-3211-6