Table 1.
Summary of DNA markers used in this study.
Table 2.
Male fertility of F1 plants in two test crosses of TA-33BB-CMS x ‘Fukkoku-ouba’.
Fig 1.
Marker analysis of s17 in F1 plants and a ‘Fukkoku-ouba’ DNA gel blot probed with the orf20-3' UTR.
(A) Agarose gel electrophoresis of s17 cleaved amplified fragments derived from representative F1 plants of the cross TA-33BB-CMS x ‘Fukkoku-ouba’ #2 and a TA-33BB-CMS plant. Male fertility of each plant is indicated as S, completely sterile, or N, normal. Size markers are shown on the left (kbp). (B) Agarose gel electrophoresis of s17 markers derived from TA-33BB-CMS and ‘Fukkoku-ouba’ plants. Size markers are shown on the left (kbp). (C) A DNA gel blot containing two genomic DNAs of ‘Fukkoku-ouba’ plants probed with the orf20-3' UTR. Size markers are shown on the left (kbp).
Fig 2.
Marker analysis of o7 in F1 plants.
Agarose gel electrophoresis of o7 amplified fragments derived from representative F1 plants of the cross TA-33BB-CMS x ‘Fukkoku-ouba’ #2 and TA-33BB-CMS x ‘Fukkoku-ouba’ #1, and a TA-33BB-CMS plant. Male fertility of each plant is indicated as S, completely sterile, or N, normal. Size markers are shown on the left (kbp).
Table 3.
Segregation of o7 marker types and male fertility in TA-33BB-CMS x ‘Fukkoku-ouba’ #2.
Table 4.
Segregation of male fertility and the o7- and s17-marker-types in the B1 and B2 populations.
Table 5.
Segregation of o7 marker types in an F2 population.
Table 6.
Segregation of s17 marker types in an F2 population.
Table 7.
Marker types and male fertility in F2 population.
Fig 3.
Comparison of the genomic organization between ‘Fukkoku-ouba’ rf1 and NK-198 Rf1.
‘Fukkoku-ouba’ has a single copy of an orf20-like gene whereas homologs are clustered in NK-198. Boxes indicate genes (introns are not shown). Brackets denote the most similar regions. A single nucleotide substitution in exon 2 of orf20fukkoku is shown by a lollipop. The scale bar is shown below. The NK-198 Rf1 sequences correspond to DDBJ/EMBL/GenBank accession numbers AB646135 and AB646133.
Fig 4.
Immunoblot analysis of transgenic suspension cells expressing FLAG-fused orf20fukkoku.
(A) Total cellular proteins were separated by SDS-PAGE and probed with αFLAG or αCOXI. Lanes 1 to 6 are transgenic cell lines expressing FLAG-fused orf20fukkoku. Lane M and lane N are the size marker (that does not react with αFLAG or αCOXI) and non-transgenic cells, respectively. Molecular masses are shown on the right. (B) Mitochondria were electrophoresed by BN-PAGE and probed with αpreSATP6. Lanes 1–3, lanes 4–5, and lane 7 are transgenic cell lines expressing FLAG-fused orf20fukkoku, FLAG-fused orf20NK-198, and FLAG-fused orf20L, respectively. Lane N is the non-transgenic cells control. Size markers are shown on the left. An arrow indicates the position of the 200-kDa signal band.
Fig 5.
Protein complexes containing preSATP6 in anthers.
(A) Anthers of a plant heterozygous for ‘Fukkoku-ouba’ rf1 (lane 1), TA-33BB-CMS (lane 2), and NK-198 (lane 3) were subjected to BN-PAGE, and the protein complexes were blotted onto membranes and probed with αpreSATP6. Size markers are shown on the left. Exposure time was 1 min. (B) The same blot as panel A but exposed for 2 hr. (C) The same blot as panel A but probed with αCOXI. (D) Anthers of a plant heterozygous for ‘Fukkoku-ouba’ rf1 (lane 1), TA-33BB-CMS (lane 2), and NK-198 (lane 3) were subjected to SDS-PAGE and αpreSATP6 was used for immunodetection.
Table 8.
Summary of qRT-PCR (mean ± SD) (n = 3).