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Mutations in the mitochondrial orf108 render Moricandia arvensis restorer ineffective in restoring male fertility to Brassica oxyrrhina-based cytoplasmic male sterile line of B. juncea

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Abstract

We reported earlier an evolutionarily conserved mitochondrial orf108 causing cytoplasmic male sterility (CMS) in four alloplasmic lines of Brassica juncea. B. oxyrrhina also carries orf108, but male sterility of B. oxyrrhina-based B. juncea CMS lines is not restored by the Moricandia arvensis restorer that rescues other orf108-containing B. juncea CMS lines. To understand this discrepancy, we characterized B. oxyrrhina-based B. juncea CMS line (oxy-cms) to identify the mitochondrial gene associated with male sterility. Examination of expression patterns of 26 mitochondrial genes in CMS and male fertile (oxy-camp, i.e. amphidiploid B. oxyrrhina × B. rapa, and euplasmic B. juncea) lines revealed polymorphic transcript patterns for six genes: atp1, atp4, ccmfn2, cox3, nad4L and orf108. Detailed analysis showed that orf108 is linked and co-transcribed with atp1. Comparison of orf108-atp1 transcripts in male sterile and male fertile flowers revealed that B. oxyrrhina restorer cleaves orf108-atp1 at a different location within the orf108 coding region as compared with the M. arvensis restorer. A total of eleven SNPs were detected within the orf108 coding region. One of these SNPs was located at the site where M. arvensis restorer processes the orf108-atp1 transcript suggesting that it could be critical for cleavage of orf108-atp1 transcript by the M. arvensis restorer.

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Author contribution

VN conducted most of the experiments; SKS and VK assisted in transcript mapping; AW assisted in preparation of Southern blots; SRB generated the materials; SRB, KRSS and PK planned the experiment; SRB, VN and PK wrote the manuscript.

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Author notes

  1. Pankaj Kumar

    Present address: Biomedical Sciences Research Complex, North Haugh, University of St. Andrews, St. Andrews, Fife, KY16 9ST, Scotland, UK

Authors and Affiliations

  1. ICAR-National Research Centre on Plant Biotechnology, New Delhi, 110012, India

    Vasupalli Naresh, Sunil Kumar Singh, Anshul Watts, Vajinder Kumar & Shripad Ramachandra Bhat

  2. Acharya Nagarjuna University, Guntur, Andhra Pradesh, 522510, India

    K. R. S. Sambasiva Rao

Authors
  1. Vasupalli Naresh
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  2. Sunil Kumar Singh
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  3. Anshul Watts
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  4. Pankaj Kumar
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  5. Vajinder Kumar
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  6. K. R. S. Sambasiva Rao
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  7. Shripad Ramachandra Bhat
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Correspondence to Shripad Ramachandra Bhat.

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Supplementary material 1 (DOCX 112 kb)

Suppl. Fig. 1

Alignment of protein sequences of atp1 of B. oxyrrhina and M. arvensis. Identical alignments are boxed. Upward arrow indicates the amino acids have same physicochemical properties (JPEG 123 kb)

Suppl. Fig. 2

Alignment of ORF108 of B. oxyrrhina and M. arvensis. Identical alignments are boxed (JPEG 26 kb)

Suppl. Fig. 3

Verification of co-transcription of atp4 and nad4L in B. juncea (1), oxy-cms (2) and oxy-camp (3). Gel photograph of amplicons obtained after RT-PCR and PCR with the primers atp4 F and nad4L R. Amplicon size is indicated in kb (JPEG 9 kb)

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Naresh, V., Singh, S.K., Watts, A. et al. Mutations in the mitochondrial orf108 render Moricandia arvensis restorer ineffective in restoring male fertility to Brassica oxyrrhina-based cytoplasmic male sterile line of B. juncea . Mol Breeding 36, 67 (2016). https://doi.org/10.1007/s11032-016-0489-4

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  • DOI: https://doi.org/10.1007/s11032-016-0489-4

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