Completion of the mitochondrial genome sequence of onion (Allium cepa L.) containing the CMS-S male-sterile cytoplasm and identification of an independent event of the ccmF N gene split

Curr Genet. 2016 Nov;62(4):873-885. doi: 10.1007/s00294-016-0595-1. Epub 2016 Mar 26.

Abstract

Cytoplasmic male-sterility (CMS) conferred by the CMS-S cytoplasm has been most commonly used for onion (Allium cepa L.) F1 hybrid seed production. We first report the complete mitochondrial genome sequence containing CMS-S cytoplasm in this study. Initially, seven contigs were de novo assembled from 150-bp paired-end raw reads produced from the total genomic DNA using the Illumina NextSeq500 platform. These contigs were connected into a single circular genome consisting of 316,363 bp (GenBank accession: KU318712) by PCR amplification. Although all 24 core protein-coding genes were present, no ribosomal protein-coding genes, except rps12, were identified in the onion mitochondrial genome. Unusual trans-splicing of the cox2 gene was verified, and the cox1 gene was identified as part of the chimeric orf725 gene, which is a candidate gene responsible for inducing CMS. In addition to orf725, two small chimeric genes were identified, but no transcripts were detected for these two open reading frames. Thirteen chloroplast-derived sequences, with sizes of 126-13,986 bp, were identified in the intergenic regions. Almost 10 % of the onion mitochondrial genome was composed of repeat sequences. The vast majority of repeats were short repeats of <100 base pairs. Interestingly, the gene encoding ccmFN was split into two genes. The ccmF N gene split is first identified outside the Brassicaceae family. The breakpoint in the onion ccmF N gene was different from that of other Brassicaceae species. This split of the ccmF N gene was also present in 30 other Allium species. The complete onion mitochondrial genome sequence reported in this study would be fundamental information for elucidation of onion CMS evolution.

Keywords: Complete mitochondrial genome sequence; Cytoplasmic male-sterility; Next-generation sequencing; Onion (Allium cepa L.); ccmF N gene split.

MeSH terms

  • Base Sequence
  • Cluster Analysis
  • Computational Biology / methods
  • Evolution, Molecular
  • Genes, Plant*
  • Genome, Mitochondrial*
  • Genomics*
  • High-Throughput Nucleotide Sequencing
  • Molecular Sequence Annotation
  • Onions / genetics*
  • Open Reading Frames
  • Phylogeny
  • Transcription, Genetic