Abstract

BackgroundSpecies within the angiosperm genus Silene contain the largest mitochondrial genomes ever identified. The enormity of these genomes (up to 11 Mb in size) appears to be the result of increased non-coding DNA, which represents >99 % of the genome content. These genomes are also fragmented into dozens of circular-mapping chromosomes, some of which contain no identifiable genes, raising questions about if and how these ‘empty’ chromosomes are maintained by selection. To assess the possibility that they contain novel and unannotated functional elements, we have performed RNA-seq to analyze the mitochondrial transcriptome of Silene noctiflora.ResultsWe identified regions of high transcript abundance in almost every chromosome in the mitochondrial genome including those that lack any annotated genes. In some cases, these transcribed regions exhibited higher expression levels than some core mitochondrial protein-coding genes. We also identified RNA editing sites throughout the genome, including 97 sites that were outside of protein-coding gene sequences and found in pseudogenes, introns, UTRs, and transcribed intergenic regions. Unlike in protein-coding sequences, however, most of these RNA editing sites were only edited at intermediate frequencies. Finally, analysis of mitochondrial small RNAs indicated that most were likely degradation products from longer transcripts, but we did identify candidates for functional small RNAs that mapped to intergenic regions and were not associated with longer RNA transcripts.ConclusionsOur findings demonstrate transcriptional activity in many localized regions within the extensive intergenic sequence content in the S. noctiflora mitochondrial genome, supporting the possibility that the genome contains previously unidentified functional elements. However, transcription by itself is not proof of functional importance, and we discuss evidence that some of the observed transcription and post-transcriptional modifications are non-adaptive. Therefore, further investigations are required to determine whether any of the identified transcribed regions have played a functional role in the proliferation and maintenance of the enormous non-coding regions in Silene mitochondrial genomes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-2155-3) contains supplementary material, which is available to authorized users.

Highlights

  • Species within the angiosperm genus Silene contain the largest mitochondrial genomes ever identified

  • Some species in this genus have massive mitochondrial genomes (7– 11 Mb), in which more than 99 % of the genome content is identified as intergenic sequence (IGS) [9, 22]

  • After trimming and quality/length filtering, an average of 34.8 % mRNA-seq reads from the two replicated mitochondrial-enriched libraries could be mapped to the reference mitochondrial genome

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Summary

Introduction

Species within the angiosperm genus Silene contain the largest mitochondrial genomes ever identified. The enormity of these genomes (up to 11 Mb in size) appears to be the result of increased non-coding DNA, which represents >99 % of the genome content These genomes are fragmented into dozens of circular-mapping chromosomes, some of which contain no identifiable genes, raising questions about if and how these ‘empty’ chromosomes are maintained by selection. The longstanding debate about the meaning and existence of “junk DNA” has come under renewed scrutiny in light of recent and controversial claims that 80 % of the human genome is functional [1,2,3,4,5] While most of this debate has focused on the nucleus, the evolution of non-coding sequence content in mitochondrial genomes has been of great interest, in plants in which mitochondrial genomes are large and variable in size relative to other eukaryotes and have low gene densities [6,7,8,9,10,11,12]. We found that many of the seemingly empty chromosomes are shared across populations, raising the possibility that they are conserved by natural selection and contain some type of unidentified functional elements

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