Reconstructing the plant mitochondrial genome for marker discovery: a case study using Pinus

Donnelly, Kevin; Cottrell, Joan; Ennos, Richard A.; Vendramin, Giovanni Giuseppe; A'Hara, Stuart; King, Sarah; Perry, Annika; Wachowiak, Witold; Cavers, Stephen. 2017 Reconstructing the plant mitochondrial genome for marker discovery: a case study using Pinus. Molecular Ecology Resources, 17 (5). 943-954.

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Whole-genome-shotgun (WGS) sequencing of total genomic DNA was used to recover ~1 Mbp of novel mitochondrial (mtDNA) sequence from Pinus sylvestris (L.) and three members of the closely related Pinus mugo species complex. DNA was extracted from megagametophyte tissue from six mother trees from locations across Europe, and 100-bp paired-end sequencing was performed on the Illumina HiSeq platform. Candidate mtDNA sequences were identified by their size and coverage characteristics, and by comparison with published plant mitochondrial genomes. Novel variants were identified, and primers targeting these loci were trialled on a set of 28 individuals from across Europe. In total, 31 SNP loci were successfully resequenced, characterizing 15 unique haplotypes. This approach offers a cost-effective means of developing marker resources for mitochondrial genomes in other plant species where reference sequences are unavailable.

Item Type: Publication - Article
Digital Object Identifier (DOI):
UKCEH and CEH Sections/Science Areas: Watt
ISSN: 1755-098X
Additional Keywords: high-throughput sequencing, marker discovery, mitochondrion, phylogeography, Pinus
NORA Subject Terms: Biology and Microbiology
Date made live: 19 Apr 2017 14:29 +0 (UTC)

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