Molecular Ecology Resources
Molecular Ecology Resources
4 years ago

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

Kevin Donnelly, Annika Perry, Stuart A'Hara, Richard A. Ennos, Stephen Cavers, Joan Cottrell, Giovanni Giuseppe Vendramin, Sarah King, Witold Wachowiak
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.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1111/1755-0998.12646

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