Hybridisation in bluebells (Hyacinthoides spec.) Using next-generation sequencing to reconstruct a natural hybrid zone in Spain
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Hybridisation is a common evolutionary process that can arise in primary or secondary contact. Gene ow and/or reproductive isolation between hybridising taxa can be explored in hybrid zones. Therefore, a (homoploid) hybrid zone in north-west Spain between Hyacinthoides non-scripta and H. hispanica was studied. The centre occurs west to east across the Galicio-Duero Mountains with H. non-scripta distributed north, and H. hispanica south of the centre. The hybrids' genome sizes and phenotypes represented a range of intermediate states between their parents. Crossing and seed germination experiments revealed a low inter-speci c barrier, and the hybrids showed similarly good tness. Genome wide markers for large genome species were designed from transcriptomes. Diagnostic SNPs between H. non-scripta and H. hispanica were targeted and re-sequenced with multiplexing PCR. Coalescence analyses suggested a Pleistocene origin of parapatric speciation between H. non-scripta and H. hispanica. These results are supported by shared inter-speci c polymorphisms, the lack of recent hybrid generations and of parental individuals in sympatry. Di erential introgression patterns between the organellar and nuclear genomes revealed that formerly H. hispanica ranged further north but was swamped by H. non-scripta alleles. Asymmetric hybridisation was reasoned by absence of backcrosses between northern hybrids to H. non-scripta, but presence between southern hybrids and H. hispanica. Combining these results, a southwards movement of the hybrid zone centre caused by climate change (and adaptive introgression), or inter-speci c di erences in owering time was suggested. Cline patterns revealed cyto-nuclear incompatibilities, which could evolve through divergent adaptation of the organelle to climate and a delayed selection on nuclear inter-acting loci. Both species are in secondary contact in the UK due to recent introduction(s) of H. hispanica and garden variants, which is considered to cause genetic pollution of native H. non-scripta. Therefore, a conservation study is in progress, in which this diagnostic marker system for bluebells is applied.
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