Integrated chloroplast genomics and whole-genome resequencing reveals demographic history and selection signatures of black walnuts.

Elucidating the impacts of demographic history and genomic selection on species evolution is a central topic in phylogeography and evolutionary biology. Black walnuts (Juglans section Rhysocaryon) are native nut trees of the NEW WORLD, with a broad distribution ranging from southern Canada to northern Argentina. The demographic history and genomic dynamics of Rhysocaryon species remain poorly understood. Here, we employed population genomics and chloroplast data to construct a high-density map of genomic variation across 108 Rhysocaryon accessions. Despite gene introgression, these accessions were clearly delimited into four groups. Evolutionary scenarios analysis showed that the diversification of black walnuts might have occurred approximately 28.74 million years ago during the late Oligocene, with the clade comprising Juglans hindsii and Juglans californica diverging earliest. The gene introgression and hybridization analysis indicated that Juglans microcarpa might be a hybrid descendant of Juglans nigra and J. hindsii. As the climate oscillated, these ancestral populations kept diverging, laying the basis for their colonization of South America. Quaternary climatic oscillations also exerted a profound influence on black walnut population size, which exhibited sensitive fluctuations in response to alternation of glacial and interglacial periods. The selection sweeps analysis unveiled highly divergent genomic regions in the economic species J. nigra, which were associated with development, reproduction, disease resistance, and stress tolerance. The genes WRKY41 and ERF012 were identified as potential drivers of J. nigra's adaptation. Our findings illuminated the demographic history and selective signatures of black walnuts, thereby providing a genetic foundation for future breeding, conservation, and genomic studies.