Pangenome graphs are revolutionising evolutionary and population genomics by moving beyond linear reference genomes to represent the full spectrum of sequence diversity within and across species. This review traces the field’s progression from reference-augmented graphs to assembly-based, alignment-first approaches that capture complex structural variation with reduced bias. We examine key strategies for graph construction, genotyping and implementing graph-aware tools in functional genomics, including transcriptomics and epigenomics. While much of the work to date has focused on humans, diverse and structurally complex plant genomes pose unique challenges that require further methodological innovation. Key bottlenecks – including visualisation, scalability and integration with multi-omic data – persist. By outlining trade-offs among current tools and emphasising the need for rigorous evaluation frameworks, we argue that progress will depend on community-driven efforts to unify graph construction, genotyping and interpretation. Despite technical hurdles, pangenome graphs offer a powerful foundation for more inclusive evolutionary and population genomics.

Third-generation long-read sequencing is transforming plant genomics. Oxford Nanopore Technologies and Pacific Biosciences are offering competing long-read sequencing technologies and enable plant scientists to investigate even large and complex plant genomes. Sequencing projects can be conducted by single research groups and sequences of smaller plant genomes can be completed within days. This also resulted in an increased investigation of genomes from multiple species in large scale to address fundamental questions associated with the origin and evolution of land plants. Increased accessibility of sequencing devices and user-friendly software allows more researchers to get involved in genomics. Current challenges are accurately resolving diploid or polyploid genome sequences and better accounting for the intra-specific diversity by switching from the use of single reference genome sequences to a pangenome graph.