Transforming ash genomics: Creating a pangenome to understand ash dieback resistance

Project leads  Dr Laura Kelly, Dr Daniel Wood, Dr Mohammad Vatanparast | Royal Botanic Gardens, Kew

Lead organisation  Royal Botanic Gardens, Kew

Collaborators  Dr Richard Whittet | Forest Research; Prof Levi Yant, Dr Emma Curran | University of Nottingham

Project status  Complete (April 2022 – March 2025)

Project funding  £448,000

Research outcome  Adaptation

Ash
Context

The European ash tree (Fraxinus excelsior) is one of the most common woodland trees in the UK, but over the last decade has suffered severe damage from an invasive fungus called Hymenoscyphus fraxineus which causes ash dieback disease.

Previous research has shown that a small percentage of ash trees have resistance to the disease resistance which can be passed to offspring and we have some understanding of the genomic basis for this. However, a more complete understanding of the basis of resistance is required to be able to accurately predict the health of individual trees.

Most studies of genomic variation look at single points on the genome that differ between individuals. This project will explore genomic structural variation, which may be important for resistance to ash dieback, but which is difficult to assess.

An improved understanding of the genomic basis of ash dieback resistance will allow selection of more resilient trees for breeding programmes and woodland restoration.

Research aims and objectives
Aim

To transform our understanding of the genomic basis of ash dieback resistance by using the latest developments in genome sequencing and analysis to discover structural variants and construct a pangenome – a reference genome incorporating information from diverse individuals rather than based on a single individual, as is the case with traditional approaches.

Objectives
  • Construct a high-quality ash pangenome capturing common structural variants.
  • Use this new resource to identify structural variants associated with ash dieback resistance.
  • Improve prediction of ash dieback resistance of individual trees by incorporating structural variant data.

We will use long-read sequencing data to construct a pangenome and identify structural variants across the genome. The pangenome will be used to find structural variants in individuals from both planted trials and a natural woodland, and to identify variants associated with ash dieback resistance.

Tissue collection and genome sequencing: Samples will be collected from resistant and susceptible individuals from screening trials and mature trees in natural woodland.

Pan-genome construction: Long-read sequence data will be mapped to an improved reference genome to identify variants and construct an annotated pangenome graph that shows the locations of genes and structural variants.

Identification of structural variants associated with ash dieback resistance: We will map sequenced DNA from the samples to the new pangenome to find individual differences. Structural variants significantly associated with ash dieback resistance will be identified and we will establish whether accounting for structural variants results in more accurate genomic prediction of individual ash dieback resistance.

Expected outcomes
  • A high-quality pangenome for European ash.
  • A collection of structural variants associated with ash dieback resistance.
  • A genomic prediction model for ash dieback resistance incorporating structural variants.
  • Publications describing these results.

Evidence of ash dieback infection.

Glossary
Genome sequencing

The process of determining the complete DNA sequence of a tree’s genome. This includes identifying the order of all the nucleotides (A, T, C, and G) that make up its genetic material.

Genomic basis

The underlying genetic features, such as DNA sequences, genes, and structural variants, that contribute to resistance or susceptibility to disease.

Genomic structural variation

Where entire regions of DNA are inverted, duplicated, deleted or moved elsewhere in the genome in some individuals.

Genomic variation

The differences in DNA sequences among individual trees, which can influence their susceptibility or resistance to disease.

Long-read sequencing DNA

DNA that is sequenced in sections that average thousands of base pairs in length, rather than the more commonly used short-read.

Pangenome

The complete set of genes found across all individuals of a species.

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