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A systems genetics approach to identify candidate genes driving salicinoid diversity in Populus tremula
Umeå University, Faculty of Science and Technology, Department of Plant Physiology. (Nathaniel Street)
2025 (English)Doctoral thesis, comprehensive summary (Other academic)Alternative title
Systemgenetik som verktyg för att identifiera generna som styr salicinoid-diversitet i Populus tremula (Swedish)
Abstract [en]

Trees have evolved an impressive array of strategies to cope with the challenges of having a long and sessile life. Not only must they withstand a fluctuating climate, but they also face instantaneous pressures from herbivores and other attackers. To protect themselves, plants can produce defence compounds, many of which are highly specialised and taxon specific. Within the Salicaceae family, a key group of such defence compounds are the salicinoid phenolic glycosides (SPGs). Many structural variants of SPGs have been identified, in which acyl groups (e.g., cinnamoyl, benzoyl, and acetyl) are common. Some of these SPGs can have toxic and deterrent effects against attackers, and a few are known for their medicinal properties in humans. However, the biological function of most SPGs in planta remains unclear, and the causal enzymes for the majority of SPGs are yet to be identified. 

The aim of this thesis was to uncover the genetic basis of SPG biosynthesis in European aspen (Populus tremula L.) and to determine the extent of ontogenic and organ-specific variation among individuals. To achieve this, SPG variation within a collection of natural aspen, the Swedish aspen (SwAsp) collection, was investigated using an integrative multi-omic approach. By analysing the metabolome and transcriptome of multiple leaf ages from aspen individuals with varying levels of cinnamoyl and acetylated SPGs, a set of candidate transferases and novel putative SPGs were identified. These analyses further suggested that young leaf tissue is a highly active site of SPG biosynthesis, compared with mature leaves.  

To extend this analysis, we performed genome-wide association studies on transcriptomic and metabolomic data from leaf buds to identify genomic regions associated with variation in SPG abundance and gene expression. These data were integrated into a systems genetics network, visualising the intricate relationship between candidate genes and the diversity of SPGs. Among the candidates, an acyltransferase was highly associated with both acetyl- and cinnamoyl-SPGs. Heterologous expression assays in Escherichia coli (E. coli) confirmed its acetylation activity. In line with these findings, overexpression of the gene in planta led to increased levels of acetyl-SPGs, suggesting acetylation activity of the enzyme.

In summary, these results have enhanced our understanding of SPG biosynthesis and provide a foundation for future studies aimed at elucidating the in planta function of the remaining candidate genes. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2025. , p. 107
Keywords [en]
aspen, Populus tremula, systems genetics, GWAS, eQTL, metabolomics, specialised metabolites, salicinoid phenolic glycosides, chemotype, liquid chromatography-mass spectrometry, transcriptomics, RNA-Seq
National Category
Bioinformatics and Computational Biology Genetics and Genomics Plant Biotechnology
Identifiers
URN: urn:nbn:se:umu:diva-247549ISBN: 978-91-8070-866-1 (print)ISBN: 978-91-8070-867-8 (electronic)OAI: oai:DiVA.org:umu-247549DiVA, id: diva2:2021228
Public defence
2026-01-09, KBE301-Lilla hörsalen, KBC-huset, Umeå, 13:00 (English)
Opponent
Supervisors
Available from: 2025-12-19 Created: 2025-12-12 Last updated: 2025-12-12Bibliographically approved
List of papers
1. An improved chromosome-scale genome assembly and population genetics resource for populus tremula
Open this publication in new window or tab >>An improved chromosome-scale genome assembly and population genetics resource for populus tremula
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2024 (English)In: Physiologia Plantarum, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 176, no 5, article id e14511Article in journal (Refereed) Published
Abstract [en]

Aspen (Populus tremula L.) is a keystone species and a model system for forest tree genomics. We present an updated resource comprising a chromosome-scale assem- bly, population genetics and genomics data. Using the resource, we explore the genetic basis of natural variation in leaf size and shape, traits with complex genetic architecture.

We generated the genome assembly using long-read sequencing, optical and high-density genetic maps. We conducted whole-genome resequencing of the Umeå Aspen (UmAsp) collection. Using the assembly and re-sequencing data from the UmAsp, Swedish Aspen (SwAsp) and Scottish Aspen (ScotAsp) collections we performed genome-wide association analyses (GWAS) using Single Nucleotide Polymorphisms (SNPs) for 26 leaf physiognomy phenotypes. We conducted Assay of Transposase Accessible Chromatin sequencing (ATAC-Seq), identified genomic regions of accessible chromatin, and subset SNPs to these regions, improving the GWAS detection rate. We identified candidate long non-coding RNAs in leaf samples, quantified their expression in an updated co-expression network, and used this to explore the functions of candidate genes identified from the GWAS.

A GWAS found SNP associations for seven traits. The associated SNPs were in or near genes annotated with developmental functions, which represent candidates for further study. Of particular interest was a !177-kbp region harbouring associations with several leaf phenotypes in ScotAsp.

We have incorporated the assembly, population genetics, genomics, and GWAS data into the PlantGenIE.org web resource, including updating existing genomics data to the new genome version, to enable easy exploration and visualisation. We provide all raw and processed data to facilitate reuse in future studies.

Place, publisher, year, edition, pages
John Wiley & Sons, 2024
Keywords
genome assembly, natural selection, co-expression, population genetics, Populus, aspen, GWAS, leaf physiognomy, leaf shape, leaf size, genetic architecture, ATAC-Seq, lncRNA
National Category
Bioinformatics and Computational Biology Genetics and Genomics
Identifiers
urn:nbn:se:umu:diva-229976 (URN)10.1111/ppl.14511 (DOI)001313686100001 ()39279509 (PubMedID)2-s2.0-85204093798 (Scopus ID)
Funder
Swedish Research Council, 2019-05476Swedish Research Council Formas, 2018-01644Vinnova, S111416L0710
Note

Supplementary figures and appendixes under Supporting information on article web page. 

Available from: 2024-09-23 Created: 2024-09-23 Last updated: 2025-12-12Bibliographically approved
2. A metabolomics and transcriptomics resource for identifying candidate genes in the biosynthesis of specialised metabolites in Populus tremula
Open this publication in new window or tab >>A metabolomics and transcriptomics resource for identifying candidate genes in the biosynthesis of specialised metabolites in Populus tremula
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2025 (English)In: Physiologia Plantarum, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 177, no 5, article id e70567Article in journal (Refereed) Published
Abstract [en]

This study aims to identify candidate genes involved in the biosynthesis of salicinoid phenolic glycosides (SPGs), a group of specialised metabolites characteristic of the Salicaceae family. While the integration of multi-omics data represents a powerful approach to link genes encoding enzymes and their regulatory factors to metabolite biosynthesis, suitable multi-omics data resources are scarce. We present a comprehensive dataset comprising untargeted liquid chromatography–mass spectrometry (LC–MS) and mRNA-sequencing data from various organs of European aspen (Populus tremula L.) and from genotypes that produce contrasting sets of SPGs. We present a reproducible pipeline for the analysis of the LC–MS data, including predicted annotation of potential novel SPGs. We demonstrate the utility of the resource by identifying candidate genes involved in the biosynthesis of SPGs with a cinnamoyl moiety. By integrating gene and metabolite differential analyses with a gene co-expression network, we identified two HXXXD-type acyltransferase genes and one UDP-glucosyltransferase gene as candidates for future downstream characterisation. The combined gene expression and metabolomics resource is integrated into PlantGenIE.org to facilitate easy access and data mining. All raw data are available in public databases, and all data and results files are available at an associated Figshare repository.

Place, publisher, year, edition, pages
John Wiley & Sons, 2025
Keywords
aspen, biosynthesis, chemotype, liquid chromatography–mass spectrometry (LC–MS), metabolomics, phenolic glycosides, Populus tremula, RNA-Seq, salicinoid, specialised metabolite
National Category
Bioinformatics and Computational Biology
Identifiers
urn:nbn:se:umu:diva-245717 (URN)10.1111/ppl.70567 (DOI)001591085400001 ()41063380 (PubMedID)2-s2.0-105018295556 (Scopus ID)
Available from: 2025-10-23 Created: 2025-10-23 Last updated: 2025-12-12Bibliographically approved
3. A systems genetics study reveals novel candidate genes for salicinoid biosynthesis in aspen
Open this publication in new window or tab >>A systems genetics study reveals novel candidate genes for salicinoid biosynthesis in aspen
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(English)Manuscript (preprint) (Other academic)
National Category
Bioinformatics and Computational Biology
Identifiers
urn:nbn:se:umu:diva-247544 (URN)
Available from: 2025-12-12 Created: 2025-12-12 Last updated: 2025-12-12

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