Identification and functional characterization of BAHD acyltransferases associated with anthocyanin acylation in blueberry

Abstract

Blueberry is promoted as a super food with several health properties derived from chlorogenic acid and anthocyanin. Previous studies indicated that anthocyanin acylation and the content of chlorogenic acid could affect their level of absorption and biological activity. In this study, a genome-wide association study was performed to identify loci associated with anthocyanin and chlorogenic acid and characterize the candidate genes controlling anthocyanin acylation. Two stable loci controlling anthocyanin acylation and glucose specific glycosylation were confirmed on chromosomes 2 and 4, respectively, while no stable loci associated with chlorogenic acid were identified. Two acyl-CoA acyltransferases named VcBAHD-AT1 and VcBAHD-AT4 were identified as best candidate genes controlling anthocyanin acylation. Interestingly, the two genes clustered in acyl-CoA acyltransferases clade III, a clade that is not commonly associated with anthocyanin acylation. A virus-induced gene silencing approach optimized for silencing VcBAHD-AT1 and VcBAHD-AT4 in the whole blueberry fruits, confirmed the role of these two genes in anthocyanin acylation. Overall, this study establishes the foundation to develop a molecular marker to select for higher acylated anthocyanin and delivered a method for rapid functional characterization of genes associated with other fruit related traits in blueberry. Also, the study adds evidence that during the evolution of acyl-CoA acyltransferases multiple routes led to the emergence and/or fixation of the anthocyanin acyltransferase activity. These outcomes advance knowledge about the genes controlling anthocyanin acylation in blueberries and that extend to other plants. Selecting new blueberry cultivars with higher acylated anthocyanin levels could potentially increase absorption of this health-related bioactive.

Acknowledgements

This research was supported by the Foundation for Food and Agriculture Research (FFAR) under award number 534667. MI, MAL, MFM, and MAA were also supported by the United States Department of Agriculture National Institute of Food and Agriculture, Hatch project 1008691, and the National Institute of Food and Agriculture, United States Department of Agriculture, under award number 2019-51181-30015, project ‘VacciniumCAP: Leveraging genetic and genomic resources to enable development of blueberry and cranberry cultivars with improved fruit quality attributes’. Partial funding for this project were also provided by the "NCSU Plant Breeding Consortium Small Grants Program", enabled by resources from licensing of improved varieties.

Author contributions

MI, MFM, and MAA conceptualized the project and wrote the original draft. MFM, MAA, MHG, and RS performed experiments and formal analysis. NB provided material. MAL, MF, CDK, and MI acquired funding and supervised the study. APD, DC, and RVE provided inputs on BAHD acyltransferase analysis. All authors edited the article and approved the submitted version.

Data availability

Phenotypic data used for QTL mapping were made available in Table S6. Genotypic data used for QTL analysis can be found on the Genome Database for Vaccinium at https://www.vaccinium.org/publication_datasets Accession # GDV24002.

Conflict of interest statement

The authors declare that they have no conflict of interest.

Supplementary Data

Supplementary data is available at Horticulture Research online.

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