AUTHOR=Salami Maryam , Heidari Bahram , Batley Jacqueline , Wang Jin , Tan Xiao-Li , Richards Christopher , Tan Helin 

TITLE=Integration of genome-wide association studies, metabolomics, and transcriptomics reveals phenolic acid- and flavonoid-associated genes and their regulatory elements under drought stress in rapeseed flowers

JOURNAL=Frontiers in Plant Science

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1249142

DOI=10.3389/fpls.2023.1249142

ISSN=1664-462X

ABSTRACT=<sec><title>Introduction</title><p>Biochemical and metabolic processes help plants tolerate the adverse effects of drought. In plants accumulating bioactive compounds, understanding the genetic control of the biosynthesis of biochemical pathways helps the discovery of candidate gene (CG)–metabolite relationships.</p></sec><sec><title>Methods</title><p>The metabolic profile of flowers in 119 rapeseed (<italic>Brassica napus</italic>) accessions was assessed over two irrigation treatments, one a well-watered (WW) condition and the other a drought stress (DS) regime. We integrated information gained from 52,157 single-nucleotide polymorphism (SNP) markers, metabolites, and transcriptomes to identify linked SNPs and CGs responsible for the genetic control of flower phenolic compounds and regulatory elements.</p></sec><sec><title>Results</title><p>In a genome-wide association study (GWAS), of the SNPs tested, 29,310 SNPs were qualified to assess the population structure and linkage disequilibrium (LD), of which several SNPs for radical scavenging activity (RSA) and total flavanol content (TFLC) were common between the two irrigation conditions and pleiotropic SNPs were found for chlorogenic and coumaric acids content. The principal component analysis (PCA) and stepwise regression showed that chlorogenic acid and epicatechin in WW and myricetin in DS conditions were the most important components for RSA. The hierarchical cluster analysis (HCA) showed that vanillic acid, myricetin, gallic acid, and catechin were closely associated in both irrigation conditions. Analysis of GWAS showed that 60 CGs were identified, of which 18 were involved in stress-induced pathways, phenylpropanoid pathway, and flavonoid modifications. Of the CGs, <italic>PAL1</italic>, <italic>CHI</italic>, <italic>UGT89B1</italic>, <italic>FLS3</italic>, <italic>CCR1</italic>, and <italic>CYP75B137</italic> contributed to flavonoid biosynthetic pathways. The results of RNA sequencing (RNA-seq) revealed that the transcript levels of <italic>PAL</italic>, <italic>CHI</italic>, and <italic>CYP75B137</italic> known as early flavonoid biosynthesis-related genes and <italic>FLS3</italic>, <italic>CCR1</italic>, and <italic>UGT89B1</italic> related to the later stages were increased during drought conditions. The transcription factors (TFs) <italic>NAC035</italic> and <italic>ERF119</italic> related to flavonoids and phenolic acids were upregulated under drought conditions.</p></sec><sec><title>Discussion</title><p>These findings expand our knowledge on the response mechanisms to DS, particularly regarding the regulation of key phenolic biosynthetic genes in rapeseed. Our data also provided specific linked SNPs for marker-assisted selection (MAS) programs and CGs as resources toward realizing metabolomics-associated breeding of rapeseed.</p></sec>