Profiling Flavonoid Metabolites in Brassica Seed Coats

Seed coats were harvested at 20, 30, and 40 DAF from individual plants of ZY821, RNAi-BnA09myb47a, GH06, and BnA09MYB47a-overexpressing lines. The raw flavonoids were extracted from the flash-frozen fresh seed coats (100 mg fresh weight), and subjected to UPLC − HESI − MS/MS analysis^{25 (link),37 (link)}. Flavonoid metabolites were identified by retention times, mass spectrometry, and public information^{25 (link),35 (link)–37 (link)}. The contents of flavonoid metabolites were calculated using the standard curves of available standards: epicatechin, quercetin, kaempferol, and isorhamnetin (Sigma, Aldrich, Shanghai, China). Metabolomics analysis was also performed using Compounds Discoverer 3.0 software (Thermo Fisher Scientific, CA, USA). The Raw MS data have been deposited in MetaboLights under accession number MTBLS6703 (https://www.ebi.ac.uk/metabolights/search). Chemical structure depiction was retrieved from the PubChem database (https://pubchem.ncbi.nlm.nih.gov). All analyses were conducted in triplicate, and the values were represented as means ± SD of three replicates. Statistical significance was based on Tukey’s test, and P ≤ 0.05 was considered to be statistically significant.

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Qu C., Zhu M., Hu R., Niu Y., Chen S., Zhao H., Li C., Wang Z., Yin N., Sun F., Chen Z., Shen S., Shang G., Zhou Y., Yan X., Wei L., Liu L., Yi B., Lian J., Li J., Tang Z., Liang Y., Xu X., Wang R., Yin J., Wan H., Du H., Qian W., Chai Y., Zhou Q., He Y., Zhong S., Qiu X., Yu H., Lam H.M., Lu K., Fu F, & Li J. (2023). Comparative genomic analyses reveal the genetic basis of the yellow-seed trait in Brassica napus. Nature Communications, 14, 5194.

Publication 2023

epicatechin quercetin kaempferol isorhamnetin Compounds Discoverer 3.0 software

Epicatechin Flavonoid Isorhamnetin Kaempferol Mass spectrometry Ms ms Plants Quercetin Retention Rnai

Corresponding Organization : Agriculture and Agri-Food Canada

Other organizations : Southwest University, Chinese University of Hong Kong, National University of Singapore, Qinghai University, Huazhong Agricultural University, University of Saskatchewan

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Variable analysis

independent variables

Genotypes: ZY821, RNAi-BnA09myb47a, GH06, and BnA09MYB47a-overexpressing lines
Developmental stages: 20, 30, and 40 DAF (days after flowering)

dependent variables

Flavonoid metabolite contents: epicatechin, quercetin, kaempferol, and isorhamnetin

control variables

Fresh weight of seed coats: 100 mg
Extraction method: flash-frozen fresh seed coats
Analytical method: UPLC-HESI-MS/MS
Identification of flavonoid metabolites: retention times, mass spectrometry, and public information
Quantification of flavonoid metabolites: standard curves of available standards

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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