Author Identifier (ORCID)
Tawfeek Khedr: https://orcid.org/0000-0003-4628-0768
Utpal Bose: https://orcid.org/0000-0003-1156-6504
Mitchell G. Nye-Wood: https://orcid.org/0000-0001-9560-9916
Angéla Juhász: https://orcid.org/0000-0002-4317-2027
Michelle Colgrave: https://orcid.org/0000-0001-8463-805X
Abstract
Quinolizidine alkaloids (QAs) are toxic secondary metabolites in narrow-leafed lupin that defend against herbivores but constrain food and feed use. We developed a single-sample multiomics workflow for simultaneous extraction and quantitative analysis of QAs and proteins from individual leaves using urea/thiourea/CHAPS extraction with acetone precipitation, validated for QA quantification by LC–MS/MS. Data-independent acquisition proteomics of eight lupin accessions (four bitter, four sweet) identified 4437 proteins, with 183 differing significantly between bitter and sweet groups. Bitter accessions were enriched in lysine biosynthesis and stress-response pathways, whereas sweet accessions showed enrichment in phenylpropanoid and other defense-related processes. Coexpression network analysis linked protein modules to total and individual QA levels, capturing known QA-related proteins and revealing candidate enzymes and transporters. This workflow advances molecular understanding of QA regulation and provides candidate protein markers and targets to support breeding of low-alkaloid varieties for food and feed applications.
Keywords
food safety, LC-MS/MS, Lupinus angustifolius, Lupinusspecies, quinolizidine alkaloids, secondary metabolites
Document Type
Journal Article
Date of Publication
5-6-2026
Volume
74
Issue
17
PubMed ID
42024454
Publication Title
Journal of Agricultural and Food Chemistry
Publisher
American Chemical Society
School
Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science / School of Science
RAS ID
99201
Funding Information
This work was supported by Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science (CE200100012).
Grant Number
ARC Number : CE200100012
Grant Link
https://dataportal.arc.gov.au/NCGP/Web/Grant/Grants#/20/1//CE200100012/
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
First Page
14092
Last Page
14101
Comments
Khedr, T., Gao, L., Kamphuis, L. G., Bose, U., Nye-Wood, M. G., Juhász, A., & Colgrave, M. L. (2026). Proteomics and alkaloid signature of narrow leaf lupin leaves reveal key enzymes in quinolizidine alkaloid biosynthesis and transportation. Journal of Agricultural and Food Chemistry, 74(17), 14092–14101. https://doi.org/10.1021/acs.jafc.6c01210