Jaegle, Benjamin, Uroic, Miran Kalle, Holtkotte, Xu, Lucas, Christina, Termath, Andreas Ole, Schmalz, Hans-Guenther ORCID: 0000-0003-0489-1827, Bucher, Marcel ORCID: 0000-0003-1680-9413, Hoecker, Ute, Huelskamp, Martin and Schrader, Andrea ORCID: 0000-0002-3879-7057 (2016). A fast and simple LC-MS-based characterization of the flavonoid biosynthesis pathway for few seed(ling)s. BMC Plant Biol., 16. LONDON: BMC. ISSN 1471-2229

Full text not available from this repository.

Abstract

Background: (Pro)anthocyanidins are synthesized by the flavonoid biosynthesis pathway with multi-layered regulatory control. Methods for the analysis of the flavonoid composition in plants are well established for different purposes. However, they typically compromise either on speed or on depth of analysis. Results: In this work we combined and optimized different protocols to enable the analysis of the flavonoid biosynthesis pathway with as little as possible biological material. We chose core substances of this metabolic pathway that serve as a fingerprint to recognize alterations in the main branches of the pathway. We used a simplified sample preparation, two deuterated internal standards, a short and efficient LC separation, highly sensitive detection with tandem MS in multiple reaction monitoring (MRM) mode and hydrolytic release of the core substances to reduce complexity. The method was optimized for Arabidopsis thaliana seeds and seedlings. We demonstrate that one Col-0 seed/seedling is sufficient to obtain a fingerprint of the core substances of the flavonoid biosynthesis pathway. For comparative analysis of different genotypes, we suggest the use of 10 seed(lings). The analysis of Arabidopsis thaliana mutants affecting steps in the pathway revealed foreseen and unexpected alterations of the pathway. For example, HY5 was found to differentially regulate kaempferol in seeds vs. seedlings. Furthermore, our results suggest that COP1 is a master regulator of flavonoid biosynthesis in seedlings but not of flavonoid deposition in seeds. Conclusions: When sample numbers are high and the plant material is limited, this method effectively facilitates metabolic fingerprinting with one seed(ling), revealing shifts and differences in the pathway. Moreover the combination of extracted non-hydrolysed, extracted hydrolysed and non-extracted hydrolysed samples proved useful to deduce the class of derivative from which the individual flavonoids have been released.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Jaegle, BenjaminUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Uroic, Miran KalleUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Holtkotte, XuUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Lucas, ChristinaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Termath, Andreas OleUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schmalz, Hans-GuentherUNSPECIFIEDorcid.org/0000-0003-0489-1827UNSPECIFIED
Bucher, MarcelUNSPECIFIEDorcid.org/0000-0003-1680-9413UNSPECIFIED
Hoecker, UteUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Huelskamp, MartinUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schrader, AndreaUNSPECIFIEDorcid.org/0000-0002-3879-7057UNSPECIFIED
URN: urn:nbn:de:hbz:38-265387
DOI: 10.1186/s12870-016-0880-7
Journal or Publication Title: BMC Plant Biol.
Volume: 16
Date: 2016
Publisher: BMC
Place of Publication: LONDON
ISSN: 1471-2229
Language: English
Faculty: Faculty of Mathematics and Natural Sciences
Divisions: Faculty of Mathematics and Natural Sciences > Department of Biology > Botanical Institute
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
MYB TRANSCRIPTION FACTORS; FUNCTIONAL GENOMICS; GENE ENCODES; ANTHOCYANIN BIOSYNTHESIS; DOMAIN PROTEIN; ARABIDOPSIS; LIGHT; EXPRESSION; COP1; IDENTIFICATIONMultiple languages
Plant SciencesMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/26538

Downloads

Downloads per month over past year

Altmetric

Export

Actions (login required)

View Item View Item