Abstract
Flavonol rhamnosides including kaempferitrin (i.e., kaempferol 3-O-α-rhamnoside-7-O-α-rhamnoside) occur throughout the plant kingdom. Mechanisms governing flavonol rhamnoside biosynthesis are established, whereas degradative processes occurring in plants are relatively unknown. Here, we investigated the catabolic events affecting kaempferitrin status in the rosette leaves of Arabidopsis thaliana L. Heynh. (Arabidopsis) and Raphanus sativus L. (radish), respectively, in response to developmental senescence and postharvest handling. On a per plant basis, losses of several kaempferol rhamnosides including kaempferitrin were apparent in senescing leaves of Arabidopsis during development and postharvest radish stored at 5 °C. Conversely, small pools of kaempferol 7-O-α-rhamnoside (K7R), kaempferol 3-O-α-rhamnoside (K3R), and kaempferol built up in senescing leaves of both species. Evidence is provided for ⍺-rhamnosidase activities targeting the 7-O-α-rhamnoside of kaempferitrin and K7R in rosette leaves of both species. An HPLC analysis of in vitro assays of clarified leaf extracts prepared from developing Arabidopsis and postharvest radish determined that these metabolic shifts were coincident with respective 237% and 645% increases in kaempferitrin 7-O-⍺-rhamnosidase activity. Lower activity rates were apparent when these ⍺-rhamnosidase assays were performed with K7R. A radish ⍺-rhamnosidase containing peak eluting from a DEAE-Sepharose Fast Flow column hydrolyzed various 7-O-rhamnosylated flavonols, as well as kaempferol 3-O-β-glucoside. Together it is apparent that the catabolism of 7-O-α-rhamnosylated kaempferol metabolites in senescing plant leaves is associated with a flavonol 7-O-α-rhamnoside-utilizing α-rhamnosidase.
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All data generated or analysed during this study are included in this published article and its supplementary information files.
Change history
25 July 2022
A Correction to this paper has been published: https://doi.org/10.1007/s00425-022-03964-6
Abbreviations
- BGLU:
-
β-Glucosidase
- K3R:
-
Kaempferol 3-O-α-rhamnoside
- K7R:
-
Kaempferol 7-O-α-rhamnoside
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Acknowledgements
The authors are grateful to Dr. Mary Ruth McDonald and Kevin D Vander Kooi at the University of Guelph Muck Crops Research Station (Holland Marsh, ON) for cultivating radish plants. We thank Michael Mucci and Leane Illman at the University of Guelph’s Phytotron facility for technical assistance with the cultivation of Arabidopsis in environment-controlled growth chambers, to Honghui Zhu at the Guelph Research and Development Centre (Agriculture and Agri-Food Canada) for technical assistance with UPLC-MS/MS analysis of radish leaf metabolites, and to Michelle Edwards at the Ontario Agricultural College (University of Guelph) for guidance on statistical analysis.
Funding
The authors would like to gratefully acknowledge funding from the Natural Sciences and Engineering Research Council (NSERC) of Canada for a Discovery grant (GGB), and for an NSERC Alexander Graham Bell Canada Graduate Scholarship (NU) and NSERC Undergraduate Summer Research Assistantship (LCM).
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Unterlander, N., Mats, L., McGary, L.C. et al. Kaempferol rhamnoside catabolism in rosette leaves of senescing Arabidopsis and postharvest stored radish. Planta 256, 36 (2022). https://doi.org/10.1007/s00425-022-03949-5
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DOI: https://doi.org/10.1007/s00425-022-03949-5