Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 61, Issue 31, Pages 7421-7430Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jf401122d
Keywords
Trifolium pratense L.; red clover; leaf nodule; polyphenol oxidase; PPO; gene expression; enzyme localization; o-diphenol substrate; Raman microspectrometry
Funding
- BBSRC, UK
- Biotechnology and Biological Sciences Research Council [BBS/E/W/10963A01D, BBS/E/W/00003134D] Funding Source: researchfish
- BBSRC [BBS/E/W/00003134D, BBS/E/W/10963A01D] Funding Source: UKRI
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Polyphenol oxidase (PPO) genes and their corresponding enzyme activities occur in many plants; natural PPO substrates and enzyme/substrate localization are less well characterized. Leaf and root PPO activities in Arabidopsis and five legumes were compared with those of high-PPO red clover (Trifolium pratense L.). Red clover PPO enzyme activity decreased leaves > stem > nodules > peduncle = petiole > embryo; PPO1 and PPO4 genes were expressed early in leaf emergence, whereas PPO4 and PPO5 predominated in mature leaves. PPO1 was expressed in embryos and nodules. PPO substrates, phaselic acid and clovamide, were detected in leaves, and clovamide was detected in nodules. Phaselic acid and clovamide, along with caffeic and chlorogenic acids, were suitable substrates for PPO1, PPO4, and PPO5 genes expressed in alfalfa (Medicago sativa L.) leaves. PPO enzyme presence and activity were colocalized in leaves and nodules by cytochemistry. Substrates and PPO activity were localized in developing squashed cell layer of nodules, suggesting PPO may have a developmental role in nodules.
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