Novel roles for the polyphenol oxidase enzyme in secondary metabolism and the regulation of cell death in walnut.
The enzyme polyphenol oxidase (PPO) catalyzes the oxidation of phenolic compounds into highly reactive quinones. Polymerization of PPO-derived quinones causes the postharvest browning of cut or bruised fruit, but the native physiological functions of PPOs in undamaged, intact plant cells are not well understood. Walnut (Juglans regia) produces a rich array of phenolic compounds and possesses a single PPO enzyme, rendering it an ideal model to study PPO. We generated a series of PPO-silenced transgenic walnut lines that display less than 5% of wild-type PPO activity. Strikingly, the PPO-silenced plants developed spontaneous necrotic lesions on their leaves in the absence of pathogen challenge (i.e. a lesion mimic phenotype). To gain a clearer perspective on the potential functions of PPO and its possible connection to cell death, we compared the leaf transcriptomes and metabolomes of wild-type and PPO-silenced plants. Silencing of PPO caused major alterations in the metabolism of phenolic compounds and their derivatives (e.g. coumaric acid and catechin) and in the expression of phenylpropanoid pathway genes. Several observed metabolic changes point to a direct role for PPO in the metabolism of tyrosine and in the biosynthesis of the hydroxycoumarin esculetin in vivo. In addition, PPO-silenced plants displayed massive (9-fold) increases in the tyrosine-derived metabolite tyramine, whose exogenous application elicits cell death in walnut and several other plant species. Overall, these results suggest that PPO plays a novel and fundamental role in secondary metabolism and acts as an indirect regulator of cell death in walnut.
Department of Biological Sciences, California State University, San Marcos, California 92096.
Soha
S
Araji
Theresa A
TA
Grammer
Ross
R
Gertzen
Stephen D
SD
Anderson
Maja
M
Mikulic-Petkovsek
Robert
R
Veberic
My L
ML
Phu
Anita
A
Solar
Charles A
CA
Leslie
Abhaya M
AM
Dandekar
Matthew A
MA
Escobar
D004156:Catechol Oxidase / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002934:Cinnamates / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D031324:Juglans / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D055432:Metabolomics; D010641:Phenotype; D010936:Plant Extracts / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D020005:Propanols / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D064210:Secondary Metabolism / Q000187:drug effects / Q000235:genetics; D013379:Substrate Specificity / Q000187:drug effects; D014439:Tyramine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D002934:Cinnamates; D010936:Plant Extracts; D010940:Plant Proteins; D020005:Propanols; D012333:RNA, Messenger; C439395:1-phenylpropanol; D004156:Catechol Oxidase; D014439:Tyramine