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PMID:10998053
| Citation |
Schaffrath, U, Zabbai, F and Dudler, R (2000) Characterization of RCI-1, a chloroplastic rice lipoxygenase whose synthesis is induced by chemical plant resistance activators. Eur. J. Biochem. 267:5935-42 |
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| Abstract |
A full-length lipoxygenase cDNA (RCI-1) has been cloned from rice (Oryza sativa) whose corresponding transcripts accumulate in response to treatment of the plants with chemical inducers of acquired resistance such as benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH), 2,6-dichloroisonicotinic acid (INA), and probenazole. In contrast, RCI-1 transcript levels did not increase after inoculation with compatible and incompatible races of the rice blast fungus Magnaporthe grisea and the nonhost pathogen Pseudomonas syringae pv. syringae. RCI-1 transcript levels also increased after exogenous application of jasmonic acid, but not upon wounding. Dose-response and time course experiments revealed a similar pattern of transcript accumulation and lipoxygenase activity in BTH-treated rice leaves. Enzymatic analysis of recombinant RCI-1 protein produced in Escherichia coli revealed that 13-hydroperoxy-octadecanoic acids were the predominant reaction products when either linoleic or linolenic acid used as a substrate. The RCI-1 sequence features a putative chloroplast targeting sequence at its N-terminus. Indeed, a protein consisting of the putative chloroplast transit peptide fused to green fluorescent protein was exclusively localized in chloroplasts, indicating that RCI-1 is a chloroplastic enzyme. |
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| Keywords |
Amino Acid Sequence; Chloroplasts/enzymology; DNA, Complementary/genetics; DNA, Plant/genetics; Enzyme Induction/drug effects; Gene Expression Regulation, Plant/drug effects; Immunity, Innate; Isonicotinic Acids/pharmacology; Linoleic Acid/metabolism; Lipoxygenase/biosynthesis; Lipoxygenase/genetics; Lipoxygenase/isolation & purification; Magnaporthe/physiology; Molecular Sequence Data; Oryza sativa/drug effects; Oryza sativa/enzymology; Oryza sativa/genetics; Oryza sativa/microbiology; Plant Diseases/genetics; Plant Diseases/microbiology; Pseudomonas/physiology; Recombinant Fusion Proteins/metabolism; Sequence Alignment; Sequence Homology, Amino Acid; Thiadiazoles/pharmacology; Thiazoles/pharmacology; Transcription, Genetic; alpha-Linolenic Acid/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
enables |
GO:0016165: linoleate 13S-lipoxygenase activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
|
part_of |
GO:0009507: chloroplast |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0016165: lipoxygenase activity |
ECO:0000314: |
F |
Crude Escherichia coli extracts containing RCI-1 recombinant protein were incubated with either linoleic or linolenic acid as a substrate. Lipoxygenase activity was measured photspectrometrically. Products analysed by HPLC. Table 1 identifies the 13-hydroperoxy-octadecanoic acid products formed as a result of lipoxygenase activity. |
complete | ||||
| GO:0009507: chloroplast |
ECO:0000314: |
C |
Figure 2 |
complete | ||||
See also
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