Molecular analysis of a null mutant for pea (Pisum sativum L.) seed lipoxygenase-2 |
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Authors: | Forster Colette North Helen Afzal Naureen Domoney Claire Hornostaj Andrzej Robinson David S. Casey Rod |
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Affiliation: | (1) John Innes Centre, Colney Lane, Norwich Research Park, Norwich, NR4 7UH, UK;(2) Laboratoire de Biologie Cellulaire, INRA, 78026 Versailles, France;(3) Procter Department of Food Science, University of Leeds, Leeds, LS2 9JT, UK |
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Abstract: | A mutant line of Pisum fulvum was identified that lacked seed lipoxygenase-2 (LOX-2). The mutant phenotype was introgressed into a standard Pisum sativum cv. Birte to provide near-isogenic lines with or without seed LOX-2. Genetic analyses showed the mutation to behave as a single, recessive Mendelian gene. Northern and dot-blot analyses showed a large reduction in LOX-2 mRNA from developing seeds of the LOX-2-null mutant. A restriction fragment length polymorphism associated with the 5 end of the LOX-2 gene(s) co-segregated with the null phenotype, indicating that the reduction of LOX-2 mRNA was neither a consequence of deletion of the LOX genes nor a consequence of the action of a genetically distant regulatory gene. Analysis of the 5 -flanking sequences of LOX-2 genes from Birte and the near-isogenic LOX-2-null mutant revealed a number of insertions, deletions and substitutions within the promoter from the LOX-2-null mutant that could be responsible for the null phenotype. Incubation of crude seed LOX preparations from Birte and the LOX-2-null mutant showed that the latter generated relatively less 13-hydroperoxides and also produced relatively more hydroxy- and ketoacid compounds that have implications for the fresh-frozen pea industry. |
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Keywords: | lipoxygenase null mutant pea (Pisum sativum) seed |
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