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Mikhailo V. Kolomiets, Richard J. Gladon and David J. Hannapel

Due to apparent participation of plant lipoxygenases (LOXs) in the biosynthetic pathways for jasmonic acid, methyl jasmonate, traumatin, and several C-6 volatile compounds, LOXs are believed to have a role in senescence, plant growth and development, and wound- and pathogen-induced defense responses. Multiple functions that are ascribed to this enzyme family are in accordance with the heterogeneity of LOX isozyme forms. It is possible that different LOX isoforms may be involved in different physiological processes. In our search for a gene that encodes a LOX isozyme form specifically involved in potato defense responses against pests and pathogens, we have screened an abscisic acid-induced potato leaf cDNA library, and we have isolated, sequenced, and characterized a cDNA clone that we have designated POTLX-3. The high sequence homology of our cDNA clone to other reported plant LOX genes provided evidence that POTLX-3 is a lipoxygenase. This cDNA clone represents a novel potato LOX gene in that it shares the least nucleotide and amino acid sequence homology to other isolated potato LOX genes. Northern analysis indicated that POTLX-3 transcripts did not accumulate in untreated potato leaves, but it was highly induced by treatment with physiological levels of ethylene. Northern analysis also was performed to study whether the POTLX-3 mRNA accumulation could be induced by other plant hormones that affect expression of the other plant LOX and defense-related genes. Treatment of potato leaves with methyl jasmonate, abscisic acid, gibberellic acid, auxin (NAA), and cytokinin (BA) did not induce POTLX-3 gene expression. Because the pattern of POTLX-3 gene expression is similar to that of pathogenesis-related (PR) proteins, especially the PR-1 and PR-5 groups, we suspect that POTLX-3 may be involved specifically in ethylene-induced defense responses against pathogens.

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Mikhailo V. Kolomiets, David J. Hannapel and Richard J. Gladon

Plant lipoxygenases (LOXs) (linoleate:oxygen oxidoreductase, EC 1.13.11.12) catalyze the oxygenation of polyunsaturated fatty acids such as linolenic and linoleic acids. Some of the final products of LOX-catalyzed reactions are traumatin, jasmonic acid (JA), methyl jasmonate (MJ), and C-6 volatile compounds, and they serve hormone-like regulatory and defense-related roles in plants. Recently, it has been proposed that LOXs play a role in potato tuberization processes because JA, MJ, and structurally similar tuberonic acid and tuberonic acid glycoside have been shown to be tuber-inducing substances. In order to study possible lipoxygenase involvement in potato tuberization, we have isolated, sequenced, and characterized the expression pattern of two cDNA clones, designated POTLX-1 and POTLX-2, that represent similar, but distinct, LOX genes. Within the scope of our experiments, northern hybridization studies with mRNA extracted from various organs of `Superior' potato plants indicated that the expression of these two genes is restricted to developing tubers and roots only. Moreover, there is a positive correlation between POTLX-1 and POTLX-2 mRNA accumulation and the stage of potato tuber development, and this implicates LOX in tuberization processes. Accumulation of their transcripts was not detected in leaves, flowers, stems, shoot tips, or axillary buds. These results indicate that the isozyme forms encoded by these two genes are tuber-specific, and they are good candidates to study LOX involvement in potato tuberization processes. Treatment of potato leaves with abscisic acid, MJ, gibberellic acid, auxin (NAA), and cytokinin (BA) did not trigger transcriptional activation of either of these genes.

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Richard J. Gladon, David J. Hannapel, Mikhailo V. Kolomiets, Mikhailo V. Andriyenko, Petro V. Kondratenko and Volodymyr Kopan