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Publications - Stress and Develop Biology

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Publications

Eschen-Lippold, L.; Altmann, S.; Gebhardt, C.; Göbel, C.; Feussner, I.; Rosahl, S.; Oxylipins are not required for R gene-mediated resistance in potato Eur. J. Plant Pathol. 127, 437-442, (2010) DOI: 10.1007/s10658-010-9621-1

The role of 9- and 13-lipoxygenase-derived oxylipins for race-cultivar-specific resistance in potato was analyzed by expressing RNA interference constructs against oxylipin biosynthetic genes in transgenic potato plants carrying the resistance gene R1 against Phytophthora infestans. Down-regulation of 9-lipoxygenase expression resulted in highly reduced levels of 9-hydroxyoctadecatrienoic acid after treatment with the pathogen-associated molecular pattern Pep-13. However, neither 9-lipoxygenase nor 9-divinyl ether synthase RNAi plants exhibited alterations in their resistance to P. infestans. Similarly, successful down-regulation of transcript accumulation of the 13-lipoxygenase pathway genes encoding allene oxide cyclase, 12-oxophytodienoic acid reductase 3 and the jasmonic acid receptor coronatine-insensitive 1 resulted in highly reduced levels of jasmonic acid after Pep-13 treatment. Race-cultivar-specific resistance, however, was not lost in these plants. Our results suggest that neither 9-lipoxygenase-derived oxylipins nor jasmonic acid are required for R-gene-mediated resistance in potato. Importantly, in tobacco, the silencing of 9-lipoxygenase expression was previously demonstrated to suppress race-cultivar-specific resistance. Thus, we conclude a differential requirement of oxylipins for R-gene-mediated resistance in different solanaceous plants.
Publications

Andreou, A.-Z.; Hornung, E.; Kunze, S.; Rosahl, S.; Feussner, I.; On the Substrate Binding of Linoleate 9-Lipoxygenases Lipids 44, 207-215, (2009) DOI: 10.1007/s11745-008-3264-4

Lipoxygenases (LOX; linoleate:oxygen oxidoreductase EC 1.13.11.12) consist of a class of enzymes that catalyze the regio‐ and stereo specific dioxygenation of polyunsaturated fatty acids. Here we characterize two proteins that belong to the less studied class of 9‐LOXs, Solanum tuberosum StLOX1 and Arabidopsis thaliana AtLOX1. The proteins were recombinantly expressed in E. coli and the product specificity of the enzymes was tested against different fatty acid substrates. Both enzymes showed high specificity against all tested C18 fatty acids and produced (9S )‐hydroperoxides. However, incubation of the C20 fatty acid arachidonic acid with AtLOX1 gave a mixture of racemic hydroperoxides. On the other hand, with StLOX1 we observed the formation of a mixture of products among which the (5S )‐hydroperoxy eicosatetraenoic acid (5S‐ H(P)ETE) was the most abundant. Esterified fatty acids were no substrates. We used site directed mutagenesis to modify a conserved valine residue in the active site of StLOX1 and examine the importance of space within the active site, which has been shown to play a role in determining the positional specificity. The Val576Phe mutant still catalyzed the formation of (9S )‐hydroperoxides with C18 fatty acids, while it exhibited altered specificity against arachidonic acid and produced mainly (11S )‐H(P)ETE. These data confirm the model that in case of linoleate 9‐LOX binding of the substrate takes place with the carboxyl‐group first.
Publications

Eschen-Lippold, L.; Rothe, G.; Stumpe, M.; Göbel, C.; Feussner, I.; Rosahl, S.; Reduction of divinyl ether-containing polyunsaturated fatty acids in transgenic potato plants Phytochemistry 68, 797-801, (2007) DOI: 10.1016/j.phytochem.2006.12.010

Oxygenated polyunsaturated fatty acids synthesized via the lipoxygenase pathway play a role in plant responses to pathogen attack. In solanaceous plants, the preferential stimulation of the 9-lipoxygenase pathway in response to pathogen infection leads to the formation of the divinyl ether-containing polyunsaturated fatty acids colneleic and colnelenic acid, as well as hydroxy and trihydroxy polyunsaturated fatty acids. To functionally assess the role of divinyl ethers, transgenic potato plants were generated which express an RNA interference construct directed against the pathogen-inducible 9-divinyl ether synthase. Efficient reduction of 9-divinyl ether synthase transcript accumulation correlated with reduced levels of colneleic and colnelenic acid. However, in response to infection with virulent Phytophthora infestans, the causal agent of late blight disease, no significant differences in pathogen biomass could be detected suggesting that the levels of antimicrobial divinyl ethers are not critical for defense against Phytophthora infestans in a compatible interaction.
Publications

NICKSTADT, A.; THOMMA, B. P. H. J.; Feussner, I.; Kangasjärvi, J.; ZEIER, J.; LOEFFLER, C.; Scheel, D.; BERGER, S.; The jasmonate-insensitive mutant jin1 shows increased resistance to biotrophic as well as necrotrophic pathogens Mol. Plant Pathol. 5, 425-434, (2004) DOI: 10.1111/j.1364-3703.2004.00242.x

Jasmonic acid and related oxylipin compounds are plant signalling molecules that are involved in the response to pathogens, insects, wounding and ozone. To explore further the role of jasmonates in stress signal transduction, the response of two jasmonate‐signalling mutants, jin1 and jin4 , to pathogens and ozone was analysed in this study. Upon treatment with the biotrophic bacterial pathogen Pseudomonas syringae , endogenous jasmonate levels increased in jin1 and jin4 similar to wild‐type, demonstrating that these mutants are not defective in jasmonate biosynthesis. Jin1 but not jin4 is more resistant to P. syringae and this higher resistance is accompanied by higher levels of salicylic acid. Jin1 is also more resistant to the necrotrophic fungal pathogen Botrytis cinerea and shows wild‐type sensitivity to ozone whereas jin4 is more susceptible to B. cinerea and ozone. These results indicate that the mutations in jin1 and jin4 affect different branches of the jasmonate signalling pathway. Additionally, in this combination of phenotypes, jin1 is unique among all other jasmonate‐related mutants described thus far. These data also provide support for a crosstalk between the jasmonate and salicylate pathways.
Books and chapters

Rosahl, S.; Feussner, I.; Oxylipins 329-354, (2004)

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Publications

Göbel, C.; Feussner, I.; Rosahl, S.; Lipid Peroxidation during the Hypersensitive Response in Potato in the Absence of 9-Lipoxygenases J. Biol. Chem. 278, 52834-52840, (2003) DOI: 10.1074/jbc.M310833200

Hypersensitive cell death is an important defense reaction of plants to pathogen infection and is accompanied by lipid peroxidation processes. These may occur non-enzymatically by the action of reactive oxygen species or may be catalyzed by enzymes such as α-dioxygenases, lipoxygenases, or peroxidases. Correlative data showing increases in 9-lipoxygenase products in hyper-sensitively reacting cells have so far suggested that a large part of lipid peroxidation is mediated by a specific set of 9-lipoxygenases. To address the significance of 9-lipoxygenases for this type of pathogen response in potato, RNA interference constructs of a specific pathogen-induced potato 9-lipoxygenase were transferred to potato plants. Significantly reduced 9-lipoxygenase transcript levels were observed in transgenic plants after pathogen treatment. In addition, 9-lipoxygenase activity was hardly detectable, and levels of 9-lipoxygenase-derived oxylipins were reduced up to 12-fold after pathogen infection. In contrast to wild type plants, high levels of non-enzymatically as well as 13-lipoxygenase-derived oxylipins were present in 9-lipoxygenase-deficient plants. From this we conclude that during the normal hypersensitive response in potato, lipid peroxidation may occur as a controlled and directed process that is facilitated by the action of a specific 9-lipoxygenase. If 9-lipoxygenase-mediated formation of hydroperoxides is repressed, autoxidative lipid peroxidation processes and 13-lipoxygenase-mediated oxylipins synthesis become prominent. The unaltered timing and extent of necrosis formation suggests that the origin of lipid hydroperoxides does not influence pathogen-induced cell death in potato.
Publications

Landgraf, P.; Feussner, I.; Hunger, A.; Scheel, D.; Rosahl, S.; Systemic Accumulation of 12-oxo-phytodienoic Acid in SAR-induced Potato Plants Eur. J. Plant Pathol. 108, 279-283, (2002) DOI: 10.1023/A:1015132615650

In potato plants induced for systemic resistance by infiltration with Pseudomonas syringae pv. maculicola, 12-oxo-phytodienoic acid (OPDA) accumulated in infiltrated leaves as well as in non-treated leaves of infected plants. In contrast, jasmonic acid (JA) levels increased only in infiltrated leaves, suggesting that the biosynthetic precursor of JA, OPDA, might play a role in systemic acquired resistance.
Publications

Göbel, C.; Feussner, I.; Hamberg, M.; Rosahl, S.; Oxylipin profiling in pathogen-infected potato leaves BBA-Mol. Cell Biol. Lipids 1584, 55-64, (2002) DOI: 10.1016/S1388-1981(02)00268-8

Plants respond to pathogen attack with a multicomponent defense response. Synthesis of oxylipins via the lipoxygenase (LOX) pathway appears to be an important factor for establishment of resistance in a number of pathosystems. In potato cells, pathogen-derived elicitors preferentially stimulate the 9-LOX-dependent metabolism of polyunsaturated fatty acids (PUFAs). Here we show by oxylipin profiling that potato plants react to pathogen infection with increases in the amounts of the 9-LOX-derived 9,10,11- and 9,12,13-trihydroxy derivatives of linolenic acid (LnA), the divinyl ethers colnelenic acid (CnA) and colneleic acid (CA) as well as 9-hydroxy linolenic acid. Accumulation of these compounds is faster and more pronounced during the interaction of potato with the phytopathogenic bacterium Pseudomonas syringae pv. maculicola, which does not lead to disease, compared to the infection of potato with Phytophthora infestans, the causal agent of late blight disease. Jasmonic acid (JA), a 13-LOX-derived oxylipin, accumulates in potato leaves after infiltration with P. syringae pv. maculicola, but not after infection with P. infestans.
Publications

Stumpe, M.; Kandzia, R.; Göbel, C.; Rosahl, S.; Feussner, I.; A pathogen-inducible divinyl ether synthase (CYP74D) from elicitor-treated potato suspension cells FEBS Lett. 507, 371-376, (2001) DOI: 10.1016/S0014-5793(01)03019-8

In elicitor-treated potato cells, 9-lipoxygenase-derived oxylipins accumulate with the divinyl ether colneleic acid as the major metabolite. Here, the identification of a potato cDNA is described, whose predicted amino acid sequence corresponds to divinyl ether synthases, belonging to the recently identified new P450 subfamily CYP74D. The recombinant protein was expressed in Escherichia coli and shown to metabolize 9-hydroperoxy linoleic acid to colneleic acid at pH 6.5. This fatty acid derivative has been implicated in functioning as a plant antimicrobial compound. RNA blot analyses revealed accumulation of divinyl ether synthase transcripts both upon infiltration of potato leaves with Pseudomonas syringae and after infection with Phytophthora infestans.
Publications

Göbel, C.; Feussner, I.; Schmidt, A.; Scheel, D.; Sanchez-Serrano, J.; Hamberg, M.; Rosahl, S.; Oxylipin Profiling Reveals the Preferential Stimulation of the 9-Lipoxygenase Pathway in Elicitor-treated Potato Cells J. Biol. Chem. 276, 6267-6273, (2001) DOI: 10.1074/jbc.M008606200

Lipoxygenases are key enzymes in the synthesis of oxylipins and play an important role in the response of plants to wounding and pathogen attack. In cultured potato cells treated with elicitor from Phytophthora infestans, the causal agent of late blight disease, transcripts encoding a linoleate 9-lipoxygenase and a linoleate 13-lipoxygenase accumulate. However, lipoxygenase activity assays and oxylipin profiling revealed only increased 9-lipoxygenase activity and formation of products derived therefrom, such as 9-hydroxy octadecadienoic acid and colneleic acid. Furthermore, the 9-lipoxygenase products 9(S),10(S),11(R)-trihydroxy-12(Z)-octadecenoic and 9(S),10(S),11(R)-trihydroxy-12(Z),15(Z)-octadecadienoic acid were identified as novel, elicitor-inducible oxylipins in potato, suggesting a role of these compounds in the defense response against pathogen attack. Neither 13-lipoxygenase activity nor 13-lipoxygenase products were detected in higher amounts in potato cells after elicitation. Thus, formation of products by the 9-lipoxygenase pathway, including the enzymes hydroperoxide reductase, divinyl ether synthase, and epoxy alcohol synthase, is preferentially stimulated in cultured potato cells in response to treatment with P. infestanselicitor. Moreover, elicitor-induced accumulation of desaturase transcripts and increased phospholipase A2 activity after elicitor treatment suggest that substrates for the lipoxygenase pathway might be provided by de novo synthesis and subsequent release from lipids of the endomembrane system.
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