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Lipoxygenases (LOXs) catalyze hydroperoxidation of polyunsaturated fatty acids (PUFAs) to form structurally and functionally diverse oxylipins. Precise physiological and biochemical functions of individual members of plant multigene LOX families are largely unknown. Herein we report on molecular and biochemical characterization of two closely related maize 9-lipoxygenase paralogs, ZmLOX4 and ZmLOX5. Recombinant ZmLOX5 protein displayed clear 9-LOX regio-specificity at both neutral and slightly alkaline pH. The genes were differentially expressed in various maize organs and tissues as well as in response to diverse stress treatments. The transcripts of ZmLOX4 accumulated predominantly in roots and shoot apical meristem, whereas ZmLOX5 was expressed in most tested aboveground organs. Both genes were not expressed in untreated leaves, but displayed differential induction by defense-related hormones. While ZmLOX4 was only induced by jasmonic acid (JA), the transcripts of ZmLOX5 were increased in response to JA and salicylic acid treatments. ZmLOX5 was transiently induced both locally and systemically by wounding, which was accompanied by increased levels of 9-oxylipins, and fall armyworm herbivory, suggesting a putative role for this gene in defense against insects. Surprisingly, despite of moderate JA- and wound-inducibility of ZmLOX4, the gene was not responsive to insect herbivory. These results suggest that the two genes may have distinct roles in maize adaptation to diverse biotic and abiotic stresses. Both paralogs were similarly induced by virulent and avirulent strains of the fungal leaf pathogen Cochliobolus carbonum. Putative physiological roles for the two genes are discussed in the context of their biochemical and molecular properties.

Plant and fungal lipoxygenases (LOX) catalyze the oxidation of polyunsaturated fatty acids, creating fatty-acid hydroperoxides (oxylipins). Fungal oxylipins are required for normal fungal development and secondary metabolism, and plant host-derived oxylipins interfere with these processes in fungi, presumably by signal mimicry. The maize LOX gene ZmLOX3 has been implicated previously in seed-Aspergillus interactions, so we tested the interactions of a mutant maize line (lox3-4, in which ZmLOX3 is disrupted) with the mycotoxigenic seed-infecting fungi Aspergillus flavus and Aspergillus nidulans. The lox3-4 mutant was more susceptible than wild-type maize to both Aspergillus species. All strains of A. flavus and A. nidulans produced more conidia and aflatoxin (or the precursor sterigmatocystin) on lox3-4 kernels than on wild-type kernels, in vitro and under field conditions. Although oxylipins did not differ detectably between A. flavus-infected kernels of the lox3-4 and wild-type (WT) maize, oxylipin precursors (free fatty acids) and a downstream metabolite (jasmonic acid) accumulated to greater levels in lox3-4 than in WT kernels. The increased resistance of the lox3-4 mutant to other fungal pathogens (Fusarium, Colletotrichum, Cochliobolus, and Exserohilum spp.) is in sharp contrast to results described herein for Aspergillus spp., suggesting that outcomes of LOX-governed host-pathogen interactions are pathogen-specific.

We have previously shown that the beneficial filamentous fungus Trichoderma virens secretes the highly effective hydrophobin-like elicitor Sm1 that induces systemic disease resistance in the dicot cotton (Gossypium hirsutum). In this study we tested whether colonization of roots by T. virens can induce systemic protection against a foliar pathogen in the monocot maize (Zea mays), and we further demonstrated the importance of Sm1 during maize-fungal interactions using a functional genomics approach. Maize seedlings were inoculated with T. virens Gv29-8 wild type and transformants in which SM1 was disrupted or constitutively overexpressed in a hydroponic system or in soil-grown maize seedlings challenged with the pathogen Colletotrichum graminicola. We show that similar to dicot plants, colonization of maize roots by T. virens induces systemic protection of the leaves inoculated with C. graminicola. This protection was associated with notable induction of jasmonic acid- and green leaf volatile-biosynthetic genes. Neither deletion nor overexpression of SM1 affected normal growth or development of T. virens, conidial germination, production of gliotoxin, hyphal coiling, hydrophobicity, or the ability to colonize maize roots. Plant bioassays showed that maize grown with SM1-deletion strains exhibited the same levels of systemic protection as non-Trichoderma-treated plants. Moreover, deletion and overexpression of SM1 resulted in significantly reduced and enhanced levels of disease protection, respectively, compared to the wild type. These data together indicate that T. virens is able to effectively activate systemic disease protection in maize and that the functional Sm1 elicitor is required for this activity.

Plant lipoxygenases (LOXs) are a functionally diverse class of dioxygenases implicated in physiological processes such as growth, senescence, and stress-related responses. LOXs incorporate oxygen into their fatty acid substrates and produce hydroperoxide fatty acids that are precursors of jasmonic acid and related compounds. Here, we report the involvement of the tuber-associated LOXs, designated the Lox1 class, in the control of tuber growth. RNA hybridization analysis showed that the accumulation of Lox1 class transcripts was restricted to developing tubers, stolons, and roots and that mRNA accumulation correlated positively with tuber initiation and growth. In situ hybridization showed that Lox1 class transcripts accumulated in the apical and subapical regions of the newly formed tuber, specifically in the vascular tissue of the perimedullary region, the site of the most active cell growth during tuber enlargement. Suppression mutants produced by expressing antisense coding sequence of a specific tuber LOX, designated POTLX-1, exhibited a significant reduction in LOX activity in stolons and tubers. The suppression of LOX activity correlated with reduced tuber yield, decreased average tuber size, and a disruption of tuber formation. Our results indicate that the pathway initiated by the expression of the Lox1 class genes of potato is involved in the regulation of tuber enlargement.

Lipoxygenase (LOX) activity has been identified consistently during pathogen-induced defense responses. Here we report the involvement of a specific leaf LOX gene of potato (Solanum tuberosum), designated POTLX-3 (GenBank/EMBL accession no. U60202), in defense responses against pathogens. The sequence of POTLX-3 does not match any other LOX genes of potato and has the greatest match to a tobacco LOX gene that contributes to a resistance mechanism against Phytophthora parasitica var nicotianae. POTLX-3 transcript accumulation was not detected in untreated, healthy potato organs or in wounded mature leaves. POTLX-3 mRNA accumulation was induced in potato leaves treated with ethylene or methyl jasmonate or infected with either virulent or avirulent strains of Phytophthora infestans, the causal agent of late blight. During the resistance response, POTLX-3 was induced within 6 hours, increased steadily through 24 hours, and its mRNA continued to accumulate for a week after inoculation. In contrast, when a plant was susceptible to P. infestans, induction of mRNA accumulation in response to inoculation was inconsistent and delayed. LOX activity assayed during an incompatible interaction in leaves peaked 3 days earlier than during a compatible interaction. POTLX-3 mRNA accumulation also was induced during hypersensitive response development caused by the incompatible pathogen Pseudomonas syringae pv phaseolicola. Our results show that POTLX-3 may be involved specifically in defense responses against pathogen infection.