Changes in the initial phase of lipid peroxidation induced by elicitor from Phytophthora infestans in Solanum species

The initial phase of the lipid peroxidation process in leaves of Solanum nigrum var. gigantea, Solanum tuberosum cv Bzura and clone H-8105, which represent non-host resistance, field resistance and susceptibility, respectively, against Phytophthora infestans, was investigated. Based on quantitative and qualitative high-performance liquid chromatography (HPLC) analyses of free and esterified fatty acid hydroperoxides (FAHs), we characterized the lipid peroxidation process induced by the pathogen-derived elicitor, culture filtrate (CF), in leaves of the studied genotypes. In all plants, FAHs generated due to 13-lipoxygenase (LOX) action dominated over those from the non-enzymatic pathway. The FAHs derived from 9-LOX activity were found only in CF-treated leaves of the non-host resistant S. nigrum. However, experiments in vitro and in planta with exogenous linoleic acid (LA) as a substrate for LOX revealed high constitutive activity of 9-LOX in all genotypes, which increased in response to CF treatment. The time course changes in polyunsaturated fatty acid (PUFA) pools in the total lipid fractions as well as the degree of their oxidation suggested that CF-induced PUFA peroxidation was enhanced mostly in S. nigrum, less so in Bzura and least in the susceptible clone H-8105. The obtained results are discussed in light of the overall biochemical cell status of plants in the studied interactions.     

Fatty acid hydroperoxides and H2O2 in the execution of hypersensitive cell death in tobacco leaves

We initially compared lipid peroxidation profiles in tobacco (Nicotiana tabacum) leaves during different cell death events. An upstream oxylipin assay was used to discriminate reactive oxygen species (ROS)-mediated lipid peroxidation from 9- and 13-lipoxygenase (LOX)-dependent lipid peroxidation. Free radical-mediated membrane peroxidation was measured during H(2)O(2)-dependent cell death in leaves of catalase-deficient plants. Taking advantage of these transgenic plants, we demonstrate that, under light conditions, H(2)O(2) plays an essential role in the execution of cell death triggered by an elicitor, cryptogein, which provokes a similar ROS-mediated lipid peroxidation. Under dark conditions, however, cell death induction by cryptogein was independent of H(2)O(2) and accompanied by products of the 9-LOX pathway. In the hypersensitive response induced by the avirulent pathogen Pseudomonas syringae pv syringae, both 9-LOX and oxidative processes operated concurrently, with ROS-mediated lipid peroxidation prevailing in the light. Our results demonstrate, therefore, the tight interplay between H(2)O(2) and lipid hydroperoxides and underscore the importance of light during the hypersensitive response.     

A novel patatin-like protein from cotton plant,GhPat1, is co-expressed with GhLox1 during <Emphasis Type="Italic">Xanthomonas campestris</Emphasis>-mediated hypersensitive cell death

In cotton plant, Xanthomonas-induced hypersensitive response (HR) is accompanied by a lipid peroxidation process involving a 9-lipoxygenase (LOX), GhLox1. Initiation of this oxidative metabolism implies the release of the LOX substrates, or polyunsaturated fatty acids. Since patatin-like proteins (PLPs) are likely candidates for mediating the latter step, we searched for genes encoding such enzymes, identified and cloned one of them that we named GhPat1. Biochemical and molecular studies showed that GhPat1 expression was up-regulated during the incompatible interaction, prior to the onset of the corresponding galactolipase activity and cell death symptoms in tissues. Protein sequence analysis and modelling also revealed that GhPat1 catalytic amino acids and fold were conserved across plant PLPs. Based on these results and our previous work (Jalloul et al. in Plant J 32:1-12, 2002), a role for GhPat1, in synergy with GhLox1, during HR-specific lipid peroxidation is discussed.     

The 9-lipoxygenase GhLOX1 gene is associated with the hypersensitive reaction of cotton Gossypium hirsutum to Xanthomonas campestris pv malvacearum.

Hypersensitive reaction (HR) cell death of cotton to the incompatible race 18 from Xanthomonas campestris pathovar malvacearum (Xcm) is associated with 9S-lipoxygenase activity (LOX) responsible for lipid peroxidation. Here, we report the cloning of cotton (Gossypium hirsutum L.) LOX gene (GhLOX1) and the sequencing of its promoter. GhLOX1 was found to be highly expressed during Xcm induced HR. Sequence analysis showed that GhLOX1 is a putative 9-LOX, and GhLOX1 promoter contains SA and JA responsive elements. Investigation on LOX signalisation on cotyledons infiltrated with salicylic acid (SA), or incubated with methyl-jasmonate (MeJA) revealed that both treatments induced LOX activity and GhLOX1 gene expression. HR-like symptoms were observed when LOX substrates were then injected in treated (MeJA and SA) cotyledons or when Xcm compatible race 20 was inoculated on MeJA treated cotyledons. Together these results support the fact that GhLOX1 encodes a 9 LOX whose activity would be involved in cell death during cotton HR.     

A 13‐lipoxygenase is Expressed Early in the Hypersensitive Reaction of Cotton Plants to Xanthomonas campestris pv. malvacearum

Lipoxygenases (LOXs) are enzymes responsible for lipid peroxidation processes during plant defence responses to pathogen infection. Jasmonates are lipid‐derived signals that mediate plant stress responses with chloroplastic LOXs implicated in the biosynthesis of oxylipins like jasmonic acid (JA). Hypersensitive reaction (HR) cell death of cotton to the incompatible race 18 of Xanthomonas campestris pathovar malvacearum (Xcm) is associated with 9S‐lipoxygenase activity and expression of a 9‐LOX GhLOX1. Here, we report the cloning of cotton (Gossypium hirsutum L.) LOX gene GhLOX2. Sequence analysis showed that GhLOX2 is a putative 13‐LOX with a chloroplast‐transit peptide in the amino acid terminus. GhLOX2 was found to be significantly expressed in the first hour of Xcm‐induced HR. Investigation into LOX signalization on cotyledons incubated with methyl‐jasmonate (MeJA) or infiltrated with salicylic acid (SA) or ethylene (ET) revealed that the first two treatments induced GhLOX2 gene expression. Our results show that GhLOX2 gene expression occurred at the stage of the HR prior biochemical events previously highlighted. The role that GhLOX2 may have in the defence strategy of cotton to Xcm is discussed regarding the HR.     

The upstream oxylipin profile of Arabidopsis thaliana: a tool to scan for oxidative stresses

Various physiological imbalances lead to reactive oxygen species (ROS) overproduction and/or increases in lipoxygenase (LOX) activities, both events ending in lipid peroxidation of polyunsaturated fatty acids (PUFAs). Besides the quantification of such a process, the development of tools is necessary in order to allow the identification of the primary cause of its development and localization. A biochemical method assessing 9 LOX, 13 LOX and ROS-mediated peroxidation of membrane-bound and free PUFAs has been improved. The assay is based on the analysis of hydroxy fatty acids derived from PUFA hydroperoxides by both the straight and chiral phase high-performance liquid chromatography. Besides the upstream products of peroxidation of the 18:2 and 18:3 PUFAs, products coming from the 16:3 were characterized and their steady-state level quantified. Moreover, the observation that the relative amounts of the ROS-mediated peroxidation isomers of 18:3 were constant in leaves allowed us to circumvent the chiral analyses for the discrimination and quantification of 9 LOX, 13 LOX and ROS-mediated processes in routine experiments. The methodology has been successfully applied to decipher lipid peroxidation in Arabidopsis leaves submitted to biotic and abiotic stresses. We provide evidence of the relative timing of enzymatic and non-enzymatic lipid peroxidation processes. The 13 LOX pathway is activated early whatever the nature of the stress, leading to the peroxidation of chloroplast lipids. Under cadmium stress, the 9 LOX pathway added to the 13 LOX one. ROS-mediated peroxidation was mainly driven by light and always appeared as a late process.     

A tale of two lipids: Lipid unsaturation commands ferroptosis sensitivity

Membrane lipids play important roles in the regulation of cell fate, including the execution of ferroptosis. Ferroptosis is a non-apoptotic cell death mechanism defined by iron-dependent membrane lipid peroxidation. Phospholipids containing polyunsaturated fatty acids (PUFAs) are highly vulnerable to peroxidation and are essential for ferroptosis execution. By contrast, the incorporation of less oxidizable monounsaturated fatty acids (MUFAs) in membrane phospholipids protects cells from ferroptosis. The enzymes and pathways that govern PUFA and MUFA metabolism therefore play a critical role in determining cellular sensitivity to ferroptosis. Here, we review three lipid metabolic processes—fatty acid biosynthesis, ether lipid biosynthesis, and phospholipid remodeling—that can govern ferroptosis sensitivity by regulating the balance of PUFAs and MUFAs in membrane phospholipids.     

Adducts of oxylipin electrophiles to glutathione reflect a 13 specificity of the downstream lipoxygenase pathway in the tobacco hypersensitive response

The response to reactive electrophile species (RES) is now considered as part of the plant response to pathogen and insect attacks. Thanks to a previously established high-performance liquid chromatography tandem mass spectrometry methodology, we have investigated the production of oxylipin RES adducts to glutathione (GSH) during the hypersensitive response (HR) of plants. We have observed that RES conjugation to GSH in tobacco (Nicotiana tabacum) leaves is facile and nonspecific. In cryptogein-elicited tobacco leaves, we show that the oxylipin RES adducts to GSH are produced in correlation with GSH consumption, increase in glutathione S-transferase activity, and the appearance of the cell death symptoms. In this model, the adducts arise mainly from the downstream 13 lipoxygenase (LOX) metabolism, although the induced 9 LOX pathway leads massively to the accumulation of upstream metabolites. The main adducts were obtained from 2-hexenal and 12-oxo-phytodienoic acid. They accumulate transiently as 1-hexanol-3-GSH, a reduced adduct, and 12-oxo-phytodienoic acid-GSH, respectively. RES conjugation does not initiate cell death but explains part of the GSH depletion that accompanies HR cell death. The nature of these GSH conjugates shows the key role played by the 13 LOX pathway in RES signaling in the tobacco HR.     

Lipid profiling of the Arabidopsis hypersensitive response reveals specific lipid peroxidation and fragmentation processes: biogenesis of pimelic and azelaic Acid

Lipid peroxidation (LPO) is induced by a variety of abiotic and biotic stresses. Although LPO is involved in diverse signaling processes, little is known about the oxidation mechanisms and major lipid targets. A systematic lipidomics analysis of LPO in the interaction of Arabidopsis (Arabidopsis thaliana) with Pseudomonas syringae revealed that LPO is predominantly confined to plastid lipids comprising galactolipid and triacylglyceride species and precedes programmed cell death. Singlet oxygen was identified as the major cause of lipid oxidation under basal conditions, while a 13-lipoxygenase (LOX2) and free radical-catalyzed lipid oxidation substantially contribute to the increase upon pathogen infection. Analysis of lox2 mutants revealed that LOX2 is essential for enzymatic membrane peroxidation but not for the pathogen-induced free jasmonate production. Despite massive oxidative modification of plastid lipids, levels of nonoxidized lipids dramatically increased after infection. Pathogen infection also induced an accumulation of fragmented lipids. Analysis of mutants defective in 9-lipoxygenases and LOX2 showed that galactolipid fragmentation is independent of LOXs. We provide strong in vivo evidence for a free radical-catalyzed galactolipid fragmentation mechanism responsible for the formation of the essential biotin precursor pimelic acid as well as of azelaic acid, which was previously postulated to prime the immune response of Arabidopsis. Our results suggest that azelaic acid is a general marker for LPO rather than a general immune signal. The proposed fragmentation mechanism rationalizes the pathogen-induced radical amplification and formation of electrophile signals such as phytoprostanes, malondialdehyde, and hexenal in plastids.     

The Pepper 9-Lipoxygenase Gene CaLOX1 Functions in Defense and Cell Death Responses to Microbial Pathogens

Lipoxygenases (LOXs) are crucial for lipid peroxidation processes during plant defense responses to pathogen infection. A pepper (Capsicum annuum) 9-LOX gene, CaLOX1, which encodes a 9-specific lipoxygenase, was isolated from pepper leaves. Recombinant CaLOX1 protein expressed in Escherichia coli catalyzed the hydroperoxidation of linoleic acid, with a K_m value of 113. 9 μm. Expression of CaLOX1 was differentially induced in pepper leaves not only during Xanthomonas campestris pv vesicatoria (Xcv) infection but also after exposure to abiotic elicitors. Transient expression of CaLOX1 in pepper leaves induced the cell death phenotype and defense responses. CaLOX1-silenced pepper plants were more susceptible to Xcv and Colletotrichum coccodes infection, which was accompanied by reduced expression of defense-related genes, lowered lipid peroxidation, as well as decreased reactive oxygen species and lowered salicylic acid accumulation. Infection with Xcv, especially in an incompatible interaction, rapidly stimulated LOX activity in unsilenced, but not CaLOX1-silenced, pepper leaves. Furthermore, overexpression of CaLOX1 in Arabidopsis (Arabidopsis thaliana) conferred enhanced resistance to Pseudomonas syringae pv tomato, Hyaloperonospora arabidopsidis, and Alternaria brassicicola. In contrast, mutation of the Arabidopsis CaLOX1 ortholog AtLOX1 significantly increased susceptibility to these three pathogens. Together, these results suggest that CaLOX1 and AtLOX1 positively regulate defense and cell death responses to microbial pathogens.To effectively combat invasion by microbial pathogens, plants activate distinct defense responses that are specifically effective. Despite the presence of plant immune systems, many pathogens can evade or suppress host defense mechanisms. Lipoxygenase (LOX) pathways are crucial for lipid peroxidation processes during plant defense responses to pathogen infection (Casey and Hughes, 2004). Plant LOXs are key enzymes involved in the generation of fatty acid derivatives in oxylipin metabolism.LOXs comprise a family of non-heme-iron-containing fatty acid dioxygenases, which are ubiquitous in plants and animals (Brash, 1999). LOXs catalyze the conversion of polyunsaturated fatty acids such as linoleic acid into hydroperoxides that are in turn converted to oxylipins. These primary products, which may cause oxidative damage to plant membranes during the hypersensitive response (HR; Slusarenko, 1996), are enzymatically metabolized into traumatin, jasmonic acid (JA), and methyl jasmonate (MeJA). These latter compounds are involved in diverse physiological functions in plant growth and development, senescence, and stress responses. Plant LOXs can be classified as 9-LOXs or 13-LOXs according to the position at which oxygen is incorporated into linoleic acid or linolenic acid, the most important substrates for LOX catalysis in plants (Feussner and Wasternack, 2002). LOX enzymatic activity initiates the different biosynthetic pathways that result in the accumulation of distinct oxylipins. The most understood functional aspects of oxylipin pathways have come mainly from studies of JA produced through the action of 13-LOXs but not 9-LOXs. The metabolism of 13-LOX has been described in tobacco (Nicotiana tabacum) leaves infected by an avirulent strain of Pseudomonas syringae pv phaseolicola (Kenton et al., 1999). During bacterial infection, JA accumulates in tobacco leaves prior to cell death (Kenton et al., 1999). The level of LOX activity and gene expression also increases in tobacco plants during infection with Phytophthora parasitica var nicotianae (Christophe et al., 1996; Rancé et al., 1998). However, the defense-related functions of 9-LOXs are not fully understood. Both 9-LOXs and oxidative processes are proposed to be involved in the HR of tobacco induced by the avirulent pathogen Pseudomonas syringae pv syringae (Montillet et al., 2005). The production of free fatty acid hydroperoxides via the 9-LOX pathway in tobacco is crucial for hypersensitive cell death induced by cryptogein, a purified protein from Phytophthora cryptogea (Rusterucci et al., 1999). The function of LOXs in defense against pathogens is likely to be related to the synthesis of fatty acid hydroperoxides and of volatile products with signaling functions (Rusterucci et al., 1999) and antimicrobial activity (Croft et al., 1993; Weber et al., 1999). Gao et al. (2007) recently suggested that oxylipin metabolism mediated by a specific 9-LOX, ZmLOX3, may be involved in fungal pathogenesis in maize (Zea mays). ZmLOX3 loss-of-function mutants are susceptible to Aspergillus flavus and Aspergillus nidulans infection (Gao et al., 2009).LOX activity may initiate the synthesis of signal molecules or induce structural and metabolic changes in the cell, ultimately leading to cell death that has been termed the HR (Maccarrone et al., 2001). Plant cell death occurs during various phases of development, senescence, and responses to abiotic and biotic stresses, and in particular, in response to pathogen invasion (Morel and Dangl, 1997). Activation of LOXs in plants may be involved in cell death induced by pathogens (Buonaurio and Servili, 1999; Rusterucci et al., 1999). The induction of HR-like cell death by the activation of the 9-LOX-encoding gene GhLOX1 was shown in cotton (Gossypium hirsutum) plants during Xanthomonas campestris pv malvacearum infection (Marmey et al., 2007). LOX activity increases in parallel with the induction of HR symptoms in tobacco; however, in compatible interactions, LOX activity is delayed and reaches much lower levels (Montillet et al., 2002). In cotton, high LOX activity supports cell death during X. campestris pv malvacearum infection (Sayegh-Alhamdia et al., 2008). The HR, an important defense reaction of plants to pathogen infection, is accompanied by lipid peroxidation processes. In particular, 9-LOX-dependent lipid peroxidation operates during cryptogein-induced HR in tobacco leaves (Rusterucci et al., 1999). In potato (Solanum tuberosum), lipid peroxidation occurs as a controlled and directed process that is facilitated by the action of a specific 9-LOX during the HR (Göbel et al., 2003; Montillet et al., 2005). GhLOX1 is associated with salicylic acid (SA) accumulation during the HR of cotton to X. campestris pv malvacearum (Marmey et al., 2007).The bacterial plant pathogen Xanthomonas campestris pv vesicatoria (Xcv) is the causative agent of bacterial spot disease on pepper (Capsicum annuum) and tomato (Solanum lycopersicum) plants. To identify genes involved in the HR-based innate immune response in pepper, we have isolated and functionally characterized defense-related genes encoding PR1 (for pathogenesis-related protein 1; Kim and Hwang, 2000; Hong and Hwang, 2005), chitinase (Hong et al., 2000), chitin-binding protein (Lee et al., 2001), thionin (Lee et al., 2000), SAR 8.2 (Lee and Hwang, 2003), peroxidase (Choi et al., 2007), and menthone reductase (Choi et al., 2008) from pepper leaves infected with the Xcv avirulent strain Bv5-4a. In this study, we used a cDNA macroarray method (Jung and Hwang, 2000) to isolate a novel pepper gene, CaLOX1, which encodes a 9-LOX and is specifically induced by avirulent Xcv infection of pepper leaves. The purified CaLOX1 protein was expressed in Escherichia coli and investigated for LOX activity. Virus-induced gene silencing (VIGS) is a widely used, powerful technique for reverse genetics. VIGS vectors derived from the Tobacco rattle virus (TRV) are the most popular for VIGS. Recently, a VIGS method was established for the functional characterization of defense-related genes in pepper (Baulcombe, 1999; Burch-Smith et al., 2006; Choi et al., 2007; Chung et al., 2007). Here, we analyzed the effect of CaLOX1 loss of function during pathogen infection using TRV-based VIGS of the CaLOX1 gene. Arabidopsis (Arabidopsis thaliana) plants that constitutively overexpressed CaLOX1 were also examined to determine the gain-of-function phenotype of CaLOX1 in plant defense. We further functionally characterized the Arabidopsis mutants lox1-1 and lox1-2, which have T-DNA insertions in AtLOX1, a putative CaLOX1 ortholog. Analysis of the function of CaLOX1 in pepper and Arabidopsis plants provided insight into the role of CaLOX1 expression in defense responses and the hypersensitive cell death of plants following pathogen invasion.     

Comparative analysis of induction pattern of programmed cell death and defense-related responses during hypersensitive cell death and development of bacterial necrotic leaf spots in eggplant

Pseudomonas cichorii causes necrotic leaf spots (NLS), while Pseudomonas syringae pv. tabaci induces a hypersensitive response (HR) in eggplant. P. cichorii induced cell death at 9 h after inoculation (HAI), reaching a maximum of around 24–30 HAI. On the other hand, cell death was induced 6 HAI with P. syringae pv. tabaci, reaching a maximum of around 12–18 HAI. Superoxide generation was observed in eggplant inoculated with both bacteria. DNA fragmentation, cytochrome c release into the cytosol and expression of defense-related genes such as PR-1 and hsr203J was also induced by inoculation with both bacteria, but these plant reactions were more rapidly induced in eggplant inoculated with P. syringae pv. tabaci rather than those with P. cichorii. Lipid peroxidation and induction of lipoxygenase (LOX) was drastically induced in eggplant inoculated with P. syringae pv. tabaci compared to P. cichorii-inoculated eggplant. Pharmacological studies showed that induction of the cell death, and the NLS or the HR in response to both bacteria was commonly associated with de novo protein synthesis, reactive oxygen species and caspase III-like protease. Interestingly, involvement of lipid peroxidation, LOX, serine protease, and DNase differed between induction of NLS and HR. These results suggest that programmed cell death might be closely associated not only with the HR but also NLS. However, there may be differences not only in the induction kinetics and level of plant responses but also in the infection-related responses between HR and NLS.     

Polyamine metabolism and lipoxygenase activity during Fusarium oxysporum f. sp. ricini -Castor interaction

Polyamine oxidase and lipoxygenase enzymes are key players for hyper sensitive reaction (HR) during incompatible interaction of host-pathogen. Thus, the role of lipoxygenase and polyamines was studied in the wilt pathogen infected and non infected tissues of resistant and susceptible genotypes of castor at 0 days after infection (DAI), 5 DAI and 10 DAI (30 days after sowing). The lipoxygenase (LOX) and polyamine oxidase (PAO) activities were higher in the incompatible interaction at all the stages of analysis. The constitutive level of malondyaldehyde (MDA) content, a product of lipid peroxidation was higher in susceptible genotypes (VP-1 and VI-9), while induced level was higher in resistant genotypes (48–1 and SKP-84) at 5 DAI and 10 DAI . Polyamine profiling using HPTLC showed higher spermidine and spermine content in resistant genotypes at 10 DAI. Furthermore, spermidine was detected only in the roots of resistant genotypes at 10 DAI. These results suggest the role of high titers of polyamines, LOX and PAO in disease resistance possibly through HR induction.     

Enzymatic and non-enzymatic lipid peroxidation in leaf development.

Enzymatic and non-enzymatic lipid peroxidation has been implicated in programmed cell death, which is a major process of leaf senescence. To test this hypothesis we developed a high-performance liquid chromatography (HPLC) method for a simultaneous analysis of the major hydro(pero)xy polyenoic fatty acids. Quantities of lipid peroxidation products in leaves of different stages of development including natural senescence indicated a strong increase in the level of oxygenated polyenoic fatty acids (PUFAs) during the late stages of leaf senescence. Comprehensive structural elucidation of the oxygenation products by means of HPLC, gas chromatography/mass spectrometry and (1)H nuclear magnetic resonance suggested a non-enzymatic origin. However, in some cases a small share of specifically oxidized PUFAs was identified suggesting involvement of lipid peroxidizing enzymes. To inspect the possible role of enzymatic lipid peroxidation in leaf senescence, we analyzed the abundance of lipoxygenases (LOXs) in rosette leaves of Arabidopsis. LOXs and their product (9Z,11E,13S,15Z)-13-hydroperoxy-9,11,15-octadecatrienoic acid were exclusively detected in young green leaves. In contrast, in senescing leaves the specific LOX products were overlaid by large amounts of stereo-random lipid peroxidation products originating from non-enzymatic oxidation. These data indicate a limited contribution of LOXs to total lipid peroxidation, and a dominant role of non-enzymatic lipid peroxidation in late stages of leaf development.     

The function of very long chain polyunsaturated fatty acids in the pineal gland

The mammalian pineal gland is a prominent secretory organ with a high metabolic activity. Melatonin (N-acetyl-5-methoxytryptamine), the main secretory product of the pineal gland, efficiently scavenges both the hydroxyl and peroxyl radicals counteracting lipid peroxidation in biological membranes. Approximately 25% of the total fatty acids present in the rat pineal lipids are represented by arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3). These very long chain polyunsaturated fatty acids play important roles in the pineal gland. In addition to the production of melatonin, the mammalian pineal gland is able of convert these polyunsaturated fatty acids into bioactive lipid mediators. Lipoxygenation is the principal lipoxygenase (LOX) activity observed in the rat pineal gland. Lipoxygenation in the pineal gland is exceptional because no other brain regions express significant LOX activities under normal physiological conditions. The rat pineal gland expresses both 12- and 15-lipoxygenase (LOX) activities, producing 12- and 15-hydroperoxyeicosatetraenoic acid (12- and 15-HpETE) from arachidonic acid and 14- and 17-hydroxydocosahexaenoic acid (14- and 17-HdoHE) from docosahexaenoic acid, respectively. The rat pineal also produces hepoxilins via LOX pathways. The hepoxilins are bioactive epoxy-hydroxy products of the arachidonic acid metabolism via the 12S-lipoxygenase (12S-LOX) pathway. The two key pineal biochemical functions, lipoxygenation and melatonin synthesis, may be synergistically regulated by the status of n-3 essential fatty acids.     

Self-consuming innate immunity in Arabidopsis

Programmed cell death (PCD) associated with the pathogen-induced hypersensitive response (HR) is a hallmark of plant innate immunity. HR PCD is triggered upon recognition of pathogen effector molecules by host immune receptors either directly or indirectly via effector modulation of host targets. However, it has been unclear by which molecular mechanisms plants execute PCD during innate immune responses. We recently examined HR PCD in autophagy-deficient Arabidopsis knockout mutants (atg) and find that PCD conditioned by one class of plant innate immune receptors is suppressed in atg mutants. Intriguingly, HR triggered by another class of immune receptors with different genetic requirements is not compromised, indicating that only a specific subset of immune receptors engage the autophagy pathway for HR execution. Thus, our work provides a primary example of autophagic cell death associated with innate immune responses in eukaryotes as well as of pro-death functions for the autophagy pathway in plants.     

Effect of cadmium and temperature on the lipoxygenase activity in barley root tip

An analysis of different cell fractions isolated from barley roots revealed that lipoxygenase (LOX) activity occurred both extra- and intracellulary. Cadmium (Cd)-induced LOX activity was observed in the fraction containing cell walls, plasma membrane and the cytoplasm. High temperature-induced root growth inhibition and elevated LOX activity did not induce lipid peroxidation. In contrast, Cd inhibited root growth and caused both enhanced lipid peroxidation and elevated LOX activity at each of the temperatures analyzed. Spatial distribution studies revealed that the patterns of apoplastic LOX activity were different from those of cytoplasmic activity. Cd-induced intracellular LOX activity increased equally along the barley root tip, while Cd-induced apoplastic LOX activity was associated mainly with the differentiation zone of the barley root tip. Our results suggest the involvement of Cd-induced LOX activity in the premature differentiation of the barley root tip during Cd stress. We hypothesize that the role of LOX in plant metabolic processes in the root may depend on the level of reactive oxygen species in the roots: at physiological concentrations of ROS, LOX may be involved in the processes of root growth, while at the elevated harmful concentrations of ROS induced by different stress conditions, it may be involved in root growth inhibition through ectopic differentiation.