Interaction of Acinetobacter sp. RIT 592 induces the production of broad-spectrum antibiotics in Exiguobacterium sp. RIT 594.

Antimicrobial resistance (AMR) is one of the most alarming global public health challenges of the 21st century. Over 3 million antimicrobial-resistant infections occur in the United States annually, with nearly 50,000 cases being fatal. Innovations in drug discovery methods and platforms are crucial to identify novel antibiotics to combat AMR. We present the isolation and characterization of potentially novel antibiotic lead compounds produced by the cross-feeding of two rhizosphere bacteria, Acinetobacter sp. RIT 592 and Exiguobacterium sp. RIT 594. We used solid-phase extraction (SPE) followed by liquid chromatography (LC) to enrich antibiotic extracts and subsequently mass spectrometry (MS) analysis of collected fractions for compound structure identification and characterization. The MS data were processed through the Global Natural Product Social Molecular Networking (GNPS) database. The supernatant from RIT 592 induced RIT 594 to produce a cocktail of antimicrobial compounds active against Gram-positive and negative bacteria. The GNPS analysis indicated compounds with known antimicrobial activity in the bioactive samples, including oligopeptides and their derivatives. This work emphasizes the utility of microbial community-based platforms to discover novel clinically relevant secondary metabolites. Future work includes further structural characterization and antibiotic activity evaluation of the individual compounds against pathogenic multidrug-resistant (MDR) bacteria.

Oxylipin biosynthetic gene families of Cannabis sativa.

Cannabis sativa is a global multi-billion-dollar cash crop with numerous industrial uses, including in medicine and recreation where its value is largely owed to the production of pharmacological and psychoactive metabolites known as cannabinoids. Often underappreciated in this role, the lipoxygenase (LOX)-derived green leaf volatiles (GLVs), also known as the scent of cut grass, are the hypothetical origin of hexanoic acid, the initial substrate for cannabinoid biosynthesis. The LOX pathway is best known as the primary source of plant oxylipins, molecules analogous to the eicosanoids from mammalian systems. These molecules are a group of chemically and functionally diverse fatty acid-derived signals that govern nearly all biological processes including plant defense and development. The interaction between oxylipin and cannabinoid biosynthetic pathways remains to be explored. Despite their unique importance in this crop, there has not been a comprehensive investigation focusing on the genes responsible for oxylipin biosynthesis in any Cannabis species. This study documents the first genome-wide catalogue of the Cannabis sativa oxylipin biosynthetic genes and identified 21 LOX, five allene oxide synthases (AOS), three allene oxide cyclases (AOC), one hydroperoxide lyase (HPL), and five 12-oxo-phytodienoic acid reductases (OPR). Gene collinearity analysis found chromosomal regions containing several isoforms maintained across Cannabis, Arabidopsis, and tomato. Promoter, expression, weighted co-expression genetic network, and functional enrichment analysis provide evidence of tissue- and cultivar-specific transcription and roles for distinct isoforms in oxylipin and cannabinoid biosynthesis. This knowledge facilitates future targeted approaches towards Cannabis crop improvement and for the manipulation of cannabinoid metabolism.

Transgenic Soybeans Expressing Phosphatidylinositol-3-Phosphate-Binding Proteins Show Enhanced Resistance Against the Oomycete Pathogen Phytophthora sojae.

Oomycete and fungal pathogens cause billions of dollars of damage to crops worldwide annually. Therefore, there remains a need for broad-spectrum resistance genes, especially ones that target pathogens but do not interfere with colonization by beneficial microbes. Motivated by evidence suggesting that phosphatidylinositol-3-phosphate (PI3P) may be involved in the delivery of some oomycete and fungal virulence effector proteins, we created stable transgenic soybean plants that express and secrete two different PI3P-binding proteins, GmPH1 and VAM7, in an effort to interfere with effector delivery and confer resistance. Soybean plants expressing the two PI3P-binding proteins exhibited reduced infection by the oomycete pathogen Phytophthora sojae compared to control lines. Measurements of nodulation by nitrogen-fixing mutualistic bacterium Bradyrhizobium japonicum, which does not produce PI3P, revealed that the two lines with the highest levels of GmPH1 transcripts exhibited reductions in nodulation and in benefits from nodulation. Transcriptome and plant hormone measurements were made of soybean lines with the highest transcript levels of GmPH1 and VAM7, as well as controls, following P. sojae- or mock-inoculation. The results revealed increased levels of infection-associated transcripts in the transgenic lines, compared to controls, even prior to P. sojae infection, suggesting that the plants were primed for increased defense. The lines with reduced nodulation exhibited elevated levels of jasmonate-isoleucine and of transcripts of a JAR1 ortholog encoding jasmonate-isoleucine synthetase. However, lines expressing VAM7 transgenes exhibited normal nodulation and no increases in jasmonate-isoleucine. Overall, together with previously published data from cacao and from P. sojae transformants, the data suggest that secretion of PI3P-binding proteins may confer disease resistance through a variety of mechanisms.

Maize biochemistry in response to root herbivory was mediated by domestication, spread, and breeding.

MAIN CONCLUSION: With domestication, northward spread, and breeding, maize defence against root-herbivores relied on induced defences, decreasing levels of phytohormones involved in resistance, and increasing levels of a phytohormone involved in tolerance. We addressed whether a suite of maize (Zea mays mays) phytohormones and metabolites involved in herbivore defence were mediated by three successive processes: domestication, spread to North America, and modern breeding. With those processes, and following theoretical predictions, we expected to find: a change in defence strategy from reliance on induced defences to reliance on constitutive defences; decreasing levels of phytohormones involved in herbivore resistance, and; increasing levels of a phytohormone involved in herbivore tolerance. We tested those predictions by comparing phytohormone levels in seedlings exposed to root herbivory by Diabrotica virgifera virgifera among four plant types encompassing those processes: the maize ancestor Balsas teosinte (Zea mays parviglumis), Mexican maize landraces, USA maize landraces, and USA inbred maize cultivars. With domestication, maize transitioned from reliance on induced defences in teosinte to reliance on constitutive defences in maize, as predicted. One subset of metabolites putatively involved in herbivory defence (13-oxylipins) was suppressed with domestication, as predicted, though another was enhanced (9-oxylipins), and both were variably affected by spread and breeding. A phytohormone (indole-3-acetic acid) involved in tolerance was enhanced with domestication, and with spread and breeding, as predicted. These changes are consistent with documented changes in herbivory resistance and tolerance, and occurred coincidentally with cultivation in increasingly resource-rich environments, i.e., from wild to highly enriched agricultural environments. We concluded that herbivore defence evolution in crops may be mediated by processes spanning thousands of generations, e.g., domestication and spread, as well as by processes spanning tens of generations, e.g., breeding and agricultural intensification.

Fusarium verticillioides Induces Maize-Derived Ethylene to Promote Virulence by Engaging Fungal G-Protein Signaling.

Seed maceration and contamination with mycotoxin fumonisin inflicted by Fusarium verticillioides is a major disease concern for maize producers worldwide. Meta-analyses of quantitative trait loci for Fusarium ear rot resistance uncovered several ethylene (ET) biosynthesis and signaling genes within them, implicating ET in maize interactions with F. verticillioides. We tested this hypothesis using maize knockout mutants of the 1-aminocyclopropane-1-carboxylate (ACC) synthases ZmACS2 and ZmACS6. Infected wild-type seed emitted five-fold higher ET levels compared with controls, whereas ET was abolished in the acs2 and acs6 single and double mutants. The mutants supported reduced fungal biomass, conidia, and fumonisin content. Normal susceptibility was restored in the acs6 mutant with exogenous treatment of ET precursor ACC. Subsequently, we showed that fungal G-protein signaling is required for virulence via induction of maize-produced ET. F. verticillioides Gß subunit and two regulators of G-protein signaling mutants displayed reduced seed colonization and decreased ET levels. These defects were rescued by exogenous application of ACC. We concluded that pathogen-induced ET facilitates F. verticillioides colonization of seed, and, in turn, host ET production is manipulated via G-protein signaling of F. verticillioides to facilitate pathogenesis.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Amino acid-derived defense metabolites from plants: A potential source to facilitate novel antimicrobial development.

For millennia, humanity has relied on plants for its medicines, and modern pharmacology continues to reexamine and mine plant metabolites for novel compounds and to guide improvements in biological activity, bioavailability, and chemical stability. The critical problem of antibiotic resistance and increasing exposure to viral and parasitic diseases has spurred renewed interest into drug treatments for infectious diseases. In this context, an urgent revival of natural product discovery is globally underway with special attention directed toward the numerous and chemically diverse plant defensive compounds such as phytoalexins and phytoanticipins that combat herbivores, microbial pathogens, or competing plants. Moreover, advancements in "omics," chemistry, and heterologous expression systems have facilitated the purification and characterization of plant metabolites and the identification of possible therapeutic targets. In this review, we describe several important amino acid-derived classes of plant defensive compounds, including antimicrobial peptides (e.g., defensins, thionins, and knottins), alkaloids, nonproteogenic amino acids, and phenylpropanoids as potential drug leads, examining their mechanisms of action, therapeutic targets, and structure-function relationships. Given their potent antibacterial, antifungal, antiparasitic, and antiviral properties, which can be superior to existing drugs, phytoalexins and phytoanticipins are an excellent resource to facilitate the rational design and development of antimicrobial drugs.

Differential Evolution of α-Glucan Water Dikinase (GWD) in Plants.

The alpha-glucan water dikinase (GWD) enzyme catalyzes starch phosphorylation, an integral step in transitory starch degradation. The high phosphate content in stored starch has great industrial value, due to its physio-chemical properties making it more versatile, although the phosphate content of stored starch varies depending on the botanical source. In this study, we used various computational approaches to gain insights into the evolution of the GWD protein in 48 plant species with possible roles in enzyme function and alteration of phosphate content in their stored starch. Our analyses identified deleterious mutations, particularly in the highly conserved 5 aromatic amino acid residues in the dual tandem carbohydrate binding modules (CBM-45) of GWD protein in C. zofingiensis, G. hirsutum, A. protothecoides, P. miliaceum, and C. reinhardtii. These findings will inform experimental designs for simultaneous repression of genes coding for GWD and the predicted interacting proteins to elucidate the role this enzyme plays in starch degradation. Our results reveal significant diversity in the evolution of GWD enzyme across plant species, which may be evolutionarily advantageous according to the varying needs for phosphorylated stored starch between plants and environments.

Relative contribution of LOX10, green leaf volatiles and JA to wound-induced local and systemic oxylipin and hormone signature in Zea mays (maize).

Green leaf volatiles (GLVs) and jasmonates (JAs) are the best-characterized groups of fatty acid-derived oxylipin signals that regulate wound-associated defenses. Beyond these two major groups of defense signals, plants produce an array of oxylipins in response to wounding, which possess potent signaling and/or insecticidal activities. In this study, we assessed the relative contribution of JAs and GLVs to wound-induced systemic signaling and the associated regulation of oxylipins in local and systemic tissues of maize (Zea mays). For this, we utilized GLV- and JA-deficient mutants, lox10 single and opr7opr8 double mutants, respectively, and profiled oxylipins in untreated leaves and roots, and in locally wounded and systemic leaves. In contrast to the studies in dicots, no systemic induction of JAs was observed in maize. Instead, a JA precursor, 12-OPDA, as well as ketols and C12/13 oxo-acids derived from 13-lipoxygenases (LOXs), were preferentially induced in both locally wounded and systemic unwounded leaves. Several 9-LOX-derived oxylipins (9-oxylipins) including hydroxides and ketones were also significantly induced locally. JA and JA-isoleucine (JA-Ile) were rapidly induced within 0.5 h, and were followed by a second increase in local tissue 4 h after wounding. GLV-deficient lox10 mutants displayed reduced levels of most 13-oxylipins, and elevated levels of several 9-oxylipins and the a-dioxygenase (DOX) product, 2-HOD. lox10 mutants were completely devoid of C6 volatiles and their C12 counterparts, and greatly decreased in C5 volatiles and their C13 oxo-acid counterparts. Thus, in addition to being the sole LOX isoform providing substrate for GLV synthesis, LOX10 is a major 13-LOX that provides substrate to several LOX branches that produce an array of 13-oxylipin products, including C5 volatiles. Interestingly, the rapid JA and JA-Ile increase at 0.5-2 h post-wounding was only moderately affected by the LOX10 mutation, while significantly reduced levels were observed at 4 h post-wounding. Combined with the previous findings that GLVs activate JA biosynthesis, these results suggest that both LOX10-derived substrates and/or GLVs are involved in the large second phase of JA synthesis proximal to the wound. Analyses of opr7opr8 mutants revealed that wound-induced oxylipin responses were positively regulated by JA signaling. The local and systemic accumulation of SA was not altered in the two mutants. Collectively, our results identified a subset of oxylipins strongly induced in wounded and systemic leaves, but their impact on insect defenses remain elusive. The lack of systemic induction of JAs points to substantial difference between systemic wound responses in studied dicots and maize. Our results show that GLV-deficiency and reduced JA in lox10 mutants had a greater impact on wound-induced local and systemic tissue oxylipin responses compared to the solely JA-deficient opr7opr8 double mutants. This suggests that GLVs or other LOX10-derived products heavily contribute to overall basal and wound-induced oxylipin responses. The specific roles of the GLV- and/or JA-dependent oxylipins in wound responses and defense remain to be further investigated by a combination of multiple orders of oxylipin-deficient mutants.

Oxylipins Other Than Jasmonic Acid Are Xylem-Resident Signals Regulating Systemic Resistance Induced by Trichoderma virens in Maize.

Multiple long-distance signals have been identified for pathogen-induced systemic acquired resistance, but mobile signals for symbiont-induced systemic resistance (ISR) are less well understood. We used ISR-positive and -negative mutants of maize (Zea mays) and the beneficial fungus Trichoderma virens and identified 12-oxo-phytodienoic acid (12-OPDA) and α-ketol of octadecadienoic acid (KODA) as important ISR signals. We show that a maize 13-lipoxygenase mutant, lox10, colonized by the wild-type T. virens (TvWT) lacked ISR response against Colletotrichum graminicola but instead displayed induced systemic susceptibility. Oxylipin profiling of xylem sap from T. virens-treated plants revealed that 12-OPDA and KODA levels correlated with ISR. Transfusing sap supplemented with 12-OPDA or KODA increased receiver plant resistance in a dose-dependent manner, with 12-OPDA restoring ISR of lox10 plants treated with TvWT or T. virens Δsm1, a mutant unable to induce ISR. Unexpectedly, jasmonic acid (JA) was not involved, as the JA-deficient opr7 opr8 mutant plants retained the capacity for T. virens-induced ISR. Transcriptome analysis of TvWT-treated maize B73 revealed upregulation of 12-OPDA biosynthesis and OPDA-responsive genes but downregulation of JA biosynthesis and JA response genes. We propose a model that differential regulation of 12-OPDA and JA in response to T. virens colonization results in ISR induction.

Rosette core fungal resistance in Arabidopsis thaliana.

MAIN CONCLUSION: Unlike rosette leaves, the mature Arabidopsis rosette core can display full resistance to Botrytis cinerea revealing the importance for spatial and developmental aspects of plant fungal resistance. Arabidopsis thaliana is a model host to investigate plant defense against fungi. However, many of the reports investigating Arabidopsis fungal defense against the necrotrophic fungus, Botrytis cinerea, utilize rosette leaves as host tissue. Here we report organ-dependent differences in B. cinerea resistance of Arabidopsis. Although wild-type Arabidopsis rosette leaves mount a jasmonate-dependent defense that slows fungal growth, this defense is incapable of resisting fungal devastation. In contrast, as the fungus spreads through infected leaf petioles towards the plant center, or rosette core, there is a jasmonate- and age-dependent fungal penetration blockage into the rosette core. We report evidence for induced and preformed resistance in the rosette core, as direct rosette core inoculation can also result in resistance, but at a lower penetrance relative to infections that approach the core from infected leaf petioles. The Arabidopsis rosette core displays a distinct transcriptome relative to other plant organs, and BLADE ON PETIOLE (BOP) transcripts are abundant in the rosette core. The BOP genes, with known roles in abscission zone formation, are required for full Arabidopsis rosette core B. cinerea resistance, suggesting a possible role for BOP-dependent modifications that may help to restrict fungal susceptibility of the rosette core. Finally, we demonstrate that cabbage and cauliflower, common Brassicaceae crops, also display leaf susceptibility and rosette core resistance to B. cinerea that can involve leaf abscission. Thus, spatial and developmental aspects of plant host resistance play critical roles in resistance to necrotrophic fungal pathogens and are important to our understanding of plant defense mechanisms.

CIRCADIAN CLOCK-ASSOCIATED1 Controls Resistance to Aphids by Altering Indole Glucosinolate Production.

CIRCADIAN CLOCK-ASSOCIATED1 (CCA1), a well-known central circadian clock regulator, coordinates plant responses to environmental challenges. Its daily rhythmic expression in Arabidopsis (Arabidopsis thaliana) confers host resistance to the caterpillar Trichoplusia ni However, it is unclear whether CCA1 plays a role in defense against phloem sap-feeding aphids. In this study, we showed that green peach aphid (Myzus persicae) displayed an intrinsic circadian feeding rhythm. Under constant light, wild-type Columbia-0 (Col-0) Arabidopsis plants coentrained with aphids in the same light/dark cycles exhibited greater antixenotic activity than plants preentrained in the opposite cycle from the aphids. Consistently, circadian mutants cca1-1, cca1-11, lhy-21, ztl-1, ztl-4, and lux-2 suffered more severe damage than Col-0 plants when infested by aphids, suggesting that the Arabidopsis circadian clock plays a defensive role. However, the arrhythmic CCA1 overexpression line (CCA1-OX) displayed strong antixenotic and antibiotic activities despite its loss of circadian regulation. Aphids feeding on CCA1-OX plants exhibited lower reproduction and smaller body size and weight than those on Col-0. Apparently, CCA1 regulates both clock-dependent and -independent defense responses. Systematic investigation based on bioinformatics analyses indicated that resistance to aphids in CCA1-OX plants was due primarily to heightened basal indole glucosinolate levels. Interestingly, aphid feeding induced alternatively spliced intron-retaining CCA1a/b transcripts, which are normally expressed at low levels, whereas expression of the major fully spliced CCA1 transcript remained largely unchanged. We hypothesize that posttranscriptional modulation of CCA1 expression upon aphid infestation maximizes the potential of circadian-mediated defense and stress tolerance while ensuring normal plant development.

Bioprospecting Plant Growth-Promoting Rhizobacteria That Mitigate Drought Stress in Grasses.

This study reports the application of a novel bioprospecting procedure designed to screen plant growth-promoting rhizobacteria (PGPR) capable of rapidly colonizing the rhizosphere and mitigating drought stress in multiple hosts. Two PGPR strains were isolated by this bioprospecting screening assay and identified as Bacillus sp. (12D6) and Enterobacter sp. (16i). When inoculated into the rhizospheres of wheat (Triticum aestivum) and maize (Zea mays) seedlings, these PGPR resulted in delays in the onset of plant drought symptoms. The plant phenotype responding to drought stress was associated with alterations in root system architecture. In wheat, both PGPR isolates significantly increased root branching, and Bacillus sp. (12D6), in particular, increased root length, when compared to the control. In maize, both PGPR isolates significantly increased root length, root surface area and number of tips when compared to the control. Enterobacter sp. (16i) exhibited greater effects in root length, diameter and branching when compared to Bacillus sp. (12D6) or the control. In vitro phytohormone profiling of PGPR pellets and filtrates using LC/MS demonstrated that both PGPR strains produced and excreted indole-3-acetic acid (IAA) and salicylic acid (SA) when compared to other phytohormones. The positive effects of PGPR inoculation occurred concurrently with the onset of water deficit, demonstrating the potential of the PGPR identified from this bioprospecting pipeline for use in crop production systems under drought stress.

Oxylipins from both pathogen and host antagonize jasmonic acid-mediated defence via the 9-lipoxygenase pathway in Fusarium verticillioides infection of maize.

Oxylipins are a newly emerging group of signals that serve defence roles or promote virulence. To identify specific host and fungal genes and oxylipins governing the interactions between maize and Fusarium verticillioides, maize wild-type and lipoxygenase3 (lox3) mutant were inoculated with either F. verticillioides wild-type or linoleate-diol-synthase 1-deleted mutant (ΔFvlds1D). The results showed that lox3 mutants were more resistant to F. verticillioides. The reduced colonization on lox3 was associated with reduced fumonisin production and with a stronger and earlier induction of ZmLOX4, ZmLOX5 and ZmLOX12. In addition to the reported defence function of ZmLOX12, we showed that lox4 and lox5 mutants were more susceptible to F. verticillioides and possessed decreased jasmonate levels during infection, suggesting that these genes are essential for jasmonic acid (JA)-mediated defence. Oxylipin profiling revealed a dramatic reduction in fungal linoleate diol synthase 1 (LDS1)-derived oxylipins, especially 8-HpODE (8-hydroperoxyoctadecenoic acid), in infected lox3 kernels, indicating the importance of this molecule in virulence. Collectively, we make the following conclusions: (1) LOX3 is a major susceptibility factor induced by fungal LDS1-derived oxylipins to suppress JA-stimulating 9-LOXs; (2) LOX3-mediated signalling promotes the biosynthesis of virulence-promoting oxylipins in the fungus; and (3) both fungal LDS1- and host LOX3-produced oxylipins are essential for the normal infection and colonization processes of maize seed by F. verticillioides.

Seed-Derived Ethylene Facilitates Colonization but Not Aflatoxin Production by Aspergillus flavus in Maize.

Ethylene (ET) emitted by plant tissues has been broadly reported to play important roles in plant development, response to environmental stresses and defense against certain pathogens. Recent evidence obtained from using in vitro fungal cultures exposed to ET suggested that exogenous ET may regulate the production of aflatoxin by Aspergilli. However, the function of endogenous, seed-derived ET has not been explored. In this study, we found that the maize lipoxygenase lox3 mutant, previously reported to be susceptible to Aspergillus spp., emitted greater levels of ET upon A. flavus infection, suggesting the potential involvement of endogenous ET in the susceptibility of maize to A. flavus. Supporting this idea, both colonization and conidiation of A. flavus were reduced in wild-type (WT) kernels treated with AgNO3, an ET synthesis inhibitor. There was no ET emission from non-viable kernels colonized by A. flavus, suggesting that living seed but not the fungus itself was the primary source of ET released upon infection with A. flavus. The kernels of acs2 and acs6, two ET biosynthetic mutants carrying Mutator transposons in the ACC synthase genes, ACS2 and ACS6, respectively, displayed enhanced seed colonization and conidiation, but not the levels of aflatoxin, upon infection with A. flavus. Surprisingly, both acs2 and acs6 mutant kernels emitted greater levels of ET in response to infection by A. flavus as compared with WT seed. The increased ET in single mutants was found to be due to overexpression of functional ACS genes in response to A. flavus infection. Collectively, these findings suggested that ET emitted by infected seed facilitates colonization by A. flavus but not aflatoxin production.

In vivo diagnostics of early abiotic plant stress response via Raman spectroscopy.

Development of a phenotyping platform capable of noninvasive biochemical sensing could offer researchers, breeders, and producers a tool for precise response detection. In particular, the ability to measure plant stress in vivo responses is becoming increasingly important. In this work, a Raman spectroscopic technique is developed for high-throughput stress phenotyping of plants. We show the early (within 48 h) in vivo detection of plant stress responses. Coleus (Plectranthus scutellarioides) plants were subjected to four common abiotic stress conditions individually: high soil salinity, drought, chilling exposure, and light saturation. Plants were examined poststress induction in vivo, and changes in the concentration levels of the reactive oxygen-scavenging pigments were observed by Raman microscopic and remote spectroscopic systems. The molecular concentration changes were further validated by commonly accepted chemical extraction (destructive) methods. Raman spectroscopy also allows simultaneous interrogation of various pigments in plants. For example, we found a unique negative correlation in concentration levels of anthocyanins and carotenoids, which clearly indicates that plant stress response is fine-tuned to protect against stress-induced damages. This precision spectroscopic technique holds promise for the future development of high-throughput screening for plant phenotyping and the quantification of biologically or commercially relevant molecules, such as antioxidants and pigments.

Synthesis and Functions of Jasmonates in Maize.

Of the over 600 oxylipins present in all plants, the phytohormone jasmonic acid (JA) remains the best understood in terms of its biosynthesis, function and signaling. Much like their eicosanoid analogues in mammalian system, evidence is growing for the role of the other oxylipins in diverse physiological processes. JA serves as the model plant oxylipin species and regulates defense and development. For several decades, the biology of JA has been characterized in a few dicot species, yet the function of JA in monocots has only recently begun to be elucidated. In this work, the synthesis and function of JA in maize is presented from the perspective of oxylipin biology. The maize genes responsible for catalyzing the reactions in the JA biosynthesis are clarified and described. Recent studies into the function of JA in maize defense against insect herbivory, pathogens and its role in growth and development are highlighted. Additionally, a list of JA-responsive genes is presented for use as biological markers for improving future investigations into JA signaling in maize.

Two closely related members of Arabidopsis 13-lipoxygenases (13-LOXs), LOX3 and LOX4, reveal distinct functions in response to plant-parasitic nematode infection.

The responses of two closely related members of Arabidopsis 13-lipoxygenases (13-LOXs), LOX3 and LOX4, to infection by the sedentary nematodes root-knot nematode (Meloidogyne javanica) and cyst nematode (Heterodera schachtii) were analysed in transgenic Arabidopsis seedlings. The tissue localization of LOX3 and LOX4 gene expression using ß-glucuronidase (GUS) reporter gene constructs showed local induction of LOX3 expression when second-stage juveniles reached the vascular bundle and during the early stages of plant-nematode interaction through gall and syncytia formation. Thin sections of nematode-infested knots indicated LOX3 expression in mature giant cells, and high expression in neighbouring cells and those surrounding the female body. LOX4 promoter was also activated by nematode infection, although the GUS signal weakened as infection and disease progressed. Homozygous insertion mutants lacking LOX3 were less susceptible than wild-type plants to root-knot nematode infection, as reflected by a decrease in female counts. Conversely, deficiency in LOX4 function led to a marked increase in females and egg mass number and in the female to male ratio of M. javanica and H. schachtii, respectively. The susceptibility of lox4 mutants was accompanied by increased expression of allene oxide synthase, allene oxide cyclase and ethylene-responsive transcription factor 4, and the accumulation of jasmonic acid, measured in the roots of lox4 mutants. This response was not found in lox3 mutants. Taken together, our results reveal that LOX4 and LOX3 interfere differentially with distinct metabolic and signalling pathways, and that LOX4 plays a major role in controlling plant defence against nematode infection.

Root-expressed maize lipoxygenase 3 negatively regulates induced systemic resistance to Colletotrichum graminicola in shoots.

We have previously reported that disruption of a maize root-expressed 9-lipoxygenase (9-LOX) gene, ZmLOX3, results in dramatic increase in resistance to diverse leaf and stalk pathogens. Despite evident economic significance of these findings, the mechanism behind this increased resistance remained elusive. In this study, we found that increased resistance of the lox3-4 mutants is due to constitutive activation of induced systemic resistance (ISR) signaling. We showed that ZmLOX3 lacked expression in leaves in response to anthracnose leaf blight pathogen Colletotrichum graminicola, but was expressed constitutively in the roots, thus, prompting our hypothesis: the roots of lox3-4 mutants are the source of increased resistance in leaves. Supporting this hypothesis, treatment of wild-type plants (WT) with xylem sap of lox3-4 mutant induced resistance to C. graminicola to the levels comparable to those observed in lox3-4 mutant. Moreover, treating mutants with the sap collected from WT plants partially restored the susceptibility to C. graminicola. lox3-4 mutants showed primed defense responses upon infection, which included earlier and greater induction of defense-related PAL and GST genes compared to WT. In addition to the greater expression of the octadecanoid pathway genes, lox3-4 mutant responded earlier and with a greater accumulation of H2O2 in response to C. graminicola infection or treatment with alamethicin. These findings suggest that lox3-4 mutants display constitutive ISR-like signaling. In support of this idea, root colonization by Trichoderma virens strain GV29-8 induced the same level of disease resistance in WT as the treatment with the mutant sap, but had no additional resistance effect in lox3-4 mutant. While treatment with T. virens GV29 strongly and rapidly suppressed ZmLOX3 expression in hydroponically grown WT roots, T. virens Δsml mutant, which is deficient in ISR induction, was unable to suppress expression of ZmLOX3, thus, providing genetic evidence that SM1 function in ISR, at least in part, by suppressing host ZmLOX3 gene. This study and the genetic tools generated herein will allow the identification of the signals regulating the induction of resistance to aboveground attackers by beneficial soil microorganisms in the future.