Specific Defense Responses Research Articles

The rice hydroperoxide lyase OsHPL3 functions in defense responses by modulating the oxylipin pathway

As important signal molecules, jasmonates (JAs) and green leaf volatiles (GLVs) play diverse roles in plant defense responses against insect pests and pathogens. However, how plants employ their specific defense responses by modulating the levels of JA and GLVs remains unclear. Here, we describe identification of a role for the rice HPL3 gene, which encodes a hydroperoxide lyase (HPL), OsHPL3/CYP74B2, in mediating plant-specific defense responses. The loss-of-function mutant hpl3-1 produced disease-resembling lesions spreading through the whole leaves. A biochemical assay revealed that OsHPL3 possesses intrinsic HPL activity, hydrolyzing hydroperoxylinolenic acid to produce GLVs. The hpl3-1 plants exhibited enhanced induction of JA, trypsin proteinase inhibitors and other volatiles, but decreased levels of GLVs including (Z)-3-hexen-1-ol. OsHPL3 positively modulates resistance to the rice brown planthopper [BPH, Nilaparvata lugens (Stål)] but negatively modulates resistance to the rice striped stem borer [SSB, Chilo suppressalis (Walker)]. Moreover, hpl3-1 plants were more attractive to a BPH egg parasitoid, Anagrus nilaparvatae, than the wild-type, most likely as a result of increased release of BPH-induced volatiles. Interestingly, hpl3-1 plants also showed increased resistance to bacterial blight (Xanthomonas oryzae pv. oryzae). Collectively, these results indicate that OsHPL3, by affecting the levels of JA, GLVs and other volatiles, modulates rice-specific defense responses against different invaders.

Hrip1, a novel protein elicitor from necrotrophic fungus, Alternaria tenuissima, elicits cell death, expression of defence‐related genes and systemic acquired resistance in tobacco

Here, we report the identification, purification, characterization and gene cloning of a novel hypersensitive response inducing protein secreted by necrotrophic fungus, Alternaria tenuissima, designated as hypersensitive response inducing protein 1 (Hrip1). The protein caused the formation of necrotic lesions that mimic a typical hypersensitive response and apoptosis-related events including DNA laddering. The protein-encoding gene was cloned by rapid amplification of cDNA ends (RACE) method. The sequence analysis revealed that the cDNA is 495 bp in length and the open reading frame (ORF) encodes for a polypeptide of 163 amino acids with theoretical pI of 5.50 and molecular weight of 17 562.5 Da. Hrip1 induced calcium influx, medium alkalinization, activation of salicylic acid-induced protein kinase and several defence-related genes after infiltration in tobacco leaves. Cellular damage, restricted to the infiltrated zone, occurred only several hours later, at a time when expression of defence-related genes was activated. After several days, systemic acquired resistance was also induced. The tobacco plant cells that perceived the Hrip1 generated a cascade of signals acting at local, short, and long distances, and caused the coordinated expression of specific defence responses in a way similar to hypersensitivity to tobacco mosaic virus. Thus, Hrip1 represents a powerful tool to investigate further the signals and their transduction pathways involved in induced disease resistance in necrotrophic fungi.

Monoterpenoid Plant-Plant Interactions Upon Herbivory

Monoterpene production by plants is not only dependent on the plant ability to produce secretory tissues where these molecules are accumulated. All plants are able to emit monoterpenes upon stress of both biotic and abiotic origin. Biotic stress induced by herbivory triggers the activation of signaling pathways eventually leading to the expression of genes coding for enzymes responsible for monoterpene emission. The emitted monoterpenes are used to defend the plant directly (in the case of terpenoid storing plants) and indirectly (for most of all other plants). Herbivore-induced monoterpene emission is perceived as a signal by neighboring plants. This activates specific defense responses in primed plants. This minireview describes herbivore-induced monoterpene production and the effect of emitted monoterpenes on plants with particular reference to calcium signaling. Keywords: Monoterpene, herbivores, calcium signaling, plant-plant interaction, signaling pathway, volatile, calcium ions, insect mediated pollination, biotic stress, insect feeding

Biotic stress in plants: life lessons from your parents and grandparents

Biotic stress in plants: life lessons from your parents and grandparents

Participation of T Lymphocytes in the Development and Perpetuation of Rheumatoid Arthritis

The emphasis on T lymphocytes (TL) as an engine or perpetuating factor in rheumatoid arthritis (RA) has changed over time. The TL constitute the most plastic specific or adaptive immune system cells, both in their ability to recognize different antigens as in their various forms of response. Thus TL are responsible for orchestrating the most powerful and specific defensive responses, but also overall autoimmune responses, them being antigen-specific (autoantigens). However, insufficient identification of autoantigens responsible for autoimmune T-cell responses, particularly in RA, has been a major constraint to understand these responses. While rheumatoid factor and more recently the anti-CCP antibodies (antiCCP) are well known B responses as well as easy to detect in patients, this does not happen with any T cell response in RA. Some animal models illustrate, however, that it is possible to induce Tdependent arthritis in the absence of antigen-specific T cells by simply expanding or increasing the activation in a poly-specific manner. This occurs, for example, in response to genetic manipulation of factors that amplify global intracellular activation of the TCR receptor (ZAP70) or cytokine receptors (gp130).1,2 Another source of skepticism about T cells dependence in RA was the development of T immunosuppressive drugs and strategies that have proven very effective in preventing transplant rejection, but had little impact on RA therapy. The advent of effective antiTNF, paradoxically a TL activator, moved the attention to innate immune response cells. Despite these questions, two facts strongly support the importance of TL participation in both susceptibility and the development of the disease. The first is related to advances of genetic studies, particularly whole genome studies. Many other associations, whose functional significance point unequivocally to a TL response, have joined the classic association of RA and the shared epitope by different alleles of HLA class II genes. While the shared epitope is the platform for interaction between HLAantigen for presentation to T-cell receptors (TCR), other genes are known to encode elements such as receptors, or intracellular soluble factors that regulate multiple functions specific to TL.

Systemic induction of a Capsicum chinense nitrate reductase by the infection with Phytophthora capsici and defence phytohormones

The mRNA differential display technique was used to identify genes from Habanero pepper (Capsicum chinense Jacq.) seedlings whose expression is modified systemically by infection with the oomycete Phytophthora capsici L. Experiments with different oligonucleotide primer combinations revealed that no single gene was synthesised de novo. Instead, the quantitative accumulation of multiple transcripts was found. From these transcripts, levels of a nitrate reductase (Capsicum chinense nitrate reductase, CcNR), which has a high percentage of identity with other Solanaceae NRs, showed a consistent increase a few hours after inoculation (hai) with P.capsici. Reverse northern blotting revealed the existence of basal levels of CcNR transcripts in different adult tissues; however, systemic levels rose dramatically after spraying seedlings with salicylic acid (SA) and ethephon (ET) but not with methyl jasmonate (MeJa). Both P.capsici and defence phytohormones (DP) also modified NR enzymatic activity (nitrite:NAD(+) oxidoreductase; EC 1.7.1.1) with similar kinetics. Because the application of DP induced and activated the CcNR differentially, it is possible that the activity of CcNR is related to a specific host defence response.

Negative regulation of defence signalling pathways by the EDR1 protein kinase

The enhanced disease resistance 1 (edr1) mutant of Arabidopsis confers enhanced resistance to bacterial and fungal pathogens. To better understand how edr1-mediated resistance occurs, we performed transcriptome analyses on wild-type and edr1 plants inoculated with the fungal pathogen Golovinomyces cichoracearum (powdery mildew). The expression of many known and putative defence-associated genes was more rapidly induced, and to higher levels, in edr1 plants relative to the wild-type. Many of the genes with elevated expression encoded WRKY transcription factors and there was enrichment for their binding sites in promoters of the genes upregulated in edr1. Confocal microscopy of transiently expressed EDR1 protein showed that a significant fraction of EDR1 was localized to the nucleus, suggesting that EDR1 could potentially interact with transcription factors in the nucleus. Analysis of gene ontology annotations revealed that genes associated with the endomembrane system, defence, reactive oxygen species (ROS) production and protein kinases were induced early in the edr1 mutant, and that elevated expression of the endomembrane system, defence and ROS-related genes was maintained for at least 4 days after infection.

Unraveling Plant Responses to Bacterial Pathogens through Proteomics

Plant pathogenic bacteria cause diseases in important crops and seriously and negatively impact agricultural production. Therefore, an understanding of the mechanisms by which plants resist bacterial infection at the stage of the basal immune response or mount a successful specific R-dependent defense response is crucial since a better understanding of the biochemical and cellular mechanisms underlying these interactions will enable molecular and transgenic approaches to crops with increased biotic resistance. In recent years, proteomics has been used to gain in-depth understanding of many aspects of the host defense against pathogens and has allowed monitoring differences in abundance of proteins as well as posttranscriptional and posttranslational processes, protein activation/inactivation, and turnover. Proteomics also offers a window to study protein trafficking and routes of communication between organelles. Here, we summarize and discuss current progress in proteomics of the basal and specific host defense responses elicited by bacterial pathogens.

Current trends in the evolutionary ecology of plant defence

Summary1. In this essay I summarize current trends in the evolutionary ecology of plant defence, while advocating for approaches that integrate community ecology with specific tests of classic evolutionary hypotheses. Several conclusions emerge.2. The microevolution of defence is perhaps best studied by reciprocal transplant experiments of differentiated plant populations while simultaneously manipulating the presence of the herbivore(s) hypothesized to be the agent(s) of natural selection.3.Although there is continued interest in the costs of defence, I argue that some empirical approaches to estimating costs (e.g. genetic engineering) may provide limited insight into evolutionary processes.4. Essentially all plants employ several different lines of defence against herbivory. It is thus time to abandon searching for single silver bullet traits and the simple trade‐off model (where traits are arbitrarily expected to negatively covary across genotypes or species). We still know very little about which trait combinations are most effective and have repeatedly evolved together. Thus, some of our prominent theories (e.g. a predicted trade‐off between direct and indirect defence) need to be revised.5. Studies of the macroevolution of plant defence are enjoying renewed interest due to available phylogenies and analytical methods. Although general trends are not currently surmisable, we will soon have strong case studies evaluating both biotic and abiotic drivers of convergent evolution in defence strategies and the role of defence evolution in the adaptive radiation of plant lineages.6. The evolution of specificity is proposed as a final frontier in understanding complexity in plant–herbivore interactions. Although it is abundantly clear that plants can deploy highly specific defensive responses that are differentially perceived by herbivore species, how such responses evolve and are physiologically regulated remains an important gap. Relatively straightforward methodologies are now available to close the loop between plant perception of herbivory, hormonal responses, and production of defensive end‐products across genotype or species.

Effects of elevated ultraviolet-B radiation on a plant–herbivore interaction

Enhanced ultraviolet-B (UV-B) radiation may have multiple effects on both plants and animals and affect plant-herbivore interactions directly and indirectly by inducing changes in host plant quality. In this study, we examined combined effects of UV-B and herbivory on the defence of the mountain birch (Betula pubescens ssp. czerepanovii) and also the effects of enhanced UV-B radiation on a geometrid with an outbreak cycle: the autumnal moth (Epirrita autumnata). We established an experiment mimicking ozone depletion of 30% (a relevant level when simulating ozone depletion above Northern Lapland). Both arctic species responded only slightly to the enhanced level of UV-B radiation, which may indicate that these species are already adapted to a broader range of UV-B radiation. UV-B exposure slightly induced the accumulation of myricetin glycosides but had no significant effect on the contents of quercetin or kaempferol derivatives. Mountain birch seedlings responded more efficiently to herbivory wounding than to enhanced UV-B exposure. Herbivory induced the activities of foliar oxidases that had earlier been shown to impair both feeding and growth of moth larvae. In contrast, the contents of foliar phenolics did not show the same response in different clones, except for a decrease in the contents of tannin precursors. The induction of foliar phenoloxidase activities is a specific defence response of mountain birches against insect herbivory. To conclude, our results do not support the hypothesis that the outbreak cycle of the autumnal moth can be explained by the cycles of solar activity and UV-B.

New insights into the early biochemical activation of jasmonic acid biosynthesis in leaves

In plants, herbivore attack elicits the rapid accumulation of jasmonic acid (JA) which results from the activation of constitutively expressed biosynthetic enzymes. The molecular mechanisms controlling the activation of JA biosynthesis remain largely unknown however new research has elucidated some of the early regulatory components involved in this process. Nicotiana attenuata plants, a wild tobacco species, responds to fatty acid amino acid conjuguates (FAC) elicitors in the oral secretion of its natural herbivore, Manduca sexta, by triggering specific defense and tolerance responses against it; all of the defense responses known to date require the amplification of the wound-induced JA increase. We recently demonstrated that this FAC-elicited JA burst requires an increased flux of free linolenic acid (18:3) likely originating from the activation of a plastidial glycerolipase (GLA1) which is activated by an abundant FAC found in insect oral secretions, N-linolenoyl-glutamate (18:3-Glu). The lack of accumulation of free 18:3 after elicitation suggests a tight physical association between GLA1 and LOX3 in N. attenuata leaves. In addition, the salicylate-induced protein kinase (SIPK) and the nonexpressor of PR-1 (NPR1) participate in this activation mechanism that controls the supply of 18:3. In contrast, the wound-induced protein kinase (WIPK) does not but instead regulates the conversion of 13(S)-hydroperoxy-18:3 into 12-oxo-phytodienoic acid (OPDA). These results open new perspectives on the complex network of signals and regulatory components inducing the JA biosynthetic pathway.

Broad-Spectrum Activity against Bacterial Mastitis Pathogens and Activation of Mammary Epithelial Cells Support a Protective Role of Neutrophil Cathelicidins in Bovine Mastitis

Cathelicidins are peptide components of the innate immune system of mammals. Apart from exerting a direct antibiotic activity, they can also trigger specific defense responses in the host. Their roles in various pathophysiological conditions have been studied, but there is a lack of published information on their expression and activities in the context of mastitis. The aims of this study were to investigate the expression of the bovine cathelicidins BMAP-27, BMAP-28, Bac5, and indolicidin in healthy and infected mammary tissue and in lipopolysaccharide (LPS)-treated cells, to determine their activities against bacteria isolated from bovine mastitis, and to examine their potentials to trigger defense responses in bovine mammary cells. The genes were found to be upregulated in LPS-stimulated neutrophils, but not in infected quarters or epithelial cells. All peptides showed a variably broad spectrum of activity against 28 bacterial isolates from bovine mastitis (MIC values, 0.5 to 32 microM), some of which were antibiotic resistant. The activity of each peptide was significantly enhanced when it was pairwise tested with the other peptides, reaching the synergy threshold when indolicidin was present. The bactericidal activity was sensitive to milk components; BMAP-27 and -28 were highly effective in mastitic bovine milk and inhibited in milk from healthy cows. Both peptides were also active in whey and in blood serum and triggered the expression of tumor necrosis factor alpha (TNF-alpha) in bovine mammary epithelial cells. Our results indicate multiple roles for the bovine cathelicidins in mastitis, with complementary and mutually enhanced antimicrobial activities against causative pathogens and the capacity to activate host cells.

UV-B impact on aphid performance mediated by plant quality and plant changes induced by aphids

Plants face various abiotic and biotic environmental factors and therefore need to adjust their phenotypic traits on several levels. UV-B radiation is believed to impact herbivorous insects via host plant changes. Plant responses to abiotic challenges (UV-B radiation) and their interaction with two aphid species were explored in a multifactor approach. Broccoli plants [Brassica oleracea L. convar. botrytis (L.), Brassicaceae] were grown in two differently covered greenhouses, transmitting either 80% (high UV-B) or 4% (low UV-B) of ambient UV-B. Three-week-old plants were infested with either specialist cabbage aphids [Brevicoryne brassicae (L.), Sternorrhyncha, Aphididae] or generalist green peach aphids [Myzus persicae (Sulzer), Sternorrhyncha, Aphididae]. Plants grown under high-UV-B intensities were smaller and had higher flavonoid concentrations. Furthermore, these plants had reduced cuticular wax coverage, whereas amino acid concentrations of the phloem sap were little influenced by different UV-B intensities. Cabbage aphids reproduced less on plants grown under high UV-B than on plants grown under low UV-B, whereas reproduction of green peach aphids in both plant light sources was equally poor. These results are likely related to the different specialisation-dependent sensitivities of the two species. The aphids also affected plant chemistry. High numbers of cabbage aphid progeny on low-UV-B plants led to decreased indolyl glucosinolate concentrations. The induced change in these glucosinolates may depend on an infestation threshold. UV-B radiation considerably impacts plant traits and subsequently affects specialist phloem-feeding aphids, whereas aphid growth forces broccoli to generate specific defence responses.

Differential gene expression in whiteflyBemisia tabaci‐infested tomato (Solanum lycopersicum) plants at progressing developmental stages of the insect's life cycle

A suppression-subtractive-hybridization (SSH) strategy was used to identify genes whose expression was modified in response to virus-free whitefly Bemisia tabaci (Bt, biotype A) infestation in tomato (Solanum lycopersicum) plants. Thus, forward and reverse SSH gene libraries were generated at four points in the whitefly's life cycle, namely at (1) 2 days (adult feeding and oviposition: phase I); (2) 7 days (mobile crawler stage: phase II); (3) 12 days (second to third instar nymphal transition: phase III) and (4) 18 days (fourth instar nymphal stage: phase IV). The 169 genes with altered expression (up and downregulated) that were identified in the eight generated SSH libraries, together with 75 additional genes that were selected on the basis of their involvement in resistance responses against phytofagous insects and pathogens, were printed on a Nexterion(®) Slide MPX 16 to monitor their pattern of expression at the above phases. The results indicated that Bt infestation in tomato led to distinctive phase-specific expression/repression patterns of several genes associated predominantly with photosynthesis, senescence, secondary metabolism and (a)biotic stress. Most of the gene expression modifications were detected in phase III, coinciding with intense larval feeding, whereas fewer changes were detected in phases I and IV. These results complement previously reported gene expression profiles in Bt-infested tomato and Arabidopisis, and support and expand the opinion that Bt infestation leads to the downregulation of specific defense responses in addition to those controlled by jasmonic acid.

BIOCONTROL OF PLANT DISEASES: PRODUCTION AND TESTING OF NOVEL ELICITORS INDUCING PLANT DEFENSE RESPONSE

The need of slowing down the incidence of fungal infections in the strawberry production sector and the urge for natural means to control these pathogens have led us to consider the production of natural compounds from plant extracts capable of inducing a specific defence response (elicitors). New elicitors obtained by a biotechnological production process of polysaccharide libraries followed by a quick molecular analysis and screening of these compounds are currently being studied. These natural means against fungal diseases avoid or minimize the application of chemical treatments, thus promoting the ecological feature of the farming, which makes the product more competitive on the international market.

Virulence mechanisms of Moraxella in the pathogenesis of infection

Moraxella catarrhalis is an emerging human-specific pathogen responsible for upper and lower respiratory tract infections. Understanding the events in the complex pathogenesis and underlying mechanisms during M. catarrhalis infection is a key to the development of novel therapeutics and vaccines. Several novel findings have been reported on Moraxella pathogenesis and, in parts, explain how the species stands as a commensal in preschool children and survives in the host. Molecular structures for different adhesins in addition to target ligands with respect to signalling and invasion have been defined. Evasion of the complement system allows Moraxella to survive in the mucosa and by neutralizing [alpha]1-antichymotrypsin the protease activity is increased, resulting in tissue destruction and thus promotion of bacterial attachment. Moraxella-dependent cell activation via immunoglobulin D in addition to toll-like receptors and specific epithelial cell inhibition by cross-linking of carcinoembryonic antigen-related cell adhesion molecule-1 in the early innate immune response and, finally, the ability of M. catarrhalis to form biofilms are other specific research areas of interest. Recent advances have allowed a more detailed picture of the processes involved in bacteria-host cell interactions, the cause of inflammatory processes and specific host defense responses against the intriguing species Moraxella.

β‐Aminobutyric Acid‐induced Resistance in Cucumber against Biotrophic and Necrotrophic Pathogens

AbstractThe non‐protein amino acid β‐aminobutyric acid (BABA) protects plants against a wide range of pathogens. Protection of cucumber plants by BABA depends on the potentiation of pathogen specific defence responses. To contribute to the analyses of the mode of action of BABA, we established a protocol for a fast and reproducible leaf disc assay to evaluate the effect of this chemical compound on cucumbers infected with either the biotrophic downy mildew fungus Pseudoperonospora cubensis, or the necrotrophic microbial pathogen Colletotrichum lagenarium. Accumulation of callose could be found in interactions with both pathogens after BABA‐treatment. Furthermore, a localized rapid cell death and the production of reactive oxygen intermediates were detected after downy mildew attack. In contrast to this, degenerated primary hyphae were found in BABA induced tissue after inoculation with C. lagenarium.

Mécanismes de défense du nématodeC. elegans

The nematode Caenorhabditis elegans has evolved as a powerful invertebrate model to study innate immunity to pathogens. C. elegans possesses inducible defence mechanisms to protect itself from pathogenic attack, mainly by the production of antimicrobial effector molecules. Its innate immune system is under the control of a surprisingly complex network of evolutionary conserved signalling pathways, which are activated depending on the pathogen, suggesting that C. elegans is able to mount a specific defence response to different pathogens. In this review we will introduce the worm's immune system and discuss the different signalling pathways that regulate its response to bacterial pathogens which mainly infect C. elegans by an oral route and by invading its intestine, before focusing our attention on the resistance of C. elegans to a natural occurring fungal -pathogen that infects the worm by invading its -epidermis.

Phenolic metabolites in the resistance of northern forest trees to pathogens — past experiences and future prospects

Phenolic metabolites are frequently implicated in chemical defense mechanisms against pathogens in woody plants. However, tree breeding programmes for resistance to pathogens and practical tree-protection applications based on these compounds seem to be scarce. To identify gaps in our current knowledge of this subject, we explored some of the recent literature on the involvement of phenolic metabolites in the resistance of northern forest trees (Pinus, Picea, Betula, Populus, and Salix spp.) to pathogens. Although it is evident that the phenolic metabolism of trees is often activated by pathogen attacks, few studies have convincingly established that this induction is due to a specific defense response that is capable of stopping the invading pathogen. The role of constitutive phenolics in the resistance of trees to pathogens has also remained unclear. In future studies, the importance of phenolics in oxidative stress, cell homeostasis and tolerance, and the spatial and temporal localization of phenolics in relation to invading pathogens should be more carefully acknowledged. Possibilities for future studies using advanced methods (e.g., metabolic profiling, confocal laser scanning microscopy, and use of modified tree genotypes) are discussed.

Accumulation and depuration of cyanobacterial toxin nodularin and biomarker responses in the mussel Mytilus edulis

Blue mussels (Mytilus edulis) were exposed to an extract made of natural cyanobacterial mixture containing toxic cyanobacterium Nodularia spumigena (70-110 microg nodularin l(-1), 24-h exposure followed by 144-h depuration period in clean water). Toxin concentration increased from initial 400 to 1100 mg kg(-1) after 24-h exposure, measured by liquid chromatography/mass spectrometry (LC/MS). Acetylcholinesterase activity (AChE), a biomarker of direct neurotoxic effects, showed inhibition after 12 and 24h exposure but returned to control level during the depuration period. Catalase (CAT) activity, an indicator of oxidative stress, showed significantly elevated levels in exposed mussels but only 72 h after the end of the exposure. No change in the activity of glutathione-S-transferase (GST) involved in conjugation reactions could be observed. A gradual yet incomplete elimination of nodularin (from 1100 to 600 mg kg(-1)) was observed during the depuration period, and the tissue levels were 30% lower in clean water after 24 h. The observed increase in oxidative stress indicated by elevated CAT activity is likely connected to detoxification reactions leading to the production of reactive oxygen species, including an apparent time lag in this specific enzymatic defence response. That no change in GST activity was observed suggests that this enzyme is not significantly involved in the detoxification process of nodularin-containing cyanobacterial extract in M. edulis.