Stimulate Defense Responses Research Articles

Nano-enhanced defense: Titanium-enriched Alginate–Bentonite coating augments Bacillus amyloliquefaciens D203 efficacy against Magnaporthe oryzae in Kenyan rice cultivation

Rice blast disease, caused by Magnaporthe oryzae, poses a significant threat to global rice production, necessitating the development of effective and sustainable management strategies. Biological control using beneficial microbes like Bacillus amyloliquefaciens has emerged as a promising approach due to its ability to enhance plant resistance and reduce disease incidence. Nano-encapsulation of bacteria, which involves embedding beneficial microbes within nanomaterials, offers a novel method to improve the stability, survival, and efficacy of these biocontrol agents. This study evaluated the capacity of encapsulated Bacillus amyloliquefaciens D203, embedded within an alginate-bentonite coating infused with titanium nanoparticles (TNs), to stimulate defense responses in rice seedlings challenged by the Magnaporthe oryzae the causal agent of rice blast disease. Encapsulation was achieved using the extrusion technique, with some modifications. Using a completely randomized design, the experiment was conducted in a greenhouse, with four treatments replicated four times. The experiment used the popular Kenyan rice variety "BASMATI 370”. The study investigated the impact of strain D203 on the incidence, severity, and area under disease progress curves related to M. oryzae, as well as the expression of defense-related enzymes. The results demonstrated that rice plants derived from seeds coated with the D203 encapsulated B. amyloliquefaciens strain exhibited higher levels of defense-related enzyme expression, including peroxidase (POD), phenylalanine ammonia-lyase (PAL), superoxide dismutase (SOD) and catalase (CAT), compared to controls. In addition, the incidence and severity of the disease were markedly lower in plants treated with encapsulated B. amyloliquefaciens compared to controls, sometimes paralleling the efficacy of hexaconazole treatment. These findings suggest that the encapsulation of strain D203 has the potential to enhance resistance against rice blast disease by inducing systemic resistance through the production of antioxidant enzymes.

Transcriptomic data reveals the dynamics of terpenoids biosynthetic pathway of fenugreek

Medicinal plants are rich sources for treating various diseases due their bioactive secondary metabolites. Fenugreek (Trigonella foenum-graecum) is one of the medicinal plants traditionally used in human nutrition and medicine which contains an active substance, called diosgenin, with anticancer properties. Biosynthesis of this important anticancer compound in fenugreek can be enhanced using eliciting agents which involves in manipulation of metabolite and biochemical pathways stimulating defense responses. Methyl jasmonate elicitor was used to increase diosgenin biosynthesis in fenugreek plants. However, the molecular mechanism and gene expression profiles underlying diosgening accumulation remain unexplored. In the current study we performed an extensive analysis of publicly available RNA-sequencing datasets to elucidate the biosynthesis and expression profile of fenugreek plants treated with methyl jasmonate. For this purpose, seven read datasets of methyl jasmonate treated plants were obtained that were covering several post-treatment time points (6–120 h). Transcriptomics analysis revealed upregulation of several key genes involved in diosgenein biosynthetic pathway including Squalene synthase (SQS) as the first committed step in diosgenin biosynthesis as well as Squalene Epoxidase (SEP) and Cycloartenol Synthase (CAS) upon methyl jasmonate application. Bioinformatics analysis, including gene ontology enrichment and pathway analysis, further supported the involvement of these genes in diosgenin biosynthesis. The bioinformatics analysis led to a comprehensive validation, with expression profiling across three different fenugreek populations treated with the same methyl jasmonate application. Initially, key genes like SQS, SEP, and CAS showed upregulation, followed by later upregulation of Δ24, suggesting dynamic pathway regulation. Real-time PCR confirmed consistent upregulation of SQS and SEP, peaking at 72 h. Additionally, candidate genes Δ24 and SMT1 highlighted roles in directing metabolic flux towards diosgenin biosynthesis. This integrated approach validates the bioinformatics findings and elucidates fenugreek’s molecular response to methyl jasmonate elicitation, offering insights for enhancing diosgenin yield. The assembled transcripts and gene expression profiles are deposited in the Zenodo open repository at https://doi.org/10.5281/zenodo.8155183.

An apoptosis-inducing factor controls programmed cell death and laccase expression during fungal interactions

Apoptotic-like programmed cell death (PCD) is one of the main strategies for fungi to resist environmental stresses and maintain homeostasis. The apoptosis-inducing factor (AIF) has been shown in different fungi to trigger PCD through upregulating reactive oxygen species (ROS). This study identified a mitochondrial localized AIF homolog, CcAIF1, from Coprinopsis cinerea monokaryon Okayama 7. Heterologous overexpression of CcAIF1 in Saccharomyces cerevisiae caused apoptotic-like PCD of the yeast cells. Ccaif1 was increased in transcription when C. cinerea interacted with Gongronella sp. w5, accompanied by typical apoptotic-like PCD in C. cinerea, including phosphatidylserine externalization and DNA fragmentation. Decreased mycelial ROS levels were observed in Ccaif1 silenced C. cinerea transformants during cocultivation, as well as reduction of the apoptotic levels, mycelial growth, and asexual sporulation. By comparison, Ccaif1 overexpression led to the opposite phenotypes. Moreover, the transcription and expression levels of laccase Lcc9 decreased by Ccaif1 silencing but increased firmly in Ccaif1 overexpression C. cinerea transformants in coculture. Thus, in conjunction with our previous report that intracellular ROS act as signal molecules to stimulate defense responses, we conclude that CcAIF1 is a regulator of ROS to promote apoptotic-like PCD and laccase expression in fungal-fungal interactions. In an axenic culture of C. cinerea, CcAIF1 overexpression and H2O2 stimulation together increased laccase secretion with multiplied production yield. The expression of two other normally silent isozymes, Lcc8 and Lcc13, was unexpectedly triggered along with Lcc9.Key points• Mitochondrial CcAIF1 induces PCD during fungal-fungal interactions• CcAIF1 is a regulator of ROS to trigger the expression of Lcc9 for defense• CcAIF1 overexpression and H2O2 stimulation dramatically increase laccase production

Harnessing Chlorophyll Fluorescence for Phenotyping Analysis of Wild and Cultivated Tomato for High Photochemical Efficiency under Water Deficit for Climate Change Resilience

Fluctuations of the weather conditions, due to global climate change, greatly influence plant growth and development, eventually affecting crop yield and quality, but also plant survival. Since water shortage is one of the key risks for the future of agriculture, exploring the capability of crop species to grow with limited water is therefore fundamental. By using chlorophyll fluorescence analysis, we evaluated the responses of wild tomato accession Solanum pennellii LA0716, Solanum lycopersicum cv. Μ82, the introgression line IL12-4 (from cv. M82 Χ LA0716), and the Greek tomato cultivars cv. Santorini and cv. Zakinthos, to moderate drought stress (MoDS) and severe drought stress (SDS), in order to identify the minimum irrigation level for efficient photosynthetic performance. Agronomic traits (plant height, number of leaves and root/shoot biomass), relative water content (RWC), and lipid peroxidation, were also measured. Under almost 50% deficit irrigation, S. pennellii exhibited an enhanced photosynthetic function by displaying a hormetic response of electron transport rate (ETR), due to an increased fraction of open reaction centers, it is suggested to be activated by the low increase of reactive oxygen species (ROS). A low increase of ROS is regarded to be beneficial by stimulating defense responses and also triggering a more oxidized redox state of quinone A (QA), corresponding in S. pennellii under 50% deficit irrigation, to the lowest stomatal opening, resulting in reduction of water loss. Solanumpennellii was the most tolerant to drought, as it was expected, and could manage to have an adequate photochemical function with almost 30% water regime of well-watered plants. With 50% deficit irrigation, cv. Μ82 and cv. Santorini did not show any difference in photochemical efficiency to control plants and are recommended to be cultivated under deficit irrigation as an effective strategy to enhance agricultural sustainability under a global climate change. We conclude that instead of the previously used Fv/Fm ratio, the redox state of QA, as it can be estimated by the chlorophyll fluorescence parameter 1 - qL, is a better indicator to evaluate photosynthetic efficiency and select drought tolerant cultivars under deficit irrigation.

Delonix Regia Bark Alleviated Inflammation by Stimulating Defense Responses in Rats

ABSTRACT The methanol extract of Delonix regia bark (DMeOH) was investigated for its anti-inflammatory property in kappa-carrageenan (Carr)-induced hind paw edema of rats. DMeOH was orally administered and evaluated as a measure of edema, tissue destruction, and neutrophil migration to the inflammation site. The pro-inflammatory mediators, p-p65 subunit of NFκB, interleukin (IL)-1β, cycloxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) were analyzed. Catalase (CAT) and superoxide dismutase (SOD) enzyme activity assays and malondialdehye (MDA) were quantified. The LC-MS fingerprint of the extract identified 10 polyphenols, of which five were structurally characterized based on their [M-H]− peak, UV-vis absorption compared to published data. Anti-inflammatory studies revealed that pre-treatment with DMeOH (0.5 g kg−1) prior to Carr-injection inhibited paw edema abating tissue destruction with decreased neutrophil infiltration. Carr-injection resulted in increased NO(•-) production, iNOS transcript and p-p65 subunit of NF-κB protein. DMeOH feed inhibited NO(•-) production and iNOS transcript levels. A decreased expression of NF-κB p-p65 subunit, COX-2 and IL-1β at 0.5 g kg−1 b.w oral feed suggest that DMeOH exerted its anti-inflammatory property by inhibiting NFκB activation. An increase in the antioxidant enzyme activities of CAT (78%) and SOD (49%) with reduced MDA levels was recorded.

Foliar application of liquiritin protects Chinese flowering cabbage against cucumber mosaic virus and increases health-promoting compounds

ABSTRACT Decades of research have revealed notable similarities between the immune systems of the plant and animal kingdoms. Liquiritin has long been used to stimulate the body immunity in animals against an array of diseases. Considering the homology of some induced immune responses between animals and plants, we examined the effects of exogenously applied liquiritin to stimulate defense responses in Chinese flowering cabbage plants against cucumber mosaic virus (CMV) infection under greenhouse and field conditions. Foliar application of liquiritin (200 ppm) effectively suppressed the development of CMV symptoms by not less than 40% compared with the control in cabbage plants in both greenhouse and field trials along with the significant increases in the marketable yield and nutritional quality of cabbage. Liquiritin application enhanced the production of phenolic compounds and different defense-related enzymes in treated plants. Moreover, quantitative real-time PCR analysis revealed that liquiritin significantly up-regulated the expression of different defense-related genes upon pathogen inoculation, indicating an induction of the salicylic acid-mediated defense system. Collectively, the findings of this study indicate that liquiritin can effectively control CMV in cabbage plants.

Exogenous dopamine and overexpression of the dopamine synthase gene MdTYDC alleviated apple replant disease.

Apple replant disease (ARD) is a soil-borne disease that leads to economic losses due to reduced plant growth and diminished fruit yields. Dopamine is involved in interactions between plants and pathogens. However, it remains unclear whether dopamine can directly stimulate defense responses to ARD. In this study, an exogenous dopamine treatment and dopamine synthetase MdTYDC (tyrosine decarboxylase) transgenic plants were used to verify the role of dopamine in treating ARD. First, 2-year-old apple trees (Malus domestica cv. Fuji), grafted onto rootstock M26, were grown in replant soils. The addition of dopamine (100μM) to the soil promoted seedling growth and changed the accumulation of mineral elements in plants in replant soils. Such supplementation improved the activity of invertase, urease, proteinase and phosphatase under replant conditions. Sequencing analysis of 16S rDNA and internal transcribed spacer (ITS) rDNA revealed that dopamine had a slight influence on bacterial diversity but had an obvious effect on the fungal diversity in replant soils. The application of dopamine to replant soil changed the composition of bacterial and fungal communities. Second, overexpression of MdTYDC in apple plants alleviated the effects of ARD. MdTYDC transgenic lines exhibited mitigated ARD through inhibited degradation of photosynthetic pigment, maintaining the stability of photosystems I and II and improving the antioxidant system. Furthermore, overexpression of MdTYDC improved arbuscular mycorrhizal fungi colonization by improving the accumulation of soluble sugars under replant conditions. Together, these results demonstrated that dopamine enhances the tolerance of apples to ARD.

Elicitation: A biotechnological tool for enhanced production of secondary metabolites in hairy root cultures.

Elicitation is a possible aid to overcome various difficulties associated with the large-scale production of most commercially important bioactive secondary metabolites from wild and cultivated plants, undifferentiated or differentiated cultures. Secondary metabolite accumulation in vitro or their efflux in culture medium has been elicited in the undifferentiated or differentiated tissue cultures of several plant species by the application of a low concentration of biotic and abiotic elicitors in the last three decades. Hairy root cultures are preferred for the application of elicitation due to their genetic and biosynthetic stability, high growth rate in growth regulator-free media, and production consistence in response to elicitor treatment. Elicitors act as signal, recognized by elicitor-specific receptors on the plant cell membrane and stimulate defense responses during elicitation resulting in increased synthesis and accumulation of secondary metabolites. Optimization of various parameters, such as elicitor type, concentration, duration of exposure, and treatment schedule is essential for the effectiveness of the elicitation strategies. Combined application of different elicitors, integration of precursor feeding, or replenishment of medium or in situ product recovery from the roots/liquid medium with the elicitor treatment have showed improved accumulation of secondary metabolites due to their synergistic effect. This is a comprehensive review about the progress in the elicitation approach to hairy root cultures from 2010 to 2019 and the information provided is valuable and will be of interest for scientists working in this area of plant biotechnology.

Grape VOCs Response to Postharvest Short-Term Ozone Treatments.

Ozone has been recently recognized as an efficient sanitizing agent in wine industry because of its powerful oxidizing properties. Furthermore, postharvest treatments of grapes with ozone can stimulate defense responses by synthetizing secondary metabolites against oxidative stress. In this study, the effect of postharvest short-term ozone treatments was assessed for the first time on free and glycosylated volatile organic compounds (VOCs) of winegrapes. Two different ozone concentrations (30 and 60 μL/L) and exposure times (24 and 48 h) were investigated just after treatment (fresh grapes) and after partial dehydration up to 20% weight loss (withered grapes). The study was carried out on Moscato bianco winegrapes (Vitis vinifera L.) due to the importance of terpenes in white aromatic cultivars to produce high quality wines. The results obtained showed that short-term ozone treatment caused a significant decrease in total contents of free VOCs in fresh grapes, mainly due to terpenes. Among them, linalool, geraniol, and nerol are the major aromatic markers of Moscato bianco grapes. Ozone entailed a significant decrease of free linalool contents in fresh grapes, the less stressful ozone treatment showing the smaller linalool degradation. However, the stronger and longer ozone treatment induced the synthesis of this compound probably in response to higher abiotic stress. Instead, significant changes were not observed in geraniol and nerol contents in fresh grapes. This last ozone treatment also reduced the loss of free linalool by water loss in withered grapes even though total VOCs and terpenes remained relatively stable. Furthermore, ozone seems to promote the synthesis of free (+)-4-carene and 4-terpineol in withered grapes under certain treatment conditions. Regarding glycosylated compounds, total VOCs and terpenes were less sensitive to ozone. Our findings highlight that ozone can be used as sanitizing agent in aromatic grape varieties prior to winemaking without affecting sharply the aromatic profile of fresh grapes and even improving it in withered grapes.

Genome-wide transcriptome analysis and identification of benzothiadiazole-induced genes and pathways potentially associated with defense response in banana

BackgroundBananas (Musa spp.) are the most important fruit crops worldwide due to their high nutrition value. Fusarium wilt of banana, caused by fungal pathogen Fusarium oxysporum f. sp. cubense tropical race 4 (Foc 4), is considered as the most destructive disease in the world and results in extensive damage leading to productivity loss. The widespread use of plant resistance inducers (PRIs), such as benzothiadiazole (BTH), is a novel strategy to stimulate defense responses in banana plants to protect against pathogens infection. The recent focus on the crop defense against fungal infections has led to a renewed interest on understanding the molecular mechanisms of specific PRIs-mediated resistance. This transcriptome study aimed to identify genes that are associated with BTH-induced resistance. Patterns of gene expression in the leaves and roots of BTH-sprayed banana plants were studied using RNA-Seq.ResultsIn this study, 18 RNA-Seq libraries from BTH-sprayed and untreated leaves and roots of the Cavendish plants, the most widely grown banana cultivar, were used for studying the transcriptional basis of BTH-related resistance. Comparative analyses have revealed that 6689 and 3624 differentially expressed genes were identified in leaves and roots, respectively, as compared to the control. Approximately 80% of these genes were differentially expressed in a tissue-specific manner. Further analysis showed that signaling perception and transduction, transcription factors, disease resistant proteins, plant hormones and cell wall organization-related genes were stimulated by BTH treatment, especially in roots. Interestingly, the ethylene and auxin biosynthesis and response genes were found to be up-regulated in leaves and roots, respectively, suggesting a choice among BTH-responsive phytohormone regulation.ConclusionsOur data suggests a role for BTH in enhancing banana plant defense responses to Foc 4 infection, and demonstrates that BTH selectively affect biological processes associated with plant defenses. The genes identified in the study could be further studied and exploited to develop Foc 4-resistant banana varieties.

Effects of carrageenan as elicitor to stimulate defense responses of basil against Cuscuta campestris Yunck

AbstractCuscuta campestrisis a holostemparasitic plant that obtains its resources from its hosts. Sweet basil is an important commercial plant, widely cultivated in many countries. It is a common host forC. campestris. Generally,C. campestrishas negative effect on the growth of infected plants and its infestation is difficult to control. Therefore, environmental friendly control ofC. campestrisseems to be useful. In this work, the relationship betweenC. campestrisand its host, sweet basil, and effects of κ-carrageenan on protection againstC. campestrisand suppression of its invasion were studied. Basil was sprayed with a solution of carrageenan at a final concentration of 1 g L−1, once a week, 3 times in total. Infection of basil withC. campestriswas performed 2 days after the last carrageenan treatment and the plants were collected two weeks afterC. campestrisattachment. In this study, phenylalanine ammonia-lyase activity (PAL), phenolic, flavonoids and antioxidant content increased remarkably in the basil plants parasitized withC. campestris, and therefore it seems that the parasitic plant induced a defense response in the host plants. Treatment with carrageenan significantly increased shoot length and leaf area of basil and decreasedC. campestrisinfestation by about 26%. Carrageenan treatment caused a significant increase in PAL activity, phenols, antioxidant and lignin content in basil. Thus, the present observation suggested the phenylpropanoid pathway was activated and defense responses were stimulated. Our results showed that carrageenan spraying induced beneficial effects in plants, corresponding to growth stimulation and defense compound synthesis. Thus carrageenan treatment is recommended as a natural biostimulator for the protection of plants againstC. campestris.

Chitosan enhances resistance in rubber tree (Hevea brasiliensis), through the induction of abscisic acid (ABA)

Chitosan is considered as a natural biodegradable compound that has potential to control plant diseases. In this work, we investigated the effect of chitosan in stimulating defense responses in rubber tree against Phytophthora palmivora. Foliar spraying of either 0.2% (w/v) chitosan or 50 μM abscisic acid (ABA) were efficient in the reduction of disease severity in P. palmivora infected rubber tree. In addition, the increases of enzyme activities, i.e. catalase, peroxidase, polyphenol oxidase and phenylalanine ammonia lyase, the expression of HbPR1, HbGLU, HbASI and HbCAT genes and the deposition of callose and lignin were observed in chitosan- or ABA-treated rubber tree leaves. Besides, a partial cDNA sequence for 9-cis-epoxycarotenoid dioxygenase (NCED), a key enzyme involved in ABA biosynthesis was firstly isolated from rubber tree using RT-PCR (designated HbNCED). The HbNCED gene fragment comprised of 753 bp and the deduced HbNCED protein showed 94% similarity to the NCED5 from Ricinus communis. Moreover, the up-regulation of HbNCED gene by exogenous chitosan was correlated with the induction of endogenous ABA measured by HPLC. Our results suggested that the exogenous chitosan could effectively reduce disease severity through the up-regulation of defense-related genes and ABA-biosynthesis gene, which led to the activation of defense-related proteins and ABA level in rubber tree.

Extracellular ATP Acts on Jasmonate Signaling to Reinforce Plant Defense.

Damaged cells send various signals to stimulate defense responses. Recent identification and genetic studies of the plant purinoceptor, P2K1 (also known as DORN1), have demonstrated that extracellular ATP is a signal involved in plant stress responses, including wounding, perhaps to evoke plant defense. However, it remains largely unknown how extracellular ATP induces plant defense responses. Here, we demonstrate that extracellular ATP induces plant defense mediated through activation of the intracellular signaling of jasmonate (JA), a well-characterized defense hormone. In Arabidopsis (Arabidopsis thaliana) leaves, ATP pretreatment induced resistance against the necrotrophic fungus, Botrytis cinerea The induced resistance was enhanced in the P2K1 receptor overexpression line, but reduced in the receptor mutant, dorn1-3 Mining the transcriptome data revealed that ATP induces a set of JA-induced genes. In addition, the P2K1-associated coexpression network contains defense-related genes, including those encoding jasmonate ZIM-domain (JAZ) proteins, which play key roles as repressors of JA signaling. We examined whether extracellular ATP impacts the stability of JAZ1 in Arabidopsis. The results showed that the JAZ1 stability decreased in response to ATP addition in a proteasome-dependent manner. This reduction required intracellular signaling via second messengers-cytosolic calcium, reactive oxygen species, and nitric oxide. Interestingly, the ATP-induced JAZ1 degradation was attenuated in the JA receptor mutant, coi1, but not in the JA biosynthesis mutant, aos, or upon addition of JA biosynthesis inhibitors. Immunoprecipitation analysis demonstrated that ATP increases the interaction between COI1 and JAZ1, suggesting direct cross talk between extracellular ATP and JA in intracellular signaling events. Taken together, these results suggest that extracellular ATP signaling directly impacts the JA signaling pathway to maximize plant defense responses.

Site-directed Mutagenesis Shows the Significance of Interactions with Phospholipids and the G-protein OsYchF1 for the Physiological Functions of the Rice GTPase-activating Protein 1 (OsGAP1)

The C2 domain is one of the most diverse phospholipid-binding domains mediating cellular signaling. One group of C2-domain proteins are plant-specific and are characterized by their small sizes and simple structures. We have previously reported that a member of this group, OsGAP1, is able to alleviate salt stress and stimulate defense responses, and bind to both phospholipids and an unconventional G-protein, OsYchF1. Here we solved the crystal structure of OsGAP1 to a resolution of 1.63 Å. Using site-directed mutagenesis, we successfully differentiated between the clusters of surface residues that are required for binding to phospholipids versus OsYchF1, which, in turn, is critical for its role in stimulating defense responses. On the other hand, the ability to alleviate salt stress by OsGAP1 is dependent only on its ability to bind OsYchF1 and is independent of its phospholipid-binding activity.

Chitosan application improves resistance to Fusarium circinatum in Pinus patula

Fusarium circinatum is the causal agent for the disease, pitch canker, in Pinus patula. Commercial forestry incurs large economic losses from the pathogen, primarily as a result of post-planting mortality resulting in increased re-establishment costs. One means of enhancing defense is through pretreatment of seedlings with chemicals or biologically derived compounds that stimulate defense responses; a process collectively known as induced resistance. We compared the efficiency of ten inducers in improving defense against F. circinatum in P. patula seedlings. Chitosan (10mg/ml) was effective in reducing and delaying disease symptoms of pitch canker in seedlings. Under both nursery and greenhouse conditions, chitosan application resulted in reduced lesion lengths in treated plants compared to non-treated plants over a period of six weeks (p<0.05, Kruskal–Wallis). Reverse transcription-quantitative PCR expression analysis revealed that the reduction in lesion size in treated seedlings was accompanied by a four-fold increase in transcript abundance of the phenylalanine ammonia lyase transcript, which encodes an enzyme involved in the first committed step of the phenylpropanoid pathway. We suggest that the application of chitosan as part of an integrated management strategy, be further investigated for an effective approach to induce resistance in P. patula seedlings against F. circinatum.

Seaweed polysaccharides and derived oligosaccharides stimulate defense responses and protection against pathogens in plants.

Plants interact with the environment by sensing “non-self” molecules called elicitors derived from pathogens or other sources. These molecules bind to specific receptors located in the plasma membrane and trigger defense responses leading to protection against pathogens. In particular, it has been shown that cell wall and storage polysaccharides from green, brown and red seaweeds (marine macroalgae) corresponding to ulvans, alginates, fucans, laminarin and carrageenans can trigger defense responses in plants enhancing protection against pathogens. In addition, oligosaccharides obtained by depolymerization of seaweed polysaccharides also induce protection against viral, fungal and bacterial infections in plants. In particular, most seaweed polysaccharides and derived oligosaccharides trigger an initial oxidative burst at local level and the activation of salicylic (SA), jasmonic acid (JA) and/or ethylene signaling pathways at systemic level. The activation of these signaling pathways leads to an increased expression of genes encoding: (i) Pathogenesis-Related (PR) proteins with antifungal and antibacterial activities; (ii) defense enzymes such as pheylalanine ammonia lyase (PAL) and lipoxygenase (LOX) which determine accumulation of phenylpropanoid compounds (PPCs) and oxylipins with antiviral, antifugal and antibacterial activities and iii) enzymes involved in synthesis of terpenes, terpenoids and/or alkaloids having antimicrobial activities. Thus, seaweed polysaccharides and their derived oligosaccharides induced the accumulation of proteins and compounds with antimicrobial activities that determine, at least in part, the enhanced protection against pathogens in plants.

Protein Elicitor PemG1 fromMagnaporthe griseaInduces Systemic Acquired Resistance (SAR) in Plants

Elicitors can stimulate defense responses in plants and have become a popular strategy in plant disease control. Previously, we isolated a novel protein elicitor, PemG1, from Magnaporthe grisea. In the present study, PemG1 protein expressed in and purified from Escherichia coli improved resistance of rice and Arabidopsis to bacterial infection, induced transient expression of pathogenesis-related (PR) genes, and increased accumulation of hydrogen peroxide in rice. The effects of PemG1 on disease resistance and PR gene expression were mobilized systemically throughout the rice plant and persisted for more than 28 days. PemG1-induced accumulation of OsPR-1a in rice was prevented by the calcium channel blockers LaCl₃, BAPTA, EGTA, W7, and TFP. Arabidopsis mutants that are insensitive to jasmonic acid (JA) and ethylene showed increased resistance to bacterial infection after PemG1 treatment but PemG1 did not affect resistance of mutants with an impaired salicylic acid (SA) transduction pathway. In rice, PemG1 induced overexpressions of the SA signal-related genes (OsEDS1, OsPAL1, and OsNH1) but not the JA pathway-related genes (OsLOX2 and OsAOS2). Our findings reveal that PemG1 protein can function as an activator of plant disease resistance, and the PemG1-mediated systemic acquired resistance is modulated by SA- and Ca(2+)-related signaling pathways.

Implications of oligomeric forms of POD-1 and POD-2 proteins isolated from cell walls of the biocontrol agent Pythium oligandrum in relation to their ability to induce defense reactions in tomato

The cell wall protein fraction (CWP) isolated from the biocontrol agent Pythium oligandrum induces defense reactions in tomato. CWP contains two novel elicitin-like proteins, POD-1 and POD-2, both with seven cysteines. To determine the essential structure in the defense-eliciting components of CWP, five fractions (F1, F2, F3, F4 and F5) were fractionated from CWP using cation chromatography and their components and disulfide bond compositions were analyzed. The expression levels of three defense-related genes ( PR-6, LeCAS and PR-2b) were determined in tomato roots treated with each of the five fractions. Of the five fractions, F4 containing a heterohexamer of POD-1 and POD-2, and F5 containing a homohexamer of POD-1, both with disulfide bonds formed between all cysteine residues, induced the expression of three genes. F4 treatment also induced the accumulation of ethylene in tomato. The predicted three-dimensional structures of POD-1 and POD-2, and the results of SEC and MALDI-TOF MS analyses suggest that F4 consists of three POD-1 and POD-2 disulfide-bonded heterodimers that interleave into a hexameric ring through noncovalent association. These results suggest that this structure, which F5 also appears to form, is essential for stimulating defense responses in tomato.

Expression of an elicitor-encoding gene from Magnaporthe grisea enhances resistance against blast disease in transgenic rice

Elicitors are molecules that stimulate defense responses in plants. Previously, an elicitor-encoding gene, named pemG1, was isolated from Magnaporthe grisea. To assess the function of pemG1 in rice (Oryza sativa L. cv. Nipponbare), the gene was cloned under a constitutive maize ubiquitin promoter and introduced into Nipponbare cultivar. The resultant plants showed stable integration and constitutive expression of the pemG1 gene. The expression of defense-related gene for phenylalanine ammonia-lyase was triggered and proline content was also increased in pemG1-expressing plants. The pemG1-expressing plants showed enhanced resistance against rice blast after inoculation with M. grisea spores, suggesting that the pemG1 expression enhances disease resistance in transgenic rice.

Cellulose Binding Domains of a Phytophthora Cell Wall Protein Are Novel Pathogen-Associated Molecular Patterns

The cellulose binding elicitor lectin (CBEL) from Phytophthora parasitica nicotianae contains two cellulose binding domains (CBDs) belonging to the Carbohydrate Binding Module1 family, which is found almost exclusively in fungi. The mechanism by which CBEL is perceived by the host plant remains unknown. The role of CBDs in eliciting activity was investigated using modified versions of the protein produced in Escherichia coli or synthesized in planta through the potato virus X expression system. Recombinant CBEL produced by E. coli elicited necrotic lesions and defense gene expression when injected into tobacco (Nicotiana tabacum) leaves. CBEL production in planta induced necrosis. Site-directed mutagenesis on aromatic amino acid residues located within the CBDs as well as leaf infiltration assays using mutated and truncated recombinant proteins confirmed the importance of intact CBDs to induce defense responses. Tobacco and Arabidopsis thaliana leaf infiltration assays using synthetic peptides showed that the CBDs of CBEL are essential and sufficient to stimulate defense responses. Moreover, CBEL elicits a transient variation of cytosolic calcium levels in tobacco cells but not in protoplasts. These results define CBDs as a novel class of molecular patterns in oomycetes that are targeted by the innate immune system of plants and might act through interaction with the cell wall.