Identification of an ISR‐related metabolite produced by rhizobacterium Klebsiella oxytoca C1036 active against soft‐rot disease pathogen in tobacco

BACKGROUND: Klebsiella oxytoca C1036 (C1036) causes induced systemic resistance (ISR) activity against the soft‐rot pathogen Pectobacterium carotovorum subsp. carotovorum SCC1 (SCC1). However, microbial metabolites from C1036 involved in ISR activity remain unknown. The present study was performed to identify an ISR‐related metabolite produced by C1036. RESULTS: The supernatants of C1036 cultures grown on Luria‐Bertani medium were subjected to solvent extraction, repeated column chromatography and preparative liquid chromatography for isolation of an ISR‐related metabolite. High‐resolution mass spectrometer analysis of the isolated metabolite indicated a C₉H₁₅O₃N compound with a mass of 185.11. Low‐resolution mass spectrometer analysis of the metabolite showed a molecular ion peak at 185 and its fragment ions at 84 and 56. Nuclear magnetic resonance spectrometer analyses characterised all protons and carbons of the isolated metabolite. Based on the data, the isolated metabolite was determined to be butyl 2‐pyrrolidone‐5‐carboxylate (BPC). BPC at 12 mM significantly suppressed the disease symptoms in ISR bioassays against SCC1. CONCLUSION: This is the first report identifying BPC as an ISR‐related metabolite produced by C1036. C1036 may play a role in promoting plant growth because it produces ISR‐related metabolites against the plant pathogen SCC1. Copyright © 2009 Society of Chemical Industry     

Suppressive effect of abscisic acid on systemic acquired resistance in tobacco plants

Recent studies have indicated that the phytohormone abscisic acid (ABA), induced in response to a variety of environmental stresses, plays an important role in modulating diverse plant–pathogen interactions. In Arabidopsis thaliana, we previously clarified that ABA suppressed the induction of systemic acquired resistance (SAR), a plant defense system induced by pathogen infection through salicylic acid (SA) accumulation. We investigated the generality of this suppressive effect by ABA on SAR using tobacco plants. For SAR induction, we used 1,2-benzisothiazole-3(2H)-one 1,1-dioxide (BIT) and benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) that activate upstream and downstream of SA in the SAR signaling pathway, respectively. Wild-type tobacco plants treated with BIT or BTH exhibited enhanced disease resistance against Tobacco mosaic virus (TMV) and tobacco wildfire bacterium, Pseudomonas syringae pv. tabaci (Pst), however, which was suppressed by pretreatment of plants with ABA. Pretreatment with ABA also suppressed the expression of SAR-marker genes by BIT and BTH, indicating that ABA suppressed the induction of SAR. ABA suppressed BTH-induced disease resistance and pathogenesis-related (PR) gene expression in NahG-transgenic plants that are unable to accumulate SA. The accumulation of SA in wild-type plants after BIT treatment was also suppressed by pretreatment with ABA. These data suggest that ABA suppresses both upstream and downstream of SA in the SAR signaling pathway in tobacco.     

Heritability Of Rhizobacteria-mediated Induced Systemic Resistance And Basal Resistance In Arabidopsis

Selected strains of non-pathogenic rhizobacteria have the ability to trigger an induced systemic resistance (ISR) response in plants. In Arabidopsis, rhizobacteria-mediated ISR has been extensively studied, using Pseudomonas fluorescens WCS417r as the inducing agent and P. syringae pv. tomato DC3000 (Pst) as the challenging pathogen. To investigate how far expression of ISR depends on the level of basal resistance, 10 different Arabidopsis ecotypes were screened for their potential to express WCS417r-mediated ISR and basal resistance against Pst. Two Arabidopsis ecotypes, RLD and Wassilewskija (Ws), were found to be blocked in their ability to express ISR. This ISR-noninducible phenotype correlated with a relatively low level of basal resistance against Pst. Genetic analysis of crosses between the ISR-inducible ecotypes Columbia (Col) and Landsberg erecta (Ler), on the one hand, and the non-inducible ecotypes RLD and Ws, on the other hand, revealed that ISR inducibility and basal resistance against Pst were inherited as monogenic dominant traits that are genetically linked. Neither ISR inducibility, nor basal resistance against Pst was complemented in the F₁ progeny of a cross between RLD and Ws, indicating that both ecotypes are affected in the same locus. This locus, designated ISR1, was mapped between markers Ein3 and GL1 on chromosome III. Interestingly, ecotypes RLD and Ws also failed to express ISR against the oomycetous pathogen Peronospora parasitica, but they were not affected in their level of basal resistance against this pathogen. Thus, the ISR1 locus controls the expression of ISR against different pathogens but basal resistance only against Pst and not against P. parasitica. Like ecotypes RLD and Ws, ethylene-insensitive mutants showed the isr1 phenotype in that they were unable to express WCS417r-mediated ISR and show enhanced susceptibility to Pst infection. Analysis of ethylene responsiveness of RLD and Ws revealed that both ecotypes exhibit reduced sensitivity to ethylene. Therefore, it is proposed that the Arabidopsis ISR1 locus encodes a component of the ethylene-response pathway that plays an important role in ethylene-dependent resistance mechanisms.     

Contribution of both lignin content and sinapyl monomer to disease resistance in tobacco

As a major component of the cell wall, lignin has been suggested to play an important role in the plant defence response to various pathogens. However, how lignin is involved in plant pathogen interaction is still unclear. Here, a series of transgenic tobacco lines were cultivated with a range of differences in lignin content and composition. Evaluation of pathogen resistance in these plants indicated that lower total lignin content aggravated the severity of tobacco black shank and bacterial wilt diseases, while increased sinapyl lignin (S) alleviated the disease symptoms. The regression analysis indicated both lignin content and S lignin were positively correlated with disease resistance. These two factors had additive effects, exhibiting stronger correlation with disease resistance when they were combined. Neither guaiacyl lignin (G) nor S/G ratio showed close correlation with disease resistance. The expression of pathogenesis‐related protein genes PR2 and PR3 was induced after pathogen inoculation. However, the up‐regulation of PR2 and PR3 was not associated with a disease resistance‐induced increase in lignin content. These data collectively suggest that both total lignin content and S lignin are main factors that contribute to the basic defence response in tobacco.     

Induction of Systemic Resistance Against Bacterial Wilt in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> by Fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> spp

The ability of selected strains of fluorescent Pseudomonas spp. to cause induced systemic resistance (ISR) in Eucalyptus urophylla against bacterial wilt caused by Ralstonia solanacearum was investigated. Four of the five strains used can produce salicylic acid (SA) in vitro and, therefore, chemical SA, that is known to induce resistance in many plant species, was used as a reference treatment. Whereas a soil drench with SA did induce systemic resistance in E. urophylla, infiltration of SA into leaves did not. None of the fluorescent Pseudomonas spp. strains caused ISR against bacterial wilt when applied to the soil, but two strains, P. putida WCS358r and P. fluorescens WCS374r triggered ISR when infiltrated into two lower leaves 3–7 days before challenge inoculation. A mutant of strain WCS358r defective in the biosynthesis of the fluorescent siderophore pseudobactin, did not cause ISR, while the purified siderophore of WCS358r did, suggesting that pseudobactin358 is the ISR determinant of WCS358. A siderophore-minus mutant of WCS374r induced the same level of disease resistance as its parental strain, but the purified siderophore induced resistance as well, indicating that both the siderophore and another, unknown, inducing determinant(s) of WCS374r can trigger ISR in Eucalyptus. A possible role of WCS374r-produced SA remains uncertain. Transformation of a siderophore-minus mutant of WCS358 with the SA biosynthetic gene cluster from WCS374 did not enable this transformant to cause ISR in E. urophylla.     

Enhanced protection against two major fungal pathogens of groundnut, <Emphasis Type="Italic">Cercospora arachidicola</Emphasis> and <Emphasis Type="Italic">Aspergillus flavus</Emphasis> in transgenic groundnut over-expressing a tobacco β 1–3 glucanase

Groundnut is an important oilseed crop of the Indian subcontinent. Yield losses due to fungal diseases are enormous in the cultivation of this crop. Over-expression of PR proteins leads to increased resistance to pathogenic fungi in several crops. The PR protein glucanase hydrolyses a major cell-wall component, glucan, of pathogenic fungi and acts as a plant defense barrier. We report in this paper, overexpression of a tobacco glucanase in transgenic groundnut and its resistance towards Cercospora arachidicola and Aspergillus flavus. PCR, Southern and Northern hybridization confirmed stable integration and expression of the glucanase gene in groundnut transgenics. When screened for resistance against Cercospora arachidicola the transgenics showed not only reduction in the number of spots but also delay in the onset of disease. Resistance was also demonstrated against one another important pathogen of groundnut, Aspergillus flavus. The transgenics not only resisted hyphal spread but also did not accumulate aflatoxin in the seeds. The results demonstrate the potential of a PR protein from a heterologous source in developing fungal disease resistant groundnut.     

Dimethyl disulfide is an induced systemic resistance elicitor produced by Bacillus cereus C1L

BACKGROUND: Bacillus cereus C1L is a plant growth‐promoting rhizobacterium and can elicit induced systemic resistance (ISR) in plants against necrotrophic pathogens. However, little is known about ISR elicitors produced by B. cereus C1L, and no ISR elicitor has been identified and characterised. Therefore, the objective of this study is to identify volatile ISR elicitor(s) produced by B. cereus C1L. RESULTS: The volatile metabolites produced by B. cereus C1L were extracted, separated and identified by solid‐phase microextraction, gas chromatography and mass spectrometry. Dimethyl disulfide (DMDS) was the only separated metabolite being determined. Afterwards, application of DMDS by means of soil drench significantly protected tobacco and corn plants against Botrytis cinerea and Cochliobolus heterostrophus, respectively, under greenhouse conditions. The results reveal that DMDS could play an important role in ISR by B. cereus C1L. CONCLUSION: This is the first report of DMDS as an elicitor produced by an ISR‐eliciting B. cereus strain and its ability to suppress plant fungal diseases under greenhouse conditions. It is suggested that DMDS has potential for practical use in controlling plant foliar diseases besides soil fumigation. Copyright © 2012 Society of Chemical Industry     

内生细菌EBS05对烟草诱导抗性的信号转导途径研究

 樟树内生枯草芽胞杆菌EBS05是一株对多种植物病原菌具有较强拮抗活性,并能诱导烟草系统抗性的生防菌株。本文以缺失Surfactin A合成相关基因的突变菌株EBS05^T为材料,研究了内生细菌EBS05对烟草诱导系统抗性的激发子及其信号转导途径。结果表明,菌株EBS05产生的Surfactin A是诱导烟草对TMV系统抗性的有效激发子;Surfactin A诱导处理后,SA信号转导途径下游的关键调节基因NPR1首先被激活,并持续超量表达,进而触发PR1b和PR1a基因持续超量表达,表明Surfactin A诱导烟草对TMV的系统抗性是通过激活SA信号转导途径实现的。同时,Surfactin A诱导处理后24~72 h,JA/ET信号转导途径调节基因PDF1.2被激活,且超量表达,表明在Surfactin A诱导烟草对TMV系统抗性的信号转导过程中,可能存在SA信号途径和JA/ET信号途径的交叉协同作用。     

Comparison of induced resistance activated by benzothiadiazole, (2R,3R)‐butanediol and an isoparaffin mixture against anthracnose of Nicotiana benthamiana

Resistance in Nicotiana benthamiana against anthracnose caused by the hemibiotrophic fungus Colletotrichum orbiculare was activated by benzothiadiazole (BTH), (2R,3R)­butanediol or PC1, an isoparaffin‐based mixture. In inoculation experiments, BTH, (2R,3R)­butanediol and PC1 reduced the number of lesions per leaf area caused by C. orbiculare by 98%, 77% and 81%, respectively. Foliar application of BTH induced expression of genes for the acidic pathogenesis‐related (PR) proteins, NbPR‐1a, NbPR‐3Q and acidic NbPR‐5. In contrast, soil application of (2R,3R)­butanediol or PC1 primed expression of genes for the basic PR proteins, NbPRb‐1b, basic NbPR‐2 and NbPR‐5dB. These results are consistent with the activation of salicylic‐acid‐dependent systemic acquired resistance (SAR) by BTH and that of jasmonate/ethylene‐dependent induced systemic resistance (ISR) by (2R,3R)­butanediol or PC1, and show that (2R,3R)­butanediol and PC1 can affect gene expression similarly to plant growth‐promoting rhizobacteria. However, the effects of (2R,3R)‐butanediol and PC1 were not identical. In addition to priming, (2R,3R)‐butanediol induced expression of basic NbPR‐2, whereas PC1 treatment induced expression of both NbPRb‐1b and basic NbPR‐2. Although a number of microbial products, such as (2R,3R)­butanediol, have been shown to produce ISR, this is the first demonstration that an isoparaffin‐based mixture, not derived from a microorganism, can produce ISR.     

Assessment of resistance pathways induced in Arabidopsis thaliana by hypovirulent Rhizoctonia spp. isolates

Certain hypovirulent Rhizoctonia isolates effectively protect plants against well-known important pathogens among Rhizoctonia isolates as well as against other pathogens. The modes of action involved in this protection include resistance induced in plants by colonization with hypovirulent Rhizoctonia isolates. The qualifications of hypovirulent isolates (efficient protection, rapid growth, effective colonization of the plants, and easy application in the field) provide a significant potential for the development of a commercial microbial preparation for application as biological control agents. Understanding of the modes of action involved in protection is important for improving the various aspects of development and application of such preparations. The hypothesis of the present study is that resistance pathways such as systemic acquired resistance (SAR), induced systemic resistance (ISR), and phytoalexins are induced in plants colonized by the protective hypovirulent Rhizoctonia isolates and are involved in the protection of these plants against pathogenic Rhizoctonia. Changes in protection levels of Arabidopsis thaliana mutants defective in defense-related genes (npr1-1, npr1-2, ndr1-1, npr1-2/ndr1-1, cim6, wrky70.1, snc1, and pbs3-1) and colonized with the hypovirulent Rhizoctonia isolates compared with that of the wild type (wt) plants colonized with the same isolates confirmed the involvement of induced resistance in the protection of the plants against pathogenic Rhizoctonia spp., although protection levels of mutants constantly expressing SAR genes (snc1 and cim6) were lower than that of wt plants. Plant colonization by hypovirulent Rhizoctonia isolates induced elevated expression levels of the following genes: PR5 (SAR), PDF1.2, LOX2, LOX1, CORI3 (ISR), and PAD3 (phytoalexin production), which indicated that all of these pathways were induced in the hypovirulent-colonized plants. When SAR or ISR were induced separately in plants after application of the chemical inducers Bion and methyl jasmonate, respectively, only ISR activation resulted in a higher protection level against the pathogen, although the protection was minor. In conclusion, plant colonization with the protective hypovirulent Rhizoctonia isolates significantly induced genes involved in the SAR, ISR, and phytoalexin production pathways. In the studied system, SAR probably did not play a major role in the mode of protection against pathogenic Rhizoctonia spp.; however, it may play a more significant role in protection against other pathogens.     

Application of Rhizobacteria for Induced Resistance

This article provides a review of experiments conducted over a six-year period to develop a biological control system for insect-transmitted diseases in vegetables based on induced systemic resistance (ISR) mediated by plant growth-promoting rhizobacteria (PGPR). Initial experiments investigated the factors involved in treatment with PGPR led to ISR to bacterial wilt disease in cucumber caused by Erwinia tracheiphila. Results demonstrated that PGPR-ISR against bacterial wilt and feeding by the cucumber beetle vectors of E. trachiphiela were associated with reduced concentrations of cucurbitacin, a secondary plant metabolite and powerful beetle feeding stimulant. In other experiments, PGPR induced resistance against bacterial wilt in the absence of the beetle vectors, suggesting that PGPR-ISR protects cucumber against bacterial wilt not only by reducing beetle feeding and transmission of the pathogen, but also through the induction of other plant defense mechanisms after the pathogen has been introduced into the plant. Additional greenhouse and field experiments are described in which PGPR strains were selected for ISR against cucumber mosaic virus (CMV) and tomato mottle virus (ToMoV). Although results varied from year to year, field-grown tomatoes treated with PGPR demonstrated a reduction in the development of disease symptoms, and often a reduction in the incidence of viral infection and an increase in tomato yield. Recent efforts on commercial development of PGPR are described in which biological preparations containing industrial formulated spores of PGPR plus chitosan were formulated and evaluated for use in a transplant soil mix system for developing plants that can withstand disease attack after transplanting in the field.     

Local and systemic resistance to fungal pathogens triggered by an AVR9-mediated hypersensitive response in tomato and oilseed rape carrying the Cf-9 resistance gene

Tomato and transgenic oilseed rape plants expressing the Cf-9 resistance gene develop a hypersensitive response (HR) after injection of the corresponding Avr9 gene product. It was investigated whether induction of a HR conferred resistance to different fungal pathogens in tomato and oilseed rape. Induction of an AVR9 mediated HR at the pathogen infection site delayed the development of the biotrophs Oidium lycopersicum in tomato and Erysiphe polygoni in oilseed rape, but enhanced the development of the necrotrophs Botrytis cinerea and Alternaria solani in tomato and Sclerotinia sclerotiorum in oilseed rape. Interestingly, delayed fungal disease development was observed in plant tissues surrounding the HR lesion regardless of whether a necrotrophic or biotrophic pathogen was used. In tomato, AVR9 injection induced systemic expression of PR1, PR2 and PR3 defence genes but did not induce systemic resistance to O. lycopersicum, B. cinerea or A. solani. In oilseed rape, AVR9 injection temporarily induced systemic resistance to Leptosphaeria maculans and E. polygoni, but did not induce detectable systemic expression of PR1, PR2 or Cxc750. These results give new insights into the potential uses of an induced HR to engineer disease resistance.     

The Relationship Between Pathogen-induced Systemic Resistance (ISR) and Multigenic (horizontal) Resistance in Plants

Plants have developed mechanisms to successfully co-exist in the presence of pathogenic organisms. Some interactions between plants and pathogens are based on recognition of specific elicitor molecules from avirulent pathogen races (avr gene products), which is described in the gene-for-gene resistance theory. Another type of resistance, multigenic (horizontal) resistance, is a less well-studied phenomenon that depends upon multiple genes in the plant host. All plants possess resistance mechamisms which can be induced upon pre-treatment of plants with a variety of organisms or compounds. This general phenomenon is known as induced systemic resistance (ISR). At least in some plant species, ISR depends on the timely accumulation of multiple gene products, such as hydrolytic enzymes, peroxidases or other gene products related to plant defences. The pre-treatment of plants with an inducing organism or compound appears to incite the plant to mount an effective defense response upon subsequent encounters with pathogens, converting what would have been a compatible interaction to an incompatible one. Our studies in three plant–pathogen systems clearly document that multigenic-resistant plants constitutively express specific isozymes of hydrolytic enzymes that release cell wall elicitors, which in turn may activate other defense mechanisms. ISR induces constitutive accumulation of these and other gene products prior to challenge. ISR is known to function against multiple organisms, and there is no specificity observed in the accumulation patterns of defense-related gene products when ISR is induced. It is therefore hypothesized that the constitutive accumulation of specific isozymes of hydrolytic enzymes, or other defense related gene products, is an integral part of both multigenic resistance and the phenomenon of ISR. Further, plants in which ISR has been activated appear to move from a latent resistance state to one in which a multigenic, non-specific form of resistance is active.     

Induction of mutants of Fusarium oxysporum f. sp. lycopersici with altered virulence

Mutants ofFusarium oxysporum f. sp.lycopersici were obtained by UV irradiation. The mutants of race 1 and race 2 caused disease symptoms on plants with resistance genes against the corresponding wild type strains. Mutants of race 1 of the pathogen were stable, whereas mutants of race 2 lost the ability to cause disease symptoms in plants carrying the 1–2 resistance gene, after prolonged maintenenance on potato dextrose agar. Mutants of race 1 resembled race 2 in pathogenicity and they were vegetatively compatible with race 2, but no longer with race 1. These results suggest that the isolated strains with an altered virulence pattern have mutations in loci involved in avirulence.     

Constitutive expression of a phenylalanine ammonia-lyase gene from Stylosanthes humilis in transgenic tobacco leads to enhanced disease resistance but impaired plant growth

Transgenic tobacco plants expressing a phenylalanine ammonia-lyase cDNA (ShPAL), isolated from Stylosanthes humilis, under the control of the 35S promoter of the cauliflower mosaic virus were produced to test the effect of high level PAL expression on disease resistance. The transgenic plants showed up to eight-fold PAL activity and were slowed in growth and flowering relative to non-transgenic controls which have segregated out the transgene. The expression of the ShPAL transgene and elevated PAL levels were correlated and stably inherited. In T₁ and T₂ tobacco plants with increased PAL activity, lesion expansion was significantly reduced by up to 55% on stems inoculated with the Oomycete pathogen Phytophthora parasitica pv. nicotianae. Lesion area was significantly reduced by up to 50% on leaves inoculated with the fungal pathogen Cercospora nicotianae. This study provides further evidence that PAL has a role in plant defence.     

枯草芽孢杆菌OKB105产生的surfactin防治烟草花叶病毒病及其机理研究

本文研究枯草芽孢杆菌产生的脂肽化合物表面活性素surfactin对烟草花叶病毒病的防治效果,并检测烟草防卫相关基因的表达情况以解析抗病机理。利用6 mol/LHCl沉淀枯草芽孢杆菌OKB105的去细胞培养液,通过甲醇抽提,Sephadex LH20层析柱获得脂肽纯化物。经MALDI-TOF-MS和血平板溶血试验检测表明,纯化物中仅含有表面活性素surfactin。将表面活性素配成0.01 g/L溶液,处理烟草并接种TMV。结果表明, surfactin对烟草花叶病毒病有显著的防治效果,病斑抑制率为70.9%。通过Real-time PCR检测烟草的防卫相关基因的表达情况结果发现,烟草SA信号通路的PR1、PR3和PR5,以及ET通路的ETR1表达量显著提高,说明surfactin诱导了烟草依赖于SA和ET信号通路的诱导系统抗性（induced systemic resistance, ISR）反应。     

Induced resistance to Verticillium longisporum in Brassica napus by β‐aminobutyric acid

A preinoculative soil drench application of 0·5 mm β‐aminobutyric acid (BABA) significantly inhibited colonization of oilseed rape (Brassica napus, susceptible cultivar Falcon) by Verticillium longisporum and also prevented plant stunting caused by the pathogen. To better understand the defence responses induced by BABA, the presence of occlusions in the plant hypocotyl, levels of salicylic acid (SA) and hydrogen peroxide (H₂O₂), phenylalanine ammonia lyase (PAL) activity and expression of PR‐1 and PDF1.2 genes were examined. Transverse sections through the hypocotyl region of BABA‐treated plants showed clear vessels surrounded by phenol‐storing cells, in contrast to numerous obstructed vessels in water‐treated plants, in response to the pathogen. A significant increase in SA levels was observed in the hypocotyls of both water‐ and BABA‐treated plants in response to the pathogen; however, SA levels were unrelated to disease resistance. The level of H₂O₂ decreased in both treatments in response to the pathogen. A significant increase in PAL activity was observed in hypocotyl tissues of BABA‐treated plants. The expression patterns of PR‐1 and PDF1.2 were similar in the two treatments in response to the pathogen, indicating no involvement of these genes in resistance. The results indicate a similar organ specificity of the plant hypocotyl for chemically induced internal resistance as for genotype‐related resistance, two phenomena which, however, are based on contrasting cytological responses in the vascular tissues. Nonetheless, evidence is provided that the activity of the phenylpropanoid pathway plays a crucial role in both types of resistance.     

Systemic resistance to bacterial leaf speck pathogen in Arabidopsis thaliana induced by the culture filtrate of a plant growth-promoting fungus (PGPF) Phoma sp. GS8-1

The culture filtrate (CF) from the plant growth-promoting fungus Phoma sp. GS8-1 was found to induce systemic resistance in Arabidopsis thaliana against the bacterial leaf speck pathogen Pseudomonas syringae pv. tomato DC3000 (Pst), and the underlying mechanism was studied. Roots of A. thaliana were treated with CF from GS8-1, and plants expressed a clear resistance to subsequent Pst infection; disease severity was reduced, and proliferation of pathogen was suppressed. Various mutants of A. thaliana were used to test whether the CF induced resistance through one of the known signaling pathways: salicylic acid (SA), jasmonic acid (JA), and ethylene (ET). The CF was fully protective against Pst in Arabidopsis mutants jar1 and ein2 similar to wild-type plants. However, its efficacy was reduced in plants containing transgene NahG. Examination of systemic gene expression revealed that CF modulates the expression of SA-inducible PR-1, PR-2 and PR-5 genes, the JA/ET-inducible ChitB gene, and the ET-inducible Hel gene. Moreover, the expression of these genes was further enhanced upon subsequent stimulation after attack by Pst. Our data suggest that in addition to a partial requirement for SA, the signals JA and ET may also play a role in defense signaling in Arabidopsis.