Methyl jasmonate and methyl salicylate, but not cis-jasmone, evoke defenses against infection of Arabidopsis thaliana by Orobanche aegyptiaca

The mechanism by which some plant species develop resistance to the root parasite, broomrape ( Orobanche aegyptiaca ), is still not clear. Resistance to other pathogens can be induced by methyl jasmonate and systemic acquired resistance can be induced by treatment with salicylic acid, while cis -jasmone can act as a signaling molecule in plant–insect interactions. The three compounds studied, methyl jasmonate, cis -jasmone, and methyl salicylate, were applied to Arabidopsis thaliana seedlings that were then transferred to Nunc cell culture plates and exposed to the germinating seeds of O. aegyptiaca . The number of infections of the roots of single seedlings of A. thaliana was then quantified. Exposure for 24 h to very low concentrations of methyl jasmonate or methyl salicylate, which were then removed, effectively induced resistance to infection of A. thaliana by O. aegyptiaca , reducing attachment and tubercle formation by 90%. cis -Jasmone was far less effective in inducing a similar resistance to infection. These results support the view that methyl jasmonate can induce almost full resistance to infection by broomrape. The fact that such resistance is not observed under normal conditions of infection supports the idea that the root parasite does not evoke the full defensive response in the host plant.     

Rhizobacteria-mediated Induced Systemic Resistance: Triggering, Signalling and Expression

Selected strains of rhizosphere bacteria reduce disease by activating a resistance mechanism in the plant named rhizobacteria-mediated induced systemic resistance (ISR). Rhizobacteria-mediated ISR resembles pathogen-induced systemic acquired resistance (SAR) in that both types of induced resistance render uninfected plant parts more resistant towards a broad spectrum of plant pathogens. Some rhizobacteria trigger the salicylic acid (SA)-dependent SAR pathway by producing SA at the root surface. In other cases, rhizobacteria trigger a different signalling pathway that does not require SA. The existence of a SA-independent ISR pathway has been demonstrated in Arabidopsis thaliana. In contrast to pathogen-induced SAR, ISR induced by Pseudomonas fluorescens WCS417r is independent of SA accumulation and pathogenesis-related (PR) gene activation but, instead, requires responsiveness to the plant hormones jasmonic acid (JA) and ethylene. Mutant analyses showed that ISR follows a novel signalling pathway in which components from the JA and ethylene response are successively engaged to trigger a defensive state that, like SAR, is controlled by the regulatory factor NPR1. Interestingly, simultaneous activation of both the JA/ethylene-dependent ISR pathway and the SA-dependent SAR pathway results in an enhanced level of protection. Thus combining both types of induced resistance provides an attractive tool for the improvement of disease control. This review focuses on the current status of our research on triggering, signalling, and expression of rhizobacteria-mediated ISR in Arabidopsis.     

Effects of pre‐ and post‐treatment with ethephon on gum formation of peach gummosis caused by Lasiodiplodia theobromae

Peach gummosis, caused by Botryosphaeria spp. fungi, is the process of gum accumulation and exudation in plants. Ethephon (2‐chloroethylphosphonic acid) has profound effects on plants, including enhanced production of secondary metabolites and regulation of plant diseases. This study investigates the effects of application of ethephon before and after inoculation with Lasiodiplodia theobromae on gum formation. Gum formation was promoted by ethephon treatment prior to pathogen inoculation, but inhibited by ethephon applied after the pathogen. The inhibitory effect was counteracted by 1‐methylcyclopropane, which is an ethylene signal inhibitor. 1‐methylcyclopropane also promoted gum formation. Exposure of three isolates of Botryosphaeria to ethephon inhibited mycelial growth. Both treatment methods increased the sugar content at 12 and 24 h post‐inoculation (hpi). However, the sucrose, glucose and fructose contents were significantly higher in shoots with ethephon post‐treatment (application of ethephon after the pathogen inoculation) than those in shoots with ethephon pre‐treatment (application of ethephon prior to pathogen inoculation) at 48 and 72 hpi. The expression of two putative senescence‐related genes, SEN2 and SEN4, were significantly enhanced in pre‐ and post‐treated shoots with ethephon at 24, 48 and 72 hpi. Ethephon application also up‐regulated expression of the pathogenesis‐related protein PR4 while down‐regulating PR1a and PR10. The results show that ethephon has a dual function in regulating gum formation by affecting both the peach shoots and the pathogen.     

心叶烟NPR1基因全长cDNA序列的克隆及表达分析

 NPR1(non-expressor of pathogenesis-related gene 1)基因在拟南芥系统获得抗性中起着关键作用,可调控拟南芥植株广谱抗性的发生。本文报道了从心叶烟中克隆NPR1同源基因(NgNPR1)及其表达特性的研究结果。NgNPR1 cDNA全长2253 bp,编码588个氨基酸。将NgNPR1基因组全长与cDNA序列进行比对发现,NgNPR1基因组DNA含有4个外显子和3个内含子。Southern杂交分析表明,在心叶烟基因组中NgNPR1为单拷贝基因。采用绿色荧光蛋白在洋葱表皮瞬时表达的试验,证明了NgNPR1蛋白在水杨酸诱导时会从细胞质转运到细胞核中。Northern杂交分析发现,NgNPR1基因可以被与植物抗病相关的信号分子如水杨酸、茉莉酸甲酯、过氧化氢和乙烯所诱导。进一步研究发现,植物病原物如赤星病菌、青枯病菌和烟草花叶病毒对心叶烟植株的侵染也会使NgNPR1表达量增加。这些结果表明,NgNPR1基因在心叶烟植株抵御病原物侵染过程中可能起着重要作用。     

Exogenous application of melatonin improves plant resistance to virus infection

In this study, melatonin (MEL)-mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti-virus infection activity to 37.4% in virus-infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT-PCR and virus titres measured by dot-ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT-PCR, in virus-infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H₂O₂) in the virus-infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL-induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT-PCR and virus titres analysed by dot-ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL-mediated plant resistance to TMV is proposed. MEL-mediated plant resistance to viruses provides a new avenue to control plant viral diseases.     

The induction of pathogenesis-related proteins by pathogens and specific chemicals

Pathogenesis-related proteins (PRs) are induced in tobacco and other plant species by both biotic and abiotic agents, comprising necrotizing and non-necrotizing viruses, viroids, fungi, bacteria, specific physiological conditions and a variety of chemicals. Both ethephon and the natural precursor of ethylene, 1-aminocyclopropane-1-carboxylic acid, are good inducers and induction under all conditions investigated so far appears to be mediated by ethylene, except treatment with benzoic acid or its derivatives salicylic acid, aspirin, and 2,6-dihydroxybenzoic acid. Whereas the production of ethylene appears to result from a general reaction to stress, the mechanism by whicho-hydroxylated benzoic acids induced PRs is different. In ‘Samsun NN’ tobacco, tobacco mosaic virus (TMV) and ethephon induced PRs in both the treated and the untreated leaves at 20°C, but not at 32°C. However, salicylic acid induces PRs only in the treated leaves, but is as effective at 32°C as it is at 20°C. It has been proposed, therefore, that ethylene leads to the temperature-sensitive synthesis of a, presumably aromatic, compound that mimics the action of salicylic acid and functions as the natural inducer of PRs. The induction of PR 1a and 1b by salicylic acid or ethephon is blocked by cycloheximide but not by actinomycin D, whereas their accumulation upon TMV infection is inhibited up to 50% by actinomycin D. Actinomycin D similarly inhibits ethylene production in TMV-infected tobacco, supporting a role of ethylene in the induction of PRs in tobacco and indicating that ethylene acts by regulating the translation of the PR-mRNAs constitutively present but not translated in non-stimulated plants.     

Up‐regulation of PR1 and less disruption of hormone and sucrose metabolism in roots is associated with lower susceptibility to ‘Candidatus Liberibacter asiaticus’

Huanglongbing (HLB), caused by ‘Candidatus Liberibacter asiaticus’ (Las), is a devastating disease of citrus trees in Florida. Previous work showed that the rootstock cultivar Cleopatra mandarin (Citrus reticulata) has a higher population of Las in roots than Swingle citrumelo (C. paradisi × Poncirus trifoliata). Las reduced fibrous root biomass and sucrose content in Cleopatra mandarin more than in Swingle citrumelo. To understand the mechanisms for susceptibility to Las infection, sucrose and hormone metabolism status were evaluated in Cleopatra mandarin and Swingle citrumelo. In fibrous roots of Cleopatra mandarin, higher expression of genes related to sucrose cleavage was consistent with lower sucrose content compared to noninoculated seedlings at 5 weeks post‐root trimming (wpt). In fibrous roots of Swingle citrumelo, both sucrose content and gene expression related to sucrose cleavage were less disrupted by Las infection compared to Cleopatra mandarin at 5 wpt. Genes associated with salicylic acid (SA), ethylene (ET) and abscisic acid (ABA) synthesis, and ABA signalling, phospholipases D (PLD), and phospholipase A2 (PLA2) were activated by Las infection at 5 wpt in Cleopatra mandarin. Expression of downstream effectors of SA, i.e. NPR1, WRKY70 and PR1, did not change in Cleopatra mandarin, suggesting inhibition of the response to SA by the elevation of ABA, ET and PLD. In contrast, the up‐regulation of PR1, lower response of sucrose metabolism genes and down‐regulation of biosynthesis of phytohormones indicates that Swingle citrumelo activates a more effective defence against this biotrophic pathogen than Cleopatra mandarin.     

Costs and trade-offs associated with induced resistance

Plants resist attack by pathogens and herbivorous insects through constitutive and inducible defences. Based on differences in signalling pathways and spectra of effectiveness, different types of induced resistance have been defined. Systemic acquired resistance (SAR) occurs in distal plant parts following localized infection by a necrotizing pathogen. It is controlled by a signalling pathway that depends upon the accumulation of salicylic acid (SA) and the regulatory protein NPR1. In contrast, induced systemic resistance (ISR) is induced by selected strains of non-pathogenic plant growth promoting bacteria (PGPR). ISR functions independently of SA, but requires NPR1 and is regulated by jasmonic acid (JA) and ethylene (ET). It is generally believed that induced resistance evolved to save energy under pathogen or insect-free conditions, although costs still arise when defences are activated following attack. Costs can arise from the allocation of resources to defence and away from plant growth and development, and there are also ecological costs which result from trade-offs between induced resistance and the plant's interaction with beneficial organisms e.g. mycorrhizal fungi. To date, few studies have examined the costs and trade-offs associated with induced resistance to pathogens. There is a clear need for long-term studies of costs and trade-offs associated with induced resistance in crops under commercial conditions. Without such information, the potential offered by induced resistance is unlikely to be realized.     

Growth environment and seedling age affect constitutive and inducible defence in radiata pine

There is increasing interest in induced resistance for crop protection. This study investigated effects of growth environment on inducible defences in developing radiata pine (Pinus radiata) seedlings. Seedlings were grown in the presence or absence of Trichoderma in either a temperature-controlled growth room or in a greenhouse. After 3, 5 or 7 months, seedlings were sprayed with 2.25 mm methyl jasmonate (MJ), 1 week before quantification of monoterpenes and total phenolics. The magnitude of induction of monoterpenes and total phenolics by MJ was inversely proportional to constitutive content, was greater in the greenhouse than the growth room and was greater at 3 months than at 7 months. This may indicate a cost-saving strategy to limit defence induction once constitutive defence reaches a certain threshold. Growth room seedlings expressed greater constitutive resistance to terminal crook (Colletotrichum acutatum) and to diplodia dieback (Diplodia sapinea) than greenhouse plants, consistent with the relative differences in constitutive defence chemistry. Indeed, while MJ induced resistance to diplodia dieback in both environments, there was no difference in terminal crook incidence (4.1%) between MJ-treated and untreated seedlings in the growth room. By the end of the study, growth room seedlings had c. 70% more β-pinene in stems and 140% more total phenolics in needles than greenhouse plants but were 40% smaller, indicating a defence–growth trade-off. Trichoderma did not have a significant effect on defence chemistry or on pathogen resistance. These results demonstrate the potential to manipulate defence in radiata pine by modification of the growth environment.     

Ethylene signalling is essential for the resistance of Nicotiana attenuata against Alternaria alternata and phytoalexin scopoletin biosynthesis

The phytohormone ethylene plays an important role in plant defence responses to pathogen attack. When infected by the necrotrophic fungal pathogen Alternaria alternata (tobacco pathotype), which causes severe diseases in Nicotiana species, the wild tobacco plant Nicotiana attenuata accumulates a high amount of the jasmonate (JA)‐dependent phytoalexin scopoletin to defend itself against this fungal pathogen. However, it is still not known whether ethylene signalling is also involved in scopoletin biosynthesis and the resistance of N. attenuata. After infection, ethylene biosynthetic genes were highly elicited. Furthermore, plants strongly impaired in ethylene biosynthesis or perception had dramatically decreased scopoletin levels, and these plants became more susceptible to the fungus, while A. alternata‐elicited JA levels were increased, indicating that the decreased defence responses were not due to lower JA levels. Thus, it is concluded that after infection, ethylene signalling is activated together with JA signalling in N. attenuata plants and this subsequently regulates scopoletin biosynthesis and plant resistance.     

Priming as a Mechanism in Induced Systemic Resistance of Plants

Induced systemic resistance is a plant defence state that is associated with an enhanced ability – the so-called priming – to resist pathogen attack by stronger activation of cellular defence responses. So far, however, priming has not been widely appreciated when studying induced plant disease resistance. During the past several years, it has been demonstrated that pre-treatment of cultured parsley cells with inducers of systemic resistance, salicylic acid or a benzothiadiazole, leads to the direct activation of a set of defence-related genes and also primes the cells for stronger elicitation of another set of defence genes including those encoding phenylalanine ammonia-lyase. From these results, it was concluded that the resistance inducers have at least a dual role in plant defence-gene activation. When elucidating whether priming plays a role in induced systemic resistance of Arabidopsis, pre-treating plants with benzothiadiazole was found to augment the subsequent activation of phenylalanine ammonia-lyase genes by Pseudomonas infection, wounding and osmotic stress and also to enhance wound/osmotic stress-induced callose production. The augmentation of phenylalanine ammonia-lyase gene activation or/and callose deposition was not seen in the Arabidopsis non-expresser of pathogenesis-related genes1 mutant which is compromised in induced resistance, while it was present, without benzothiadiazole pre-treatment, in the constitutive expresser of pr genes1 and 5 mutants in which induced resistance is constitutive. Together these studies point to priming as an important cellular mechanism in induced systemic resistance of plants which requires the intact non-expresser of pathogenesis-related genes1 gene.