Unraveling regulatory networks in plant defense using microarrays

DNA microarrays are being used to comprehensively examine gene expression networks during the plant defense response that is triggered when a plant encounters a pathogen or an elicitor molecule. In addition to identifying new genes induced during defense, these studies are providing new insights into the complex pathways governing defense gene regulation.     

ASK1 physically interacts with COI1 and is required for male fertility in Arabidopsis

Jasmonates are a new class of plant hormones that play important roles in plant development and plant defense. The COI1 gene was previously shown to be required for jasmonate-regulated plant fertility and defense. We demonstrated for the first time that COI1 interacts with the Arabidopsis SKP1-LIKE1 (ASK1) to form a complex that is required for jasmonate action in planta. Functional analysis by antisense strategy showed that ASK1 is involved in male fertility.     

Evidence for an Important Role of WRKY DNA Binding Proteins in the Regulation of NPR1 Gene Expression

The Arabidopsis NPR1 gene is a positive regulator of inducible plant disease resistance. Expression of NPR1 is induced by pathogen infection or treatment with defense-inducing compounds such as salicylic acid (SA). Transgenic plants overexpressing NPR1 exhibit enhanced resistance to a broad spectrum of microbial pathogens, whereas plants underexpressing the gene are more susceptible to pathogen infection. These results suggest that regulation of NPR1 gene expression is important for the activation of plant defense responses. In the present study, we report the identification of W-box sequences in the promoter region of the NPR1 gene that are recognized specifically by SA-induced WRKY DNA binding proteins from Arabidopsis. Mutations in these W-box sequences abolished their recognition by WRKY DNA binding proteins, rendered the promoter unable to activate a downstream reporter gene, and compromised the ability of NPR1 to complement npr1 mutants for SA-induced defense gene expression and disease resistance. These results provide strong evidence that certain WRKY genes act upstream of NPR1 and positively regulate its expression during the activation of plant defense responses. Consistent with this model, we found that SA-induced expression of a number of WRKY genes was independent of NPR1.     

Diversity of plant–animal interactions: Possibilities for a new plant defense indicator value?

The interactions between herbivores and plants are of general interest in ecology. Even though the extensive research carried out during the last decades has culminated in many theories, additional studies are necessary to validate these findings. In particular, the hypotheses dealing with the complex interrelations of plant defense mechanisms and herbivores continue to be debated.In this paper, we develop a new indicator value that quantifies the defense mechanisms of Central European woody plants against large mammalian herbivores. The indicator value is based on three plant-specific traits: chemical defense (toxic compounds, digestion inhibitors), mechanical defense and leaf size. Our validation of the newly established indicator shows that evergreen woody plants have a significantly higher indicator value than deciduous woody plants. Moreover, plant defense is correlated with growth height: woody plants growing in the browsing zone preferred by large mammalian herbivores have significantly higher levels of defense compared with woody plants capable of growth high above the reach of large herbivores.We conclude that the new plant defense indicator value is a valuable tool for the validation of existing hypotheses and habitat calibration on a statistical basis. The quantification of plant mechanisms of defense against large herbivores produces a significantly better understanding of the multifaceted nature of plant–animal interactions and should contribute positively to future studies.     

ASK1 physically interacts with COI1 and is required for male fertility inArabidopsis

Jasmonates are a new class of plant hormones that play important roles in plant development and plant defense. TheCOI1 gene was previously shown to be required for jasmonate-regulated plant fertility and defense. We demonstrated for the first time that COI1 interacts with theArabidopsis SKP1-LIKE1 (ASK1) to form a complex that is required for jasmonate action inplanta. Functional analysis by antisense strategy showed thatASK1 is involved in male fertility.     

ASK1 physically interacts with COI1 and is required for male fertility in<Emphasis Type="Italic">Arabidopsis</Emphasis>

Jasmonates are a new class of plant hormones that play important roles in plant development and plant defense. TheCOI1 gene was previously shown to be required for jasmonate-regulated plant fertility and defense. We demonstrated for the first time that COI1 interacts with theArabidopsis SKP1-LIKE1 (ASK1) to form a complex that is required for jasmonate action inplanta. Functional analysis by antisense strategy showed thatASK1 is involved in male fertility.     

Structural variants in the soybean genome localize to clusters of biotic stress-response genes

Genome-wide structural and gene content variations are hypothesized to drive important phenotypic variation within a species. Structural and gene content variations were assessed among four soybean (Glycine max) genotypes using array hybridization and targeted resequencing. Many chromosomes exhibited relatively low rates of structural variation (SV) among genotypes. However, several regions exhibited both copy number and presence-absence variation, the most prominent found on chromosomes 3, 6, 7, 16, and 18. Interestingly, the regions most enriched for SV were specifically localized to gene-rich regions that harbor clustered multigene families. The most abundant classes of gene families associated with these regions were the nucleotide-binding and receptor-like protein classes, both of which are important for plant biotic defense. The colocalization of SV with plant defense response signal transduction pathways provides insight into the mechanisms of soybean resistance gene evolution and may inform the development of new approaches to resistance gene cloning.     

hsr203J, a tobacco gene whose activation is rapid, highly localized and specific for incompatible plant/pathogen interactions

A novel plant defense gene, hsr203J, whose corresponding mRNA accumulates preferentially during the incompatible interaction of tobacco (Nicotiana tabacum L.) with a pathogenic bacterium, Pseudomonas solanacearum, has been isolated and sequenced. No sequence homology of the putative product of this gene has been found in data bases. Evidence is presented here that the hsr203J gene promoter, when fused to the GUS reporter gene, is selectively expressed in response to the hypersensitive response (HR)-inducing bacteria in tobacco protoplasts and that the sequences responsible for this response are contained within 1.4 kb of the 5′ noncoding region. The temporal and spatial patterns of hsr203J activation in leaves and roots inoculated with P. solanacearum indicate that the hsr 203J promoter exhibits a rapid (3–6 h post-inoculation) and high level of induction only in plant cells inoculated with the HR-inducing bacterial isolate. In addition, this gene promoter which does not respond to various stress conditions and is only very weakly induced during compatible interactions, is strongly dependent on hrp (hypersensitive response and pathogenicity) genes of P. solanacearum. These data indicate that the hsr 203J gene promoter exhibits new and original characteristics of activation with regard to the plant defense genes studied so far; its spatial and temporal program of activation together with its specific induction during the HR underline the importance of this gene as a molecular tool for studying the establishment and regulation of the HR.     

Hydrogen peroxide generation by the pepper extracellular peroxidase CaPO2 activates local and systemic cell death and defense response to bacterial pathogens

Reactive oxygen species (ROS) are responsible for mediating cellular defense responses in plants. Controversy has existed over the origin of ROS in plant defense. We have isolated a novel extracellular peroxidase gene, CaPO2, from pepper (Capsicum annuum). Local or systemic expression of CaPO2 is induced in pepper by avirulent Xanthomonas campestris pv vesicatoria (Xcv) infection. We examined the function of the CaPO2 gene in plant defense using the virus-induced gene silencing technique and gain-of-function transgenic plants. CaPO2-silenced pepper plants were highly susceptible to Xcv infection. Virus-induced gene silencing of the CaPO2 gene also compromised hydrogen peroxide (H(2)O(2)) accumulation and hypersensitive cell death in leaves, both locally and systemically, during avirulent Xcv infection. In contrast, overexpression of CaPO2 in Arabidopsis (Arabidopsis thaliana) conferred enhanced disease resistance accompanied by cell death, H(2)O(2) accumulation, and PR gene induction. In CaPO2-overexpression Arabidopsis leaves infected by Pseudomonas syringae pv tomato, H(2)O(2) generation was sensitive to potassium cyanide (a peroxidase inhibitor) but insensitive to diphenylene iodonium (an NADPH oxidase inhibitor), suggesting that H(2)O(2) generation depends on peroxidase in Arabidopsis. Together, these results indicate that the CaPO2 peroxidase is involved in ROS generation, both locally and systemically, to activate cell death and PR gene induction during the defense response to pathogen invasion.     

Gene expression profile changes in cotton root and hypocotyl tissues in response to infection with Fusarium oxysporum f. sp. vasinfectum

Microarray analysis of large-scale temporal and tissue-specific plant gene expression changes occurring during a susceptible plant-pathogen interaction revealed different gene expression profile changes in cotton root and hypocotyl tissues. In hypocotyl tissues infected with Fusarium oxysporum f. sp. vasinfectum, increased expression of defense-related genes was observed, whereas few changes in the expression levels of defense-related genes were found in infected root tissues. In infected roots, more plant genes were repressed than were induced, especially at the earlier stages of infection. Although many known cotton defense responses were identified, including induction of pathogenesis-related genes and gossypol biosynthesis genes, potential new defense responses also were identified, such as the biosynthesis of lignans. Many of the stress-related gene responses were common to both tissues. The repression of drought-responsive proteins such as aquaporins in both roots and hypocotyls represents a previously unreported response of a host to pathogen attack that may be specific to vascular wilt diseases. Gene expression results implicated the phytohormones ethylene and auxin in the disease process. Biochemical analysis of hormone level changes supported this observation.     

Genes expressed in Ascochyta rabiei-inoculated chickpea plants and elicited cell cultures as detected by differential cDNA-hybridization

In response to the exogenous application of elicitors and attempted invasion by pathogens, plants exhibit a wide range of defense reactions. To understand the defense mechanisms at the level of gene activation and deactivation, differential screenings were performed to isolate cDNA clones which are differentially expressed in pathogen-inoculated resistant chickpea plants and elicitor-treated cell cultures. A plenty of genes were isolated and arranged in 5 groups, namely defense-related pathways, signal transduction pathways, regulation of gene expression, catabolic pathways and primary metabolism. Most of these genes were activated although several genes were also found to be suppressed. We discuss the plausible functions of cDNA products in plant defense responses. The cDNAs provide a variety of tools to investigate molecular mechanisms of defense responses and clearly reflect the massive genomic and metabolic changes which occur during manifestation of antimicrobial defense.     

植物抗病反应的信号传导网络

植物由抗病基因介导的防卫过程存在一系列生理生化和分子生物学反应,这些反应从病原菌侵染点开始的超敏反应(HR)并延伸到远处组织的系统抗性或获得性抗性(SAR),受制于一种信号传导网络的调控。这个信号系统由抗病蛋白和病原菌非毒性蛋白在一种配体-受体的互作模式下激发,并由信号分子H2O2,NO和系统信号分子SA,JA和乙烯和通过关键调控基因传递和放大,最终诱导一系列防卫反应基因的表达和代谢的变化而产生抗性。植物防卫信号的产生有类似于动物免疫系统因子的介导,并可由非寄主病原菌或诱导子诱发。这些信号途径所产生的广谱抗性为植物抗病基因工程的应用奠定了基础。     

Pyrrolizidine alkaloid biosynthesis,evolution of a pathway in plant secondary metabolism

The system of pyrrolizidine alkaloids has proven to be a powerful system for studying the evolution of a biosynthetic pathway in plant secondary metabolism. Pyrrolizidine alkaloids are typical plant secondary products produced by the plant as a defense against herbivores. The first specific enzyme, homospermidine synthase, has been shown to have evolved by duplication of the gene encoding deoxyhypusine synthase, which is involved in primary metabolism. Despite the identical function of homospermidine synthase for pyrrolizidine alkaloid biosynthesis in the various plant lineages, this gene duplication has occurred several times independently during angiosperm evolution. After duplication, these gene copies diverged with respect to gene function and regulation. In the diverse plant lineages producing pyrrolizidine alkaloids, homospermidine synthase has been shown to be expressed in a variety of tissues, suggesting that the regulatory elements were recruited individually after the duplication of the structural gene. The molecular, kinetic, and expression data of this system are discussed with respect to current models of gene and pathway evolution.     

Regulatory mechanisms of host plant defense responses to arbuscular mycorrhiza

Arbuscular mycorrhizal (AM) fungi colonize the roots of over 80% of terrestrial plant species, forming mutually beneficial symbioses. During the colonization process, symbiotic partners recognize each other, and undergo observable morphological and physiological changes; indicating that symbiosis formation involves multiple factors that are finely regulated. Sometimes host plants generate a transient, weak, defense response. This response and its down-regulation play a very important role in the development of AM symbioses. Although AM fungi can infect a wide range of host root tissues, which host defense may play a crucial role is hypothesized from the fact that hyphal expansion is only observed in the root cortex.
We discuss five defense mechanisms. (1) The degradation of exogenous elicitors. The host’s weak defense response may be due to the degradation of the exogenous elicitor chitin, or the prevention of release of an endogenous inductor from the plant cell wall. (2) The inactivation of defense signal molecules. Some defense signal molecules such as hydrogen peroxidase, salicylic acid (SA), and jasmonic acid (JA), are inactivated in host plants. This helps to avoid the turn-on of defense-related genes and facilitate mycorrhizal formation. (3) The regulation of plant hormones and plant photosynthates. Plant hormone levels and plant photosynthate metabolism both change during AM colonization. These mechanisms need further exploration. (4) Changes in levels of phosphorous (P), and (iso)flavonoids. High P levels can induce some defense genes to express hydrogen peroxidase, chitinase, and glucanase. These gene products can repress colonization by AM fungi. The plant defense response regulatory effect for different (iso)flavonoids varies, and their levels are regulated by P. (5) The suppressed expression of symbiotic genes. Some symbiosis-related genes inhibit plant defense responses, but it is still unclear which mechanisms underlie gene regulation. We provide here a theoretical basis for research into AM symbiosis that may promote study of host plant resistance and the mechanisms of symbiosis formation.
We provide a deeper insight into the signal transduction pathways of mycorrhization that will aid understanding and analysis of plant defense mechanisms in the AM context. The on-going development of genome sequencing technology will contribute greatly to the detailed study of symbiosis-related genes, and pathogenesis-related protein genes. These related genes may be induced to express corresponding proteins, be repressed, postpone expression or even shutdown, or both may work together to form symbioses. Elucidation of these features will help us understand the roles that plant defenses play in mycorrhizal formation; providing an unprecedented opportunity for research into mycorrhizal molecular biology and the interaction of symbiotic partners, and allowing the underlying mechanisms to be gradually uncovered.     

Oligogalacturonide-mediated induction of a gene involved in jasmonic acid synthesis in response to the cell-wall-degrading enzymes of the plant pathogen Erwinia carotovora.

Identification of Arabidopsis thaliana genes responsive to plant cell-wall-degrading enzymes of Erwinia carotovora subsp. carotovora led to the isolation of a cDNA clone with high sequence homology to the gene for allene oxide synthase, an enzyme involved in the biosynthesis of jasmonates. Expression of the corresponding gene was induced by the extracellular enzymes from this pathogen as well as by treatment with methyl jasmonate and short oligogalacturonides (OGAs). This suggests that OGAs are involved in the induction of the jasmonate pathway during plant defense response to E. carotovora subsp. carotovora attack.