Sheng Yang He

JAZ repressor proteins are targets of the SCFCOI1 complex during jasmonate signalling

Jasmonate and related signalling compounds have a crucial role in both host immunity and development in plants, but the molecular details of the signalling mechanism are poorly understood. Here we identify members of the jasmonate ZIM-domain (JAZ) protein family as key regulators of jasmonate signalling. JAZ1 protein acts to repress transcription of jasmonate-responsive genes. Jasmonate treatment causes JAZ1 degradation and this degradation is dependent on activities of the SCFCOI1 ubiquitin ligase and the 26S proteasome. Furthermore, the jasmonoyl–isoleucine (JA–Ile) conjugate, but not other jasmonate-derivatives such as jasmonate, 12-oxo-phytodienoic acid, or methyl-jasmonate, promotes physical interaction between COI1 and JAZ1 proteins in the absence of other plant proteins. Our results suggest a model in which jasmonate ligands promote the binding of the SCFCOI1 ubiquitin ligase to and subsequent degradation of the JAZ1 repressor protein, and implicate the SCFCOI1–JAZ1 protein complex as a site of perception of the plant hormone JA–Ile.

Plant Stomata Function in Innate Immunity against Bacterial Invasion

Microbial entry into host tissue is a critical first step in causing infection in animals and plants. In plants, it has been assumed that microscopic surface openings, such as stomata, serve as passive ports of bacterial entry during infection. Surprisingly, we found that stomatal closure is part of a plant innate immune response to restrict bacterial invasion. Stomatal guard cells of Arabidopsis perceive bacterial surface molecules, which requires the FLS2 receptor, production of nitric oxide, and the guard-cell-specific OST1 kinase. To circumvent this innate immune response, plant pathogenic bacteria have evolved specific virulence factors to effectively cause stomatal reopening as an important pathogenesis strategy. We provide evidence that supports a model in which stomata, as part of an integral innate immune system, act as a barrier against bacterial infection.

Jasmonate perception by inositol-phosphate-potentiated COI1–JAZ co-receptor

Jasmonates are a family of plant hormones that regulate plant growth, development and responses to stress. The F-box protein CORONATINE INSENSITIVE 1 (COI1) mediates jasmonate signalling by promoting hormone-dependent ubiquitylation and degradation of transcriptional repressor JAZ proteins. Despite its importance, the mechanism of jasmonate perception remains unclear. Here we present structural and pharmacological data to show that the true Arabidopsis jasmonate receptor is a complex of both COI1 and JAZ. COI1 contains an open pocket that recognizes the bioactive hormone (3R,7S)-jasmonoyl-l-isoleucine (JA-Ile) with high specificity. High-affinity hormone binding requires a bipartite JAZ degron sequence consisting of a conserved α-helix for COI1 docking and a loop region to trap the hormone in its binding pocket. In addition, we identify a third critical component of the jasmonate co-receptor complex, inositol pentakisphosphate, which interacts with both COI1 and JAZ adjacent to the ligand. Our results unravel the mechanism of jasmonate perception and highlight the ability of F-box proteins to evolve as multi-component signalling hubs.

Innate immunity in plants : an arms race between pattern recognition receptors in plants and effectors in microbial pathogens

For many years, research on a suite of plant defense responses that begin when plants are exposed to general microbial elicitors was underappreciated, for a good reason: There has been no critical experimental demonstration of their importance in mediating plant resistance during pathogen infection. Today, these microbial elicitors are named pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) and the plant responses are known as PAMP-triggered immunity (PTI). Recent studies provide an elegant explanation for the difficulty of demonstrating the role of PTI in plant disease resistance. It turns out that the important contribution of PTI to disease resistance is masked by pathogen virulence effectors that have evolved to suppress it.

COI1 is a critical component of a receptor for jasmonate and the bacterial virulence factor coronatine

Jasmonate (JA) is a lipid-derived hormone that regulates diverse aspects of plant immunity and development. An amino acid-conjugated form of JA, jasmonoyl–isoleucine (JA–Ile), stimulates binding of the F-box protein coronatine-insensitive 1 (COI1) to, and subsequent ubiquitin-dependent degradation of, jasmonate ZIM domain (JAZ) proteins that repress transcription of JA-responsive genes. The virulence factor coronatine (COR), which is produced by plant pathogenic strains of Pseudomonas syringae, suppresses host defense responses by activating JA signaling in a COI1-dependent manner. Although previous data indicate that COR acts as a molecular mimic of JA–Ile, the mechanism by which JA–Ile and COR are perceived by plant cells remains unknown. Here, we show that interaction of tomato COI1 with divergent members of the JAZ family is highly specific for JA–Ile and structurally related JA conjugates and that COR is ≈1,000-fold more active than JA–Ile in promoting this interaction in vitro. JA–Ile competes for binding of COR to COI1–JAZ complexes, demonstrating that COR and JA–Ile are recognized by the same receptor. Binding of COR to the COI1–JAZ complex requires COI1 and is severely impaired by a point mutation in the putative ligand-binding pocket of COI1. Finally, we show that the C-terminal region of JAZ3 containing the highly conserved Jas motif is necessary and sufficient for hormone-induced COI1–JAZ interaction. These findings demonstrate that COI1 is a critical component of the JA receptor and that COR exerts its virulence effects by functioning as a potent agonist of this receptor system.

A Pseudomonas syringae type III effector suppresses cell wall-based extracellular defense in susceptible Arabidopsis plants

Bacterial effector proteins secreted through the type III secretion system (TTSS) play a crucial role in causing plant and human diseases. Although the ability of type III effectors to trigger defense responses in resistant plants is well understood, the disease-promoting functions of type III effectors in susceptible plants are largely enigmatic. Previous microscopic studies suggest that in susceptible plants the TTSS of plant-pathogenic bacteria transports suppressors of a cell wall-based plant defense activated by the TTSS-defective hrp mutant bacteria. However, the identity of such suppressors has remained elusive. We discovered that the Pseudomonas syringae TTSS down-regulated the expression of a set of Arabidopsis genes encoding putatively secreted cell wall and defense proteins in a salicylic acid-independent manner. Transgenic expression of AvrPto repressed a similar set of host genes, compromised defense-related callose deposition in the host cell wall, and permitted substantial multiplication of an hrp mutant. AvrPto is therefore one of the long postulated suppressors of an salicylic acid-independent, cell wall-based defense that is aimed at hrp mutant bacteria.

Role of Stomata in Plant Innate Immunity and Foliar Bacterial Diseases

Pathogen entry into host tissue is a critical first step in causing infection. For foliar bacterial plant pathogens, natural surface openings, such as stomata, are important entry sites. Historically, these surface openings have been considered as passive portals of entry for plant pathogenic bacteria. However, recent studies have shown that stomata can play an active role in limiting bacterial invasion as part of the plant innate immune system. As a counter-defense, the plant pathogen Pseudomonas syringae pv. tomato DC3000 uses the virulence factor coronatine to actively open stomata. In nature, many foliar bacterial disease outbreaks require high humidity, rain, or storms, which could favor stomatal opening and/or bypass stomatal defense by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomatal closure and opening could fill gaps in our understanding of bacterial pathogenesis, disease epidemiology, and microbiology of the phyllosphere.

The Arabidopsis thaliana-pseudomonas syringae interaction.

Pseudomonas syringae is a Gram-negative, rod-shaped bacterium with polar flagella (Figure 1; Agrios, 1997). Strains of P. syringae collectively infect a wide variety of plants. Different strains of P. syringae, however, are known for their diverse and host-specific interactions with plants (Hirano and Upper, 2000). A specific strain may be assigned to one of at least 40 pathovars based on its host range among different plant species (Gardan et al., 1999) and then further assigned to a race based on differential interactions among cultivars of the host plant. Understanding the molecular basis of this high level of host specificity has been a driving force in using P. syringae as a model for the study of host-pathogen interactions. In crop fields, infected seeds are often an important source of primary inoculum in P. syringae diseases, and epiphytic bacterial growth on leaf surfaces often precedes disease development (Hirano and Upper, 2000). P. syringae enters the host tissues (usually leaves) through wounds or natural openings such as stomata, and in a susceptible plant it multiplies to high population levels in intercellular spaces. Infected leaves show water-soaked patches, which eventually become necrotic. Depending on P. syringae strains, necrotic lesions may be surrounded by diffuse chlorosis. Some strains of P. syringae also cause cankers and galls (Agrios, 1997). In resistant plants, on the other hand, P. syringae triggers the hypersensitive response (HR), a rapid, defense-associated death of plant cells in contact with the pathogen (Klement, 1963; Klement et al., 1964; Bent, 1996; Greenberg, 1996; Dangl et al., 1996; Hammond-Kosack and Jones, 1997). In this situation, P. syringae fails to multiply to high population levels and causes no disease symptoms. Figure 1. A transmission electron microscope image of Pseudomonas syringae pv. tomato DC3000. Note that DC3000 produces polar flagella (15 nm in diameter) and a few Hrp pili (8 nm in diameter). The flagella and Hrp pili are indicated with arrows. Flagella enable ... In the late 1980s, several strains belonging to pathovars tomato, maculicola, pisi, and atropurpurea of Pseudomonas syringae were discovered to infect the model plant Arabidopsis thaliana (reviewed by Crute et al., 1994). The establishment of the Arabidopsis-P. syringae pathosystem triggered a period of highly productive research that has contributed to the elucidation of the fascinating mechanisms underlying plant recognition of pathogens, signal transduction pathways controlling plant defense responses, host susceptibility, and pathogen virulence and avirulence determinants. In this chapter we trace the discovery of this pathosystem, overview the most salient aspects of this interaction, and point out the gaps in our knowledge. At the end of this chapter we will also provide a glossary of relevant pathology-related technical terms (Appendix I), a list of people who are studying this interaction so readers can seek help if they have further questions about the Arabidopsis-P. syringae interaction (Appendix II), and several experimental procedures commonly used in the study of the Arabidopsis-P. syringae interaction (Appendix III).

Mitogen-Activated Protein Kinases 3 and 6 Are Required for Full Priming of Stress Responses in Arabidopsis thaliana

In plants and animals, induced resistance (IR) to biotic and abiotic stress is associated with priming of cells for faster and stronger activation of defense responses. It has been hypothesized that cell priming involves accumulation of latent signaling components that are not used until challenge exposure to stress. However, the identity of such signaling components has remained elusive. Here, we show that during development of chemically induced resistance in Arabidopsis thaliana, priming is associated with accumulation of mRNA and inactive proteins of mitogen-activated protein kinases (MPKs), MPK3 and MPK6. Upon challenge exposure to biotic or abiotic stress, these two enzymes were more strongly activated in primed plants than in nonprimed plants. This elevated activation was linked to enhanced defense gene expression and development of IR. Strong elicitation of stress-induced MPK3 and MPK6 activity is also seen in the constitutive priming mutant edr1, while activity was attenuated in the priming-deficient npr1 mutant. Moreover, priming of defense gene expression and IR were lost or reduced in mpk3 or mpk6 mutants. Our findings argue that prestress deposition of the signaling components MPK3 and MPK6 is a critical step in priming plants for full induction of defense responses during IR.

Suppression of the microRNA pathway by bacterial effector proteins.

Plants and animals sense pathogen-associated molecular patterns (PAMPs) and in turn differentially regulate a subset of microRNAs (miRNAs). However, the extent to which the miRNA pathway contributes to innate immunity remains unknown. Here, we show that miRNA-deficient mutants of Arabidopsis partly restore growth of a type III secretion-defective mutant of Pseudomonas syringae. These mutants also sustained growth of nonpathogenic Pseudomonas fluorescens and Escherichia coli strains, implicating miRNAs as key components of plant basal defense. Accordingly, we have identified P. syringae effectors that suppress transcriptional activation of some PAMP-responsive miRNAs or miRNA biogenesis, stability, or activity. These results provide evidence that, like viruses, bacteria have evolved to suppress RNA silencing to cause disease.