\u3ci\u3ePseudomonas syringae\u3c/i\u3e type III effectors: Targets and roles in plant immunity

Pseudomonas syringae is a Gram-negative bacterial pathogen that infects many crops. A central virulence strategy P. syringae uses to successfully infect plants is the injection of type III effector proteins (T3Es) into plant cells through a type IIII protein secretion system (T3SS). The T3SS is a molecular syringe found in many Gram-negative bacterial pathogens of plants and animals that transport T3Es from the bacterial cytosol into eukaryotic cells. T3Es disrupt host processes in the plant immune system required to restrict pathogen ingress. The plant innate immune system is divided in two branches, pathogen-associated molecular patterns (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). The first branch recognizes conserved molecules found in microbes, known as PAMPs, and the second has the capacity to recognize injected T3Es. T3Es can suppress both PTI and ETI allowing P. syringae to circumvent the plant immune system and multiply in plant tissue. The majority of T3Es plant targets, their enzymatic activity and the mechanism of suppression of plant immunity are not known. P. syringae pv. tomato (Pto) DC3000 injects about 35 T3Es into plant cells. In this study I characterized two T3Es from Pto DC3000. Firstly, I focused on the T3E HopD1. HopD1 suppresses plant immunity associated with ETI but not PTI, suggesting that HopD1 was acquired later in the co-evolution of the pathogen and plant. HopD1 is targeted to the endoplasmic reticulum of plant cells where it interacts with the Arabidopsis NAC transcription factor NTL9. HopD1’s function in virulence involves the inhibition of NTL9-regulated genes during ETI. Secondly, I focused on the T3E HopA1. This T3E exists in two classes, which I found are recognized differently in plants. HopA1 suppresses PTI and its structure resembles phosphothreonine lyases form animal pathogens. The putative active site of HopA1 was identified and I found that site-directed mutations in the active site abrogated HopA1-dependent phenotypes. HopA1 localizes mainly to plasma membrane of plant cells where it interacts with the Arabidopsis type 2C phosphatases PLL4 and PLL5. These phosphatases play roles in plant immunity as negative regulators and HopA1 likely prevents their deregulation preventing induction of the plant immune system. Adviser: James R. Alfan

Determinar la presencia de geminivirus y fitoplasmas en tomate en Guatemala, El Salvador, Honduras y Nicaragua

96 p.Las infecciones virales en cultivos hortícolas en Centro América son el principal problema en la producción. En las últimas dos décadas se ha abusado de los agroquímicos para controlar a los insectos vectores, creando problemas de resistencia, contaminación ambiental y daños a la salud humana. Asimismo, en los últimos años, el cultivo de papa (Solanum tuberosum L.) ha sido afectado por una nueva enfermedad denominada “punta morada de la papa”, causada por un fitoplasma que disminuye la calidad de los tubérculos. Desde el 2000 se han promovido campañas en México y Centro América para hacer aplicaciones de antibióticos en cultivos de tomate, ya que los daños se atribuyen a fitoplasmas (susceptibles a antibióticos), que no han podido ser controlados por los insecticidas tradicionales utilizados para manejar infecciones virales. Estas recomendaciones se han hecho sin ningún análisis previo para establecer la etiología de estas enfermedades

Plant Pathogen Effectors: Cellular Probes Interfering with Plant Defenses in Spatial and Temporal Manners

Plants possess large arsenals of immune receptors capable of recognizing all pathogen classes. To cause disease, pathogenic organisms must be able to overcome physical barriers, suppress or evade immune perception, and derive nutrients from host tissues. Consequently, to facilitate some of these processes, pathogens secrete effector proteins that promote colonization. This review covers recent advances in the field of effector biology, focusing on conserved cellular processes targeted by effectors from diverse pathogens. The ability of effectors to facilitate pathogen entry into the host interior, suppress plant immune perception, and alter host physiology for pathogen benefit is discussed. Pathogens also deploy effectors in a spatial and temporal manner, depending on infection stage. Recent advances have also enhanced our understanding of effectors acting in specific plant organs and tissues. Effectors are excellent cellular probes that facilitate insight into biological processes as well as key points of vulnerability in plant immune signaling networks

Regulated Disorder: Posttranslational Modifications Control the RIN4 Plant Immune Signaling Hub.

RIN4 is an intensively studied immune regulator in Arabidopsis and is involved in perception of microbial features outside and bacterial effectors inside plant cells. Furthermore, RIN4 is conserved in land plants and is targeted for posttranslational modifications by several virulence proteins from the bacterial pathogen Pseudomonas syringae. Despite the important roles of RIN4 in plant immune responses, its molecular function is not known. RIN4 is an intrinsically disordered protein (IDP), except at regions where pathogen-induced posttranslational modifications take place. IDP act as hubs for protein complex formation due to their ability to bind to multiple client proteins and, thus, are important players in signal transduction pathways. RIN4 is known to associate with multiple proteins involved in immunity, likely acting as an immune-signaling hub for the formation of distinct protein complexes. Genetically, RIN4 is a negative regulator of immunity, but diverse posttranslational modifications can either enhance its negative regulatory function or, on the contrary, render it a potent immune activator. In this review, we describe the structural domains of RIN4 proteins, their intrinsically disordered regions, posttranslational modifications, and highlight the implications that these features have on RIN4 function. In addition, we will discuss the potential role of plasma membrane subdomains in mediating RIN4 protein complex formations

Plant-Pathogen Effectors: Cellular Probes Interfering with Plant Defenses in Spatial and Temporal Manners

Plants possess large arsenals of immune receptors capable of recognizing all pathogen classes. To cause disease, pathogenic organisms must be able to overcome physical barriers, suppress or evade immune perception, and derive nutrients from host tissues. Consequently, to facilitate some of these processes, pathogens secrete effector proteins that promote colonization. This review covers recent advances in the field of effector biology, focusing on conserved cellular processes targeted by effectors from diverse pathogens. The ability of effectors to facilitate pathogen entry into the host interior, suppress plant immune perception, and alter host physiology for pathogen benefit is discussed. Pathogens also deploy effectors in a spatial and temporal manner, depending on infection stage. Recent advances have also enhanced our understanding of effectors acting in specific plant organs and tissues. Effectors are excellent cellular probes that facilitate insight into biological processes as well as key points of vulnerability in plant immune signaling networks

The Pseudomonas syringaetype III effector HopD1 suppresses effector-triggered immunity, localizes to the endoplasmic reticulum, and targets the Arabidopsis transcription factor NTL9

Pseudomonas syringae type III effectors are known to suppress plant immunity to promote bacterial virulence. However, the activities and targets of these effectors are not well understood. We used genetic, molecular, and cell biology methods to characterize the activities, localization, and target of the HopD1 type III effector in Arabidopsis. HopD1 contributes to P. syringae virulence in Arabidopsis and reduces effector-triggered immunity (ETI) responses but not pathogen-associated molecular pattern-triggered immunity (PTI) responses. Plants expressing HopD1 supported increased growth of ETI-inducing P. syringae strains compared with wild-type Arabidopsis. We show that HopD1 interacts with the membrane-tethered Arabidopsis transcription factor NTL9 and demonstrate that this interaction occurs at the endoplasmic reticulum (ER). A P. syringae hopD1 mutant and ETI-inducing P. syringae strains exhibited enhanced growth on Arabidopsis ntl9 mutant plants. Conversely, growth of P. syringae strains was reduced in plants expressing a constitutively active NTL9 derivative, indicating that NTL9 is a positive regulator of plant immunity. Furthermore, HopD1 inhibited the induction of NTL9-regulated genes during ETI but not PTI. HopD1 contributes to P. syringae virulence in part by targeting NTL9, resulting in the suppression of ETI responses but not PTI responses and the promotion of plant pathogenicity

XAP5 CIRCADIAN TIMEKEEPER Affects Both DNA Damage Responses and Immune Signaling in Arabidopsis

Numerous links have been reported between immune response and DNA damage repair pathways in both plants and animals but the precise nature of the relationship between these fundamental processes is not entirely clear. Here, we report that XAP5 CIRCADIAN TIMEKEEPER (XCT), a protein highly conserved across eukaryotes, acts as a negative regulator of immunity in Arabidopsis thaliana and plays a positive role in responses to DNA damaging radiation. We find xct mutants have enhanced resistance to infection by a virulent bacterial pathogen, Pseudomonas syringae pv. tomato DC3000, and are hyper-responsive to the defense-activating hormone salicylic acid (SA) when compared to wild-type. Unlike most mutants with constitutive effector-triggered immunity (ETI), xct plants do not have increased levels of SA and retain enhanced immunity at elevated temperatures. Genetic analysis indicates XCT acts independently of NONEXPRESSOR OF PATHOGENESIS RELATED GENES1 (NPR1), which encodes a known SA receptor. Since DNA damage has been reported to potentiate immune responses, we next investigated the DNA damage response in our mutants. We found xct seedlings to be hypersensitive to UV-C and γ radiation and deficient in phosphorylation of the histone variant H2A.X, one of the earliest known responses to DNA damage. These data demonstrate that loss of XCT causes a defect in an early step of the DNA damage response pathway. Together, our data suggest that alterations in DNA damage response pathways may underlie the enhanced immunity seen in xct mutants

Effectors from a Bacterial Vector-Borne Pathogen Exhibit Diverse Subcellular Localization, Expression Profiles, and Manipulation of Plant Defense

Climate change is predicted to increase the prevalence of vector-borne disease due to expansion of insect populations. 'Candidatus Liberibacter solanacearum' is a phloem-limited pathogen associated with multiple economically important diseases in solanaceous crops. Little is known about the strategies and pathogenicity factors 'Ca. L. solanacearum' uses to colonize its vector and host. We determined the 'Ca. L. solanacearum' effector repertoire by predicting proteins secreted by the general secretory pathway across four different 'Ca. L. solanacearum' haplotypes, investigated effector localization in planta, and profiled effector expression in the vector and host. The localization of 'Ca. L. solanacearum' effectors in Nicotiana spp. revealed diverse eukaryotic subcellular targets. The majority of tested effectors were unable to suppress plant immune responses, indicating they possess unique activities. Expression profiling in tomato and the psyllid Bactericera cockerelli indicated 'Ca. L. solanacearum' differentially interacts with its host and vector and can switch effector expression in response to these environments. This study reveals 'Ca. L. solanacearum' effectors possess complex expression patterns, target diverse host organelles and the majority are unable to suppress host immune responses. A mechanistic understanding of 'Ca. L. solanacearum' effector function will reveal novel targets and provide insight into phloem biology. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license