Host Specificity Controlled by and Effector Genes in the Finger Millet Blast Pathogen in Eastern Africa.

, a devastating pathogen of finger millet (), secretes effector molecules during infection to manipulate host immunity. This study determined the presence of avirulence effector genes and in 221 blast isolates from eastern Africa. Most Ethiopian isolates carried both and .

Disruption of the Interfacial Membrane Leads to Effector Re-location and Lifestyle Switch During Rice Blast Disease.

To cause the devastating rice blast disease, the hemibiotrophic fungus produces invasive hyphae (IH) that are enclosed in a plant-derived interfacial membrane, known as the extra-invasive hyphal membrane (EIHM), in living rice cells. Little is known about when the EIHM is disrupted and how the disruption contributes to blast disease. Here we show that the disruption of the EIHM correlates with the hyphal growth stage in first-invaded susceptible rice cells.

Subcellular three-dimensional imaging deep through multicellular thick samples by structured illumination microscopy and adaptive optics.

Structured Illumination Microscopy enables live imaging with sub-diffraction resolution. Unfortunately, optical aberrations can lead to loss of resolution and artifacts in Structured Illumination Microscopy rendering the technique unusable in samples thicker than a single cell. Here we report on the combination of Adaptive Optics and Structured Illumination Microscopy enabling imaging with 150 nm lateral and 570 nm axial resolution at a depth of 80 µm through Caenorhabditis elegans.

Tandem DNA repeats contain cis-regulatory sequences that activate biotrophy-specific expression of Magnaporthe effector gene PWL2.

During plant infection, fungi secrete effector proteins in coordination with distinct infection stages. Thus, the success of plant infection is determined by precise control of effector gene expression. We analysed the PWL2 effector gene of the rice blast fungus Magnaporthe oryzae to understand how effector genes are activated specifically during the early biotrophic stages of rice infection.

Role of Two Metacaspases in Development and Pathogenicity of the Rice Blast Fungus Magnaporthe oryzae.

Rice blast disease caused by is a devastating disease of cultivated rice worldwide. Infections by this fungus lead to a significant reduction in rice yields and threats to food security. To gain better insight into growth and cell death in during infection, we characterized two predicted metacaspase proteins, MoMca1 and MoMca2.

Two nuclear effectors of the rice blast fungus modulate host immunity via transcriptional reprogramming.

Pathogens utilize multiple types of effectors to modulate plant immunity. Although many apoplastic and cytoplasmic effectors have been reported, nuclear effectors have not been well characterized in fungal pathogens. Here, we characterize two nuclear effectors of the rice blast pathogen Magnaporthe oryzae.

Nup84 persists within the nuclear envelope of the rice blast fungus, Magnaporthe oryzae, during mitosis.

The arrangement of the nuclear envelope in the rice blast fungus, Magnaporthe oryzae, was previously undetermined. Here, we identified two conserved components of the nuclear envelope, a core nucleoporin, Nup84, and an inner nuclear membrane protein, Src1. Live-cell super-resolution structured illumination microscopy revealed that Nup84-tdTomato and Src1-EGFP colocalized within the nuclear envelope during interphase and that Nup84-tdTomato remained associated with the dividing nucleus.

Thiosulfinate Tolerance Is a Virulence Strategy of an Atypical Bacterial Pathogen of Onion.

Unlike most characterized bacterial plant pathogens, the broad-host-range plant pathogen Pantoea ananatis lacks both the virulence-associated type III and type II secretion systems. In the absence of these typical pathogenicity factors, P. ananatis induces necrotic symptoms and extensive cell death in onion tissue dependent on the HiVir proposed secondary metabolite synthesis gene cluster.

A strikingly-angled spindle mediates nuclear migration during colonization of rice cells infected by Magnaporthe oryzae.

To cause rice blast disease, Magnaporthe oryzae must properly organize microtubules and position nuclei during colonization of host cells. Live cell confocal imaging of fluorescently-tagged microtubules and nuclei of M. oryzae invasive hyphae reveals that microtubules form a cage-like arrangement around nuclei during interphase and that the mitotic spindle forms and mediates nuclear migration while integrity of the nuclear envelope is lost.

Visualizing the Movement of Magnaporthe oryzae Effector Proteins in Rice Cells During Infection.

During biotrophy, filamentous pathogens such as the rice blast fungus Magnaporthe oryzae deliver effector proteins into live host cells to facilitate colonization. We describe three complementary assays for visualizing M. oryzae effector translocation into the rice cytoplasm and cell-to-cell movement during infection.

A nuclear contortionist: the mitotic migration of nuclei during plant infection.

is a filamentous fungus, which causes significant destruction to cereal crops worldwide. To infect plant cells, the fungus develops specialised constricted structures such as the penetration peg and the invasive hyphal peg. Live-cell imaging of during plant infection reveals that nuclear migration occurs during intermediate mitosis, in which the nuclear envelope neither completely disassembles nor remains entirely intact.

Vacuole Dynamics in Rice Cells Invaded by the Blast Fungus Magnaporthe oryzae.

We describe a fluorescence imaging method to visualize the dynamics of the central vacuole in rice cells during invasion by the blast fungus Magnaporthe oryzae. This method utilizes the combination of confocal microscopy, rice sheath cells (optically transparent), fluorescently tagged M. oryzae (red fluorescence), and fluorescein diacetate staining (green fluorescence; visualizing vacuole dynamics).

Genome wide analysis of the transition to pathogenic lifestyles in Magnaporthales fungi.

The rice blast fungus Pyricularia oryzae (syn. Magnaporthe oryzae, Magnaporthe grisea), a member of the order Magnaporthales in the class Sordariomycetes, is an important plant pathogen and a model species for studying pathogen infection and plant-fungal interaction. In this study, we generated genome sequence data from five additional Magnaporthales fungi including non-pathogenic species, and performed comparative genome analysis of a total of 13 fungal species in the class Sordariomycetes to understand the evolutionary history of the Magnaporthales and of fungal pathogenesis.

Nuclear and structural dynamics during the establishment of a specialized effector-secreting cell by Magnaporthe oryzae in living rice cells.

Background: To cause an economically important blast disease on rice, the filamentous fungus Magnaporthe oryzae forms a specialized infection structure, called an appressorium, to penetrate host cells. Once inside host cells, the fungus produces a filamentous primary hypha that differentiates into multicellular bulbous invasive hyphae (IH), which are surrounded by a host-derived membrane. These hyphae secrete cytoplasmic effectors that enter host cells presumably via the biotrophic interfacial complex (BIC).

The appressorium of the rice blast fungus Magnaporthe oryzae remains mitotically active during post-penetration hyphal growth.

To investigate the mitotic dynamics of an appressorium, we used live-cell confocal imaging of a fluorescence-based mitotic reporter strain of Magnaporthe oryzae. We present evidence that the M. oryzae appressorium remains viable and mitotically active well after host penetration.

Mitotic stopwatch for the blast fungus Magnaporthe oryzae during invasion of rice cells.

To study nuclear dynamics of Magnaporthe oryzae, we developed a novel mitotic reporter strain with GFP-NLS (localized in nuclei during interphase but in the cytoplasm during mitosis) and H1-tdTomato (localized in nuclei throughout the cell cycle). Time-lapse confocal microscopy of the reporter strain during host cell invasion provided several new insights into nuclear division and migration in M. oryzae: (i) mitosis lasts about 5min; (ii) mitosis is semi-closed; (iii) septal pores are closed during mitosis; and (iv) a nucleus exhibits extreme constriction (approximately from 2μm to 0.

Live-cell fluorescence imaging to investigate the dynamics of plant cell death during infection by the rice blast fungus Magnaporthe oryzae.

Background: Plant cell death plays important roles during plant-pathogen interactions. To study pathogen-induced cell death, there is a need for cytological tools that allow determining not only host cell viability, but also cellular events leading to cell death with visualization of pathogen development. Here we describe a live cell imaging method to provide insights into the dynamics of cell death in rice (Oryza sativa).

Global expression profiling of transcription factor genes provides new insights into pathogenicity and stress responses in the rice blast fungus.

Because most efforts to understand the molecular mechanisms underpinning fungal pathogenicity have focused on studying the function and role of individual genes, relatively little is known about how transcriptional machineries globally regulate and coordinate the expression of a large group of genes involved in pathogenesis. Using quantitative real-time PCR, we analyzed the expression patterns of 206 transcription factor (TF) genes in the rice blast fungus Magnaporthe oryzae under 32 conditions, including multiple infection-related developmental stages and various abiotic stresses. The resulting data, which are publicly available via an online platform, provided new insights into how these TFs are regulated and potentially work together to control cellular responses to a diverse array of stimuli.

The Magnaporthe oryzae effector AvrPiz-t targets the RING E3 ubiquitin ligase APIP6 to suppress pathogen-associated molecular pattern-triggered immunity in rice.

Although the functions of a few effector proteins produced by bacterial and oomycete plant pathogens have been elucidated in recent years, information for the vast majority of pathogen effectors is still lacking, particularly for those of plant-pathogenic fungi. Here, we show that the avirulence effector AvrPiz-t from the rice blast fungus Magnaporthe oryzae preferentially accumulates in the specialized structure called the biotrophic interfacial complex and is then translocated into rice (Oryza sativa) cells. Ectopic expression of AvrPiz-t in transgenic rice suppresses the flg22- and chitin-induced generation of reactive oxygen species (ROS) and enhances susceptibility to M.

Recent advances in rice blast effector research.

To cause rice blast disease, the fungus Magnaporthe oryzae produces biotrophic invasive hyphae that secrete effectors at the host-pathogen interface. Effectors facilitate disease development, but some (avirulence effectors) also trigger the host's resistance gene-mediated hypersensitive response and block disease. The number of cloned M.

Translocation of Magnaporthe oryzae effectors into rice cells and their subsequent cell-to-cell movement.

Knowledge remains limited about how fungal pathogens that colonize living plant cells translocate effector proteins inside host cells to regulate cellular processes and neutralize defense responses. To cause the globally important rice blast disease, specialized invasive hyphae (IH) invade successive living rice (Oryza sativa) cells while enclosed in host-derived extrainvasive hyphal membrane. Using live-cell imaging, we identified a highly localized structure, the biotrophic interfacial complex (BIC), which accumulates fluorescently labeled effectors secreted by IH.

Interaction transcriptome analysis identifies Magnaporthe oryzae BAS1-4 as Biotrophy-associated secreted proteins in rice blast disease.

Biotrophic invasive hyphae (IH) of the blast fungus Magnaporthe oryzae secrete effectors to alter host defenses and cellular processes as they successively invade living rice (Oryza sativa) cells. However, few blast effectors have been identified. Indeed, understanding fungal and rice genes contributing to biotrophic invasion has been difficult because so few plant cells have encountered IH at the earliest infection stages.

The ER chaperone LHS1 is involved in asexual development and rice infection by the blast fungus Magnaporthe oryzae.

In planta secretion of fungal pathogen proteins, including effectors destined for the plant cell cytoplasm, is critical for disease progression. However, little is known about the endoplasmic reticulum (ER) secretion mechanisms used by these pathogens. To determine if normal ER function is crucial for fungal pathogenicity, Magnaporthe oryzae genes encoding proteins homologous to yeast Lhs1p and Kar2p, members of the heat shock protein 70 family in Saccharomyces cerevisiae, were cloned and characterized.

Plant pathogen culture collections: it takes a village to preserve these resources vital to the advancement of agricultural security and plant pathology.

ABSTRACT Plant pathogen culture collections are essential resources in our fight against plant disease and for connecting discoveries of the present with established knowledge of the past. However, available infrastructure in support of culture collections is in serious need of improvement, and we continually face the risk of losing many of these collections. As novel and reemerging plant pathogens threaten agriculture, their timely identification and monitoring depends on rapid access to cultures representing the known diversity of plant pathogens along with genotypic, phenotypic, and epidemiological data associated with them.

Genome organization and evolution of the AVR-Pita avirulence gene family in the Magnaporthe grisea species complex.

The avirulence (AVR) gene AVR-Pita in Magnaporthe oryzae prevents the fungus from infecting rice cultivars containing the resistance gene Pi-ta. A survey of isolates of the M. grisea species complex from diverse hosts showed that AVR-Pita is a member of a gene family, which led us to rename it to AVR-Pita1.