scholarly journals Putative Rust Fungal Effector Proteins in Infected Bean and Soybean Leaves

2016 ◽  
Vol 106 (5) ◽  
pp. 491-499 ◽  
Author(s):  
Bret Cooper ◽  
Kimberly B. Campbell ◽  
Hunter S. Beard ◽  
Wesley M. Garrett ◽  
Nazrul Islam
Keyword(s):  
Plant Cell ◽  
Crop Production ◽  
Rust Fungus ◽  
Fungal Spores ◽  
Pathogenic Fungi ◽  
Rust Fungi ◽  
Effector Proteins ◽  
Glycoside Hydrolases ◽  
Data Set ◽  
Soybean Leaves

The plant-pathogenic fungi Uromyces appendiculatus and Phakopsora pachyrhizi cause debilitating rust diseases on common bean and soybean. These rust fungi secrete effector proteins that allow them to infect plants, but their effector repertoires are not understood. The discovery of rust fungus effectors may eventually help guide decisions and actions that mitigate crop production loss. Therefore, we used mass spectrometry to identify thousands of proteins in infected beans and soybeans and in germinated fungal spores. The comparative analysis between the two helped differentiate a set of 24 U. appendiculatus proteins targeted for secretion that were specifically found in infected beans and a set of 34 U. appendiculatus proteins targeted for secretion that were found in germinated spores and infected beans. The proteins specific to infected beans included family 26 and family 76 glycoside hydrolases that may contribute to degrading plant cell walls. There were also several types of proteins with structural motifs that may aid in stabilizing the specialized fungal haustorium cell that interfaces the plant cell membrane during infection. There were 16 P. pachyrhizi proteins targeted for secretion that were found in infected soybeans, and many of these proteins resembled the U. appendiculatus proteins found in infected beans, which implies that these proteins are important to rust fungal pathology in general. This data set provides insight to the biochemical mechanisms that rust fungi use to overcome plant immune systems and to parasitize cells.

scholarly journals Suppression or Activation of Immune Responses by Predicted Secreted Proteins of the Soybean Rust Pathogen Phakopsora pachyrhizi

2018 ◽  
Vol 31 (1) ◽  
pp. 163-174 ◽  
Author(s):  
Mingsheng Qi ◽  
James P. Grayczyk ◽  
Janina M. Seitz ◽  
Youngsill Lee ◽  
Tobias I. Link ◽  
...  
Keyword(s):  
Immune Responses ◽  
Crop Production ◽  
Rust Fungus ◽  
Functional Characterization ◽  
Rust Fungi ◽  
Effector Proteins ◽  
Soybean Rust ◽  
Phakopsora Pachyrhizi ◽  
Immune Systems ◽  
Rust Pathogen

Rust fungi, such as the soybean rust pathogen Phakopsora pachyrhizi, are major threats to crop production. They form specialized haustoria that are hyphal structures intimately associated with host-plant cell membranes. These haustoria have roles in acquiring nutrients and secreting effector proteins that manipulate host immune systems. Functional characterization of effector proteins of rust fungi is important for understanding mechanisms that underlie their virulence and pathogenicity. Hundreds of candidate effector proteins have been predicted for rust pathogens, but it is not clear how to prioritize these effector candidates for further characterization. There is a need for high-throughput approaches for screening effector candidates to obtain experimental evidence for effector-like functions, such as the manipulation of host immune systems. We have focused on identifying effector candidates with immune-related functions in the soybean rust fungus P. pachyrhizi. To facilitate the screening of many P. pachyrhizi effector candidates (named PpECs), we used heterologous expression systems, including the bacterial type III secretion system, Agrobacterium infiltration, a plant virus, and a yeast strain, to establish an experimental pipeline for identifying PpECs with immune-related functions and establishing their subcellular localizations. Several PpECs were identified that could suppress or activate immune responses in nonhost Nicotiana benthamiana, N. tabacum, Arabidopsis, tomato, or pepper plants.

Use of Freeze Substitution for TEM Studies of Fungal Spores

2000 ◽  
Vol 6 (S2) ◽  
pp. 684-685 ◽  
Author(s):  
C. W. Minis ◽  
E. A. Richardson
Keyword(s):  
Gold Standard ◽  
Fungal Spores ◽  
Freeze Substitution ◽  
Pathogenic Fungi ◽  
Rust Fungi ◽  
Smut Fungi ◽  
Plant Pathogenic Fungi ◽  
Dialysis Membrane ◽  
Direct Exposure ◽  
Fungal Hyphae

Since its initial use to fix fungal hyphae (1), plunge freezing followed by freeze substitution has become the “gold standard” for TEM studies of fungal hyphae and spores. In this presentation we discuss results we have obtained using plunge freezing and freeze substitution to fix various types of spores produced by plant pathogenic fungi. Examples include basidiospores and aeciospores of rust fungi, teliospores of rust and smut fungi and conidia of a variety of ascomycetes and deuteromycetes.Generally speaking, plunge freezing followed by freeze substitution usually yields best results when spore diameter does not exceed 10-15 μm. However, in a number of cases we (2) have obtained good to excellent results with much larger spores. A key to good freezing appears to be direct exposure of spores to liquid propane during plunging. We have had best results by placing spores on small pieces of dialysis membrane and then plunging these membranes. In most cases the spores remain attached to membranes throughout the freezing, substitution, infiltration and embedment procedures.

scholarly journals Progress on Molecular Genetics and Manipulation of Rust Fungi

2020 ◽  
Vol 110 (3) ◽  
pp. 532-543 ◽  
Author(s):  
Guus Bakkeren ◽  
Les J. Szabo
Keyword(s):  
Rust Fungus ◽  
Molecular Genetic ◽  
Resistance Breeding ◽  
Pathogenic Fungi ◽  
Rust Fungi ◽  
Host Interactions ◽  
Similar Work ◽  
Biotrophic Fungi ◽  
Pathogenic Microbes ◽  
Design And Testing

Among the thousands of rust species described, many are known for their devastating effects on their hosts, which include major agriculture crops and trees. Hence, for over a century, these basidiomycete pathogenic fungi have been researched and experimented with. However, due to their biotrophic nature, they are challenging organisms to work with and, needing their hosts for propagation, represent pathosystems that are not easily experimentally accessible. Indeed, efforts to perform genetics have been few and far apart for the rust fungi, though one study performed in the 1940s was famously instrumental in formulating the gene-for-gene hypothesis describing pathogen−host interactions. By taking full advantage of the molecular genetic tools developed in the 1980s, research on many plant pathogenic microbes thrived, yet similar work on the rusts remained very challenging though not without some successes. However, the genomics era brought real breakthrough research for the biotrophic fungi and with innovative experimentation and the use of heterologous systems, molecular genetic analyses over the last 2 decades have significantly advanced our insight into the function of many rust fungus genes and their role in the interaction with their hosts. This has allowed optimizing efforts for resistance breeding and the design and testing of various novel strategies to reduce the devastating diseases they cause.

scholarly journals Xyloglucan processing machinery in Xanthomonas pathogens and its role in the transcriptional activation of virulence factors

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Plinio S. Vieira ◽  
Isabela M. Bonfim ◽  
Evandro A. Araujo ◽  
Ricardo R. Melo ◽  
Augusto R. Lima ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Transcriptional Activation ◽  
Molecular Mechanisms ◽  
Model Organism ◽  
Citrus Canker ◽  
Effector Proteins ◽  
Glycoside Hydrolases ◽  
Host Cells ◽  
System A ◽  
Enzymatic Machinery

AbstractXyloglucans are highly substituted and recalcitrant polysaccharides found in the primary cell walls of vascular plants, acting as a barrier against pathogens. Here, we reveal that the diverse and economically relevant Xanthomonas bacteria are endowed with a xyloglucan depolymerization machinery that is linked to pathogenesis. Using the citrus canker pathogen as a model organism, we show that this system encompasses distinctive glycoside hydrolases, a modular xyloglucan acetylesterase and specific membrane transporters, demonstrating that plant-associated bacteria employ distinct molecular strategies from commensal gut bacteria to cope with xyloglucans. Notably, the sugars released by this system elicit the expression of several key virulence factors, including the type III secretion system, a membrane-embedded apparatus to deliver effector proteins into the host cells. Together, these findings shed light on the molecular mechanisms underpinning the intricate enzymatic machinery of Xanthomonas to depolymerize xyloglucans and uncover a role for this system in signaling pathways driving pathogenesis.

scholarly journals The Ustilago hordei–Barley Interaction is a Versatile System for Characterization of Fungal Effectors

2021 ◽  
Vol 7 (2) ◽  
pp. 86
Author(s):  
Bilal Ökmen ◽  
Daniela Schwammbach ◽  
Guus Bakkeren ◽  
Ulla Neumann ◽  
Gunther Doehlemann
Keyword(s):  
Fungal Pathogens ◽  
Plant Pathogens ◽  
Expression System ◽  
Pathogenic Fungi ◽  
Effector Proteins ◽  
Mating Partner ◽  
Fungal Virulence ◽  
Functional Studies ◽  
Infection Structures ◽  
Ustilago Hordei

Obligate biotrophic fungal pathogens, such as Blumeria graminis and Puccinia graminis, are amongst the most devastating plant pathogens, causing dramatic yield losses in many economically important crops worldwide. However, a lack of reliable tools for the efficient genetic transformation has hampered studies into the molecular basis of their virulence or pathogenicity. In this study, we present the Ustilago hordei–barley pathosystem as a model to characterize effectors from different plant pathogenic fungi. We generate U. hordei solopathogenic strains, which form infectious filaments without the presence of a compatible mating partner. Solopathogenic strains are suitable for heterologous expression system for fungal virulence factors. A highly efficient Crispr/Cas9 gene editing system is made available for U. hordei. In addition, U. hordei infection structures during barley colonization are analyzed using transmission electron microscopy, showing that U. hordei forms intracellular infection structures sharing high similarity to haustoria formed by obligate rust and powdery mildew fungi. Thus, U. hordei has high potential as a fungal expression platform for functional studies of heterologous effector proteins in barley.

scholarly journals BAK1 Is Not a Target of the Pseudomonas syringae Effector AvrPto

2011 ◽  
Vol 24 (1) ◽  
pp. 100-107 ◽  
Author(s):  
Tingting Xiang ◽  
Na Zong ◽  
Jie Zhang ◽  
Jinfeng Chen ◽  
Mingsheng Chen ◽  
...  
Keyword(s):  
Cell Surface ◽  
Immune Responses ◽  
Pseudomonas Syringae ◽  
Plant Cell ◽  
Crucial Role ◽  
Pathogenic Bacteria ◽  
Effector Proteins ◽  
Receptor Like Kinase ◽  
Receptor Kinases ◽  
New Evidence

Plant cell surface-localized receptor kinases such as FLS2, EFR, and CERK1 play a crucial role in detecting invading pathogenic bacteria. Upon stimulation by bacterium-derived ligands, FLS2 and EFR interact with BAK1, a receptor-like kinase, to activate immune responses. A number of Pseudomonas syringae effector proteins are known to block immune responses mediated by these receptors. Previous reports suggested that both FLS2 and BAK1 could be targeted by the P. syringae effector AvrPto to inhibit plant defenses. Here, we provide new evidence further supporting that FLS2 but not BAK1 is targeted by AvrPto in plants. The AvrPto-FLS2 interaction prevented the phosphorylation of BIK1, a downstream component of the FLS2 pathway.

scholarly journals Quantification of leaf greenness and leaf spectral profile in plant diagnosis using an optical scanner

2012 ◽  
Vol 36 (3) ◽  
pp. 309-317 ◽  
Author(s):  
Ryoichi Doi
Keyword(s):  
Spectral Data ◽  
Crop Production ◽  
Response Pattern ◽  
Quadratic Growth ◽  
Spectral Profile ◽  
Growth Data ◽  
Data Set ◽  
Optical Scanner ◽  
Color Scale ◽  
Management Measures

Observation of leaf spectral profile (color) enables suitable management measures to be taken for crop production. An optical scanner was used: 1) to obtain an equation to determine the greenness of plant leaves and 2) to examine the power to discriminate among plants grown under different nutritional conditions. Sweet basil seedlings grown on vermiculite were supplemented with one-fifth-strength Hoagland solutions containing 0, 0.2, 1, 5, 20, and 50 mM NH4+. The 5 mM treatment resulted in the greatest leaf and shoot weights, indicating a quadratic growth response pattern to the NH4+ gradient. An equation involving b*, black and green to describe the greenness of leaves was provided by the spectral profiling of a color scale for rice leaves as the standard. The color scale values for the basil leaves subjected to 0.2 and 1 mM NH4+ treatments were 1.00 and 1.12, respectively. The other treatments resulted in significantly greater values of 2.25 to 2.42, again indicating a quadratic response pattern. Based on the spectral data set consisting of variables of red-green-blue and other color models and color scale values, in discriminant analysis, 81% of the plants were correctly classified into the six NH4+ treatment groups. Combining the spectral data set with the growth data set consisting of leaf and shoot weights, 92% of the plant samples were correctly classified whereas, using the growth data set, only 53% of plants were correctly classified. Therefore, the optical scanning of leaves and the use of spectral profiles helped plant diagnosis when biomass measurements were not effective.

scholarly journals The Role of Arabinogalactan Type II Degradation in Plant-Microbe Interactions

2021 ◽  
Vol 12 ◽  
Author(s):  
Maria Guadalupe Villa-Rivera ◽  
Horacio Cano-Camacho ◽  
Everardo López-Romero ◽  
María Guadalupe Zavala-Páramo
Keyword(s):  
Cell Wall ◽  
Plant Defense ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Degradation Products ◽  
Hydrolytic Enzymes ◽  
Pathogenic Fungi ◽  
Plant Interactions ◽  
Root Tips ◽  
Microbe Interactions

Arabinogalactans (AGs) are structural polysaccharides of the plant cell wall. A small proportion of the AGs are associated with hemicellulose and pectin. Furthermore, AGs are associated with proteins forming the so-called arabinogalactan proteins (AGPs), which can be found in the plant cell wall or attached through a glycosylphosphatidylinositol (GPI) anchor to the plasma membrane. AGPs are a family of highly glycosylated proteins grouped with cell wall proteins rich in hydroxyproline. These glycoproteins have important and diverse functions in plants, such as growth, cellular differentiation, signaling, and microbe-plant interactions, and several reports suggest that carbohydrate components are crucial for AGP functions. In beneficial plant-microbe interactions, AGPs attract symbiotic species of fungi or bacteria, promote the development of infectious structures and the colonization of root tips, and furthermore, these interactions can activate plant defense mechanisms. On the other hand, plants secrete and accumulate AGPs at infection sites, creating cross-links with pectin. As part of the plant cell wall degradation machinery, beneficial and pathogenic fungi and bacteria can produce the enzymes necessary for the complete depolymerization of AGs including endo-β-(1,3), β-(1,4) and β-(1,6)-galactanases, β-(1,3/1,6) galactanases, α-L-arabinofuranosidases, β-L-arabinopyranosidases, and β-D-glucuronidases. These hydrolytic enzymes are secreted during plant-pathogen interactions and could have implications for the function of AGPs. It has been proposed that AGPs could prevent infection by pathogenic microorganisms because their degradation products generated by hydrolytic enzymes of pathogens function as damage-associated molecular patterns (DAMPs) eliciting the plant defense response. In this review, we describe the structure and function of AGs and AGPs as components of the plant cell wall. Additionally, we describe the set of enzymes secreted by microorganisms to degrade AGs from AGPs and its possible implication for plant-microbe interactions.

scholarly journals Biological Function(s) and Application (s) of Pectin and Pectin Degrading Enzymes

2018 ◽  
Vol 15 (1) ◽  
pp. 87-100 ◽  
Author(s):  
Puja Chandrayan
Keyword(s):  
Cell Wall ◽  
Structure Function ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Plant Defence ◽  
Glycoside Hydrolases ◽  
Limited Information ◽  
Thermostable Enzymes ◽  
Network Cell ◽  
Degrading Enzymes

Pectin is an integral part of plant cell wall and since centuries pectin extracted from plants is widely used in food and fruit juice processing. Moreover, in last half century, the applications have also invaded into many bio-processing applications such as pharmaceutical, bioenergy, textile, paper and tea processing. In these growing industries, the use of pectinases has grown with a significant amount i.e. approximately 10 % of total global enzyme market comes from pectinases. Herein comprehensive analyses of information related to structure and function of pectin in plant cell wall as well as structural classes of pectins have been discussed. The major function of pectin is in cementing the cellulose and hemicelluloses network, cell-cell adhesion and plant defence. Keeping the wide use of pectin in food industry and growing need of environment friendly technology for pectin extraction has accelerated the demand of pectin degrading enzymes (PDEs). PDEs are from three enzyme classes: carbohydrate esterases from CE8 and CE12 family, glycoside hydrolases from GH28 family and lyases from PL1, 2, 3, 9 and 10. We have reviewed the literature related to abundance and structure-function of these abovementioned enzymes from bacteria. From the current available literature, we found very limited information is present about thermostable PDEs. Hence, in future it could be a topic of study to gain the insight about structure-function of enzymes together with the expanded role of thermostable enzymes in development of bioprocesses based on these enzymes.

scholarly journals Structural genomic applied on the rust fungus Melampsora larici-populina reveals two candidate effector proteins adopting cystine-knot and nuclear transport factor 2-like protein folds

2019 ◽  
Author(s):  
Karine de Guillen ◽  
Cécile Lorrain ◽  
Pascale Tsan ◽  
Philippe Barthe ◽  
Benjamin Petre ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Nuclear Transport ◽  
Sequence Similarity ◽  
Rust Fungus ◽  
Parasitic Infection ◽  
Effector Proteins ◽  
Host Cells ◽  
Dimensional Structure ◽  
Structural Genomic ◽  
Transport Factor

ABSTRACTRust fungi are plant pathogens that secrete an arsenal of effector proteins interfering with plant functions and promoting parasitic infection. Effectors are often species-specific, evolve rapidly, and display low sequence similarities with known proteins or domains. How rust fungal effectors function in host cells remains elusive, and biochemical and structural approaches have been scarcely used to tackle this question. In this study, we used a strategy based on recombinant protein production in Escherichia coli to study eleven candidate effectors of the leaf rust fungus Melampsora larici-populina. We successfully purified and solved the three-dimensional structure of two proteins, MLP124266 and MLP124017, using NMR spectroscopy. Although both proteins show no sequence similarity with known proteins, they exhibit structural similarities to knottin and nuclear transport factor 2-like proteins, respectively. Altogether, our findings show that sequence-unrelated effectors can adopt folds similar to known proteins, and encourage the use of biochemical and structural approaches to functionally characterize rust effector candidates.

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