scholarly journals Structural requirements of the phytoplasma effector protein SAP54 for causing homeotic transformation of floral organs

2019 ◽  
Author(s):  
Marc-Benjamin Aurin ◽  
Michael Haupt ◽  
Matthias Görlach ◽  
Florian Rümpler ◽  
Günter Theißen
Keyword(s):  
Plant Pathogens ◽  
Ornamental Plants ◽  
Effector Proteins ◽  
Plant Diseases ◽  
Scattering Data ◽  
Effector Protein ◽  
Homeotic Transformation ◽  
Structural Requirements ◽  
Floral Organs ◽  
Floral Homeotic

SummaryPhytoplasmas are intracellular bacterial plant pathogens that cause devastating diseases in crops and ornamental plants by the secretion of effector proteins. One of these effector proteins, termed SECRETED ASTER YELLOWS-WITCHES’ BROOM PROTEIN 54 (SAP54), leads to the degradation of a specific subset of floral homeotic proteins of the MIKC-type MADS-domain family via the ubiquitin-proteasome pathway. In consequence, the developing flowers show the homeotic transformation of floral organs into vegetative leaf-like structures. The molecular mechanism of SAP54 action involves physical binding to the keratin-like K-domain of MIKC-type proteins, and to some RAD23 proteins, which translocate ubiquitylated substrates to the proteasome. The structural requirements and specificity of SAP54 function are poorly understood, however. Here we report, based on biophysical and molecular biological analyses, that SAP54 folds into α-helical structures. We also show that the insertion of helix-breaking mutations disrupts correct folding of SAP54, which interferes with the ability of SAP54 to bind to its target proteins and to cause disease phenotypes in vivo. Surprisingly, dynamic light scattering data together with electrophoretic mobility shift assays suggest that SAP54 preferentially binds to multimeric complexes of MIKC-type proteins rather than to dimers or monomers of these proteins. Together with literature data this finding suggests that MIKC-type proteins and SAP54 constitute multimeric α-helical coiled-coils, possibly also involving other partners such as RAD23 proteins. Our investigations clarify the structure-function relationship of an important phytoplasma effector protein and thus may ultimately help to develop treatments against some devastating plant diseases.SIGNIFICANCE STATEMENTPhytoplasmas are bacterial plant pathogens that cause devastating diseases in crops and ornamental plants by the secretion of effector proteins such as SAP54, which leads to the degradation of some floral homeotic proteins. Our study clarifies the structural requirements of SAP54 function and illuminates the molecular mode of interaction and thus may ultimately help to develop treatments against some devastating plant diseases.

scholarly journals Structural Requirements of the Phytoplasma Effector Protein SAP54 for Causing Homeotic Transformation of Floral Organs

2020 ◽  
Vol 33 (9) ◽  
pp. 1129-1141
Author(s):  
Marc-Benjamin Aurin ◽  
Michael Haupt ◽  
Matthias Görlach ◽  
Florian Rümpler ◽  
Günter Theißen
Keyword(s):  
Plant Pathogens ◽  
Effector Proteins ◽  
Plant Diseases ◽  
Scattering Data ◽  
Effector Protein ◽  
Mobility Shift ◽  
Homeotic Transformation ◽  
Structural Requirements ◽  
Floral Organs ◽  
Specific Subset

Phytoplasmas are intracellular bacterial plant pathogens that cause devastating diseases in crops and ornamental plants by the secretion of effector proteins. One of these effector proteins, termed SECRETED ASTER YELLOWS WITCHES’ BROOM PROTEIN 54 (SAP54), leads to the degradation of a specific subset of floral homeotic proteins of the MIKC-type MADS-domain family via the ubiquitin-proteasome pathway. In consequence, the developing flowers show the homeotic transformation of floral organs into vegetative leaf-like structures. The molecular mechanism of SAP54 action involves binding to the keratin-like domain of MIKC-type proteins and to some RAD23 proteins, which translocate ubiquitylated substrates to the proteasome. The structural requirements and specificity of SAP54 function are poorly understood, however. Here, we report, based on biophysical and molecular biological analyses, that SAP54 folds into an α-helical structure. Insertion of helix-breaking mutations disrupts correct folding of SAP54 and compromises SAP54 binding to its target proteins and, concomitantly, its ability to evoke disease phenotypes in vivo. Interestingly, dynamic light scattering data together with electrophoretic mobility shift assays suggest that SAP54 preferentially binds to multimeric complexes of MIKC-type proteins rather than to dimers or monomers of these proteins. Together with data from literature, this finding suggests that MIKC-type proteins and SAP54 constitute multimeric α-helical coiled coils. Our investigations clarify the structure-function relationship of an important phytoplasma effector protein and may thus ultimately help to develop treatments against some devastating plant diseases.

scholarly journals A Novel Effector Protein of Apple Proliferation Phytoplasma Disrupts Cell Integrity of Nicotiana spp. Protoplasts

2019 ◽  
Vol 20 (18) ◽  
pp. 4613 ◽  
Author(s):  
Cecilia Mittelberger ◽  
Hagen Stellmach ◽  
Bettina Hause ◽  
Christine Kerschbamer ◽  
Katja Schlink ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Effector Proteins ◽  
Plant Diseases ◽  
Effector Protein ◽  
Insect Vector ◽  
Cell Integrity ◽  
Apple Proliferation ◽  
Disease Manifestation ◽  
Candidatus Phytoplasma Mali ◽  
Exact Function

Effector proteins play an important role in the virulence of plant pathogens such as phytoplasma, which are the causative agents of hundreds of different plant diseases. The plant hosts comprise economically relevant crops such as apples (Malus × domestica), which can be infected by ‘Candidatus Phytoplasma mali’ (P. mali), a highly genetically dynamic plant pathogen. As the result of the genetic and functional analyses in this study, a new putative P. mali effector protein was revealed. The so-called “Protein in Malus Expressed 2” (PME2), which is expressed in apples during P. mali infection but not in the insect vector, shows regional genetic differences. In a heterologous expression assay using Nicotiana benthamiana and Nicotiana occidentalis mesophyll protoplasts, translocation of both PME2 variants in the cell nucleus was observed. Overexpression of the effector protein affected cell integrity in Nicotiana spp. protoplasts, indicating a potential role of this protein in pathogenic virulence. Interestingly, the two genetic variants of PME2 differ regarding their potential to manipulate cell integrity. However, the exact function of PME2 during disease manifestation and symptom development remains to be further elucidated. Aside from the first description of the function of a novel effector of P. mali, the results of this study underline the necessity for a more comprehensive description and understanding of the genetic diversity of P. mali as an indispensable basis for a functional understanding of apple proliferation disease.

SEUSS, a member of a novel family of plant regulatory proteins, represses floral homeotic gene expression withLEUNIG

Development ◽  
2002 ◽  
Vol 129 (1) ◽  
pp. 253-263 ◽  
Author(s):  
Robert G. Franks ◽  
Chunxin Wang ◽  
Joshua Z. Levin ◽  
Zhongchi Liu
Keyword(s):  
Gene Expression ◽  
Floral Meristem ◽  
Genetic Identification ◽  
Homeotic Gene ◽  
Dimerization Domain ◽  
Homeotic Transformation ◽  
Floral Organs ◽  
Floral Homeotic Gene ◽  
Homeotic Gene Expression ◽  
Floral Homeotic

Proper regulation of homeotic gene expression is critical for pattern formation during both animal and plant development. A negative regulatory mechanism ensures that the floral homeotic gene AGAMOUS is only expressed in the center of an Arabidopsis floral meristem to specify stamen and carpel identity and to repress further proliferation of the floral meristem. We report the genetic identification and characterization of a novel gene, SEUSS, that is required in the negative regulation of AGAMOUS. Mutations in SEUSS cause ectopic and precocious expression of AGAMOUS mRNA, leading to partial homeotic transformation of floral organs in the outer two whorls. The effects of seuss mutations are most striking when combined with mutations in LEUNIG, a previously identified repressor of AGAMOUS. More complete homeotic transformation of floral organs and a greater extent of organ loss in all floral whorls were observed in the seuss leunig double mutants. By in situ hybridization and double and triple mutant analyses, we showed that this enhanced defect was caused by an enhanced ectopic and precocious expression of AGAMOUS. Using a map-based approach, we isolated the SEUSS gene and showed that it encodes a novel protein with at least two glutamine-rich domains and a highly conserved domain that shares sequence identity with the dimerization domain of the LIM-domain-binding transcription co-regulators in animals. Based on these molecular and genetic analyses, we propose that SEUSS encodes a regulator of AGAMOUS and functions together with LEUNIG.

scholarly journals Molecular mechanisms of host cell manipulation by plant pathogens

2014 ◽  
Vol 70 (a1) ◽  
pp. C801-C801
Author(s):  
Richard Hughes ◽  
Stuart King ◽  
Abbas Maqbool ◽  
Hazel McLellan ◽  
Tolga Bozkurt ◽  
...  
Keyword(s):  
Cell Death ◽  
Crop Production ◽  
Plant Pathogens ◽  
Molecular Mechanisms ◽  
Complex Structure ◽  
Effector Proteins ◽  
Plant Diseases ◽  
Cell Manipulation ◽  
Cell Physiology ◽  
Structural Studies

An estimated 15% of global crop production is lost to pre-harvest disease every year. New ways to manage plant diseases are required. A mechanistic understanding of how plant pathogens re-program their hosts to enable colonisation may provide novel genetic or chemical opportunities to interfere with disease. One notorious plant parasite is the Irish potato famine pathogen Phytophthora infestans. This pathogen remains a considerable threat to potato/tomato crops today as the agent of late blight. Plant pathogens secrete effector proteins outside of and into plant cells to suppress host defences and manipulate cell physiology. Structural studies have provided insights into effector evolution and enabled experiments to probe function [1-3]. Crystal structures of 4 Phytophthora RXLR-type effectors, which are unrelated in primary sequence, revealed similarities in the fold of these proteins. This fold was proposed to act as a stable scaffold that supports diversification of effectors. Further, molecular modelling has enabled mapping of single-site variants responsible for specialisation of a Phytophthora Cystatin-like effector, revealing how effectors can adapt to new hosts after a "host jump". Structural studies describing how RXLR-effectors interact with host targets are lacking. We have used Y2H/co-IP studies to identify host proteins that interact with P. infestans effectors PexRD2 and PexRD54. PexRD2 interacts with MAPKKKe, a component of plant immune signalling pathways, and suppressed cell death activities of this protein. We used the structure of PexRD2 to design mutants that fail to interact with MAPKKKe, and no longer suppress cell-death activities. We found that PexRD54 interacts with potato homologues of the autophagy protein ATG8. We have obtained a crystal structure for PexRD54 in the presence of ATG8. We are now using X-ray scattering to verify the complex structure in solution prior to establishing the role of this interaction during infection.

scholarly journals Phytopathogenic species of fungi and fungal-like organisms identified in plant samples delivered to the Plant Disease Clinic in 2018–2020

2021 ◽  
Vol 61 (3) ◽  
Keyword(s):  
Host Plants ◽  
Plant Disease ◽  
Plant Pathogens ◽  
Ornamental Plants ◽  
Single Species ◽  
Plant Diseases ◽  
Effective Management ◽  
Fusarium Spp ◽  
Molecular Tests ◽  
Disease Clinic

One of the conditions for effective management of farm is an access to quick diagnostics of plant pathogens in order to reduce the occurrence of plant diseases. The Plant Diseases Clinic receives samples of infected plants supplied by growers and gardeners from all over Poland. In the years 2018–2020, a total of 274 samples were tested at the Clinic for the presence of fungi and fungal-like organisms pathogenic for plants. The tests were carried out using the microscopic method, and in case of doubt, the result was confirmed by molecular tests. The most frequently studied plant was tomato (26%), followed by strawberry (9%), cucumber (5%) and tobacco, sugar beet, onion, blueberry, raspberry, lettuce, cauliflower and potato. Conifers were also a large group, such as: thujas, cypresses and pines; a total of 17 host plants. Single species of ornamental plants were very numerous, e.g. gerbera, anthurium, aster, geranium, phlox, chrysanthemum and others. The fungi of the genus Fusarium spp. constituted about 38% of infections. This was followed by Alternaria spp. (26%), Botrytis cinerea (11%) and Cladosporium sp. (10%). The remaining diseases were caused by Pythium sp., Rhizoctonia sp., Colletotrichum sp., Ulocladium sp., Pestalotia sp. and Phytophthora sp. In recent years, the greatest threat to tomatoes and strawberries has been the fungi of the Fusarium genus, and the pathogens of the Pythium genus to cucumbers.

scholarly journals Harnessing Effector-Triggered Immunity for Durable Disease Resistance

2017 ◽  
Vol 107 (8) ◽  
pp. 912-919 ◽  
Author(s):  
Meixiang Zhang ◽  
Gitta Coaker
Keyword(s):  
Disease Resistance ◽  
Plant Pathogens ◽  
Domain Architecture ◽  
Effector Proteins ◽  
Plant Diseases ◽  
Sources Of Resistance ◽  
Pathogen Effectors ◽  
Pathogen Effector ◽  
Effector Triggered Immunity ◽  
Durable Disease Resistance

Genetic control of plant diseases has traditionally included the deployment of single immune receptors with nucleotide-binding leucine-rich repeat (NLR) domain architecture. These NLRs recognize corresponding pathogen effector proteins inside plant cells, resulting in effector-triggered immunity (ETI). Although ETI triggers robust resistance, deployment of single NLRs can be rapidly overcome by pathogen populations within a single or a few growing seasons. In order to generate more durable disease resistance against devastating plant pathogens, a multitiered strategy that incorporates stacked NLRs combined with other sources of disease resistance is necessary. New genetic and genomic technologies have enabled advancements in identifying conserved pathogen effectors, isolating NLR repertoires from diverse plants, and editing plant genomes to enhance resistance. Significant advancements have also been made in understanding plant immune perception at the receptor level, which has promise for engineering new sources of resistance. Here, we discuss how to utilize recent scientific advancements in a multilayered strategy for developing more durable disease resistance.

scholarly journals The Epichloë festucae Antifungal Protein Efe-AfpA Is Also a Possible Effector Protein Required for the Interaction of the Fungus with its Host Grass Festuca rubra subsp. rubra

2021 ◽  
Vol 9 (1) ◽  
pp. 140
Author(s):  
Ruying Wang ◽  
Simin Luo ◽  
Bruce B. Clarke ◽  
Faith C. Belanger
Keyword(s):  
Disease Resistance ◽  
Insect Resistance ◽  
Effector Proteins ◽  
Antifungal Protein ◽  
Grass Species ◽  
Effector Protein ◽  
Festuca Rubra ◽  
Symbiotic Interaction ◽  
Red Fescue ◽  
Epichloë Festucae

Strong creeping red fescue (Festuca rubra subsp. rubra) is a commercially important low-maintenance turfgrass and is often naturally infected with the fungal endophyte Epichloë festucae. Epichloë spp. are endophytes of several cool-season grass species, often conferring insect resistance to the grass hosts due to the production of toxic alkaloids. In addition to insect resistance, a unique feature of the strong creeping red fescue/E. festucae symbiosis is the endophyte-mediated disease resistance to the fungal pathogen Clarireedia jacksonii, the causal agent of dollar spot disease. Such disease resistance is not a general feature of other grass/ Epichloë interactions. E. festucae isolates infecting red fescue have an antifungal protein gene Efe-afpA, whereas most other Epichloë spp. do not have a similar gene. The uniqueness of this gene suggests it may, therefore, be a component of the unique disease resistance seen in endophyte-infected red fescue. Here, we report the generation of CRISPR-Cas9 Efe-afpA gene knockouts with the goal of determining if absence of the protein in endophyte-infected Festuca rubra leads to disease susceptibility. However, it was not possible to infect plants with the knockout isolates, although infection was possible with the wild type E. festucae and with complemented isolates. This raises the interesting possibility that, in addition to having antifungal activity, the protein is required for the symbiotic interaction. The antifungal protein is a small secreted protein with high expression in planta relative to its expression in culture, all characteristics consistent with effector proteins. If Efe-AfpA is an effector protein it must be specific to certain interactions, since most Epichloë spp. do not have such a gene in their genomes.

scholarly journals Microbial antagonists against plant pathogens in Iran: A review

2020 ◽  
Vol 5 (1) ◽  
pp. 404-440 ◽  
Author(s):  
Mehrdad Alizadeh ◽  
Yalda Vasebi ◽  
Naser Safaie
Keyword(s):  
Biological Control ◽  
Chemical Control ◽  
Plant Disease ◽  
Plant Pathogens ◽  
Plant Diseases ◽  
Agricultural Products ◽  
Control Measures ◽  
Wide Range ◽  
Plant Disease Management ◽  
Published Research

AbstractThe purpose of this article was to give a comprehensive review of the published research works on biological control of different fungal, bacterial, and nematode plant diseases in Iran from 1992 to 2018. Plant pathogens cause economical loss in many agricultural products in Iran. In an attempt to prevent these serious losses, chemical control measures have usually been applied to reduce diseases in farms, gardens, and greenhouses. In recent decades, using the biological control against plant diseases has been considered as a beneficial and alternative method to chemical control due to its potential in integrated plant disease management as well as the increasing yield in an eco-friendly manner. Based on the reported studies, various species of Trichoderma, Pseudomonas, and Bacillus were the most common biocontrol agents with the ability to control the wide range of plant pathogens in Iran from lab to the greenhouse and field conditions.

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.

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