scholarly journals Robust Cre recombinase activity in the biotrophic smut fungus Ustilago maydis enables efficient conditional null mutants in planta

Genetics ◽  
2021 ◽  
Author(s):  
Antonio de la Torre ◽  
Matteo Jurca ◽  
Kai Hoffmann ◽  
Lara Schmitz ◽  
Kai Heimel ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Fungal Growth ◽  
Ustilago Maydis ◽  
Cre Recombinase ◽  
Phytopathogenic Fungus ◽  
Marker Genes ◽  
In Planta ◽  
Gene Deletions ◽  
Site Specific ◽  
Plant Pathosystems

Abstract Site-specific recombinases have been used in higher eukaryotes, especially in animals, for a broad range of applications, including chromosomal translocations, large deletions, site-specific integration, and tissue-specific as well as conditional knock-outs. The application of site-specific recombination has also been demonstrated in simple eukaryotes like fungi and protozoa. However, its use in fungal research, especially in phytopathogenic fungi, has often been limited to “recycle” the marker genes used in transformation experiments. We show that Cre recombinase can be used for conditional gene deletions in the phytopathogenic fungus Ustilago maydis. Conditional gene knock-outs can be generated via the transcriptional control of the recombinase by U. maydis promoters specifically activated during the biotrophic phase of fungal growth, enabling gene deletions at defined developmental stages inside the plant tissue. Also, we show that a tamoxifen-activated Cre-recombinase allows the tight control necessary for the induced deletion of essential genes by the addition of tamoxifen. These tools will be helpful to address the function of genes under both axenic and in planta conditions for the U. maydis-maize pathosystem and should pave the way for similar approaches in other plant pathosystems.

scholarly journals The DNA Damage Response Signaling Cascade Regulates Proliferation of the Phytopathogenic Fungus Ustilago maydis in Planta

2011 ◽  
Vol 23 (4) ◽  
pp. 1654-1665 ◽  
Author(s):  
Carmen de Sena-Tomás ◽  
Alfonso Fernández-Álvarez ◽  
William K. Holloman ◽  
José Pérez-Martín
Keyword(s):  
Dna Damage ◽  
Dna Damage Response ◽  
Ustilago Maydis ◽  
Phytopathogenic Fungus ◽  
Signaling Cascade ◽  
In Planta ◽  
Damage Response

scholarly journals Crayfish hemocytes develop along the granular cell lineage

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fang Li ◽  
Zaichao Zheng ◽  
Hongyu Li ◽  
Rongrong Fu ◽  
Limei Xu ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cell Lineage ◽  
Granular Cell ◽  
Marker Genes ◽  
Hematopoietic Tissue ◽  
Differentiated Cells ◽  
Stage Of Development ◽  
Almost All ◽  
Relationship Of

AbstractDespite the central role of hemocytes in crustacean immunity, the process of hemocyte differentiation and maturation remains unclear. In some decapods, it has been proposed that the two main types of hemocytes, granular cells (GCs) and semigranular cells (SGCs), differentiate along separate lineages. However, our current findings challenge this model. By tracking newly produced hemocytes and transplanted cells, we demonstrate that almost all the circulating hemocytes of crayfish belong to the GC lineage. SGCs and GCs may represent hemocytes of different developmental stages rather than two types of fully differentiated cells. Hemocyte precursors produced by progenitor cells differentiate in the hematopoietic tissue (HPT) for 3 ~ 4 days. Immature hemocytes are released from HPT in the form of SGCs and take 1 ~ 3 months to mature in the circulation. GCs represent the terminal stage of development. They can survive for as long as 2 months. The changes in the expression pattern of marker genes during GC differentiation support our conclusions. Further analysis of hemocyte phagocytosis indicates the existence of functionally different subpopulations. These findings may reshape our understanding of crustacean hematopoiesis and may lead to reconsideration of the roles and relationship of circulating hemocytes.

scholarly journals The Effect of Fusarium verticillioides Fumonisins on Fatty Acids, Sphingolipids, and Oxylipins in Maize Germlings

2021 ◽  
Vol 22 (5) ◽  
pp. 2435
Author(s):  
Marzia Beccaccioli ◽  
Manuel Salustri ◽  
Valeria Scala ◽  
Matteo Ludovici ◽  
Andrea Cacciotti ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fusarium Verticillioides ◽  
Fungal Growth ◽  
Defense Responses ◽  
Ceramide Synthase ◽  
Ear Rot ◽  
In Planta ◽  
Disease Symptoms ◽  
The Impact

Fusarium verticillioides causes multiple diseases of Zea mays (maize) including ear and seedling rots, contaminates seeds and seed products worldwide with toxic chemicals called fumonisins. The role of fumonisins in disease is unclear because, although they are not required for ear rot, they are required for seedling diseases. Disease symptoms may be due to the ability of fumonisins to inhibit ceramide synthase activity, the expected cause of lipids (fatty acids, oxylipins, and sphingolipids) alteration in infected plants. In this study, we explored the impact of fumonisins on fatty acid, oxylipin, and sphingolipid levels in planta and how these changes affect F. verticillioides growth in maize. The identity and levels of principal fatty acids, oxylipins, and over 50 sphingolipids were evaluated by chromatography followed by mass spectrometry in maize infected with an F. verticillioides fumonisin-producing wild-type strain and a fumonisin-deficient mutant, after different periods of growth. Plant hormones associated with defense responses, i.e., salicylic and jasmonic acid, were also evaluated. We suggest that fumonisins produced by F. verticillioides alter maize lipid metabolism, which help switch fungal growth from a relatively harmless endophyte to a destructive necrotroph.

scholarly journals Defects in Mitochondrial and Peroxisomal β-Oxidation Influence Virulence in the Maize Pathogen Ustilago maydis

2012 ◽  
Vol 11 (8) ◽  
pp. 1055-1066 ◽  
Author(s):  
Matthias Kretschmer ◽  
Jana Klose ◽  
James W. Kronstad
Keyword(s):  
Ustilago Maydis ◽  
Short Chain Fatty Acids ◽  
Phytopathogenic Fungi ◽  
Metabolic Adaptation ◽  
In Planta ◽  
Gene Encoding ◽  
Disease Symptoms ◽  
Content Type ◽  
C Terminus ◽  
Fungal Phytopathogens

ABSTRACTAn understanding of metabolic adaptation during the colonization of plants by phytopathogenic fungi is critical for developing strategies to protect crops. Lipids are abundant in plant tissues, and fungal phytopathogens in the phylum basidiomycota possess both peroxisomal and mitochondrial β-oxidation pathways to utilize this potential carbon source. Previously, we demonstrated a role for the peroxisomal β-oxidation enzyme Mfe2 in the filamentous growth, virulence, and sporulation of the maize pathogenUstilago maydis. However,mfe2mutants still caused disease symptoms, thus prompting a more detailed investigation of β-oxidation. We now demonstrate that a defect in thehad1gene encoding hydroxyacyl coenzyme A dehydrogenase for mitochondrial β-oxidation also influences virulence, although its paralog,had2, makes only a minor contribution. Additionally, we identified a gene encoding a polypeptide with similarity to the C terminus of Mfe2 and designated it Mfe2b; this gene makes a contribution to virulence only in the background of anmfe2Δ mutant. We also show that short-chain fatty acids induce cell death inU. maydisand that a block in β-oxidation leads to toxicity, likely because of the accumulation of toxic intermediates. Overall, this study reveals that β-oxidation has a complex influence on the formation of disease symptoms byU. maydisthat includes potential metabolic contributions to proliferationin plantaand an effect on virulence-related morphogenesis.

scholarly journals Expression of alternative NADH dehydrogenases ( NDH ‐2) in the phytopathogenic fungus Ustilago maydis

FEBS Open Bio ◽  
2018 ◽  
Vol 8 (8) ◽  
pp. 1267-1279 ◽  
Author(s):  
Deyamira Matuz‐Mares ◽  
Genaro Matus‐Ortega ◽  
Christian Cárdenas‐Monroy ◽  
Lucero Romero‐Aguilar ◽  
Juan Carlos Villalobos‐Rocha ◽  
...  
Keyword(s):  
Ustilago Maydis ◽  
Phytopathogenic Fungus ◽  
Nadh Dehydrogenases

scholarly journals Expression of the ToxA and PtrPf2 genes of the phytopathogenic fungus Pyrenophora tritici-repentis at the beginning of the infection process

2019 ◽  
Author(s):  
Nina V. Mironenko ◽  
Alexandra S. Orina ◽  
Nadezhda M. Kovalenko
Keyword(s):  
Transcription Factor ◽  
Wheat Variety ◽  
Infection Process ◽  
Phytopathogenic Fungus ◽  
In Planta ◽  
Effector Gene ◽  
Pyrenophora Tritici Repentis ◽  
Wheat Leaves ◽  
Necrotrophic Effector

This study shows that the necrotrophic effector gene ToxA is differentially expressed in isolates of P. tritici-repentis fungus at different time periods after inoculation of the wheat variety Glenlea which has the gene Tsn1 controlling sensitivity to the necrosis inducing toxin Ptr ToxA. Two P. tritici-repentis isolates with different ability to cause necrosis on the leaves of Glenlea variety (nec + and nec-) and with different expression level of ToxA and gene of factor transcription PtrPf2 in vitro were used for analysis. Isolates of P. tritici-repentis are characterized by the differential expression of ToxA in planta. The expression of the ToxA gene in P. tritici-repentis ToxA+ isolates significantly increased when infected the wheat leaves compared to ToxA expression results obtained in vitro. The levels of ToxA expression in both isolates differed significantly after 24, 48 and 96 hours after inoculation, however, the dynamics of the trait change over time were similar. However, the highest ToxA expression in the virulent (nec+) isolate in contrast with the avirulent (nec-) isolate was observed at a point of 48 hours. Whereas the expression of regulating transcription factor PtrPf2 in planta differed imperceptibly from expression in vitro throughout the observation period. Obviously, the role of the fungal transcription factor in regulating the effector gene expression weakens in planta, and other mechanisms regulating the expression of pathogen genes at the biotrophic stage of the disease develop.

scholarly journals Pheromone-Induced G2 Arrest in the Phytopathogenic Fungus Ustilago maydis

2003 ◽  
Vol 2 (3) ◽  
pp. 494-500 ◽  
Author(s):  
Tatiana García-Muse ◽  
Gero Steinberg ◽  
José Pérez-Martín
Keyword(s):  
Cell Cycle ◽  
Ustilago Maydis ◽  
Phytopathogenic Fungus ◽  
Microtubule Organization ◽  
Mating Response ◽  
Cycle Arrest ◽  
Synthetic Pheromone ◽  
Genes Encoding ◽  
The Relationship ◽  
Corn Smut

ABSTRACT In the corn smut fungus Ustilago maydis, pathogenic development is initiated when two compatible haploid cells fuse and form the infectious dikaryon. Mating is dependent on pheromone recognition by compatible cells. In this report, we set out to evaluate the relationship between the cell cycle and the pheromone response in U. maydis. To achieve this, we designed a haploid pheromone-responsive strain that is able to faithfully reproduce the native mating response in nutrient-rich medium. Addition of synthetic pheromone to the responsive strain induces the formation of mating structures, and this response is abolished by mutations in genes encoding components of the pheromone signal transduction cascade. After recognition of pheromone, U. maydis cells arrest the cell cycle in a postreplicative stage. Visualization of the nucleus and microtubule organization indicates that the arrest takes place at the G2 phase. Chemical-induced cell cycle arrest and release in the presence of pheromone further support this conclusion.

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