scholarly journals Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion

2017 ◽  
Author(s):  
Kristina Marton ◽  
Marko Flajšman ◽  
Sebastjan Radišek ◽  
Katarina Košmelj ◽  
Jernej Jakše ◽  
...  
Keyword(s):  
Verticillium Wilt ◽  
Sequence Similarity ◽  
Vascular Plant ◽  
Xylem Sap ◽  
Fungal Growth ◽  
Effector Proteins ◽  
Specific Gene ◽  
In Planta ◽  
Verticillium Nonalfalfae ◽  
Spatio Temporal

AbstractBackgroundThe vascular plant pathogen Verticillium nonalfalfae causes Verticillium wilt in several important crops. VnaSSP4.2 was recently discovered as a V. nonalfalfae virulence effector protein in the xylem sap of infected hop. Here, we expanded our search for candidate secreted effector proteins (CSEPs) in the V. nonalfalfae predicted secretome using a bioinformatic pipeline built on V. nonalfalfae genome data, RNA-Seq and proteomic studies of the interaction with hop.ResultsThe secretome, rich in carbohydrate active enzymes, proteases, redox proteins and proteins involved in secondary metabolism, cellular processing and signaling, includes 263 CSEPs. Several homologs of known fungal effectors (LysM, NLPs, Hce2, Cerato-platanins, Cyanovirin-N lectins, hydrophobins and CFEM domain containing proteins) and avirulence determinants in the PHI database (Avr-Pita1 and MgSM1) were found. The majority of CSEPs were non-annotated and were narrowed down to 44 top priority candidates based on their likelihood of being effectors. These were examined by spatio-temporal gene expression profiling of infected hop. Among the highest in planta expressed CSEPs, five deletion mutants were tested in pathogenicity assays. A deletion mutant of VnaUn.279, a lethal pathotype specific gene with sequence similarity to SAM-dependent methyltransferase (LaeA), had lower infectivity and showed highly reduced virulence, but no changes in morphology, fungal growth or conidiation were observed.ConclusionsSeveral putative secreted effector proteins that probably contribute to V. nonalfalfae colonization of hop were identified in this study. Among them, LaeA gene homolog was found to act as a potential novel virulence effector of V. nonalfalfae. The combined results will serve for future characterization of V. nonalfalfae effectors, which will advance our understanding of Verticillium wilt disease.

scholarly journals Verticillium dahliae effector VDAL protects MYB6 from degradation by interacting with PUB25/26 E3 ligases for enhancing Verticillium wilt resistance in Arabidopsis

2021 ◽  
Author(s):  
Zhizhong Gong ◽  
Junsheng Qi ◽  
Aifang Ma ◽  
Dingpeng Zhang ◽  
Guangxing Wang ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Plant Resistance ◽  
Plant Disease Resistance ◽  
Plant Disease ◽  
Effector Proteins ◽  
In Planta ◽  
E3 Ligases

Verticillium wilt is a severe plant disease, increasing the plant resistance to this disease is a critical challenge worldwide. Here, we report that the Verticillium dahliae (V. dahliae)-secreted Aspf2-like protein VDAL causes leaf wilting when applied to cotton leaves in vitro, but enhances the resistance to V. dahliae when overexpressed in Arabidopsis or cotton. VDAL interacts with Arabidopsis E3 ligases PUB25 and PUB26 (PUBs) and is ubiquitinated by PUBs in vitro. However, VDAL is not degraded by PUBs in planta. Besides, the pub25 pub26 shows higher resistance to V. dahliae than the wild type. PUBs interact with the transcription factor MYB6 in a yeast two-hybrid screen. MYB6 promotes plant resistance to Verticillium wilt while PUBs ubiquitinate MYB6 and mediate its degradation. VDAL competes with MYB6 for binding to PUBs, and the role of VDAL in increasing wilt disease depends on MYB6. These results suggest that plants evolute a strategy to utilize the invaded effector protein VDAL to resist the V. dahliae infection without causing a hypersensitive response. This study provides the molecular mechanism for plants increasing disease resistance when overexpressing some effector proteins, and may promote searching for more genes from pathogenic fungi or bacteria to engineer plant disease resistance.

The effects of amino acids on the growth and sporulation of Verticillium dahliae

1987 ◽  
Vol 65 (7) ◽  
pp. 1299-1302 ◽  
Author(s):  
D. R. Duncan ◽  
E. B. Himelick
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Verticillium Wilt ◽  
Verticillium Dahliae ◽  
Acer Saccharum ◽  
Sugar Maple ◽  
Xylem Sap ◽  
Amino Acid Content ◽  
Fungal Growth ◽  
Dry Weight

Conidial and dry weight production of Verticillium dahliae varied greatly depending on the amino acid used as the nitrogen source in Czapek's–Dox medium. Variable fungal growth was also noted when mixtures of amino acids were used and growth was dependent on the type and concentration of each amino acid in the mixture. Similar variation was obtained when sugar maple (Acer saccharum) sap was amended with individual amino acids. These results are examined in relation to the reported variation in the amino acid content of host xylem sap and in relation to verticillium wilt development.

scholarly journals Identification of Novel Virulence-Associated Proteins Secreted to Xylem by Verticillium nonalfalfae During Colonization of Hop Plants

2016 ◽  
Vol 29 (5) ◽  
pp. 362-373 ◽  
Author(s):  
Marko Flajsman ◽  
Stanislav Mandelc ◽  
Sebastjan Radisek ◽  
Natasa Stajner ◽  
Jernej Jakse ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Xylem Sap ◽  
Hypothetical Protein ◽  
Secreted Proteins ◽  
In Planta ◽  
Fungal Virulence ◽  
Associated Proteins ◽  
Verticillium Nonalfalfae ◽  
Identification And Characterization

Plant pathogens employ various secreted proteins to suppress host immunity for their successful host colonization. Identification and characterization of pathogen-secreted proteins can contribute to an understanding of the pathogenicity mechanism and help in disease control. We used proteomics to search for proteins secreted to xylem by the vascular pathogen Verticillium nonalfalfae during colonization of hop plants. Three highly abundant fungal proteins were identified: two enzymes, α-N-arabinofuranosidase (VnaAbf4.216) and peroxidase (VnaPRX1.1277), and one small secreted hypothetical protein (VnaSSP4.2). These are the first secreted proteins so far identified in xylem sap following infection with Verticillium spp. VnaPRX1.1277, classified as a heme-containing peroxidase from Class II, similar to other Verticillium spp. lignin-degrading peroxidases, and VnaSSP4.2, a 14-kDa cysteine-containing protein with unknown function and with a close homolog in related V. alfalfae strains, were further examined. The in planta expression of VnaPRX1.1277 and VnaSSP4.2 genes increased with the progression of colonization, implicating their role in fungal virulence. Indeed, V. nonalfalfae deletion mutants of both genes exhibited attenuated virulence on hop plants, which returned to the level of the wild-type pathogenicity in the knockout complementation lines, supporting VnaPRX1.1277 and VnaSSP4.2 as virulence factors required to promote V. nonalfalfae colonization of hop plants.

scholarly journals HopZ4 from Pseudomonas syringae, a Member of the HopZ Type III Effector Family from the YopJ Superfamily, Inhibits the Proteasome in Plants

2014 ◽  
Vol 27 (7) ◽  
pp. 611-623 ◽  
Author(s):  
Suayib Üstün ◽  
Patrick König ◽  
David S. Guttman ◽  
Frederik Börnke
Keyword(s):  
Pseudomonas Syringae ◽  
Sequence Similarity ◽  
Bacterial Species ◽  
Inhibitory Effect ◽  
Catalytic Triad ◽  
Effector Proteins ◽  
In Planta ◽  
High Sequence Similarity ◽  
Type Iii Effector ◽  
Type Iii

The YopJ family of type III effector proteins (T3E) is one of the largest and most widely distributed families of effector proteins, whose members are highly diversified in virulence functions. In the present study, HopZ4, a member of the YopJ family of T3E from the cucumber pathogen Pseudomonas syringae pv. lachrymans is described. HopZ4 shares high sequence similarity with the Xanthomonas T3E XopJ, and a functional analysis suggests a conserved virulence function between these two T3E. As has previously been shown for XopJ, HopZ4 interacts with the proteasomal subunit RPT6 in yeast and in planta to inhibit proteasome activity during infection. The inhibitory effect on the proteasome is dependent on localization of HopZ4 to the plasma membrane as well as on an intact catalytic triad of the effector protein. Furthermore, HopZ4 is able to complement loss of XopJ in Xanthomonas spp., as it prevents precocious host cell death during a compatible Xanthomonas-pepper interaction. The data presented here suggest that different bacterial species employ inhibition of the proteasome as a virulence strategy by making use of conserved T3E from the YopJ family of bacterial effector proteins.

scholarly journals Identification and Characterization of In planta–Expressed Secreted Effector Proteins from Magnaporthe oryzae That Induce Cell Death in Rice

2013 ◽  
Vol 26 (2) ◽  
pp. 191-202 ◽  
Author(s):  
Songbiao Chen ◽  
Pattavipha Songkumarn ◽  
R. C. Venu ◽  
Malali Gowda ◽  
Maria Bellizzi ◽  
...  
Keyword(s):  
Cell Death ◽  
Magnaporthe Oryzae ◽  
Massively Parallel Signature Sequencing ◽  
Expression System ◽  
Ectopic Expression ◽  
Fungal Growth ◽  
Effector Proteins ◽  
In Planta ◽  
Physiological Basis ◽  
Fungal Genes

Interactions between rice and Magnaporthe oryzae involve the recognition of cellular components and the exchange of complex molecular signals from both partners. How these interactions occur in rice cells is still elusive. We employed robust-long serial analysis of gene expression, massively parallel signature sequencing, and sequencing by synthesis to examine transcriptome profiles of infected rice leaves. A total of 6,413 in planta–expressed fungal genes, including 851 genes encoding predicted effector proteins, were identified. We used a protoplast transient expression system to assess 42 of the predicted effector proteins for the ability to induce plant cell death. Ectopic expression assays identified five novel effectors that induced host cell death only when they contained the signal peptide for secretion to the extracellular space. Four of them induced cell death in Nicotiana benthamiana. Although the five effectors are highly diverse in their sequences, the physiological basis of cell death induced by each was similar. This study demonstrates that our integrative genomic approach is effective for the identification of in planta–expressed cell death–inducing effectors from M. oryzae that may play an important role facilitating colonization and fungal growth during infection.

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.

Meiotic Recombination Between Paralogous RBCSB Genes on Sister Chromatids of Arabidopsis thaliana

Genetics ◽  
2004 ◽  
Vol 166 (2) ◽  
pp. 947-957 ◽  
Author(s):  
John G Jelesko ◽  
Kristy Carter ◽  
Whitney Thompson ◽  
Yuki Kinoshita ◽  
Wilhelm Gruissem
Keyword(s):  
Gene Cluster ◽  
Dna Sequences ◽  
Meiotic Recombination ◽  
Sequence Similarity ◽  
Specific Gene ◽  
High Sequence Similarity ◽  
Paralogous Genes ◽  
Chimeric Genes ◽  
Unequal Recombination ◽  
Sister Chromatids

Abstract Paralogous genes organized as a gene cluster can rapidly evolve by recombination between misaligned paralogs during meiosis, leading to duplications, deletions, and novel chimeric genes. To model unequal recombination within a specific gene cluster, we utilized a synthetic RBCSB gene cluster to isolate recombinant chimeric genes resulting from meiotic recombination between paralogous genes on sister chromatids. Several F1 populations hemizygous for the synthRBCSB1 gene cluster gave rise to Luc+ F2 plants at frequencies ranging from 1 to 3 × 10-6. A nonuniform distribution of recombination resolution sites resulted in the biased formation of recombinant RBCS3B/1B::LUC genes with nonchimeric exons. The positioning of approximately half of the mapped resolution sites was effectively modeled by the fractional length of identical DNA sequences. In contrast, the other mapped resolution sites fit an alternative model in which recombination resolution was stimulated by an abrupt transition from a region of relatively high sequence similarity to a region of low sequence similarity. Thus, unequal recombination between paralogous RBCSB genes on sister chromatids created an allelic series of novel chimeric genes that effectively resulted in the diversification rather than the homogenization of the synthRBCSB1 gene cluster.

scholarly journals Considerations for Verticillium Wilt Resistance Evaluation in Potato

Plant Disease ◽  
2007 ◽  
Vol 91 (4) ◽  
pp. 360-367 ◽  
Author(s):  
K. E. Frost ◽  
D. I. Rouse ◽  
S. H. Jansky
Keyword(s):  
Disease Severity ◽  
Verticillium Wilt ◽  
Yield Loss ◽  
Control Culture ◽  
In Planta ◽  
Disease Symptoms ◽  
Accuracy And Precision ◽  
Population Sizes ◽  
Pathogen Colonization ◽  
Time Accuracy

Verticillium wilt (Vw), caused by the soilborne fungi Verticillium dahliae and V. albo-atrum, is an important disease of potato (Solanum tuberosum). Host plant resistance is a promising method of Vw control. Culture-based methods that quantify the pathogen in host tissue often are used for Vw resistance screening. To evaluate the processing time, accuracy, and precision of these methods, 46 clones were planted in a field naturally infested with V. dahliae to collect data on visual disease symptoms, pathogen colonization, and yield. In 2002, disease severity explained 4.34% of the variability of yield loss, but the linear relationship between stem colonization and yield loss was not significant. In 2003, stem colonization explained 57.5% of the variability of yield loss, whereas disease severity explained 1.7% of the variability of yield loss. Correlations comparing clone ranks from repeated pathogen measurements indicated that culturing sap from individual stems or bulked stems generated more repeatable clone rankings than culturing dried stems. Clone rankings were more repeatable between years if pathogen measurements were made earlier in the growing season. The results indicate a need to characterize the effect of the environment on the relationship among pathogen population sizes in planta, disease symptoms, and yield loss.

Evolutionary rewiring of the wheat transcriptional regulatory network by lineage-specific transposable elements

2021 ◽  
pp. gr.275658.121
Author(s):  
Yuyun Zhang ◽  
Zijuan Li ◽  
Yu'e Zhang ◽  
Kande Lin ◽  
Yuan Peng ◽  
...  
Keyword(s):  
Transposable Elements ◽  
Genome Evolution ◽  
Regulatory Networks ◽  
Transcriptional Regulatory Network ◽  
Sequence Similarity ◽  
Purifying Selection ◽  
Wheat Genome ◽  
Specific Gene ◽  
High Sequence Similarity ◽  
Wheat Genome Evolution

More than 80% of the wheat genome consists of transposable elements (TEs), which act as one major driver of wheat genome evolution. However, their contributions to the regulatory evolution of wheat adaptations remain largely unclear. Here, we created genome-binding maps for 53 transcription factors (TFs) underlying environmental responses by leveraging DAP-seq in Triticum urartu, together with epigenomic profiles. Most TF-binding sites (TFBS) located distally from genes are embedded in TEs, whose functional relevance is supported by purifying selection and active epigenomic features. About 24% of the non-TE TFBS share significantly high sequence similarity with TE-embedded TFBS. These non-TE TFBS have almost no homologous sequences in non-Triticeae species and are potentially derived from Triticeae-specific TEs. The expansion of TE-derived TFBS linked to wheat-specific gene responses, suggesting TEs are an important driving force for regulatory innovations. Altogether, TEs have been significantly and continuously shaping regulatory networks related to wheat genome evolution and adaptation.

pha-4 is Ce-fkh-1, a fork head/HNF-3alpha, beta, gamma homolog that functions in organogenesis of the C. elegans pharynx

Development ◽  
1998 ◽  
Vol 125 (12) ◽  
pp. 2171-2180 ◽  
Author(s):  
J.M. Kalb ◽  
K.K. Lau ◽  
B. Goszczynski ◽  
T. Fukushige ◽  
D. Moons ◽  
...  
Keyword(s):  
Early Stage ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Ectopic Expression ◽  
Specific Gene ◽  
Gata Factor ◽  
Enhancer Element ◽  
C Elegans ◽  
Fork Head ◽  
Organ Identity

The C. elegans Ce-fkh-1 gene has been cloned on the basis of its sequence similarity to the winged-helix DNA binding domain of the Drosophila fork head and mammalian HNF-3alpha, beta, gamma genes, and mutations in the zygotically active pha-4 gene have been shown to block formation of the pharynx (and rectum) at an early stage in embryogenesis. In the present paper, we show that Ce-fkh-1 and pha-4 are the same gene. We show that PHA-4 protein is present in nuclei of essentially all pharyngeal cells, of all five cell types. PHA-4 protein first appears close to the point at which a cell lineage will produce only pharyngeal cells, independently of cell type. We show that PHA-4 binds directly to a ‘pan-pharyngeal enhancer element’ previously identified in the promoter of the pharyngeal myosin myo-2 gene; in transgenic embryos, ectopic PHA-4 activates ectopic myo-2 expression. We also show that ectopic PHA-4 can activate ectopic expression of the ceh-22 gene, a pharyngeal-specific NK-2-type homeodomain protein previously shown to bind a muscle-specific enhancer near the PHA-4 binding site in the myo-2 promoter. We propose that it is the combination of pha-4 and regulatory molecules such as ceh-22 that produces the specific gene expression patterns during pharynx development. Overall, pha-4 can be described as an ‘organ identity factor’, completely necessary for organ formation, present in all cells of the organ from the earliest stages, capable of integrating upstream developmental pathways (in this case, the two distinct pathways that produce the anterior and posterior pharynx) and participating directly in the transcriptional regulation of organ specific genes. Finally, we note that the distribution of PHA-4 protein in C. elegans embryos is remarkably similar to the distribution of the fork head protein in Drosophila embryos: high levels in the foregut/pharynx and hindgut/rectum; low levels in the gut proper. Moreover, we show that pha-4 expression in the C. elegans gut is regulated by elt-2, a C. elegans gut-specific GATA-factor and possible homolog of the Drosophila gene serpent, which influences fork head expression in the fly gut. Overall, our results provide evidence for a highly conserved pathway regulating formation of the digestive tract in all (triploblastic) metazoa.

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