Sclerotinia sclerotiorum (Lib.) de Bary is a necrotrophic fungal pathogen causing disease in a wide range of plants. This review summarizes current knowledge of mechanisms employed by the fungus to parasitize its host with emphasis on biology, physiology and molecular aspects of pathogenicity. In addition, current tools for research and strategies to combat S. sclerotiorum are discussed. Taxonomy: Sclerotinia sclerotiorum (Lib.) de Bary: kingdom Fungi, phylum Ascomycota, class Discomycetes, order Helotiales, family Sclerotiniaceae, genus Sclerotinia. Identification: Hyphae are hyaline, septate, branched and multinucleate. Mycelium may appear white to tan in culture and in planta. No asexual conidia are produced. Long-term survival is mediated through the sclerotium; a pigmented, multi-hyphal structure that can remain viable over long periods of time under unfavourable conditions for growth. Sclerotia can germinate to produce mycelia or apothecia depending on environmental conditions. Apothecia produce ascospores, which are the primary means of infection in most host plants. Host range: S. sclerotiorum is capable of colonizing over 400 plant species found worldwide. The majority of these species are dicotyledonous, although a number of agriculturally significant monocotyledonous plants are also hosts. Disease symptoms: Leaves usually have water-soaked lesions that expand rapidly and move down the petiole into the stem. Infected stems of some species will first develop dark lesions whereas the initial indication in other hosts is the appearance of water-soaked stem lesions. Lesions usually develop into necrotic tissues that subsequently develop patches of fluffy white mycelium, often with sclerotia, which is the most obvious sign of plants infected with S. sclerotiorum.

Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen.

Taxonomy: Kingdom Fungi; Phylum Ascomycota; Class Sordariomycetes; Order Hypocreales; Family Nectriaceae; genus Fusarium. Host range: Very broad at the species level. More than 120 different formae speciales have been identified based on specificity to host species belonging to a wide range of plant families. Disease symptoms: Initial symptoms of vascular wilt include vein clearing and leaf epinasty, followed by stunting, yellowing of the lower leaves, progressive wilting, defoliation and, finally, death of the plant. On fungal colonization, the vascular tissue turns brown, which is clearly visible in cross-sections of the stem. Some formae speciales are not primarily vascular pathogens, but cause foot and root rot or bulb rot. Economic importance: Can cause severe losses in many vegetables and flowers, field crops, such as cotton, and plantation crops, such as banana, date palm and oil palm. Control: Use of resistant varieties is the only practical measure for controlling the disease in the field. In glasshouses, soil sterilization can be performed. Useful websites: http://www.broad.mit.edu/annotation/genome/fusarium_group/MultiHome.html; http://www.fgsc.net/Fusarium/fushome.htm; http://www.phi-base.org/query.php

Pathogen profile update: Fusarium oxysporum

SUMMARY
Taxonomy: Vascular wilt fungus; Ascomycete although sexual stage is yet to be found. The most closely related teleomorphic group, Gibberella, is classified within the Pyrenomycetes.

 Host range: Very broad at the species level. More than 120 different formae speciales have been identified based on specificity to host species belonging to a wide range of plant families.

 Disease symptoms: Initial symptoms of vascular wilt include vein clearing and leaf epinasty, followed by stunting, yellowing of the lower leafs, progressive wilting of leaves and stem, defoliation and finally death of the plant. In cross-sections of the stem, a brown ring is evident in the area of the vascular bundles. Some formae speciales are not primarily vascular pathogens but cause foot- and rootrot or bulbrot.

 Economic importance: Causes severe losses on most vegetables and flowers, several field crops such as cotton and tobacco, plantation crops such as banana, plantain, coffee and sugarcane, and a few shade trees.

 Control: Use of resistant varieties is the only practical measure for controlling the disease in the field. Under greenhouse conditions, soil sterilization can be performed. Alternative control methods with potential for the future include soil solarization and biological control with antagonistic bacteria or fungi.

 Useful websites: http://www.fgsc.net/fus.htm, http://www-genome.wi.mit.edu/annotation/fungi/fusarium/, http://www.cbs.knaw.nl/fusarium/database.html

Fusarium oxysporum: exploring the molecular arsenal of a vascular wilt fungus

Summary The soil-borne vascular wilt fungus Fusarium oxysporum infects a wide variety of plant species by directly penetrating roots, invading the cortex and colonizing the vascular tissue. We have identified fmk1, encoding a mitogen-activated protein kinase (MAPK) of F. oxysporum that belongs to the yeast and fungal extracellular signal-regulated kinase (YERK1) subfamily. Targeted mutants of F. oxysporum f. sp. lycopersici carrying an inactivated copy of fmk1 have lost pathogenicity on tomato plants but show normal vegetative growth and conidiation in culture. Colonies of the fmk1 mutants are easily wettable, and hyphae are impaired in breaching the liquid‐air interface, suggesting defects in surface hydrophobicity. Fmk1 mutants also show reduced invasive growth on tomato fruit tissue and drastically reduced transcript levels of pl1 encoding the cell wall-degrading enzyme pectate lyase. Conidia of the mutants germinating in the tomato rhizosphere fail to differentiate penetration hyphae, resulting in greatly impaired root attachment. The orthologous MAPK gene Pmk1 from the rice leaf pathogen Magnaporthe grisea complements invasive growth and partially restores surface hydrophobicity, root attachment and pathogenicity in an fmk1 mutant. These results demonstrate that FMK1 controls several key steps in the pathogenesis of F. oxysporum and suggest a fundamentally conserved role for the corresponding MAPK pathway in soil-borne and foliar plant pathogens.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2958.2001.02307.x

A MAP kinase of the vascular wilt fungus Fusarium oxysporum is essential for root penetration and pathogenesis

Fusarium graminearum is the causal agent of the Fusarium head blight (FHB) and a destructive pathogen of cereals accounting for high grain yield losses especially on wheat and maize. Like other fungal pathogens, F. graminearum secretes various extracellular enzymes, which are hypothesized to be involved in host infection. Extracellular lipolytic activity of F. graminearum was strongly induced in culture by wheat germ oil; this allowed us to isolate, clone, and characterize a gene (FGL1) encoding a secreted lipase. Expression analysis indicated that FGL1 is induced by lipid-containing substrates and repressed by glucose. In planta, FGL1 transcription was detected 1 day post-infection of wheat spikes. The function of the FGL1 gene product was verified by specifically demonstrating lipase activity after expression in a heterologous host. Ebelactone B, a known lipase inhibitor, repressed the lipolytic activity of the enzyme. Disease severity was strongly reduced when wild-type conidia were supplemented with ebelactone B. Transformation-mediated disruption of FGL1 led to reduced extracellular lipolytic activity in culture and to reduced virulence to both wheat and maize.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-313X.2005.02377.x

A secreted lipase of Fusarium graminearum is a virulence factor required for infection of cereals.

The tomato vascular wilt pathogen Fusarium oxysporum f. sp. lycopersici produces an array of pectinolytic enzymes that may contribute to penetration and colonization of the host plant. Here we report the isolation of pg5, encoding a novel extracellular endopolygalacturonase (endoPG) that is highly conserved among different formae speciales of F. oxysporum. The putative mature pg5 product has a calculated molecular mass of 35 kDa and a pI of 8.3 and is more closely related to endoPGs from other fungal plant pathogens than to PG1, the major endoPG of F. oxysporum. Overexpression of pg5 in a bacterial heterologous system produced a 35-kDa protein with endoPG activity. Accumulation of pg5 transcript is induced by citrus pectin and D-galacturonic acid and repressed by glucose. As shown by reverse transcription-PCR, pg5 is expressed by F. oxysporum in tomato roots during the initial stages of infection. Targeted inactivation of pg5 has no detectable effect on virulence toward tomato plants.

Molecular characterization of an endopolygalacturonase from Fusarium oxysporum expressed during early stages of infection.

Role in pathogenesis of two endo-β-1,4-xylanase genes from the vascular wilt fungus Fusarium oxysporum

Two genes encoding putative family F xylanases from the tomato vascular wilt pathogen Fusarium oxysporum f.sp. lycopersici have been cloned and sequenced. The two genes, designated xyl2 and xyl3, encode proteins with calculated molecular masses of 33 and 39.3 kDa and isoelectric points of 8.9 and 6.7, respectively. The predicted amino acid sequences show significant homology to other family F xylanases. XYL3 contains a cellulose-binding domain in its N-terminal region. Southern analysis suggested that xyl2 and xyl3 homologs are also present in other formae speciales of F. oxysporum. Both genes were expressed during growth on oat spelt xylan and tomato vascular tissue in vitro. RT-PCR revealed that xyl3 is expressed in roots and in the lower stems of tomato plants infected by F. oxysporum f.sp. lycopersici throughout the whole disease cycle, whereas xyl2 is only expressed during the final stages of disease.

Two xylanase genes of the vascular wilt pathogen fusarium oxysporum are differentially expressed during infection of tomato plants

The aim of this work was to investigate the major mechanisms involved in antagonism of BO7, a novel strain of Bacillus amyloliquefaciens isolated from orchard soil, against the vascular wilt fungus Fusarium oxysporum f. sp. lycopersici (Fol). BO7 markedly reduced the incidence of Fol vascular wilt on tomato plants and displayed strong in vitro inhibitory activity against Fol. Scanning electron microscopy demonstrated the ability of BO7 cells to adhere to fungal hyphae and to efficiently colonize tomato roots. The low molecular weight fraction of BO7 culture filtrate displayed high antifungal activity against Fol, resulting in growth inhibition and dramatic alterations of hyphal morphology. Three structurally related surfactin lipopeptides, identified as the major components of the bioactive fraction, were assayed for their inhibitory activity against Fol. One of these compounds exerted a strong and uncommon antifungal activity for lipopeptides of the surfactin family, accounting for most of the inhibitory activity of the BO7 culture filtrate. Among a collection of Fol knockout isolates tested, mutants altered in cell wall structure showed increased sensitivity to the bacterial compounds. These results suggest that the fungal cell wall might have a key role in the sensitivity of Fol towards bacterial surfactins from B. amyloliquefaciens.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-3059.2011.02561.x

Role of new bacterial surfactins in the antifungal interaction between Bacillus amyloliquefaciens and Fusarium oxysporum

The biolistic particle delivery system was used to transfer the uidA gene into different garlic tissues, including regenerable calli. These tissues showed a high endogenous nuclease activity preventing exogenous DNA expression. After submerging the tissues for 24 h in a 2 mm solution of the nuclease inhibitor aurintricarboxylic acid, transient expression of the uidA gene was detected. Furthermore, vacuum infiltration of aurintricarboxylic acid into the garlic tissues significantly increased the percentage of explants showing transient expression. Among four plasmids tested, pDE4 containing the CaMV35S-uidA construct produced the best results.

A. Di Pietro

Papers

Molecular characterization of an endopolygalacturonase from Fusarium oxysporum expressed during early stages of infection.

Role in pathogenesis of two endo-β-1,4-xylanase genes from the vascular wilt fungus Fusarium oxysporum

Two xylanase genes of the vascular wilt pathogen fusarium oxysporum are differentially expressed during infection of tomato plants

Role of new bacterial surfactins in the antifungal interaction between Bacillus amyloliquefaciens and Fusarium oxysporum

Biolistic transfer and expression of a uidA reporter gene in different tissues of Allium sativum L