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Quantitative trait loci for resistance to Sclerotinia sclerotiorum and its association with a homeologous non-reciprocal transposition in Brassica napus L.

Sclerotinia stem rot, caused by fungus Sclerotinia sclerotiorum, is one of the most devastating diseases in rapeseed (Brassica napus L.). We report the identification of Quantitative trait loci (QTL) involved in the resistance to S. sclerotiorum in two segregating populations of DH lines: the HUA population, derived from a cross between a partially resistant Chinese winter line (Hua dbl2) and a susceptible European spring line (P1804); and the MS population, derived from a partially resistant French winter cultivar (Major) and a susceptible Canadian spring cultivar (Stellar). A petiole inoculation technique and two scoring methods, days to wilt (DW) and stem lesion length (SLL), were used for the resistance assessment. A total of eight genomic regions affecting resistance were detected in the HUA population, with four of these regions affecting both measures of resistance. Only one region, which affected both measurements, was detected in the MS population. Individual QTL explained 6-22% of the variance. At five of the QTL from both populations, alleles from the resistant parent contributed to the resistance. QTL on N2 from the HUA population had the highest LOD score and R (2) value and was detected for SLL in the first evaluation. The N12 resistance allele in Hua dbl2 was detected in a region containing a homeologous non-reciprocal transposition (HNRT) from the resistance-containing portion of N2. This result suggests that QTL in the N12.N2 HNRT enhanced the resistance of Hua dbl2 by increasing the dosage of resistance genes. The relationship of QTL from different genetic backgrounds and their associations with other agronomic traits are discussed.

Ascomycota↗

Serial Analysis of Gene Expression (SAGE) of Magnaporthe grisea: genes involved in appressorium formation.

Treatment with cyclic AMP (cAMP) induces appressorium formation in the phytopathogenic fungus Magnaporthe grisea, the causative agent of rice blast disease. In a search for the M. grisea genes responsible for appressorium formation and host invasion, SAGE (Serial Analysis of Gene Expression) was carried out using mRNA isolated from fungal conidia germinating in the presence and absence of cAMP. From cAMP-treated conidia 5087 tags including 2889 unique tags were isolated, whereas untreated conidia yielded 2342 unique tags out of total of 3938. cAMP treatment resulted in up- and down-regulation of genes corresponding to 57 and 53 unique tags, respectively. Upon consultation of EST/cDNA databases, 22 tags with higher representation in cAMP-treated conidia were annotated with putative gene names. Furthermore, 28 tags corresponding to cAMP-induced genes could be annotated with the help of the recently published genome sequence of M. grisea. cAMP-induced genes identified by SAGE included many genes that have not been described so far, as well as a number of genes known to be involved in pathogenicity, e.g. MPG1, MAS1 and MAC1. RT-PCR of 13 randomly selected genes confirmed the SAGE results, verifying the fidelity of the SAGE data.

Base Sequence↗

Antimicrobial peptide defenses against chytridiomycosis, an emerging infectious disease of amphibian populations.

Chytridiomycosis, an emerging infectious disease (EID) of the skin caused by the chytrid fungus, Batrachochytrium dendrobatidis, has been linked with continuing amphibian population declines in the western USA, Central America, Europe, Africa, and Australia. Genetic analysis suggests that B. dendrobatidis is a recently emerged pathogen. This article reviews the biology of this pathogenic chytrid and the evidence for chytridiomycosis as a cause of declines in amphibian populations worldwide. Data are presented to show that antimicrobial peptides, produced in granular glands of the skin and released in high concentrations into skin secretions, are highly effective in inhibiting growth of B. dendrobatidis in vitro and may provide limited protection for some species. Ongoing studies suggest a correlation between resistance to lethal infection by B. dendrobatidis and synthesis of antimicrobial peptides by the host amphibian, but further research is needed to define better the role of antimicrobial peptides in protection of amphibian populations and the effect of environmental factors upon antimicrobial peptide synthesis.

Amphibians↗

Inactivation of Colletotrichum gloeosporioides spores by high hydrostatic pressure combined with citral or lemongrass essential oil.

Anthracnose, caused by the fungus Colletotrichum gloeosporioides, is the main post-harvest disease of the papaya. Inactivation of the spores of C. gloeosporioides in saline solution by the use of high hydrostatic pressure, citral oil and lemongrass oil, alone and in combination, was studied. C. gloeosporioides spores were efficiently inhibited after a pressure treatment of 350 MPa for 30 min. When C. gloeosporioides was treated with 0.75 mg ml(-1) of citral or lemongrass oil, the pressure needed to achieve the same spore inhibition was 150 MPa. This work suggests the use of high hydrostatic pressure and plant essential oils as an alternative control for fruit diseases.

Acyclic Monoterpenes↗

Acetylenic aromatic compounds from Stereum hirsutum.

Stereum hirsutum is a one of several fungi involved in a grapevine disease called esca. From the culture medium of this fungus, four new acetylenic compounds 1-3 and 6 have been isolated and identified. Structural elucidation and biological activity are reported.

Acetylene↗

Fumonisin B(1)-induced alterations in cytokine expression and apoptosis signaling genes in mouse liver and kidney after an acute exposure.

Fumonisin B(1) (FB(1)), a carcinogenic mycotoxin produced primarily by fungus Fusarium verticillioides in corn, causes several fatal animal diseases. In mice, liver is the primary site of its toxicity. Our previous study showed that maximum induction of interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) was observed at 4 and 8 h, respectively, after an acute po FB(1) treatment. To further investigate the time-related induction of other cytokines and genes involved in apoptosis signaling, male BALB/c mice were administered orally with either saline or 25 mg/kg of FB(1) and sampled 4 or 8 h after treatment. Expression of various genes was analyzed by ribonuclease protection assay. FB(1) treatment caused increased expression of TNFalpha and interleukin (IL)-1beta in both liver and kidney, whereas IL-1alpha and IL-1 receptor antagonist (IL-1Ra) expression was induced only in the liver. Expression of TNFalpha signaling molecules, TNF receptor 55 and receptor interacting protein, was increased in liver and kidney after FB(1) treatment. Caspase 8 expression was increased only in liver with no changes in kidney with FB(1). FB(1) treatment induced expression of Fas in liver and kidney with no alterations in Fas signaling molecules, Fas ligand, Fas-associated death domain and Fas-associated protein factor. Treatment of mice with FB(1) increased the expression of B-Myc, c-Myc and Max, oncogenic transcription factors in the kidney. FB(1) toxicity caused induction of cytokine network in liver with involvement of TNFalpha signaling pathway. Increased expression of caspase 8 involved in the TNFalpha signaling pathway may contribute to the apoptosis, whereas IL-1Ra induction could contribute to the proliferating effects observed in FB(1) toxicity.

Animals↗

Bioactive metabolites from the fungus Nectria galligena, the main apple canker agent in Chile.

The phytopathogenic fungus Nectria galligena Bres. is the most common canker disease agent of hardwood trees. The terpenoids colletochlorin B, colletorin B, ilicicolin C, E, and F, as well as the phytotoxin alpha,beta-dehydrocurvularin have been isolated from liquid cultures of N. galligena obtained from the xylem of infected apple trees in central Chile. Ilicicolin C and F and alpha,beta-dehydrocurvularin were active against Pseudomonas syringae with IC50 values of 28.5, 28.5, and 14.2 microg/mL, respectively, in the same range as streptomycin and penicillin G (11 and 15 microg/mL, respectively). All of the compounds showed moderate inhibitory activity toward the enzymes acetylcholinesterase (AChE) and beta-glucuronidase. The most active enzyme inhibitors were colletochlorin B and ilicicolin C and E, with IC50 values of 30-36 microg/mL in the AChE assay and 32-43 microg/mL in the beta-glucuronidase test. All of the chlorinated compounds showed some toxicity toward human lung fibroblasts, with IC50 values in the range of 64-120 microg/mL. alpha,beta-Dehydrocurvularin proved to be the most toxic compound, showing IC50 values less than 12 microg/mL. The effect of the isolated compounds on seed germination and radicle and epicotyl growth was assessed in lettuce and millet seeds. At 100 and 200 microg/disk, alpha,beta-dehydrocurvularin significantly reduced radicle length and epicotyl growth in Lactuca sativa. This is the first report on the occurrence of colletochlorin B, colletorin B, ilicicolin C, E, and F, as well as alpha,beta-dehydrocurvularin associated to N. galligena.

Anti-Bacterial Agents↗

Single-chain fusions of two unrelated homeodomain proteins trigger pathogenicity in Ustilago maydis.

Pathogenic and sexual development of the fungus Ustilago maydis, the causal agent of corn smut disease, is regulated by heterodimerization of two unrelated homeodomain proteins bE and bW, both encoded by the multi-allelic b mating-type locus. This complex can only be formed if the two proteins are derived from different alleles. The heterodimer is believed to function as a transcriptional regulator that binds to target sites upstream of developmentally regulated genes. We have synthesized a translational fusion in which bE is tethered to bW by a designed flexible kink region. U. maydis strains expressing this synthetic b-fusion become pathogenic for corn illustrating that the single-chain fusion substitutes for the active bE/bW heterodimer. Synthetic b-fusions in which bE and bW originate from the same allele as well as fusions deleted for the dimerization domains were shown to be active while both homeodomains were required for function. Such active fusion proteins are expected to be instrumental in the identification of pathogenicity genes.

Journal Article↗

RAR1, ROR1, and the actin cytoskeleton contribute to basal resistance to Magnaporthe grisea in barley.

The fungus Magnaporthe grisea, the causal agent of rice blast disease, is a major pathogen of rice and is capable of producing epidemics on other cultivated cereals, including barley (Hordeum vulgare). We explored the requirements for basal resistance of barley against a compatible M. grisea isolate using both genetic and chemical approaches. Mutants of the RAR1 gene required for the function of major resistance gene-mediated resistance and mutants of the ROR1 and ROR2 genes required for full expression of cell-wall-penetration resistance against powdery mildew pathogens were examined for macroscopic and microscopic alterations in M. grisea growth and symptoms. RAR1 contributed to resistance in epidermis and mesophyll at different stages of fungal infection dependent on the MLO/mlo-5 status. Whereas no ROR2 effect was detected, ROR1 was found to contribute to cell-wall-penetration resistance, at least in the epidermis. Application of the actin agonist cytochalasin E promoted cell wall penetration by M. grisea in a dose-dependent manner, demonstrating an involvement of the actin cytoskeleton in penetration resistance.

Actins↗

MPG1 Encodes a Fungal Hydrophobin Involved in Surface Interactions during Infection-Related Development of Magnaporthe grisea.

The rice blast fungus expresses a pathogenicity gene, MPG1, during appressorium formation, disease symptom development, and conidiation. The MPG1 gene sequence predicts a small protein belonging to a family of fungal proteins designated hydrophobins. Using random ascospore analysis and genetic complementation, we showed that MPG1 is necessary for infection-related development of Magnaporthe grisea on rice leaves and for full pathogenicity toward susceptible rice cultivars. The protein product of MPG1 appears to interact with hydrophobic surfaces, where it may act as a developmental sensor for appressorium formation. Ultrastructural studies revealed that MPG1 directs formation of a rodlet layer on conidia composed of interwoven ~5-nm rodlets, which contributes to their surface hydrophobicity. Using combined genetic and biochemical approaches, we identified a 15-kD secreted protein with characteristics that establish it as a class I hydrophobin. The protein is able to form detergent-insoluble high molecular mass complexes, is soluble in trifluoroacetic acid, and exhibits mobility shifts after treatment with performic acid. The production of this protein is directed by MPG1.

Journal Article↗

Specific antibody can prevent fungal biofilm formation and this effect correlates with protective efficacy.

One of the most troublesome medical problems today is infection of prosthetic devices with organisms that form polysaccharide biofilms. This combined with increasing antimicrobial drug resistance is making many infectious diseases incurable. Cryptococcus neoformans is a human-pathogenic fungus that has a polysaccharide capsule and can form biofilms in prosthetic medical devices. We developed a system to study cryptococcal biofilm formation in vitro and studied the effect of antibody to the C. neoformans capsular polysaccharide on this process. C. neoformans biofilm formation was dependent on the presence of a polysaccharide capsule and correlated with the ability of capsular polysaccharide to bind the polystyrene solid support. Protective antibodies prevented biofilm formation whereas nonprotective antibodies were not effective. The mechanism of antibody action involved interference with capsular polysaccharide release from the fungal cell. In contrast, lactoferrin, an effector molecule of innate immune mechanisms, was unable to prevent fungal biofilm formation despite its efficacy against bacterial biofilms. Our results suggest a new role of adaptive humoral immunity whereby some antibodies can inhibit biofilm formation by encapsulated organisms. Vaccines that elicit antibody responses to capsular antigens and/or passive transfer of antibodies to microbial polysaccharides may be useful in preventing biofilm formation.

Antibodies, Fungal↗

Clinical isolates of Histoplasma capsulatum from Indianapolis, Indiana, have a recombining population structure.

A sample of 30 clinical isolates of Histoplasma capsulatum was analyzed to determine (i) whether genetic exchange is important in the life cycle of this fungus and (ii) whether distinct subpopulations which correlate with disease severity or host immune status exist. Eleven biallelic molecular markers were developed, with the frequency of the least common allele at each molecular locus ranging from 10 to 50%. Every isolate had a different, unique multilocus genotype. Data analysis indicated that frequent recombination occurs within the Indianapolis, Ind., population. There were no associations between isolates from the immunocompromised population or from those with different clinical manifestations of histoplasmosis.

Genes, Fungal↗

Management of Pneumocystis pneumonia in patients with inflammatory disorders.

Pneumocystis jirovecii is an atypical fungus that causes Pneumocystis pneumonia in immunocompromised patients. Underlying diseases associated with Pneumocystis pneumonia mainly consist of hematologic malignancies, solid tumors, organ transplant recipients and inflammatory disorders. Currently, inflammatory disorders represent 20% of underlying diseases. Corticosteroids are considered as a major risk factor. Recently introduced immunosuppressive drugs, such as antitumor necrosis factor monoclonal antibodies, could enhance the risk of Pneumocystis pneumonia. In patients with inflammatory disorders, lymphopenia is probably a determining factor but CD4+ T-cell count associated with the risk of Pneumocystis pneumonia remains unassessed. The diagnosis is based upon clinical, radiologic and biologic data. The identification of P. jirovecii usually requires a lower respiratory tract specimen, even if oral washes samples seem to be promising. According to recent data, immunofluorescent stains should be considered as the new gold standard, and specialized techniques such as PCR should be applied for sputum samples or oral washes. Recommendations on prophylaxis remains controversial except in patients with Wegener's granulomatosis and systemic lupus erythematosus. Cotrimoxazole is the preferred agent for prophylaxis as well as for treatment. An adjunctive corticosteroid therapy is usually prescribed despite the lack of evidence for utility in patients with inflammatory disorders. As person-to-person transmission is the most likely mode of acquiring P. jirovecii, isolation precautions should be advised.

Antifungal Agents↗

Duodenal obstruction caused by infection with Pythium insidiosum in a 12-week-old puppy.

Pythium insidiosum is an aquatic fungus-like organism that causes a serious chronic granulomatous disease called pythiosis in animals and humans in tropical and subtropical regions of the world. In North America, pythiosis is most often diagnosed in the Gulf Coast states. Early recognition of the disease is crucial to successful treatment, which includes surgical resection of granulomatous lesions and administration of antifungal agents. Despite increasing availability of diagnostic tests, intestinal pythiosis is insidious and is often not detected until lesions are extensive. Intestinal pythiosis was diagnosed in a 12-week-old puppy from South Carolina examined because of vomiting, diarrhea, and anorexia. Pythiosis was not initially suspected because of the young age of the patient and because pythiosis is uncommon in this area.

Animals↗

Co-localisation of Batrachochytrium dendrobatidis and keratin for enhanced diagnosis of chytridiomycosis in frogs.

Chytridiomycosis is a disease of post-metamorphic frogs caused by the fungus Batrachochytrium dendrobatidis and is associated with large declines in frog populations on a global scale. B. dendrobatidis is found only in the keratinised tissues, which include the mouthparts of healthy tadpoles. The epidermis of infected post-metamorphic frogs is thickened (hyperkeratosis) and the superficial layer can sometimes slough. Diagnosis is most commonly performed on stained sections of toe clips or ventral skin. Accurate interpretation can be difficult and requires a high level of expertise, particularly in infected animals exhibiting hyperkeratosis with sloughing. Misdiagnosis can occur when zoosporangia of B. dendrobatidis are shed with the superficial keratin layers. We have developed a staining protocol based on previously described methods to detect both B. dendrobatidis and keratin, to improve the sensitivity and specificity of diagnosis of chytridiomycosis by inexperienced diagnosticians.

Animals↗

Phaeohyphomycosis caused by Drechslera spicifera in a cat.

A slowly evolving subcutaneous mycosis in a 10-year-old domestic shorthair cat was found to be caused by Drechslera spicifera, the imperfect state of ascomycete Cochliobolus spicifer. The cat had circular, nodular, granulomatous lesions over its sternum. Scattered individual and small groups of septate hyphae and chlamydospores were found in histologic sections. Many of the hyphae also had bizarre dilatations. Most of the fungal elements were hyaline; a few, however, were dematiacious. Because the fungus was not organized into granules in tissue, the disease could not be classified as a mycetoma. The preferred name for infections of this type is phaeohyphomycosis.

Animals↗

[Occupational allergic asthma in greenhouses. Report of a clinical case].

The Authors report a particular case of occupational asthma observed in a greenhouse worker. They describe the diagnostic procedure followed after identification on the cultivated plants of colonies of aspergillus niger, a fungus identified as an etiologic agent of the allergic disease. The peculiarity of the causative agent is stressed and it is recommended that environmental and health prevention measures be improved for this poorly protected class of workers.

Adult↗

Cutaneous lesions of histoplasmosis with transepidermal elimination in a patient with acquired immunodeficiency syndrome.

Histoplasma capsulatum is a dimorphic pathogenic fungus that produces a variety of self-limiting disease syndromes in healthy persons, but commonly disseminates in immunocompromised hosts. Although histoplasmosis has been recently described in patients with acquired immunodeficiency syndrome, it is an uncommon finding, occurring in fewer than 0.5 percent of patients with acquired immunodeficiency syndrome. We are reporting a second case of disseminated histoplasmosis presenting as keratotic papules with transepidermal elimination of Histoplasma capsulatum. Our case demonstrates the importance of including histoplasmosis in the differential diagnosis of keratotic papules occurring in patients with acquired immunodeficiency syndrome.

Acquired Immunodeficiency Syndrome↗