NADPH Oxidase Gene, FgNoxD, Plays a Critical Role in Development and Virulence in Fusarium graminearum.

NADPH oxidase is an enzyme that generates reactive oxygen species from oxygen and NADPH and is highly conserved in eukaryotes. In Fusarium graminearum, a series of different Nox enzymes have been identified. NoxA is involved in sexual development and ascospore production and, like NoxB, also contributes to pathogenicity. Both NoxA and NoxB are regulated by the subunit NoxR, whereas NoxC is usually self-regulated by EF-hand motifs found on the enzyme. In this study, we characterized another NADPH oxidase in F. graminearum, FgNoxD. In the FgNoxD deletion mutant, vegetative growth and conidia production were reduced, while sexual development was totally abolished. The FgNoxD deletion mutant also showed reduced resistance to cell wall perturbing agents; cell membrane inhibitors; and osmotic, fungicide, cold, and extracellular oxidative stress, when compared to the wild type. Moreover, in comparison to the wild type, the FgNoxD deletion mutant exhibited reduced virulence against the host plant. The FgNoxD deletion mutant produced less deoxynivalenol than the wild type, and the Tri5 and Tri6 gene expression was also downregulated. In conclusion, our findings show that FgNoxD is involved in the survival against various stresses, conidiation, sexual development, and virulence, highlighting this enzyme as a new target to control the disease caused by F. graminearum.

Genetic Diversity of Epicoccum nigrum and its Effects on Fusarium graminearum.

Epicoccum nigrum is a saprophytic or endophytic fungus that is found worldwide. Because of the antagonist effects of E. nigrum on many plant pathogens, current studies on E. nigrum have focused on the development of biological control agents and the utilization of its various metabolites. In this study, E. nigrum was collected from a wheat field, and its genetic diversity was analyzed. Phylogenetic analyses identified 63 isolates of E. nigrum divided into seven groups, indicating a wide genetic diversity. Isolates antagonized the wheat pathogen Fusarium graminearum, and reduced disease symptoms caused by F. graminearum in wheat coleoptiles. Moreover, pretreatment of wheat coleoptiles with E. nigrum induced the upregulation of pathogen-related (PR) genes, PR1, PR2, PR3, PR5, PR9, and PR10 in wheat coleoptiles responding to F. graminearum invasion. Overall, this study indicates that E. nigrum isolates can be used as biological pathogen inhibitors applied in wheat fields.

Transcriptome analyses of the ginseng root rot pathogens Cylindrocarpon destructans and Fusarium solani to identify radicicol resistance mechanisms.

BACKGROUND: The ascomycete fungi Cylindrocarpon destructans (Cd) and Fusarium solani (Fs) cause ginseng root rot and significantly reduce the quality and yield of ginseng. Cd produces the secondary metabolite radicicol, which targets the molecular chaperone Hsp90. Fs is resistant to radicicol, whereas other fungal genera associated with ginseng disease are sensitive to it. Radicicol resistance mechanisms have not yet been elucidated. METHODS: Transcriptome analyses of Fs and Cd mycelia treated with or without radicicol were conducted using RNA-seq. All of the differentially expressed genes (DEGs) were functionally annotated using the Fusarium graminearum transcript database. In addition, deletions of two transporter genes identified by RNA-seq were created to confirm their contributions to radicicol resistance. RESULTS: Treatment with radicicol resulted in upregulation of chitin synthase and cell wall integrity genes in Fs and upregulation of nicotinamide adenine dinucleotide dehydrogenase and sugar transporter genes in Cd. Genes encoding an ATP-binding cassette transporter, an aflatoxin efflux pump, ammonium permease 1 (mep1), and nitrilase were differentially expressed in both Fs and Cd. Among these four genes, only the ABC transporter was upregulated in both Fs and Cd. The aflatoxin efflux pump and mep1 were upregulated in Cd, but downregulated in Fs, whereas nitrilase was downregulated in both Fs and Cd. CONCLUSION: The transcriptome analyses suggested radicicol resistance pathways, and deletions of the transporter genes indicated that they contribute to radicicol resistance.

Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum.

Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

Efficacy of Diphenyleneiodonium Chloride (DPIC) Against Diverse Plant Pathogens.

Many of the fungicides and antibiotics currently available against plant pathogens are of limited use due to the emergence of resistant strains. In this study, we examined the effects of diphenyleneiodonium chloride (DPIC), an inhibitor of the superoxide producing enzyme NADPH oxidase, against fungal and bacterial plant pathogens. We found that DPIC inhibits fungal spore germination and bacterial cell proliferation. In addition, we demonstrated the potent antibacterial activity of DPIC using rice heads infected with the bacterial pathogen Burkholderia glumae which causes bacterial panicle blight (BPB). We found that treatment with DPIC reduced BPB when applied during the initial flowering stage of the rice heads. These results suggest that DPIC could serve as a new and useful antimicrobial agent in agriculture.

Cooperative interactions between seed-borne bacterial and air-borne fungal pathogens on rice.

Bacterial-fungal interactions are widely found in distinct environments and contribute to ecosystem processes. Previous studies of these interactions have mostly been performed in soil, and only limited studies of aerial plant tissues have been conducted. Here we show that a seed-borne plant pathogenic bacterium, Burkholderia glumae (Bg), and an air-borne plant pathogenic fungus, Fusarium graminearum (Fg), interact to promote bacterial survival, bacterial and fungal dispersal, and disease progression on rice plants, despite the production of antifungal toxoflavin by Bg. We perform assays of toxoflavin sensitivity, RNA-seq analyses, lipid staining and measures of triacylglyceride content to show that triacylglycerides containing linolenic acid mediate resistance to reactive oxygen species that are generated in response to toxoflavin in Fg. As a result, Bg is able to physically attach to Fg to achieve rapid and expansive dispersal to enhance disease severity.

Identification of Genes Related to Fungicide Resistance in Fusarium fujikuroi.

We identified two genes related to fungicide resistance in Fusarium fujikuroi through random mutagenesis. Targeted gene deletions showed that survival factor 1 deletion resulted in higher sensitivity to fungicides, while deletion of the gene encoding F-box/WD-repeat protein increased resistance, suggesting that the genes affect fungicide resistance in different ways.

[Tissue expansion under the cicatrix].

OBJECTIVE: To investigate a more simple and effective method to repair cicatrix by tissue expansion. METHODS: The dilator with the capacity of 80 - 500 ml was implanted into the subcutaneous pocket under the cicatrix. After dilating for one to two months, the dilator was taken out and the wound surface of the cicatrix was removed. The expanded skin flap was advanced or rotated to cover the defects. The procedure was used on 203 cases. RESULTS: The dilatation was achieved successfully in all the cases, followed by cicatrix removing and repair. The incision scar was not noticeable. CONCLUSIONS: Tissue expansion under the cicatrix has the advantages of safety, less trauma and less extra incisions. It is a reasonable choice to obtain more flexible surgical designs and more economical skin flap applications. It is suitable for most of the treatment for cicatrix.

[Full face dermabrasion for acne scars].

OBJECTIVE: Dermabrasion has been of great value in plastic surgery. Dermabrasion was developed for a specific indication; however, within a very short time, the concept of dermabrasion found wide applicability. This study was to investigate the application of dermabrasion in the treatment of acne scars. METHODS: From Feb. 1996 to May 2004, a total of 110 patients with acne scar were treated with dermabrasion. RESULTS: Postoperatively, the curative results were achieved in 45 cases; good results in 40 cases and effective results in 25 cases. The study revealed that the patients at 18-46 years of age have good results. CONCLUSIONS: Dermabrasion is a good and safe technique to treat the scar of acne.

[Cleft lip nasal deformity corrected by an alar flap and the alar cartilage sling method].

OBJECTIVE: To search an effective method to correct the secondary nasal deformity. METHODS: The "spilth" tissue asymmetry to the another side on the cleft side alar is formed as a flap, which is used to drive up or reconstruct the nostril base (sill), readjust nostril size and shape. The cleft side alar cartilage lateral foot is disassociated, replaced and fixed into the normal place. RESULTS: Nineteen patients were received this operation, their nasal alar, nostril, sill, on the two sides are symmetry, and the result is good. CONCLUSIONS: The cleft side alar flap and alar cartilage sling procedure is effective to correct secondary cleft lip nasal deformity.