Control Of Fungal Plant Diseases Research Articles

The new CFEM protein CgCsa required for Fe3+ homeostasis regulates the growth, development, and pathogenicity of Colletotrichum gloeosporioides

Secreted common fungal extracellular membrane (CFEM) domain proteins have been implicated in multiple biological functions in fungi. However, it is still largely unknown whether the ferric iron (Fe3+), as an important trace element, was involved with the biological function of CFEM proteins. In this study, a new CFEM protein CgCsa, with high expression levels at the early inoculation stage on peppers by Colletotrichum gloeosporioides was investigated. Deletion of the targeted gene CgCsa revealed multiple biological roles in hyphal growth restriction, highly reduced conidial yield, delayed conidial germination, abnormal appressorium with elongated bud tubes, and significantly reduced virulence of C. gloeosporioides. Moreover, in CgCsa mutants, the expression levels of four cell wall synthesis-related genes were downregulated, and cell membrane permeability and electrical conductivity were increased. Compared to the wild-type, the CgCsa mutants downregulated expressions of iron transport-related genes, in addition, its three-dimensional structure was capable binding with iron. Increase in the Fe3+ concentration in the culture medium partially recovered the functions of ΔCgCsa mutant. This is probably the first report to show the association between CgCsa and iron homeostasis in C. gloeosporioides. The results suggest an alternative pathway for controlling plant fungal diseases by deplete their trace elements.

Goldenseal (Hydrastis canadensis L.) Extracts Inhibit the Growth of Fungal Isolates Associated with American Ginseng (Panax quinquefolius L.).

American ginseng, a highly valuable crop in North America, is susceptible to various diseases caused by fungal pathogens, including Alternaria spp., Fusarium spp., and Pestalotiopsis spp. The development of alternative control strategies that use botanicals to control fungal pathogens in American ginseng is desired as it provides multiple benefits. In this study, we isolated and identified three fungal isolates, Alternaria panax, Fusarium sporotrichioides, and Pestalotiopsis nanjingensis, from diseased American ginseng plants. Ethanolic and aqueous extracts from the roots and leaves of goldenseal were prepared, and the major alkaloid constituents were assessed via liquid chromatography-mass spectrometry (LC-MS). Next, the antifungal effects of goldenseal extracts were tested against these three fungal pathogens. Goldenseal root ethanolic extracts exhibited the most potent inhibition against fungal growth, while goldenseal root aqueous extracts and leaf ethanolic extracts showed only moderate inhibition. At 2% (m/v) concentration, goldenseal root ethanolic extracts showed an inhibition rate of 86.0%, 94.9%, and 39.1% against A. panax, F. sporotrichioides, and P. nanjingensis, respectively. The effect of goldenseal root ethanolic extracts on the mycelial morphology of fungal isolates was studied via scanning electron microscopy (SEM). The mycelia of the pathogens treated with the goldenseal root ethanolic extract displayed considerable morphological alterations. This study suggests that goldenseal extracts have the potential to be used as a botanical fungicide to control plant fungal diseases caused by A. panax, F. sporotrichioides, or P. nanjingensis.

A potential antifungal agent: Insight into the antifungal mechanism against Phomopsis sp

19 myricetin derivatives containing thiazole were designed and synthesized. Their fungicidal activities in vitro against ten species of plant pathogenic fungi were evaluated. Bioassay results indicated that some of compounds exhibited remarkable antifungal activities. Among them, Z17 showed the strongest antifungal activity against Phomopsis sp, with in vitro EC50 of 12.3 µg/mL, which was superior to those of the control drug azoxystrobin (32.2 µg/mL) and fluopyram (77.7 µg/mL). In addition, Z18 also had the inhibitory activity against the Alternaria brassicae, with an EC50 value of 32.5 µg/mL, which was much higher than azoxystrobin (49.3 µg/mL). Z17 exhibited good protective activity (62.5 %) against Phomopsis sp on kiwifruit in vivo at 200 µg/mL. The results of in vivo experiments revealed that Z18 could effectively defend against the infestation of cabbage by Alternaria brassicae, and improve the protection ability of the crop. Scanning electron microscopy (SEM) and fluorescence microscopy (FM) demonstrated that Z17 could destroy the integrity of cell membrane of pathogen Phomopsis sp, thus affecting the normal growth of mycelia. The results of the mechanism research further confirmed that the action of Z17 changed the mycelial morphology of Phomopsis sp, affected the permeability of cells, increased the leakage of cytoplasm and MDA contents. Molecular docking simulation results revealed that compound Z17 could readily bind with the active site of SDH and could be a potential SDH inhibitor. In summary, this study provides new ideas for effectively controlling plant fungal diseases and developing new green chemical pesticide products.

Design and Synthesis of Antifungal Candidates Containing Triazole Scaffold from Natural Rosin against Valsa mali for Crop Protection.

Two series of dehydroabietyl-1,2,4-triazole-4-Schiff-based derivatives were synthesized from rosin to control plant fungal diseases. In vitro evaluation and screening of the antifungal activity were performed using Valsa mali, Colletotrichum orbiculare, Fusarium graminearum, Sclerotinia sclerotiorum, and Gaeumannomyces graminis. Compound 3f showed excellent fungicidal activity against V. mali (EC50 = 0.537 μg/mL), which was significantly more effective than the positive control fluconazole (EC50 = 4.707 μg/mL). Compound 3f also had a considerable protective effect against V. mali (61.57%-92.16%), which was slightly lower than that of fluconazole (85.17-100%) at 25-100 μg/mL. Through physiological and biochemical analyses, the preliminary mode of action of compound 3f against V. mali was explored. Ultrastructural observation of mycelia showed that compound 3f hindered the growth of the mycelium and destroyed the ultrastructure of V. mali seriously. Conductivity analysis and laser scanning confocal microscope staining showed that compound 3f changed cell-membrane permeability and caused accumulation of reactive oxygen species. The enzyme activity results showed that compound 3f significantly inhibited the activity of CYP51 (59.70%), SOD (76.9%), and CAT (67.86%). Molecular docking identified strong interaction energy between compound 3f and crystal structures of CYP51 (-11.18 kcal/mol), SOD (-9.25 kcal/mol), and CAT (-8.79 kcal/mol). These results provide guidance for the discovery of natural product-based antifungal pesticide candidates.

The use of selenium for controlling plant fungal diseases and insect pests.

The selenium (Se) applications in biomedicine, agriculture, and environmental health have become great research interest in recent decades. As an essential nutrient for humans and animals, beneficial effects of Se on human health have been well documented. Although Se is not an essential element for plants, it does play important roles in improving plants' resistances to a broad of biotic and abiotic stresses. This review is focused on recent findings from studies on effects and mechanisms of Se on plant fungal diseases and insect pests. Se affects the plant resistance to fungal diseases by preventing the invasion of fungal pathogen through positively affecting plant defense to pathogens; and through negative effects on pathogen by destroying the cell membrane and cellular extensions of pathogen inside plant tissues after invasion; and changing the soil microbial community to safeguard plant cells against invading fungi. Plants, grown under Se enriched soils or treated with Se through foliar and soil applications, can metabolize Se into dimethyl selenide or dimethyl diselenide, which acts as an insect repellent compound to deter foraging and landing pests, thus providing plant mediated resistance to insect pests; moreover, Se can also lead to poisoning to some pests if toxic amounts of Se are fed, resulting in steady pest mortality, lower reproduction rate, negative effects on growth and development, thus shortening the life span of many insect pests. In present manuscript, reports are reviewed on Se-mediated plant resistance to fungal pathogens and insect pests. The future perspective of Se is also discussed on preventing the disease and pest control to protect plants from economic injuries and damages.

Potential of Alpha-(α)-Solanine as a Natural Inhibitor of Fungus Causing Leaf Spot Disease in Strawberry

Curvularia trifolii is an important pathogenic fungus that causes leaf spot disease in strawberry and other crops. Increased resistance in pathogenic fungi against chemical fungicides necessitates the search for biological alternatives to control plant fungal diseases. The present study aimed to perform transcriptome and metabolome analysis of C. trifolii fungi. We evaluated the potential of an alkaloid, namely alpha (α)-solanine, to inhibit the growth of Curvularia under in vitro conditions. Furthermore, transcriptomic and metabolomic analysis of treated C. trifolii was performed to identify the differential genes and metabolites. Results revealed that treatment with α-solanine resulted in the poor growth and development of fungal spores. The transcriptome analysis revealed that 1413 genes were differentially expressed (DEGs), among which 340 unigenes were up-regulated, 100 unigenes were down-regulated, and the rest were unaffected in treated samples. Gene ontology analysis revealed that the majority of the genes were related to oxidative stress in the fungus. Additionally, using ultra-high performance liquid chromatography-tandem mass spectrometry, we identified 455 metabolites, among which the majority of metabolites were related to lipid biosynthesis. The high number of genes related to lipid biosynthesis and reactive oxygen species revealed that α-solanine causes oxidative stress in Curvularia, leading to growth inhibition, and can be potentially used as an alternative to chemical fungicides.

BioClay™ prolongs RNA interference-mediated crop protection against Botrytis cinerea.

One of the most promising tools for the control of fungal plant diseases is spray-induced gene silencing (SIGS). In SIGS, small interfering RNA (siRNA) or double-stranded RNA (dsRNA) targeting essential or virulence-related pathogen genes are exogenously applied to plants and postharvest products to trigger RNA interference (RNAi) of the targeted genes, inhibiting fungal growth and disease. However, SIGS is limited by the unstable nature of RNA under environmental conditions. The use of layered double hydroxide or clay particles as carriers to deliver biologically active dsRNA, a formulation termed BioClay™, can enhance RNA durability on plants, prolonging its activity against pathogens. Here, we demonstrate that dsRNA delivered as BioClay can prolong protection against Botrytis cinerea, a major plant fungal pathogen, on tomato leaves and fruit and on mature chickpea plants. BioClay increased the protection window from 1 to 3 weeks on tomato leaves and from 5 to 10 days on tomato fruits, when compared with naked dsRNA. In flowering chickpea plants, BioClay provided prolonged protection for up to 4 weeks, covering the critical period of poding, whereas naked dsRNA provided limited protection. This research represents a major step forward for the adoption of SIGS as an eco-friendly alternative to traditional fungicides.

Antifungal Activity of Hyptis spicigera Methanol Leaf Extract and Flavonoid Fraction


 
 
 Control of plant fungal diseases using synthetic fungicides continue to cause major problems to human health and the entire ecosystem. The aim of this research was to investigate the phytochemical and antifungal properties of Hyptis spicigera methanol leaf extract and flavonoid fraction on Aspergillus and Fusarium species, with a view to uncovering effective bio-fungicides for development as substitutes to chemical fungicides. Phytochemical screening revealed the presence of sterols and triterpenes, cardiac glycosides, flavonoids, tannins and alkaloids. The quantitative analysis showed that saponin (690 mg/g GAE), phenolics (220 mg/g GAE) and flavonoids (140 mg/g GAE) were found to be in high concentration. The antifungal effects of H. spicigera methanol extract on F. graminearum (21 mm) was significantly (p<0.05) higher than the control fungicide (Mancozeb). Similarly, the flavonoid fraction was more effective on A. parasitic (18 mm) than the control fungicide used. The extract and fraction exhibited MIC with range 3.13-12.5 mg/mL and MFC 6.25-12.5 mg/mL, indicating promising antifungal efficacies against A. flavus and F. graminearum. Our findings have revealed that H. spicigera flavonoid rich fraction has potential for development as effective bio-fungicide to control plant fungal diseases of the Aspergillus and Fusarium species.
 
 

Isolation and identification of Amycolatopsis sp. strain 1119 with potential to improve cucumber fruit yield and induce plant defense responses in commercial greenhouse

The application of chemical fungicides is the first strategy to control plant fungal diseases. This approach is highly polluting for the environment and affects human health. The artificial introduction of beneficial rhizobacteria into the soil can be an economical and practical way to control phytopathogenic fungi in commercial greenhouses. Here, we recount the travel of a rare Actinomycete (Amycolatopsis strain 1119) from a maize field to a commercial cucumber greenhouse. Culturable bacteria from rhizosphere and bulk soils of dicot and monocot crops were isolated and screened. About 20% of the representative colonies showed Actinobacteria appearance. 106 Actinobacteria that had antagonistic activity against Phytophthora capsici and were able to produce IAA-like molecules were selected for further analysis. Two Streptomyces strains (432 and 615) and 2 Amycolatopsis strains (3513 and 1119) that showed a positive effect on plant growth in greenhouse conditions were selected to evaluate the biocontrol potential. Strains 432, 3513, 615 and 1119 controlled incidence of the damping-off by 65%, 42%, 83% and 100% respectively. Application of strain 1119 under commercial greenhouse conditions resulted in an increase in fruit yield (20%) and a decrease in fruit nitrate content (70%). Increased antioxidant enzymes activity and increased LOX and APX transcription and also, increased expression of two genes PR1-1a and GLU (SAR genes) showed that strain 1119 could induce both ISR and SAR in cucumber without pathogen exposure. Our results demonstrate that the Amycolatopsis strain 1119 has a great potential to be used as an active principle for bio-inoculant development because of the ability to improve cucumber fruit yield and induce plant defense responses in a commercial greenhouse.

Characterization and Antifungal Activity of Limonoid Constituents Isolated from Meliaceae Plants Melia dubia, Aphanamixis polystachya, and Swietenia macrophylla against Plant Pathogenic Fungi In Vitro

The plants of Meliaceae are native to tropical and subtropical regions as the Americas, west India, Southeast Asia, and Southern China. Many species of the genera Khaya, Swietenia, Aphanamixis, and Melia in this family are known as medicinal plants and have biological activities such as antiviral, antimicrobial, antifeeding, insecticidal, and cytotoxic properties. The objectives of this research are to characterize and evaluate the bioactive limonoids from several plants of Meliaceae against phytopathogenic fungi. During the search of antifungal compounds from the plants of Meliaceae, the three methanol extracts of Melia dubia, Aphanamixis polystachya, and Swietenia macrophylla were found to suppress the mycelial growth of several phytopathogenic fungi. Nine limonoids isolated from M. dubia (1–2), A. polystachya (3–5), and S. macrophylla (6–9) were evaluated, for the first time, their antifungal effectiveness against nine phytopathogenic fungi Fusarium oxysporum, Magnaporthe oryzae, Sclerotium rolfsii, Rhizoctonia solani, Alternaria spp., and Botrytis cinerea, and three oomycetes Phytophthora species. Limonoids 2, 3, 6, and 8 displayed a remarkable broad-spectrum antifungal activity against all the test fungi. Sclerotium rolfsii was highly sensitive to the four limonoids with IC50 values ranging from 79.4 to 128.0 µg/mL. Notably, chisocheton compound G (3) isolated from A. polystachya and khayanolide B (8) isolated from S. macrophylla were the most potent antifungal limonoids and exhibited a dose-dependent activity against Phytophthora species. Compounds 2 and 9 displayed moderate activity against M. oryzae. Our study results demonstrated the discovery of antifungal and lead compounds from the group of limonoids for application in the control of fungal plant diseases.

Enhanced spoVF operon increases host attachment and biocontrol ability of Bacillus subtilis for the management of Ceratocystis fimbriata in sweet potato

Bacillus subtilis is a ubiquitous environmental bacterium commonly used for the biological control of fungal plant diseases. The ability to form surface-attached microbial communities is essential for B. subtilis to colonize and protect plants from pathogen infection. In this work, it was found that enhanced spoVF operon altered the carbon metabolic pathways of B. subtilis 168, accelerating cell growth and glucose consumption, and inducing the production of antifungal metabolite dipicolinic acid (DPA). These metabolic effects significantly increased swimming motility and biofilm formation, enhancing the colonization ability and host attachment of B. subtilis. Mutant B. subtilis showed higher inhibitory activity in vivo against sweet potato pathogen Ceratocystis fimbriata in comparison with the wild-type strain. Collectively, novel insights on the key genes and pathways involved in biofilm formation and attachment of B. subtilis were revealed in this work.

Quantitative proteomics reveals the antifungal effect of canthin-6-one isolated from Ailanthus altissima against Fusarium oxysporum f. sp. cucumerinum in vitro.

Canthin-6-one, one of the main alkaloid compounds extracted from Ailanthus altissima, has recently attracted increasing interest for its antifungal activity. To evaluate the potential of canthin-6-one in controlling plant fungal diseases, we investigated the antifungal activity of canthin-6-one isolated from A. altissima against Fusarium oxysporum f. sp. cucumerinum (Foc) in vitro. The mycelial growth rate and micro-broth dilution were used to test antifungal activity. Furthermore, label-free quantitative proteomics and parallel reaction monitoring (PRM) techniques were applied to analyze the antifungal mechanism. It was found that canthin-6-one significantly inhibited the growth of Foc, and had higher inhibitory action than chlorothalonil at the same concentration. Proteomic analysis showed that the expression of 203 proteins altered significantly after canthin-6-one treatment. These differentially expressed proteins were mainly involved in amino acid biosynthesis and nitrogen metabolism pathways. These results suggest that canthin-6-one significantly interferes with the metabolism of amino acids. Therefore, it affects nitrogen nutrients and disturbs the normal physiological processes of fungi, and ultimately leads to the death of pathogens. This study provides a natural plant antifungal agent and a new perspective for the study of antifungal mechanisms.

The Development of a Phytopathogenic Fungi Control Trial: Aspergillus flavus and Aspergillus niger Infection in Jojoba Tissue Culture as a Model

After introducing the idea of using concentrations equal to or less than the minimum inhibition concentration (MIC) of some active chemical compounds for evacuating microbial cells, different types of microbes were evacuated. The original protocol was given the name sponge-like protocol and then was reduced and modified from a microorganism to another to prepare microbial ghosts for various applications such as immunological applications, drug delivery, and isolation of DNA and protein. Fungal pathogens that infect plants critically affect cost effectiveness, quality, and quantity of their production. They kill plant cells and/or cause plant stress. Plant fungal infections can originate from many sources such as infected soil, seeds, or crop debris causing diseases and quality losses around the world with billions of US dollars annually as costs of the associated productivity loss. This study focused on the application of the sponge-like protocol in protecting in vitro tissue cultures of plants against fungal pathogens. This can be useful for research purposes or may be developed to be introduced in field applications. Aspergillus flavus and Aspergillus niger infection in tissue culture of jojoba (Simmondsia chinensis (Link) Schn.) was used as a model to establish the employment of this protocol to control plant fungal diseases. The best conditions for A. flavus and A. niger ghosts production previously mapped by randomization experimental design (reduced Plackett–Burman experimental design) were used to prepare fungal ghosts. SDS, NaOH, NaHCO3, and H2O2 were used in their MIC (+1 level) or minimum growth concentration (MGC, −1 level) according to the determined optimal experimental design. The release of both of DNA and protein from the fungal cells was evaluated spectrophotometrically at 260nm and 280nm, respectively, as an indicator for cell loss of their cytoplasm. Fungal ghost cells were also examined by transmission electron microscopy. After confirming the preparation of high-quality fungal ghost cells, the same conditions were mimicked to control plant fungal infection. Jojoba grown in tissue culture was sprayed with fungal cells (about 103 CFU) as a control experiment or fungal cells followed by treatment with solution (a) represents the fungal ghost cells formation calculated critical concentration (FGCCC) of SDS, NaOH, and NaHCO3 and then treatment with solution (b) represents H2O2 FGCCC. The plant was examined on day 0 (plant grown before any infection or infection followed by treatment), day 5 (plant at day 5 after infection or infection followed by treatment), and day 10 (plant at day 10 after infection or infection followed by treatment). We observed fungal growth in case of control experiments at days 5 and 10 on the tissue culture medium, as well as plant, and the absence of any fungal growth in case of plant treated with FGCCC even after day 10. We recommend using this FGCCC in the form of chemical spraying formulation to treat the plants aiming to control different plant fungal infections in in vitro tissue culture systems or applied in field.

Comparative genomics highlights the importance of drug efflux transporters during evolution of mycoparasitism in Clonostachys subgenus Bionectria (Fungi, Ascomycota, Hypocreales).

Various strains of the mycoparasitic fungal species Clonostachys rosea are used commercially as biological control agents for the control of fungal plant diseases in agricultural crop production. Further improvements of the use and efficacy of C. rosea in biocontrol require a mechanistic understanding of the factors that determines the outcome of the interaction between C. rosea and plant pathogenic fungi. Here, we determined the genome sequences of 11 Clonostachys strains, representing five species in Clonostachys subgenus Bionectria, and performed a comparative genomic analysis with the aim to identify gene families evolving under selection for gene gains or losses. Several gene families predicted to encode proteins involved in biosynthesis of secondary metabolites, including polyketide synthases, nonribosomal peptide syntethases and cytochrome P450s, evolved under selection for gene gains (p ≤ .05) in the Bionectria subgenus lineage. This was accompanied with gene copy number increases (p ≤ .05) in ATP‐binding cassette (ABC) transporters and major facilitator superfamily (MFS) transporters predicted to contribute to drug efflux. Most Clonostachys species were also characterized by high numbers of auxiliary activity (AA) family 9 lytic polysaccharide monooxygenases, AA3 glucose–methanol–choline oxidoreductases and additional carbohydrate‐active enzyme gene families with putative activity (or binding) towards xylan and rhamnose/pectin substrates. Particular features of the C. rosea genome included expansions (p ≤ .05) of the ABC‐B4 multidrug resistance transporters, the ABC‐C5 multidrug resistance‐related transporters and the 2.A.1.3 drug:H + antiporter‐2 MFS drug resistance transporters. The ABC‐G1 pleiotropic drug resistance transporter gene abcG6 in C. rosea was induced (p ≤ .009) by exposure to the antifungal Fusarium mycotoxin zearalenone (1121‐fold) and various fungicides. Deletion of abcG6 resulted in mutants with reduced (p < .001) growth rates on media containing the fungicides boscalid, fenhexamid and iprodione. Our results emphasize the role of biosynthesis of, and protection against, secondary metabolites in Clonostachys subgenus Bionectria.

Cell Wall Biogenesis Protein Phosphatase CrSsd1 Is Required for Conidiation, Cell Wall Integrity, and Mycoparasitism in Clonostachys rosea.

Cell wall biogenesis protein phosphatases play important roles in various cellular processes in fungi. However, their functions in the widely distributed mycoparasitic fungus Clonostachys rosea remain unclear, as do their potential for controlling plant fungal diseases. Herein, the function of cell wall biogenesis protein phosphatase CrSsd1 in C. rosea 67-1 was investigated using gene disruption and complementation approaches. The gene-deficient mutant ΔCrSsd1 exhibited much lower conidiation, hyphal growth, mycoparasitic ability, and biocontrol efficacy than the wild-type (WT) strain, and it was more sensitive to sorbitol and Congo red. The results indicate that CrSsd1 is involved in fungal conidiation, osmotic stress adaptation, cell wall integrity, and mycoparasitism in C. rosea.

Trichoderma/pathogen/plant interaction in pre-harvest food security

Large losses before crop harvesting are caused by plant pathogens, such as viruses, bacteria, oomycetes, fungi, and nematodes. Among these, fungi are the major cause of losses in agriculture worldwide. Plant pathogens are still controlled through application of agrochemicals, causing human disease and impacting environmental and food security. Biological control provides a safe alternative for the control of fungal plant pathogens, because of the ability of biocontrol agents to establish in the ecosystem. Some Trichoderma spp. are considered potential agents in the control of fungal plant diseases. They can interact directly with roots, increasing plant growth, resistance to diseases, and tolerance to abiotic stress. Furthermore, Trichoderma can directly kill fungal plant pathogens by antibiosis, as well as via mycoparasitism strategies. In this review, we will discuss the interactions between Trichoderma/fungal pathogens/plants during the pre-harvest of crops. In addition, we will highlight how these interactions can influence crop production and food security. Finally, we will describe the future of crop production using antimicrobial peptides, plants carrying pathogen-derived resistance, and plantibodies.

Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09.

Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome.

Potential biocontrol actinobacteria: Rhizospheric isolates from the Argentine Pampas lowlands legumes.

Control of fungal plant diseases by using naturally occurring non-pathogenic microorganisms represents a promising approach to biocontrol agents. This study reports the isolation, characterization, and fungal antagonistic activity of actinobacteria from forage soils in the Flooding Pampa, Argentina. A total of 32 saprophytic strains of actinobacteria were obtained by different isolation methods from rhizospheric soil of Lotus tenuis growing in the Salado River Basin. Based on physiological traits, eight isolates were selected for their biocontrol-related activities such as production of lytic extracellular enzymes, siderophores, hydrogen cyanide (HCN), and antagonistic activity against Cercospora sojina, Macrophomia phaseolina, Phomopsis sp., Fusarium oxysporum, and Fusarium verticilloides. These actinobacteria strains were characterized morphologically, physiologically, and identified by using molecular techniques. The characterization of biocontrol-related activities in vitro showed positive results for exoprotease, phospholipase, fungal growth inhibition, and siderophore production. However, none of the strains was positive for the production of hydrogen cyanide (HCN). Streptomyces sp. MM140 presented the highest index for biocontrol, and appear to be promising pathogenic fungi biocontrol agents. These results show the potential capacity of actinobacteria isolated from forage soils in the Argentine Pampas lowlands as promising biocontrol agents, and their future agronomic applications.

English

The present study was initiated to (i) determine burned forest-inhabiting fungi in Zouagha, TerriBeinene, Mila and (ii) study the antagonistic activities of Trichoderma sp against Fusarium sp, Penicillium sp, Rhizoctonia sp, and Alternaria sp. Eighteen fungal strains representing six genera were isolated from soil samples obtained from the burned forest of Zouagha in the Mila region: Trichoderma sp, Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp, and Rhizopus sp. The direct antagonistic activity assays of Trichoderma sp on Potato Dextrose Agar medium (PDA) against the four fungi: Fusarium sp, Penicillium sp, Rhizoctonia sp, and Alternaria sp revealed that the fungus Trichoderma sp reduced the mycelium growth of Fusarium sp, Penicillium sp, Rhizoctonia sp and Alternaria sp to 23.13, 33.13, 33.75, and 38.31%, respectively, compared to the control after six days at room temperature. The results illustrated an inhibitory action of the antagonist Trichoderma sp characterized by slowing the mycelial growth of fungal strains. Strains of Fusarium sp, Penicillium sp, Rhizoctonia sp and Alternaria sp showed differences in the sensitivity to the antagonist. Because Trichoderma occurred more frequently in burned soils and were more antagonistic to phytopathogenic fungi in culture than isolates from unburned soils, the judicious use of fire may increase the abundance of Trichoderma isolates and their inhibitory action may be used for the control of fungal plant diseases. Key words: Fungi, burned soil, Zouagha, Antagonism, Trichoderma sp.

Control of human and plant fungal pathogens using pentaene macrolide 32, 33-didehydroroflamycoin.

The aim of this study was to address the toxicity of recently described polyene macrolide 32, 33-didehydroroflamycoin (DDHR) on a wide range of fungal pathogens and its potential to control plant fungal diseases. The antifungal activity of DDHR in vitro was examined against common human and plant pathogenic fungi using a broth microdilution assay and a disk diffusion assay. Minimum inhibitory concentrations ranged from 12·5 to 35 μg ml(-1) . A radial growth inhibition assay showed that DDHR inhibited mycelia growth, inducing mycelial necrosis and affecting sporulation. During the in vivo assay on apple fruits administration of DDHR 1 h before fungal inoculation inhibited spreading of the infection. Importantly, DDHR exhibited no phytotoxic effects on the model plant, Capsicum annum, verified by the plant growth rate and chlorophyll content. DDHR inhibits growth of various plant pathogens in vitro with the strongest activity against Alternaria alternata, Colletotrichum acutatum and Penicillium expansum, and protects apple fruits from decay. This is the first report of the inhibitory effect of DDHR on important pathogenic fungal isolates. DDHR could be a good scaffold for developing new antifungal agents for fruit and vegetable protection.