Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean

Trichoderma viride was proved as an effective biocontrol agent against two fungal pathogens, Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes, infecting soybean. During an in vitro biocontrol test, Trichoderma showed mycoparasitism and destructive control against the tested fungal pathogens. Both the pathogens significantly influence the germination and P. arrhenomanes had a severe effect (only 5% germination). The root system of the soybean plant was poorly developed due to the infection and it exerted a negative influence on the nodulation and further growth phases of the plant. During pot assay along with biocontrol activity, Trichoderma showed growth promoting action on the soybean plant. Trichoderma enhanced growth of shoot and root systems and fruit yield after 12 weeks of growth. Pythium and Fusarium infected plants treated with Trichoderma had ∼194% and 141% more height than pathogens alone. The fruit yield treated with Trichoderma was ∼66 per plant whereas the yield was only 41 for a control plant. The plants infected with Pythium and Fusarium and treated with Trichoderma had fruit yields of 43 and 53 respectively and those were 5 and 1.6 times higher than plants infected with pathogens.     

Management of fusarium basal rot (Fusarium oxysporum f. sp. cepae) on shallot through fungicidal bulb treatment

Fusarium basal rot caused by Fusarium oxysporum f.sp. cepae is an economic disease of shallot. Field experiments were conducted at Debre Zeit during 2006 and 2007 cropping seasons to determine effective fungicides and their method of application for the management of fusarium basal rot of shallot. The field was naturally infested with F. oxysporum f. sp. cepae and treatments were arranged in randomized complete block design in four replications. Five fungicides, Mirage 50 WP, Folicur 25 EC, Seed plus 30 WS, Penncozeb 80 WP and Ridomil Gold 68 WG were evaluated as seed bulb dressing and bulb dip treatments against basal rot in the field and storage. Bulb dressing with Mirage, and dip treatment in Seed plus reduced the disease incidence by 40% and 43%, respectively over control. These fungicides also resulted in a significant reduction in severity, basal rot affected cull bulbs on shallot. Bulb rot during three months of storage on concrete ground floor and on wire mesh shelves was also reduced by seed bulb treatment over control. The highest increase in yield was obtained from bulb dressing with Mirage (42%) and Seed plus (45%) and from bulb dip treatment in Seed plus (44%) over control. Fusarium basal rot caused 45% loss in yield and 12-30% of bulb loss in the storage. The study showed that basal rot of shallot can be managed effectively by seed bulb dressing or dip treatment in Mirage or Seed plus.     

Effectiveness of plant extracts on suppression of damping-off and wilt diseases of lupine (Lupinus termis Forsik)

The aim of this study was to evaluate water and organic solvent of plant extracts for protection of lupine plants against damping-off and wilt diseases caused by Fusarium oxysporum f. sp. lupini, F. oxysporum f. sp. lupini Snyder & Hansen was isolated from diseased lupine roots collected from different locations of Minia, Assiut and New Valley governorates. Water leaf extracts of Calotropis procera, Nerium oleander, Eugenia jambolana, Citrullus colocynthis, Ambrosia maritime, Acacia nilotica and Ocimum basilicum and fruit extracts of C. colocynthis, C. procera and E. jambolana reduced damping-off and wilt diseases caused by F. oxysporum f. sp. lupini. Water extracts of E. jambolana leaves, C. colocynthis fruits and N. oleander leaves were the most effective ones for controlling such diseases. In contrast, A. nilotica and O. basilicum extracts were the least effective ones. Organic solvent extracts of tested plants reduced damping-off and wilt diseases than water extracts. Butanolic and ethereal extracts were highly effective in reduction of diseases than the other tested extracts. Under field conditions, during winter growing seasons 2008/09 and 2009/10, ethereal and butanolic extracts of N. oleander and E. jambolana leaves and C. colocynthis fruits significantly reduced the percentage of wilt severity as well as improved growth parameters (plant height, number of branches, pods and seeds per plant) and increased seed index, total seed yield/hectare compared with control treatment, while protein content in seeds was not effected. In conclusion, our study demonstrated that some plant extracts can be used for biocontrol of lupine damping-off and wilt diseases.     

Effects of soil amendment with poultry manure on carnation Fusarium wilt in greenhouses in southwest Spain

In a search for alternatives to methyl bromide for controlling carnation vascular wilt caused by Fusarium oxysporum f. sp. dianthi (Fod), poultry manure plus soil solarization was studied in soil under greenhouse conditions in four 2-year experiments. These were conducted in naturally infested soil to compare the effects of this treatment with soil solarization alone and methyl bromide. Soil treatments were performed during June 2000 for Experiment 1, from July to mid-August 2002 for Experiment 2, from late July to late August 2006 for Experiment 3 and from late May to late June 2008 for Experiment 4. Additionally, a treatment with commercial poultry manure pellet plus soil solarization was included in the two latter experiments. Poultry manure caused reductions of Fod viability in soil samples at depths of 15 and 30 cm, ranging respectively from 93 to 100% and 89 to 100% for Experiments 1, 2 and 3. Carnations planted in plots treated either with poultry manure, methyl bromide or soil solarization had lower final disease incidences, smaller areas under their disease progress curves and higher yields in comparison with untreated plots in Experiments 2, 3 and 4. In Experiment 1, soil solarization was performed under suboptimal conditions, and it provided disease levels and yields similar to those of the untreated control plots. Nevertheless, under the same conditions, previous amendment of Fod-infested soil with poultry manure increased disease control over soil solarization alone, improved carnation yield and quality and also increased plant vigor, thus providing a satisfactory alternative to methyl bromide. The application of organic amendment to the same plot before every crop cycle is recommended to ensure continuous disease control, but the rates of application could be reduced to half for the third and fourth crop cycles, thereby reducing undesirable environmental effects.     

Disinfection using ozone microbubbles to inactivate Fusarium oxysporum f. sp. melonis and Pectobacterium carotovorum subsp. carotovorum

To establish a method for disinfecting hydroponic culture solutions using ozone microbubbles (OMB), we examined the disinfectant activity of OMB against Fusarium oxysporum f. sp. melonis and Pectobacterium carotovorum subsp. carotovorum in infected plant roots. OMB had a higher solubility and remained in the water for a longer period than ozone millibubbles, resulting in extremely high disinfecting activity against both phytopathogens. Furthermore, disinfectant activity and durability of OMB-treated water against both phytopathogens increased with an increase in the initial concentration of dissolved ozone. Therefore, these results suggest that OMB may be suitable for use as a new disinfectant against phytopathogens in hydroponic culture solutions.     

香蕉枯萎病菌及其粗毒素对香蕉的致病性比较

从8种培养液中筛选出能诱导香蕉枯萎病菌4号小种(Fusarium oxysporum f.sp.cubense race4)FOCAAA315菌株高产毒素的改良CzapekB培养液,FOCAAA315菌株在该培养液中培养第15d,镰刀菌酸(FA)产量高达669.2mg/L,蔗糖为诱导产毒的最佳碳源。采用伤根浸渍法接种,测定枯萎病菌和病菌发酵粗毒素对香蕉组培苗的致病性。结果表明,粗毒素接种可诱发与病原菌类似的病害症状。受侵染的植物组织在细胞水平上发生一系列形态结构和生理生化代谢的病理变化,包括细胞壁消解、侵填体堵塞气腔、粘胶质减少、淀粉量减少、细胞木质化和黄褐色胶状物堵塞导管等。香蕉枯萎病菌的粗毒素是重要的致病化学物质;利用病菌孢子悬浮液在香蕉苗期接种,可以作为室内快速鉴定香蕉品种抗病性的方法。     

Effects of crop rotation of soybean with corn on severity of sudden death syndrome and population densities of Heterodera glycines in naturally infested soil

Sudden death syndrome, caused by Fusarium virguliforme, and the soybean cyst nematode, Heterodera glycines, combined cause the highest yield losses in soybean. The objective of this study was to determine the effectiveness of corn rotated annually with soybean on reducing severity of sudden death syndrome (SDS) and if such crop rotation is beneficial to soybean root health and thus improves disease management strategies. Experiments were conducted from 2003 to 2006 through two cycles of a corn–soybean rotation on two commercial fields in Indiana. With one exception, the rotation of soybean with corn did not provide yield benefits compared to monoculture of soybean. Severity of foliar and root symptoms of SDS in rotation plots were never less than in soybean monoculture plots. At one location, soybean monoculture resulted in suppression of SDS compared to the corn–soybean rotation, while H. glycines reproduced freely. At the other location, monoculture of soybean resulted in suppressiveness against H. glycines, while SDS was limited in all treatments. The data suggest that soil suppressiveness can independently impact the pathogens that are important in SDS development. Because H. glycines can increase SDS symptoms, its suppression may also reduce severity of SDS. Current production systems consisting of yearly rotation of soybean with corn are highly vulnerable to the development of severe soil-borne disease complexes. The simple year-to-year rotation of corn and soybean is not considered sustainable. While monoculture of soybean resulted in some disease suppression in these trials, reliance on monoculture may be detrimental due to other environmental considerations beyond the scope of these trials. Including other crops may be beneficial in improving the sustainability of soybean and corn production systems.     

Effects of leaf scales of different pineapple cultivars on the epiphytic stage of Fusarium guttiforme

The fungus Fusarium guttiforme (Syn. F. subglutinans f. sp. ananas) is responsible for fusariosis, one of the main phytosanitary threats to pineapple (Ananas comosus var. comosus). A structural study comparing epidermal differences in pineapple cultivars resistant and susceptible to fusariosis was performed, relating properties of the epidermis to known susceptibility to the disease. The basal, non-chlorophylled, portions of mature leaves of pineapple plants were analyzed by light and electron microscopy. All cultivars showed common morpho-anatomic aspects characteristic of Bromeliaceae, such as scutiform scales and unstratified epidermis. However, cultivar Vitoria (resistant) had less scales than cultivars Smooth Cayenne (susceptible, intermediate severity) and Perola (susceptible, with extreme severity of fusariosis symptoms). Inoculation of conidia suspension (10⁵ conidia ml⁻¹) of the fungus F. guttiforme to leaves and harvesting 24 h later yielded numbers of viable colonies related to the density of leaf scales. This suggests that scales can act as havens for fungal conidia and favour the epiphytic stage of the fungus on pineapple plants, and are involved in the interaction of plant and pathogen. A reduction in scale numbers was related to lower infection levels and is relevant to the future breeding programme for development of new pineapple cultivars resistant to fusariosis and their involvement in integrated control strategies.     

Effect of antagonistic Fusarium spp. and of different commercial biofungicide formulations on Fusarium wilt of basil (Ocimum basilicum L.)

The ability of antagonistic Fusarium spp. to control Fusarium wilt of basil caused by F. oxysporum f. sp. basilici was tested under glasshouse conditions from 1994 to 1996. Fusarium oxysporum antagonistic strain 251/2, a protoplast fusion-derived hybrid coded FI-11 and F. moniliforme strains TF4 and TF4 RB were able to decrease (P = 0.05) the incidence of Fusarium wilt significantly. Biocontrol was consistent, especially when the antagonists were applied by mixing a talc preparation of propagules in the substrate (10⁵ CFU/ml of substrate) 1–2 weeks before sowing or at sowing. Seed coating with the biocontrol strains (10⁸ CFU/g of seed) did not generally provide sufficient disease containment, whereas the combination of substrate treatment and seed coating did not improve wilt control in comparison with substrate treatment alone. Two commercial formulations, based on antagonistic F. oxysporum strains and one containing the antagonistic strain K61 of Streptomyces griseoviridis, were generally not sufficiently effective at the tested dosages.     

Application of an emulsifiable mixture of 1,3-dichloropropene and chloropicrin against root knot nematodes and soilborne fungi for greenhouse tomatoes in Italy

A mixture of 1,3-dicloropropene 60.5% w/w and chloropicrin 33.3% w/w (Telone C35 EC) may be registered in Italy for soil drip fumigation. Five experiments on greenhouse tomatoes in Northern, Central and Southern Italy compared the effectiveness of this mixture in comparison with methyl bromide to find the optimum application rate in soils infested by Fusarium oxysporum f.sp. lycopersici, F. oxysporum f.sp. radicis lycopersici, Sclerotium rolfsii, Meloidogyne javanica and M. incognita. Its efficacy against F. oxysporum f.sp. radicis lycopersici and M. incognita was confirmed when applied to soils at 100, 200, 300 and 400 l ha⁻¹ (132.4, 268.4, 402.6 and 536.8 kg ha⁻¹) under gas-tight films with 15–45 mm of application water (900–1200 mg Telone C35 EC l⁻¹). In sandy soils, with slight F. radicis lycopersici infections and with heavy nematode (M. incognita) attacks, the mixture, drip applied at 900 mg l⁻¹ during late summer (fumigation: late summer; transplant: late-summer/autumn; last harvest: early spring), performed well up to 132.4 kg ha⁻¹ (100 l ha⁻¹). In sandy loam soils with slight F. radicis lycopersici infections and severe infections of F. lycopersici and galling nematodes (M. javanica), 268.4 kg ha⁻¹ (200 l ha⁻¹) of the mixture applied at 900 mg l⁻¹ as a drip provided yields similar to those of methyl bromide treated plots both in spring and summer cycles. In sandy loam soils, the diseases (F. lycopersici, F. radicis lycopersici) were controlled at rates 268.4 kg ha⁻¹ (containing 90 kg ha⁻¹ of chloropicrin), but the mixture was ineffective against Sclerotium rolfsii occasionally observed in sandy loam soils. In both sandy and sandy loam soils, no significant relationships were found between the rates of mixture applied (132.4, 268.4, 402.6 and 536.8 kg ha⁻¹) and the degree of nematode infestation.     

Effects of emission reduction surface seal treatments on pest control with shank-injected 1,3-dichloropropene and chloropicrin

The phase-out of methyl bromide for preplant soil fumigation has resulted in an increased reliance on combinations of 1,3-dichloropropene and chloropicrin in many annual and perennial cropping systems in California. However, these fumigants also can have negative environmental and human health consequences and considerable research has been conducted on methods to minimize emission of these products from the soil to the atmosphere. To ensure that pest control efficacy is not compromised by emission reduction techniques, this research was conducted to simultaneously evaluate the effects of several surface seal techniques on fumigant emissions and the efficacy of soil-borne pest control with a mixture of 1,3-dichloropropene + chloropicrin. Results indicated that the interaction between emission reduction techniques and pest control efficacy can be complicated. For example in the 2006 trial, some surviving nematodes were observed in plots with both high (manure plus high density polyethylene film) and intermediate (pre-irrigation) 1,3-D cumulative emissions which suggested that emission loses are not solely responsible for some pest control failures. Weed control tended to be better with plastic film treatments and worse with pre-fumigation soil moisture manipulations but was affected less than expected by intermittent water seals. Although pest control clearly was affected by surface seal techniques, especially in shallow soil layers, some differences in nematode and weed control could not be explained solely by surface seals. These results underline the complex interactions among soil moisture and other environmental factors, application techniques, and emission reducing surface seal treatments. As new techniques and technologies become available to reduce fumigant emissions, we recommend that research include pest control efficacy evaluations before any emerging techniques are required by regulators or implemented by growers.     

Studies onFusarium wilt of potatoes. 2. Leaf,sprout and tuber infection in artificial inoculations

Summary In artificial inoculations it was found thatFusarium oxysporum f.sp.tuberosi is able to penetrate through potato leaves. The fungus was isolated from the stem of infected plants from all inoculated cultivars. Tubers were completely destroyed if they were inoculated before sprouting. Not any destruction was observed if inoculation was made before young sprouts are longer than 1–3 cm; plants from these sprouts were infected. Lesioned tubers were more sensitive than unlesioned ones, and tuber rot and sprout damage were increased significantly.     

拮抗香蕉枯萎病镰刀菌木霉菌株的分离筛选

香蕉枯萎病是由尖孢镰刀菌古巴专化型引起的,目前尚缺乏有效的防治方法,木霉菌是重要的植物病害防治菌,广泛分布于自然界。木霉菌具有广泛的适应性,能杀伤多种重要的植物病原菌且作用机制多种多样。笔者从广东湛江、海南儋州等地152份土样中分离出330株木霉,通过对峙法对木霉菌进行体外拮抗作用测定及评价,选出10株对尖孢镰刀菌有较好拮抗效果的木霉菌;并测定无菌土中木霉与病原菌的种群动态变化,结果表明木霉在土壤中对病原菌有一定的抑制作用。     

玉米茎腐病病原禾谷镰孢拮抗菌筛选及分子鉴定

禾谷镰孢引起的玉米茎腐病会导致玉米产量锐减,筛选高效的拮抗菌株对防治玉米茎腐病尤为重要。采用稀释培养法从土壤中分离得到810株细菌,通过平板对峙法和16S rDNA分子测序方法进行拮抗菌的筛选和分子鉴定,共分离出禾谷镰孢拮抗菌23株,占分离细菌总数的2.84%,抑菌圈直径10.12～27.56 mm。拮抗细菌主要分布在厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)和放线菌门(Actinobacteria)3个门及芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas)、链霉菌属(Streptomyces)、不动杆菌属(Acinetobacter)和节杆菌属(Arthrobacter)5个属中,其中,芽孢杆菌属细菌数量最多且拮抗能力较强,暹罗芽孢杆菌(Bacillus siamensis)的拮抗活性最强,具有较大的生防潜力。实验分离的23株菌株对禾谷镰孢有较好的拮抗活性,在玉米茎腐病防控中具有潜在的应用价值。     

Wilt disease management and enhancement of growth and yield of Cajanus cajan (L) var. Manak by bacterial combinations amended with chemical fertilizer

Root nodulating Sinorhizobium fredii KCC5 and Pseudomonas fluorescens LPK2 were isolated from nodules of Cajanus cajan and disease suppressive soil of tomato rhizosphere, respectively. Both strains produced IAA, siderophore, solubilized insoluble phosphate, showed chitinase and β-1,3-glucanase activities, and strongly inhibited the growth of Fusarium udum. It also caused degradation and digestion of cell wall components, resulting in hyphal perforations, empty cell (halo) formation, shrinking and lysis of fungal mycelia along with significant degeneration of conidia. LPK2 produced volatile cyanogen (HCN). Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers showed a significant increase in seed germination (94%) while seed germination with co-inoculated strains (KCC5 + LPK2), KCC5 and LPK2 alone was 90, 84 and 82% respectively as compared to control 77%. After 120 days of sowing, per plant number of pods, nodules, shoot length, root length, shoot weight and root weight were greater for the combination with half dose of chemical fertilizers compared to the control. Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers resulted in an 82% increase in grain yield per hectare compared to the control. Both strains KCC5 and LPK2 led to proto-cooperation as evidenced by synergism, aggressive colonization of the roots, and enhanced growth, suggesting potential biocontrol efficacy against Fusarium wilt in C. cajan.     

Bioprospecting from Marine Sediments of New Brunswick,Canada: Exploring the Relationship between Total Bacterial Diversity and Actinobacteria Diversity

Actinomycetes are an important resource for the discovery of natural products with therapeutic properties. Bioprospecting for actinomycetes typically proceeds without a priori knowledge of the bacterial diversity present in sampled habitats. In this study, we endeavored to determine if overall bacterial diversity in marine sediments, as determined by 16S rDNA amplicon pyrosequencing, could be correlated with culturable actinomycete diversity, and thus serve as a powerful tool in guiding future bioprospecting efforts. Overall bacterial diversity was investigated in eight marine sediments from four sites in New Brunswick, Canada, resulting in over 44,000 high quality sequences (x = 5610 per sample). Analysis revealed all sites exhibited significant diversity (H’ = 5.4 to 6.7). Furthermore, statistical analysis of species level bacterial communities (D = 0.03) indicated community composition varied according to site and was strongly influenced by sediment physiochemical composition. In contrast, cultured actinomycetes (n = 466, 98.3% Streptomyces) were ubiquitously distributed among all sites and distribution was not influenced by sediment composition, suggesting that the biogeography of culturable actinomycetes does not correlate with overall bacterial diversity in the samples examined. These actinomycetes provide a resource for future secondary metabolite discovery, as exemplified by the antimicrobial activity observed from preliminary investigation.     

香蕉根际促生菌的研究展望

植物根际促生菌(plant growth-promoting rhizobacteria,PGPR)是一类具有促进作物生长并增加产量的作用,兼有抑制植物病原菌、根际有害微生物的根际微生物,作为生物肥料和生物农药的重要资源库,PGPR相关研究受到越来越多的重视。着重从PGPR的概念演变、功能机制、研究手段及应用现状等方面进行综述,并在分析香蕉根际促生菌研究现状的基础上,对香蕉PGPR研究的理论意义和现实意义提出讨论与展望。