A novel strain of Fusarium oxysporum from Germany and its potential for biocontrol of Orobanche ramosa

A newly isolated Fusarium oxysporum strain was investigated for its biocontrol potential against the root parasitic weed Orobanche ramosa (branched broomrape). The fungus was found to affect all developmental stages of the parasite. Orobanche seed germination was reduced by 40% in the presence of fungal conidia in vitro . The number of underground tubercles and shoots of the weed was reduced by the fungus by 55% compared with the control and 92% of tubercles were recorded as diseased in root chambers. In pot experiments, soil application of a granular formulation of the fungus resulted in a reduction of number and dry matter of Orobanche shoots by more than 90%. Spraying of a conidial suspension on aboveground Orobanche shoots caused the death of 75% of them within 2 weeks. Data from initial host-range experiments indicate that the isolate is very host-specific, not even attacking shoots of other Orobanche species. Because of these promising results, we conclude that the fungal isolate should be investigated under field conditions and be compared with other Fusarium isolates proposed for biocontrol of O. ramosa .     

Characteristics of alginate pellets formulated with Trichoderma and Gliocladium and their effect on the proliferation of the fungi in soil

Pelletized formulations of wheat bran or kaolin clay in an alginate gel containing conidia, chlamydospores, or fermentor biomass (FB) of several isolates of the biocontrol fungi Trichoderma spp. and Gliocladium virens were prepared. The ability of fungal propagules within the pellets to proliferate in soil was determined. Higher population densities were obtained when alginate pellets added to soil contained chlamydospores rather than condia, and bran rather than kaolin as the bulking agent. The active ingredient in pellets prepared from FB was approximately 5% biomass by weight and contained many chlamydospores. Colony-forming units (cfu) ranged from 10⁶'to 10¹⁰/g of soil after soil amendment with FB pellets of 12 Trichoderma and G. virens isolates. Population densities were high during the first 3 weeks of incubation and declined only gradually during 9 weeks. Propagules in FB pellets were more viable at 5° than at 25°C. Viability at 25°C remained high (> 70%) after 1 week, but declined to less than 10% after 24 weeks. Despite reduction in propagule viability in stored pellets, numbers of cfu formed after adding these pellets to soil were comparable with those formed from freshly prepared pellets.     

Effect of Relative Humidity on Sporulation of Fusarium oxysporum in Various Formulations and Effect of Water on Spore Movement Through Soil

ABSTRACT Fusarium oxysporum f. sp. erythroxyli is being investigated as a mycoherbicide for the narcotic plant coca. Sporulation of the fungus in seven formulations containing different organic substrates and movement of its propagules through soil were studied. The formulations were a granular wheat flour/kaolin (pesta); an extruded wheat and rice flour (C-6); and five alginate pellet products containing corn cobs, soybean hull fiber, canola meal, rice flour, or rice flour plus canola oil. Formulations were incubated at 25 degrees C for 6 weeks in desiccators with various salt solutions to provide nine relative humidities (RH), ranging from 100% (pure deionized water) to 0% (anhydrous (CaSO(4)). Hyphae of F. oxysporum f. sp. erythroxyli grew out of alginate pellets with canola meal, rice, and rice plus canola oil as early as 24 h at 100% constant RH. Alginate pellets of rice plus canola oil and granular C-6 and pesta formulations consistently produced more microconidia, macroconidia, and CFU than the other four formulations at all RH tested. The C-6 formulation produced more propagules than the other formulations at low RH (<53%). Canola meal pellets produced more spores than three other formulations when exposed to fluctuating RH (100 to 75%). The effect of percolating water on spore movement through soil was studied using a plant-pathogenic isolate of F. oxysporum f. sp. niveum. To determine the effect of water percolation on propagule movement, formulations were placed on soil columns and artificial rain was applied. In general, 10-fold fewer CFU were recovered at a 8- to 10-cm depth compared with a 0- to 2-cm depth.     

Induction of systemic resistance in banana (<Emphasis Type="Italic">Musa</Emphasis> spp.) against <Emphasis Type="Italic">Banana bunchy top virus</Emphasis> (BBTV) by combining chitin with root-colonizing <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> strain CHA0

Pseudomonas fluorescens strains CHA0 and Pf1 were investigated for their biocontrol efficacy against Banana bunchy top virus (BBTV) in banana (Musa spp.) alone and in combination with chitin under glasshouse and field conditions. Bioformulation of P. fluorescens strain CHA0 with chitin was effective in reducing the banana bunchy top disease (BBTD) incidence in banana under glasshouse and field conditions. In addition to disease control, the bioformulation increased the economic yield significantly compared to the untreated control. Increased accumulation of oxidative enzymes, peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), pathogenesis-related (PR) proteins, chitinase, β-1,3-glucanase and phenolics were observed in CHA0 bioformulation amended with chitin-treated plants challenged with BBTV under glasshouse conditions. Indirect ELISA indicated the reduction in viral antigen concentration in P. fluorescens strain CHA0 with chitin-treated banana plants corresponding to reduced disease ratings. The present study revealed that induction of defence enzymes by P. fluorescens with chitin amendment reduced the BBTD incidence and increased bunch yield in banana.     

Pathogenicity of branched broomrape (Orobanche ramosa) populations on tobacco cultivars

Parasitic weed species of the genus Orobanche are a serious threat for the production of several crops in Europe, Africa and Asia. In contrast to other broomrape species of agronomic importance, O. ramosa (branched broomrape) has a broad host range and in Europe particularly affects hemp, tobacco, tomato and, in recent times, oilseed rape. Two separate sets of experiments investigated the effect of two populations of O. ramosa on nine tobacco cultivars grown in Europe and belonging to the three major tobacco types: Virgin (flue-cured), Burley (light air-cured) and dark air-cured under standardized glasshouse conditions. The two broomrape populations were discriminated by means of polymorphic DNA fragments obtained by PCR of the intersimple sequence repeat regions (ISSRs). The Orobanche populations exhibited different levels of pathogenicity but all various tobacco cultivars were susceptible. Dark air-cured tobacco cultivars were the least susceptible to both broomrape populations. Virgin and Burley tobacco cultivars were more susceptible to one population of O. ramosa .     

Glasshouse and field evaluation of benomyl and triadimefon applied at seeding to control take-all in wheat

The effects of two fungicides, benomyl and triadimefon, applied at seeding, on the incidence and severity of take-all ( Gaeumannomyces graminis var. tritici ) of wheat were investigated in the glasshouse and in the field. In glasshouse studies with an artificially inoculated and naturally infested sandy loam, both fungicides caused significantly less disease, whereas in a field study with a clay soil and artificial inoculum, triadimefon caused significantly less disease. In the field, both fungicides were more effective when placed near the seed as granules or as pellets than when applied as seed treatments.     

Influence of a Fungus-Feeding Nematode on Growth and Biocontrol Efficacy of Trichoderma harzianum

ABSTRACT A fungivorous nematode, Aphelenchoides sp., was isolated from field soil by baiting with mycelium of the biocontrol fungus Trichoderma harzianum ThzID1, and subsequently was maintained on agar cultures of the fungus. Interactions between the nematode and the green fluorescent protein-producing transformant, T. harzianum ThzID1-M3, were investigated in both heat-treated (80 degrees C, 30 min) and untreated field soil. ThzID1-M3 was identified in soil by epifluorescence microscopy. When ThzID1-M3 was added to soil as an alginate pellet formulation, addition of the nematode (10 per gram of soil) significantly reduced radial growth and recoverable populations of the fungus, and the effect was greater in heat-treated soil than in untreated soil. Addition of ThzID1-M3 to soil pretreated with the nematode (10 per gram of soil) stimulated nematode population growth for approximately 10 to 20 days, whereas nematode populations decreased in the absence of added Trichoderma sp. When sclerotia of Sclerotinia sclerotiorum were added to soil (10 per 200 g of soil) with ThzID1-M3 (40 pellets per 200 g of soil), addition of Aphe-lenchoides sp. (2,000 per 200 g of soil) reduced the number of sclerotia colonized by ThzID1-M3. These results suggest that fungivorous nematodes may be a significant biotic constraint on activity of biocontrol fungi in the field.     

烟草赤星病菌拮抗菌解淀粉芽胞杆菌YW-2-6鉴定及生防效果

为获得有效拮抗烟草赤星病菌的生防资源,采用稀释分离法和平板对峙法从烟草根际土壤中分离筛选烟草赤星病菌拮抗菌株。通过形态学观察、生理生化特征和16S rDNA序列分析进行鉴定,并测定其对烟株的促生作用,验证菌株的产IAA能力,评价生防菌发酵液对烟草赤星病的田间防效。结果表明,从分离的65株细菌菌株中筛选到5株烟草赤星病拮抗细菌,其中菌株YW-2-6对烟草赤星病菌拮抗效果最好,抑制率为71.32%,初步鉴定菌株YW-2-6为解淀粉芽胞杆菌Bacillus amyloliquefaciens。菌株YW-2-6对小麦赤霉病菌、辣椒疫霉病菌、烟草靶斑病菌等病原菌具有显著的拮抗作用。YW-2-6发酵液对烟草赤星病防效为56.93%,且菌株YW-2-6对烟株有明显的促生作用。菌株YW-2-6具有良好的生防潜力和开发前景。     

Evaluation of potential biocontrol agents against Claviceps africana in vitro and in vivo

Undiluted culture filtrates of two commercial products of Trichoderma spp., Trichopel and Trichoflow, and two isolates of Penicillium citrinum completely inhibited the conidial germination of macroconidia of Claviceps africana , the cause of ergot or sugary disease of sorghum ( Sorghum bicolor ) in vitro . Similarly, Pseudomonas aeruginosa and Burkholderia cepacia completely inhibited macroconidial germination, with the former being more effective at high dilutions. In contrast, these bacterial isolates failed to inhibit infection in vivo in glasshouse tests with ergot-inoculated sorghum, but all fungal biocontrol agents (including an isolate of Epicoccum nigrum ) reduced the severity of disease (percentage of infected spikelets per panicle), in some cases completely inhibiting the development of ergot. In a second glasshouse trial, optimum control was achieved when the biocontrol agents were applied 3–7 days before inoculation with conidia of C. africana .     

Controlled release of isoproturon from an alginate–bentonite formulation: water release kinetics and soil mobility

The herbicide isoproturon [3‐(4‐isopropylphenyl)‐1,1‐dimethylurea] was incorporated in alginate‐based granules to obtain controlled‐release (CR) properties. The basic formulation (sodium alginate (1.87%)–isoproturon (0.67%) in water) was modified by addition of different sorbents. The effect on isoproturon release rate, modified by the incorporation of natural and acid‐treated bentonite in alginate formulation, was studied by immersion of the granules in water while shaking. The release of isoproturon was diffusion‐controlled. The time taken for 50% of the active ingredient to be released into water, T₅₀, was longer for those formulations containing added bentonite (5.98 and 7.43 days, for natural and acid‐treated (1 M H₂SO₄) bentonite, respectively) than for the preparation without bentonite (3.78 days). The mobilities of non‐formulated technical grade (98%) and formulated isoproturon were compared using soil columns. The use of alginate‐based CR formulations containing bentonite reduced isoproturon movement compared with the technical product. Sorption capacity of the soil for isoproturon was measured using batch experiments (0.29 litre kg⁻¹) and the results obtained here in agreement with those obtained under dynamic conditions. © 2000 Society of Chemical Industry     

Suppression of Dollar Spot by Hypovirulent Isolates of Sclerotinia homoeocarpa

ABSTRACT Selected hypovirulent isolates of Sclerotinia homoeocarpa were evaluated for efficacy in suppressing dollar spot of turfgrass under growth room and field conditions. Under growth room conditions, hypovirulent isolates Sh12B, Sh09B, or Sh08D of S. homoeocarpa caused 3.4 to 30.4% diseased turf in comparison to virulent isolates Sh48B and Sh14D, which caused 80.2 to 90.2% disease. In treatments that received both virulent and hypovirulent isolates, only hypovirulent isolate Sh12B significantly reduced disease as compared with the control with virulent isolates alone. In a field experiment in 1993 on swards of creeping bentgrass artificially inoculated with a virulent isolate of the pathogen, all treatments containing hypovirulent isolate Sh12B applied as a mycelial suspension, granular mix, or alginate pellets developed significantly less disease (6.3 to 20.8% diseased turf) compared with their respective formulation controls (23.8 to 31.2%). Suppression of dollar spot by treatment with mycelial suspensions of isolate Sh12B was evident up to 45 days postinoculation, and disease suppression was still significant 1 year after application when compared with the water control. Applications of hypovirulent isolate Sh09B did not reduce dollar spot in any treatments. Significant suppression of dollar spot by isolate Sh12B was also observed in the experiment conducted in 1994. In addition, suppression of dollar spot by hypovirulent isolate Sh12B was evaluated on swards with naturally occurring inoculum during 1994. Treatments with a mycelial suspension and alginate pellets of hypovirulent isolate Sh12B significantly reduced dollar spot compared with their respective formulation controls. With few exceptions, there was no statistical difference between treatments with hypovirulent isolate Sh12B and the fungicide chlorothalonil (Daconil 2787). Multiple applications of the hypovirulent isolate did not result in greater suppression of dollar spot as compared with a single application. The results indicate that hypovirulence has potential as an effective strategy for the management of dollar spot.     

Application of Trichoderma and Gliocladium in alginate pellets for control of Rhizoctonia damping-off

Alginate pellets were prepared from wet fermentor biomass of 11 isolates of Trichoderma spp. and Gliocladium virens , with wheat bran as a food base carrier. Pellets with eight of the isolates reduced survival (34-78%) of Rhizoctonia solani in infested beet seed in soil. Pellets containing a T. harzianum isolate (Th-58) and a T. hamatum isolate (TRI-4) were the most effective. All isolates significantly reduced growth of the pathogen from infested beet seed into natural soil. Populations of isolates proliferated in soil to 10⁶−10¹¹ colony-forming units/g (cfu) from propagules within the pellets. Pellets with TRI-4 reduced pathogen survival and growth (>70%) in six different soils and were effective against six R. solani isolates in a natural loamy sand. Survival of R. solani in infested beet seed was not reduced when TRI-4 pellets were added to soil 1-6 weeks before the pathogen; however, saprophytic growth was prevented. Small amounts of biomass (3.0–7.5 g wet weight) in pellets were as effective as a large amount (300 g) in suppressing the pathogen. The storage of pellets for more than 6 weeks at 5 or 25C reduced their effectiveness against R. solani. Pellets prepared with four and three of the 11 isolates prevented damping-off of cotton and sugar beet in the greenhouse, respectively.     

Effect of moisture, inoculum production, and planting substrate on disease reaction of field bindweed (Convolvulus arvensis L.) to the fungal pathogen, Phomopsis convolvulus

Phomopsis convolvulus Ormeno is being evaluated as a possible bioherbicide for field bindweed (Convolvulus arvensis L.) control. A granular barley formulation was applied pre-emergence onto the soil surface of pots containing pregerminated field bindweed seeds. Covering the pots with transparent plastic bags immediately after application increased disease incidence and resulted in up to 81% reduction in above-ground dry biomass, whereas a treatment of interrupted dew periods (8 h day^-1) for six days, resulted in only 56% biomass reduction. The size of container used for producing and for incubating the fungus granules had no significant effect on disease incidence and subsequent weed control of field bindweed. Likewise, no significant differences in efficacy were observed using inoculum that was milled once and then sieved or repeatedly milled and non-sieved. For early application dates, the use of two different planting substrates led to major differences in disease development. Pre-emergence application of inoculum on the surface of field collected soil on the same day that field bindweed seeds were planted resulted in an 81% mortality of seedlings emerging. In contrast, only 50% of emerging seedlings were killed when inoculum was applied on the surface of peat moss. Findings in this study indicate that moisture conditions and planting substrate may affect disease incidence and subsequent control of field bindweed by pre-emergence application of the selective fungal pathogen, P. convolvulus.     

The potential of nonpathogenic Fusarium oxysporum and other biological control organisms for suppressing fusarium wilt of banana

The aim of this study was to evaluate the ability of nonpathogenic F. oxysporum and Trichoderma isolates from suppressive soils in South Africa to suppress fusarium wilt of banana in the glasshouse. Several biological control agents and commercial biological control products were included in the study. The isolates were first screened in vitro on potato dextrose agar. In glasshouse evaluations, the fungal and bacterial isolates were established on banana roots before they were replanted in pathogen-infested soil, while the commercial biocontrol agents were applied as directed by the supplier. Banana plantlets were evaluated for disease development after 7 weeks. In vitro tests showed none of the nonpathogenic isolates suppressed Fusarium oxysporum f.sp. cubense ( Foc ), while slight suppression was observed with the two Trichoderma isolates. Results of the glasshouse evaluations revealed that two of the nonpathogenic F. oxysporum isolates, CAV 255 and CAV 241, reduced fusarium wilt incidence by 87·4 and 75·0%, respectively. The known biological control agent Fo47 did not suppress Foc significantly. Pseudomonas fluorescens strain WCS 417, known for its ability to suppress other fusarium wilt diseases (WCS 417), reduced disease incidence by 87·4%. These isolates should be further evaluated for potential application in the field, independently and in combination.     

生物农药“蔬得康”对西瓜细菌性果斑病的温室防效评估

通过温室试验的防效、促生、叶片中叶绿素含量和N、P、K含量的影响,多方面评估生物农药“蔬得康”（由两株芽孢杆菌和一株沙雷氏菌株组成）对西瓜细菌性果斑病的生防潜力。结果表明,生物农药“蔬得康”在温室中防效达到了40.8%,也使西瓜苗期的生物量增加了51.0%,叶绿素a、叶绿素b和叶绿素总量均明显高于清水对照组,分别提高了13.4%、46.2%和21.3%,同时也使叶片中N、P、K的含量分别提高了22.5%、36.4%和29.0%。     

Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).     

Control of Tobacco mosaic virus by PopW as a result of induced resistance in tobacco under greenhouse and field conditions

In a previous study, we isolated a new harpin protein, PopW, from the bacterium Ralstonia solanacearum ZJ3721 that can induce a hypersensitive response in tobacco, Nicotiana tabacum, leaves. In the current study, we demonstrate that, in a greenhouse experiment, PopW induced tobacco-acquired resistance against the Tobacco mosaic virus (TMV) with a biocontrol efficacy of 80.9 to 97.4% at a concentration as low as 25 μg/ml in both PopW-treated and neighboring leaves. The resistance induced by PopW is systemic acquired resistance mediated by salicylic acid, which was certified by the development of resistance being accompanied by the expression of the pathogenesis-related-1 gene (PR1) 8 h after PopW was sprayed onto the tobacco leaves. In addition, hydrogen peroxide began to accumulate 10 h after PopW spraying, peaking at 24 h with a maximum concentration of 1.97 μM/g fresh weight. The activities of phenylalanine ammonia lyase (EC4.3.1.5), polyphenoloxidase (EC1.14.18.1), and peroxidase (EC1.11.1.7) also increased, peaking at different times in the PopW-treated tobacco leaves. PopW also reduced the level of TMV disease in field trials with a biocontrol efficacy of 45.2%. Furthermore, PopW both increased tobacco yield (by 30.4 more than in control plants) and improved tobacco foliar quality, with an increase of 50.2% in the number of first-class tobacco leaves from treated compared with untreated plants. All of these results indicate that the new harpin protein PopW has the potential to be an effective biocontrol agent against TMV in tobacco.     

Alternaria eichhorniae, a biological control agent for waterhyacinth: mycoherbicidal formulation and physiological and ultrastructural host responses

The bioherbicidal efficacy of different alginate formulations of Alternaria eichhorniae 5 (isolate Ae5), a virulent Egyptian isolate, was compared on waterhyacinth (Eichhornia crassipes). The fungus was formulated as alginate pellets containing mycelium alone, mycelium plus culture filtrate or culture filtrate alone. Each formulation was applied with and without a hydrophilic humectant (Evergreen 500). These formulations were evaluated for disease incidence (DI), and disease severity (DS). Maximum DS, but not DI, was obtained with the alginate pellets of mycelium plus culture filtrate. Alginate formulations supplemented with the hydrophilic polymer were more effective in promoting disease. Physiological changes associated with the treated waterhyacinth plants were determined 3, 6 and 9 days after treatment. Waterhyacinth plants treated with alginate pellets of mycelium plus culture filtrate of Ae5 had the lowest levels of pigments, carbohydrates and relative water content. Infection of waterhyacinth with Ae5 led to a significant increase in total phenols of leaves as compared to control. Penetration of waterhyacinth leaves by the fungus occurred only through the stomata, and the invading hyphae were located in the intercellular spaces of leaf tissues. Cytological changes noted in infected cells included changes in chloroplast, nucleus and mitochondria. Invagination of the plasma membrane, particularly at plasmodesmata was also noticed in infected cells. The associations between the infection process, the physiological disorder and the ultrastructure of infected leaves are discussed.     

Effects of timing and method of application of Penicillium oxalicum on efficacy and duration of control of Fusarium wilt of tomato

The effects of timing and method of application of Penicillium oxalicum on the control of fusarium wilt of tomato were investigated. Application of P. oxalicum to tomato seedlings in seedbeds reduced disease caused by Fusarium oxysporum f.sp. lycopersici in a growth chamber by 45–49% and in glasshouse experiments by 22–69%. Disease suppression was maintained for 60–100 days after inoculation with the pathogen in the glasshouse. No disease reduction was observed in tomato plants where P. oxalicum was applied to seeds. Treatment with P. oxalicum did not affect the population of F. oxysporum f.sp. lycopersici in the rhizosphere.     

Biology and management of Phalaris minor in wheat under a rice/wheat system

Summary Germination of Phalaris minor declined with the increase in duration of imbibition in water from 30 min to 72 h at temperatures above 22 °C . Germination was reduced down to 10 cm and 2 cm soil depth by wheat straw burning in puddled and non-puddled soil, respectively, with maximum reduction near the soil surface. The dormancy of P . minor seed was not more than 60 days under field conditions. In puddled soil, 38–60% of the viable seeds of P. minor remained concentrated in the upper 5-cm layer. Germination decreased with an increase in soil depth. In total, 15% of seeds stored in the laboratory emerged from 10-cm depth, whereas seeds did not germinate below 4.2-cm depth under field conditions. Depth of emergence of P . minor was shallower in zero tillage compared with the conventional method of wheat sowing. The seeds retrieved from rice soils kept under continuous submergence for 60 days exhibited 26% and 57% loss of germination over semi-submergence and semi-wet conditions respectively. There was 100% loss of germination in 10-month-old seeds retrieved from the soil under rice-growing conditions. Plant density of P. minor was lower in zero tillage than with the conventional method of wheat sowing. Cross-ploughing in the upper 2–5 cm of soil (shallow tillage) and drill-sowing of wheat 1 week after shallow tillage reduced germination of P. minor by 44% and 37% and increased grain yield by 21% and 47% over zero-tillage and conventional methods respectively.