Secondary Metabolites Produced by a Strain of Fusarium oxysporum Used for Striga Control in West Africa

The production of fusaric acid and other toxins by a strain of Fusarium oxysporum used for control of the weed Striga hermonthica (Del.) Benth. was investigated. Culturing of the strain under optimal conditions for toxin production produced small amounts of fusaric acid and dehydrofusaric acid but no other toxins reported to be produced by some strains of F. oxysporum. Culturing of the fungus under conditions similar to those that would be used in the field produced no detectable toxins of concern for human health.     

Chlamydospore Production,Inoculation Methods and Pathogenicity of Fusarium oxysporum M12-4A,a Biocontrol for Striga hermonthica

Fusarium oxysporum isolate M12-4A is currently being evaluated for the biological control of Striga hermonthica . Inoculum production, inoculum delivery to the target, chlamydospore germination, and weed growth suppression of this weed-pathogen system were investigated. Liquid fermentation systems using organic material were evaluated for the production of large numbers of chlamydospores. A 1% sorghum straw powder (< 1 mm) substrate, exposed to black light at 21°C for 21 days, yielded 3.23 X 10 8 colony forming units (CFU) l -1 medium. A two-stage fermentation system using 5% w/v straw substrate under black light at 30°C for 14 days yielded 3.5 X 10 8 CFU l -1 medium. In vitro variations in chlamydospore germination were governed by the presence of exogenous carbon, nitrogen, and sorghum root exudates. Ammonium-nitrogen compounds and urea, in combination with glucose had a stronger stimulatory effect on chlamydospore germ tube growth than did potassium nitrate. Maximal germ tube elongation occurred when chlamydospores were exposed to urea at a C/N ratio of 10. Some mineral solutions and sorghum root exudates inhibited chlamydospore germ tube elongation; however, arabic gum, a complex polysaccharide, stimulated chlamydospore germ tube elongation and the production of secondary chlamydospores. In field trials, chlamydospore powder harvested from small-scale fermenters reduced S. hermonthica emergence by 92%. Complete inhibition of S. hermonthica emergence occurred when the chlamydospore powder was added to the soil at sowing and when sorghum seeds coated with chlamydospores were sown. Effective biological control of S. hermonthica was achieved using a simple fermentation system with sorghum straw as the inoculum growth substrate. For inoculum delivery to the farmers' fields, sorghum seeds were coated with the inoculum using arabic gum as the adhesive. This simple delivery system permits a uniform inoculation of the field as well as the proper positioning of the inoculum in the immediate environment of sorghum roots, where S. hermonthica attaches to its host. To facilitate a broad usage of F. oxysporum M12-4A for the biocontrol of S. hermonthica , we propose an inoculum production strategy based on a cottage industry model that utilizes a liquid fermentation process and inexpensive locally-available substrates including sorghum straw and arabic gum.     

Microorganisms of Striga hermonthica in Northern Ghana with Potential as Biocontrol Agents

A survey of microorganisms of Striga hermonthica Del. Benth., a root parasite of graminaceous crops, was conducted in northern Ghana in 1992. Thirteen fungal species were isolated from infected S. hermonthica plants. Fusarium spp. were the most prevalent, and were isolated from more than 90% of the S. hermonthica samples collected. Other fungi isolated were Alternaria alternata, Bipolaris specifera, Cladosporium oxysporum, Curvularia falax, Macrophomina phaseolina, Nodulisporium gregarium, Phoma sorghina and Sclerotium rolfsii.     

Fusarium oxysporum and post-emergence herbicide for the control of the parasitic plant Striga hermonthica in maize

Two trials were conducted to evaluate the efficacy of a granular mycoherbicide formulation based on Fusarium oxysporum and post-emergence herbicide for the control of the parasitic plant, Striga hermonthica in the Nigerian Savanna. Four fungal treatments were used: F. oxysporum followed by 2,4-D, F. oxysporum followed by supplementary hoe weeding, F. oxysporum followed by Triclopyr and a control (No F. oxysporum but hoe-weeded). The experiments were laid out in a split plot design with three replications in the two locations. The two varieties (Across 97 TZL and farmer's local variety) formed the main plot treatments, while the Striga fungal treatments formed the sub-plot treatments. At the Lafia location, the emergence of Striga was delayed by 7 days (46 days) as compared to the Makurdi location, which germinated earlier 39 days after sowing. Maize variety Across 97 TZL similarly delayed the time to Striga emergence when compared to the farmer's local variety. However, the different Striga control methods did not have any significant effect on the time of Striga emergence. Generally, number of maize plants infected with Striga was highest with the farmer's local variety throughout the period of observation, while in the Striga control treatments, hoe-weeded check recorded the highest; the minimum was obtained with plots treated with F. oxysporum followed by postemergence application of Triclopyr at the rate of 0.36 kg a.i./ha at 6 weeks after sowing. Highest maize grain yields were obtained at Lafia with Across 97 TZL and plots treated with F. oxysporum followed by either post-emergence 2,4-D or Triclopyr at 0.36 kg a.i./ha each. The results demonstrate the high potentiality of using F. oxysporum as a spot application at planting followed by post-emergence herbicide (2,4-D or Triclopyr) application at 6 weeks after sowing for the control of the parasitic plant S. hermonthica in the Nigerian Savanna.     

Influence of Agricultural By-products in Liquid Culture on Chlamydospore Production by the Potential Mycoherbicide Fusarium oxysporum Foxy 2

Economically feasible inoculum mass production methods are required for successful application of Fusarium oxysporum Foxy 2 as a potential mycoherbicide. Therefore, different substrates (agricultural by-products) and the factors that influence the production of spores, especially chlamydospores, of Foxy 2 were investigated in liquid cultures. The substrates tested were cotton seed cake, maize stover, wheat and triticale stillage. The presence of plant fibers in the medium of unfiltered cotton seed cake (2.5%, w/v) significantly enhanced chlamydospore, micro- and macroconidia production by 150, 185, and 300%, respectively, compared to the filtered (fiber-free) medium. Regardless of the type of substrate tested, Foxy 2 was able to produce abundant chlamydospores (0.14-2.7×10⁷ mL^-1) in all growth media. Generally, increasing the concentrations of cotton seed cake and maize stover in the medium significantly increased chlamydospore formation; however, this was not the case for wheat-based stillage. To optimize conditions required for chlamydospore production of Foxy 2, the effect of near ultra-violet (NUV) light, substrate combinations (synergism), level of agitation, incubation time and their interaction were studied. A liquid culture of 2.5% (w/v) filtered cotton seed cake, exposed to continuous NUV for 15 days, doubled the yield of chlamydospores (4.7×10⁶ mL^-1) and macroconidia (5×10⁵ mL^-1), and increased microconidial production by one-third (1×10⁸ mL^-1) compared to natural light. An apparent synergistic effect of substrate combination was observed, since the addition of 20% (v/v) of either wheat or triticale-based stillage to maize stover medium (1%, w/v) increased the number of chlamydospores produced by 16 or 12 times, respectively, compared to maize stover alone (1.4×10⁶ mL^-1), and 2 times more than either of the stillages. A significantly positive effect between a high level of agitation tested during incubation and chlamydospore production of Foxy 2 was recorded. All in all, substantial chlamydospore production of Foxy 2 (4.3×10⁷ mL^-1) was successfully achieved within 12 days of incubation in a single-step liquid fermentation through the combination of 0.5% (w/v) maize stover plus 20% (v/v) wheat-based stillage and a high level of agitation (200 rpm).     

Herbivores of Striga hermonthica in Northern Ghana and Approaches to Their Use as Biocontrol Agents

A survey of herbivores of Striga hermonthica Del. Benth., a root parasite of cereal crops, was conducted in northern Ghana in 1992. Smicronyx umbrinus Hustache (Coleoptera: Curculionidae) and Junonia orithya L. (Lepidoptera: Nymphalidae) were found to attack S. hermonthica in all areas surveyed. Preliminary observations indicate that these two insects have a great potential to cause damage. In the Shinyanga region of Tanzania, Malawi and the middle-west of Madagascar, investigations revealed that only J. orithya is common in all areas surveyed, in contrast to the situation in northern Ghana.     

Effect of Growth Medium and Method of Application of Fusarium oxysporum on Infestation of Sorghum by Striga hermonthica in Burkina Faso

A 2-year (1997–1998) study was conducted at Kouaré, Burkina Faso, to investigate effect of growth medium and application method of Fusarium oxysporum isolate 4-3-B to control Striga hermonthica. In 1997, growth medium and isolate 4-3-B delayed striga emergence by 9 days. Isolate 4-3-B reduced emerged striga number by 33% compared to treatments without Fusarium. In 1998, striga emergence was delayed by 13 days by growth medium and Fusarium. The fungus reduced the number of emerged striga by 27% and, as a result, sorghum straw and grain yields were significantly improved by 10 and 38%, respectively.     

Effect of cultural methods on Striga (Striga hermonthica (Del.)) Benth management and yield of cereals in the Savanna Zone of Nigeria: a review

Data were obtained from the research done in the Guinea Savanna (Zaria 11°11′N; 070 38′E) and Sudan Savanna (Maiduguri ?11°51′N; 13°15′E) regions of Nigeria, respectively on different cultural methods of Striga control and management. In the Guinea Savanna, trials on the effects of nitrogen on the response of resistant and susceptible upland rice varieties to Striga hermonthica infestation and the effect of resistant and susceptible varieties of maize and crop rotation on Striga infestation was carried out, while the effect of inter-cropping trap crop (Bambaranut) with resistant sorghum varieties on S. hermonthica was studied in the Sudan Savanna Zone of Nigeria. In the Guinea Savanna, it was observed that a combination of upland rice variety, Faro 40 and an application of 90 kg N/ha in the wet season and WAB 56-50 upland rice variety and 120 kg N/ha in the dry season, respectively reduced Striga infestation and produced maximum grain yield. Also, the growing of resistant variety of maize (Across 97ITZ comp. I-W) after 1 or 2 years' rotation with cowpea or soybean was observed not only to be effective in Striga control, but resulted in higher grain yield of maize. In the Sudan Savanna, the use of resistant varieties of sorghum, 1CSV1002 and 1CSV1007 intercropped with bambaranut significantly reduced Striga infestation, but the grain yield of the resistant varieties was low. From these studies, Faro 40 with 90 kg N/ha application rate and WAB56–50 with 120 kg N/ha were suitable for the management of Striga and for higher grain yield of upland rice in both wet and dry seasons, respectively, while Across 97ITZ comp. I-W, resistant maize variety and 1 or 2 years rotation with cowpea or soybean were also the best for the management of S. hermonthica and for higher maize yield in the Guinea Savanna zone. Further research needs to be carried out in the Sudan Savanna to select a high yielding resistant variety of sorghum which when intercropped with bambaranut will not only control Striga infestation but will also give high grain yield.     

Biocontrol efficacy of Trichoderma spp. against sesame wilt caused by Fusarium oxysporum f. sp. sesami

Sesame (Sesamum indicum L.) is one of the most important oilseed crops in Egypt and worldwide. It is being infected with many pathogens, among these pathogens Fusarium oxysporum f.sp. sesami (Zap.) Cast is causing severe economic losses on sesame. In this study, antagonistic capability of 24 isolates of Trichoderma spp. was assessed in vitro against F. oxysporum f.sp. sesami. Two strains; T. harzianum (T9) and T. viride (T21) were revealed to have high antagonistic effect against F. oxysporum f.sp. sesami in vitro with inhibition percentage about 70 and 67%, respectively. These two isolates proved to have high ability to control Fusarium wilt disease under greenhouse conditions. The highest reduction in disease severity was achieved with T. viride followed by T. harzianum with reduction in disease severity about 77 and 74%, respectively. This study revealed that the time of application of bioagents is a decisive factor in determining the efficacy of Trichoderma isolates to control Fusarium wilt of sesame. It was revealed that the highest reduction in the disease severity was achieved when either Trichoderma viride or T. harzianum were applied 7 days before challenging with the F. oxysporum f.sp. sesami.     

西葫芦根腐病菌拮抗细菌的防病促生作用

【目的】获取促生的同时可抑制尖孢镰刀菌(Fusarium oxysporum)的拮抗菌并明确其防治效果。【方法】平板对峙生长法测定前期分离的19株促生菌对尖孢镰刀菌的抑制作用,温室条件下接种微生物菌剂对西葫芦根腐病的防治作用;田间条件下接种复合菌剂代替部分化肥对西葫芦生长的影响。【结果】可有效拮抗尖孢镰刀菌的菌株有9株,其中,菌株FX2的抑菌活性较好,抑制率达到66.80%。在温室盆栽中,接种微生物菌剂(LHS11+FX2)对西葫芦根腐病抑制率达到57.14%。在田间试验中,微生物菌剂配施化肥对西葫芦的生物量和根系形态影响显著,以85%化肥+复合菌剂处理效果较优,其对西葫芦成熟期的产量显著提高27.13%。【结论】复合菌剂(LHS11+FX2)对西葫芦根腐病具有较好的防治作用;85%化肥+复合菌剂对西葫芦的促生作用明显,在一定程度上节约了化肥投入成本,提高了增产效益。     

An Alginate Prill Formulation of Fusarium oxysporum Schlechtend: Fr. f. sp. erythroxyli for Biocontrol of Erythroxylum coca var. coca

A rice alginate prill formulation of isolate EN - 4 of Fusarium oxysporum f . sp . erythroxyli, pathogenic to Erythroxylum coca var . coca (coca) , was evaluated in greenhouse and field studies for its ability to enhance pathogen populations in the soil and cause disease in coca . The formulation was applied to four different soil types in the greenhouse at 33 . 6 kg ha 1 . It enhanced the population of EN - 4 in each soil and most ( > 90%) of the fungal population remained in the upper 5 cm of soil during the 49 - day experiment . When applied in field experiments , the formulation enhanced the population of EN - 4 in the soil . Isolate EN - 4 was present in the upper 7 . 6 cm of soil 28 days after application at populations similar to those in the greenhouse studies (1 103 to 1 104 colony - forming units (CFUs) / g of soil) . Elevated populations of the pathogen (1 102 CFUs / g of soil) were still present in treated soils 229 days after application of the formulation . The areas used for field studies were already infested with the pathogen and typically developed high levels of fusarium wilt within 2 years of planting with coca . The formulated F. oxysporum began having a significant effect on plant death 100 - 200 days after application based on repeated measures analysis . These data suggest that a formulation of F. oxysporum f . sp . erythroxyli which enhances the incidence of fusarium wilt in coca fields can be produced using established techniques .     

Biological Control of Fusarium oxysporum f.sp. asparagi by Applying Non-pathogenic Isolates of F . oxysporum

Root rot severity of asparagus plants grown in sterilized field soil inoculated with Fusarium oxysporum f . sp . asparagi (Foa) was reduced by more than 50% when the soil was precolo nized by each of 13 non - pathogenic (np) isolates of F. oxysporum originating from asparagus roots or field soils . In a greenhouse experiment , application of six np isolates to naturally infested field soil was followed by a 23 - 49% decrease of disease severity , depending on the isolate . One of them , Fo47 originating from Fusarium suppressive soil in France , was applied to field plots infested with Foa . Foa root rot was not suppressed in asparagus plants grown for 1 year in these plots . Pathogenic and np isolates extensively colonized the root surface and isolates of both types infected the roots of asparagus plants grown in sterilized field soil , with significant differences among the np isolates . Inoculation of sterilized field soil with np isolates reduced germination of Foa chlamydospores by 43 - 64% depending on the isolate used . It is concluded that np isolates of F. oxysporum can suppress asparagus root rot caused by Foa in naturally infested field soil . The differences for root colonization capacity among the np isolates imply that selection for this trait might reveal isolates that perform better under field conditions .     

Bioherbicidal effect of fumonisin B1, a phytotoxic metabolite naturally produced by Fusarium nygamai, on parasitic weeds of the genus Striga

Fusarium nygamai has been isolated and identified from diseased Striga hermonthica plants collected from sorghum fields in the Sudan. Fumonisin B₁, produced by this fungus, was isolated and purified. In in vitro experiments, the bioherbicidal potential of this mycotoxin was evaluated against S. hermonthica and S. asiatica. Different concentrations (250, 500 and 1000 μg L^-1 of water) of fumonisin B₁, applied during the conditioning phase of the Striga seeds significantly reduced the germination of both species in a range of 19.8-32.2 and 34.5-47.6%, respectively. In addition, the length of the germ tubes was reduced and the germ tubes became brownish. Fumonisin B₁ slightly delayed the emergence of Striga when 250 mg fumonisin B₁ dissolved in 1 L of water were applied into the soil per pot before sowing sorghum. By the way of contrast, fumonisin B₁ was very phytotoxic when Striga seedlings (leaves and stems) of different heights and ages were sprayed with a concentration series of 250, 500 and 1000 μg fumonisin B₁ mL^-1 until run-off, or when plants were injured with a scalpel before fumonisin B₁ was applied into the injury at concentrations of 250 and 500 μg mL^-1. One day after application, the plants showed symptoms of wilting, first at the top and then moving downwards. Leaves turned black and stems desiccated at the point of injuries. Within 4 days after the application, 55% of the plants tested were completely destroyed. In total, at the end of the experiment, 40 days after the treatment, 85% of the treated plants were killed by fumonisin B_1, regardless of the concentration applied and the application technique used.     

Seed treatment technology: An attractive delivery system for controlling root parasitic weed Striga with mycoherbicide

Coating sorghum seeds with Fusarium oxysporum (Foxy 2) for control of the root parasitic weed Striga, appears to be an attractive option for minimizing the inoculum amount, establishing the biocontrol agent in the potential infection zone of the host plants, and offering a simple, easy and economical delivery system. Our investigations resulted in the selection of appropriate seed coating materials and a suitable type and form of fungal inoculum. The coating materials tested were arabic gum (AG10%, 20%, 40%), carboxymethylcellulose (CMC1%, 2%) and pectin (LS 440, LM-5 CS) 1%, while the fungal inoculum included microconidia and fresh and dried chlamydospores produced using different substrates. Foxy 2 survived the seed treatment processing and showed excellent viability on seeds for at least 8 months of storage after coating. In general, the performance of 40% arabic gum in combination with dried chlamydospores was the best among the other types of inoculum and coating material tested. Regardless of the type and form of inoculum and coating materials tested, Foxy 2 was able to colonize all roots, even root tips and hairs of the host (sorghum), thereby meeting important criteria of a promising candidate for controlling Striga when applied as a seed treatment. The efficacy of treated sorghum seed with Foxy 2 using different coating materials in reducing S. hermonthica infestation was evaluated in pot and root chamber trials. Foxy 2 markedly reduced Striga emergence and dry weight and increased the percentage of the diseased emerged Striga shoots. However, the efficacy of seed coating varied according to the type and form of fungal inoculum as well as coating material. Coating sorghum seed with dried chlamydospore inoculum homogenized into 20% arabic gum (as adhesive) showed the highest efficacy of 81 and 77% (i.e., percent reduction in healthy emerged Striga shoots compared to the control treatment) against Striga using either sterilized or non-sterilized soil, respectively. In root chamber bioassays, the application of Foxy 2 in combination with AG40% significantly caused disease in 77% of the germinated Striga seeds and in all tubercles after 25 days of sowing. These findings provide an optimized coating protocol as an attractive delivery system for bioherbicides for root parasitic weeds.     

Characterisation of Simple In Vitro Cultures of Striga hermonthica Suitable for Metabolic Studies

Abstract: The aim of this study is to develop simplified models for standardised screenings of xenobiotics, especially targeted against mannitol production, to control the harmful parasite, S. hermonthica. Chlorophyllous protoplasts and calli were produced from the young leaves of the parasite. Best yield from protoplast isolation was obtained when leaf segments were incubated at 30 °C, in the light, under shaking in an enzyme cocktail containing 2 % cellulase Onozuka R10, 0.1 % Pectolyase and sorbitol 1 M as the osmoticum. Oxygen exchange measurements, as well as labelling experiments with ¹⁴C-bicarbonate, emphasised a significant decrease in photosynthetic capacity of protoplasts, mannitol remaining, however, a major primary product of photosynthesis. Initiation of cell cultures was unsuccessful and instability of protoplasts prevents their standardised utilisation for herbicide screening. In contrast, globular calli produced first on MS medium containing 0.5 mg L^-1 NAA, 2.5 mg L^-1 BAP and 2 % sucrose were stable for two years, after monthly subculturing on fresh medium. Sucrose substitution by mannose in the medium did not change kinetic growth and stability. Potential autotrophy was lost for calli by increasing exogenous sugar level. Biochemical analyses and labelling experiments with ¹⁴C-bicarbonate or ¹⁴C-sucrose or -mannose showed that carbon partitioning is modified in calli, in comparison with young leaves or protoplasts, sucrose or mannose accumulation being favoured in sucrose- or mannose-fed calli, respectively. However, carbon flow towards mannitol was more marked in calli growing on high mannose medium. Stability and preservation of an active mannitol biosynthetic pathway allows planning of xenobiotic assays with calli as a simplified model for Striga hermonthica.     

Agricultural By-products as Substrates for Growth, Conidiation and Chlamydospore Formation by a Potential Mycoherbicide, Fusarium oxysporum Strain EN4

Characteristics, such as mycelial growth, conidiation and chlamydospore formation, of three distinct variants (EN4-FT, EN4-FF and EN4-S), of the potential mycoherbicide, Fusarium oxysporum, strain EN4 were studied in liquid cultures. Agricultural by-products, such as cornmeal, corn cob, soya bean hull fiber (Dietfiber-Soyfiber), cotton seed embryo flour (Pharmamedia), cane molasses yeast extract and potato dextrose medium were used as substrates. The mycelial growth and conidia and chlamydospore formation were different for the three strains and varied with the substrate used. The quantity of conidia and chlamydospores produced depended on the concentration of the substrates. Irrespective of carbon:nitrogen ratios, chlamydospores were formed readily in liquid media and in greater amount on substrates with low utilizable carbon content (aqueous extracts of soya bean hull fiber and corn cob) than on those with higher utilizable carbon content (potato dextrose broth, aqueous extracts of cotton seeds and molasses yeast extract broth). In liquid cultures, increases in the concentration of substrates high in utilizable carbon resulted in reduced chlamydospore formation; however, this was far less in those with a lower carbon content. The effect on fungal growth characteristics due to the presence or absence of plant fibers in various concentrations of unfiltered and filtered extracts of soya bean hull fiber depended on the fungal variant tested. However, it had a greater effect on viable and microconidial counts than on chlamydospore and macroconidial counts. The viable and microconidial counts were significantly higher in unfiltered than in filtered extracts. The final pH of the liquid cultures (4.1-8.5), after 10 days of growth, was affected by the fungal variant, the substrate used and by the substrate concentration. In liquid cultures with 1% soya bean hull fiber, chlamydospore formation corresponded to an increase in the pH of the medium from 6.0 to 7.7. The interaction between fungal variants, the substrate used and the substrate concentration was highly significant for all the variables tested, indicating that optimization of the production of fungal propagules is complex. Overall, for all three variants, the best substrates for optimum conidiation and chlamydospore formation were aqueous extracts of soya bean hull fiber and corn cob at concentrations of 1-2.5%.     

Growth and carbon allocation in Pennisetum typhoides infected with the parasitic angiosperm Striga hermonthica

Abstract Growth and gas exchange measurements are used in conjunction with a carbon balance model to describe the millet (Pennisetum typhoides)–witchweed (Striga hermonthica) host—parasite association. Striga hermonthica reduces the growth of millet by 28% and radically alters the architecture of infected plants. Whilst grain yield and stem dry weight are reduced (by 80 and 53%, respectively), leaf and root growth are stimulated (by 41 and 86%, respectively). The difference in production between infected and uninfected millet plants can be accounted for by two processes: first, export of carbon to the parasite (accounting for 16% of the dry weight not gained); and second, parasite-induced reductions in host photosynthesis (accounting for 84% of the dry weight not gained). Striga hermonthica is dependent on carbon exported from the host, since the plant has low rates of photosynthesis coupled with high rates of respiration. The carbon balance model suggests that in mature S. hermonthica plants parasitic on millet, 85% of the carbon is host-derived. Carbon fluxes are also estimated using δ¹³C measurements, since S. hermonthica is a C₃ plant parasitizing a C₄ host. In conjunction with gas exchange measurements, these suggest that in root, stem and leaf of S. hermonthica, 87, 70 and 49% of carbon is hostderived, respectively.