Biocontrol action mechanisms of Cryptococcus laurentii on Colletotrichum gloeosporioides of mango

Mechanisms of action and effectiveness of the antifungal yeast Cryptococcus laurentii [(Kuff.) C.E. Skinner] strain L5D, were examined against the causal agent of anthracnose Colletotrichum gloeosporioides ((Penz.) Penz. & Sacc.) in mango (Mangifera indica L.). C. laurentii showed a high antagonistic potential in vivo, with significant inhibition of anthracnose (75.88%). Different mechanisms of action were examined in C. laurentii among them competition for nutrients, specifically for sucrose (p < 0.05). Scanning Electron Microscopy (SEM) showed that the yeast biofilm adheres to the fruit and to C. gloeosporioides hyphae showing competition for space; C. laurentii was not washed off from treated Colletotrichum hyphae as observed with SEM. According to statistical analysis, only nagase and chitinase were significantly stimulated on wounded fruit. Activity of all three hydrolytic enzymes was detected in vitro but only nagase was induced by addition of autoclaved pathogen mycelium. Treated wounds with the yeast biocontrol stimulated glucanase activity and suppressed chitinase activity on fruit wounds with or without presence of autoclaved pathogen mycelium but did not affect nagase significantly (p = 0.05). Parasitic activity of yeast on pathogen was not detected.     

Induction of lignin and pathogenesis related proteins in dragon fruit plants in response to submicron chitosan dispersions

Chitosan is capable of triggering the induction of defence responses of the plants. This study was carried out to investigate in vitro and in vivo antifungal activity of submicron chitosan dispersions and also to determine the mechanism of induced resistance in dragon fruit plants by strengthening of cell wall and production of pathogenesis related proteins. Highest inhibition in radial mycelial growth (90.2%) was obtained by 600 nm droplet size at 1.0% chitosan concentration. None of the emulsifiers showed any antifungal effect during in vitro studies. Field studies showed the reduction of 22% disease incidence and 66% disease severity in anthracnose on dragon fruit plants treated with 600 nm droplet size as compared to control plants after one month of assessment. Lignin contents in cell wall of dragon fruit plants increased 2 folds in plants treated with 1.0% chitosan at 600 nm droplet size while 11 and 10 folds increase in β-1,3-glucanase and chitinase activity, respectively was also recorded in the plants inoculated with Colletotrichum gloeosporioides. The accumulation of lignin and higher level activity of pathogenesis related proteins in response to submicron chitosan dispersions may bolster in mounting effective resistance against the devastating disease of anthracnose.     

Tolerance to Fusarium wilt and anthracnose diseases and changes of antioxidative activity in mycorrhizal cyclamen

Mycorrhizal associations imply a remarkable reprogramming of functions in both plant and fungal symbionts. This consequent alteration on plant physiology has a clear impact on the plant responses to biotic stress management. As a consequence, a pot experiment was conducted to study the interactions between the arbuscular mycorrhizal fungus (AMF) Glomus fasciculatum and the two pathogens Fusarium oxysporum and Colletotrichum gloeosporioides and subsequent effect on growth, disease tolerance and the changes in antioxidative ability in cyclamen plants under growth chamber condition were investigated. At plant maturity, inoculation with F. oxysporum and C. gloeosporioides, responsible for Fusarium wilt and anthracnose of cyclamen respectively, significantly reduced shoot and root dry weights, increased both the disease incidence percentage and showed lower antioxidative activity viz. superoxide dismutase (SOD), ascorbate peroxidase (APX), ascorbic acid (AA) and polyphenol contents in plants. In contrast, the growth response and biomass production of cyclamen plants inoculated with AMF was significantly higher than the nonmycorrhizal control plants, both in the presence and absence of the pathogens. Mycorrhization enhanced plants to reduce the Fusarium wilt and anthracnose incidence compared to nonmycorrhizal controls. In every case, without and with pathogen association, plants inoculated with AMF increased the antioxidant (SOD, APX, AA and polyphenol) production compared to control plants. The results demonstrate that AMF have the ability to induce resistance against Fusarium wilt and anthracnose in cyclamen by increasing the antioxidative activity in plants, which promoted plant growth, biomass production and drastically reduced the disease incidence in cyclamen.     

Management of Eumusae leaf spot disease of banana caused by Mycosphaerella eumusae with Zimmu (Allium sativum × Allium cepa) leaf extract

Among extracts of 33 plant species screened against Mycosphaerella eumusae, the causal agent of Eumusae leaf spot disease of banana, water extract of Cassia senna, Zimmu (Interspecific hybrid between Allium cepa × Allium sativum) and Rhincanthus nasutus provided 100% inhibition of spore germination and 1.7–2.0 cm zone of inhibition of mycelial growth under in vitro conditions. The maximum efficacy of mycelial inhibition was observed with Zimmu leaf extract. When Zimmu leaf extract was tested at different concentrations (5, 10, 25, 50 and 100% w/v), all tested concentrations provided complete inhibition of mycelial growth of the pathogen. The field evaluation of Zimmu leaf extract at different concentrations in cv. Grand Naine showed that the application of the water extract of Zimmu leaf at 50% concentration (w/v) provided 55% reduction of disease severity compared to the unsprayed control. Besides, the application of Zimmu leaf extract increased the value of youngest leaf spotted-0 (up to 60.5%) as well as increased the yield of banana (up to 46.8%) as compared to control. The effect of Zimmu in increasing the value of YLS-0 and the bunch yield was comparable with the chemical fungicide Propiconazole 25% EC (0.1%). Thin layer chromatography (TLC) analysis showed that among different major compounds, two lipid compounds (LP-B1 and LP-B2) extracted using methanol had the ability to inhibit M. eumusae growth (0.7–1.5 cm zone of inhibition). The gas chromatography–mass spectrometry (GC–MS) analysis of lipid bands revealed the presence of six different lipid compounds, which may be responsible for the growth inhibition of the pathogen. Since the application of water extract of Zimmu was found to be not only effective in controlling the leaf spot disease severity but also increased the number of green leaves and yield of banana fruits, the Zimmu extract can be used effectively in integrated disease management of Eumusae leaf spot disease for enhancing banana production in an ecologically sustainable manner.     

Saccharin-induced systemic acquired resistance against rust (Phakopsora pachyrhizi) infection in soybean: Effects on growth and development

We examined the effect of saccharin on the systemic acquired resistance (SAR) response of soybean to the fungus Phakopsora pachyrhizi, the causal agent of soybean rust. Plants were grown hydroponically in half-strength Hoagland’s solution and were challenged with the pathogen 1, 5, 10 and 15 d after treatment with 3 mM saccharin applied either as a foliar spray or a root drench at the 2nd trifoliate (V3) and early reproductive (R1) stages. Plants were destructively harvested and assessed for visible rust symptoms 2 wk after inoculation. Mode of saccharin application was a significant factor influencing the severity of rust infection. Saccharin applied as a root drench was more effective than the foliar spray treatment at inducing SAR, with increased resistance observed 1 d after application. Systemic protection against rust infection was still apparent 15 d after application of saccharin as a root drench. In contrast, foliar treatment with saccharin did not increase systemic protection until 15 d after treatment. When systemic protection was induced by the application of saccharin in either manner, there was no significant reduction of plant growth, except when plants were inoculated 15 d after the saccharin application as a root drench at the R1 stage of development.     

Dry heat treatment of Andean lupin seed to reduce anthracnose infection

The potential of dry heat treatment of Andean lupin seed to reduce seed–borne infection of the anthracnose pathogen, Colletotrichum acutatum, was investigated. First, the effect of dry heat (65 °C) over duration times of 0–96 h on germination and disease incidence after germination was evaluated for artificially– and naturally–infected seed. Dry heat treatment from 8 to 96 h reduced disease incidence after germination to undetectable levels in four cultivars compared with 7.5% disease incidence after germination in seed maintained at room temperature. Moreover, heat treatments of 4–12 h showed seed germination rates that were equivalent to the non–treated control. Under greenhouse conditions, dry heat treatments for 8 or 12 h reduced transmission of the pathogen from seed by 75 or 85%, respectively and dry heat treatment increased emergence of seedlings in comparison the non-treated control. Dry heat treatment is an environmentally friendly alternative for reducing anthracnose infections in Andean lupin seed.     

Agronomic and economic assessment of intensive pest management of dry bean (Phaseolus vulgaris)

Two common production constraints of dry bean (Phaseolus vulgaris) in Ontario are annual weeds and anthracnose (caused by Colletotrichum lindemuthianum). Dry bean is not considered a competitive crop and weed interference can result in substantial yield losses, while anthracnose is considered one of the most devastating diseases in dry bean production. A study conducted in Ontario Canada, examined the effect of two herbicide programs on weed management, thiamethoxam insecticide treatment on plant enhancement and three fungicide programs on anthracnose development in a navy bean cv. ‘OAC Rex’. The premium herbicide program (s-metolachlor + imazethapyr) reduced percent weed ground cover relative to the economic herbicide program (trifluralin) in five of six locations. Thiamethoxam increased emergence and vigour at only one location, which contradicts reported benefits of thiamethoxam on plant health. The herbicide or thiamethoxam treatments did not affect anthracnose disease severity, visible seed quality, net yield or economic return. The fungicide seed treatment was often superior to the untreated control, for a number of the parameters measured. The application date of the foliar fungicide, relative to the onset of disease, varied between site-years. This dramatically influenced the fungicide’s effectiveness. Foliar fungicides increased seed quality and net economic return compared to the control when applied prior to disease development. The combination of fungicide seed treatment followed by a foliar fungicide provided the largest reduction in anthracnose severity.     

Mature watermelon vine decline: Suppression with fumigants of a soil-borne problem and association with Rhizopycnis vagum

Mature watermelon vine decline (MWVD) is a late-season disease of unknown etiology, characterized by vine collapse and discolored, much reduced root systems. While several new disorders in watermelon have been reported recently, this problem appears specific to the Midwest. To test for a biological cause and methods to suppress this disorder, soil was collected from two southern Indiana fields with histories of MWVD for microplot and greenhouse experiments. Watermelons grew best in microplots treated with methyl bromide or methyl iodide. Roots of plants growing in the untreated MWVD soil were severely necrotic; those from the fumigation treatments appeared healthy. In heating experiments, watermelon seedlings were grown in autoclaved soil mixed with 10% MWVD soil that was either untreated or heated at 40, 50, 60, 70 or 80 °C for 30 min. Watermelon plants in the untreated controls and the 40 and 50 °C treatments had reduced top dry weights with decreased fibrous roots and associated discoloration in contrast to treatments of equal to and greater than 60 °C. Representative cultures from necrotic roots were identified as Rhizopycnis vagum by morphological analysis and sequencing the ITS region of the ribosomal DNA. PCR analysis with specific primers confirmed that R. vagum was present in roots from untreated MWVD soil but not in symptomless roots. This is the first report of R. vagum associated with plants suffering from MWVD in Indiana and indicates that this fungus occurs much farther north in the U.S. than known previously.     

Biological weed control in rice with a strain of Colletotrichum gloeosporioides (Penz.) Sacc. used as a mycoherbicide

An indigenous fungal pathogen of the native leguminous weed, northern jointvetch (Aeschynomene virginica [L.] B.S.P.) was found to cause an anthracnose disease at endemic levels annually on its host in Arkansas rice and soybean fields without apparent impact on competitive ability or dispersal of the weed. The pathogen was isolated in pure culture and identified as a host-specific strain of Colletotrichum gloeosporioides, an intensively studied ‘group species’ which causes many anthracnose diseases and consists of species and strains representing wide latitudes of hosts specificity and virulence on both economic crops and wild plants in the tropical and temperate regions of the world. An understanding of anthracnose disease aetiology gained from the literature and confirmed by empirical studies with the weed and fungus revealed that the pathogen was specific and potentially devastating to its weed host, but was constrained in natural situations by poor capacity for dissemination of inoculum. Its spores, although produced in abundance, develop late, are sticky and dispersed mainly by splashing rain or insects, mechanisms that are suppressed on annual weed plants interspersed with annual crop plants. For the fungus to be effective as a biological control agent (i.e. as a mycoherbicide), complete coverage of a field with inoculum at appropriate times and levels to compensate for the pathogen's poor capacity for dissemination would be required. A multidisciplinary team of plant pathologists, weed scientists, veterinary scientists and fermentation scientists investigated the practical potential of the mycoherbicide concept with this host-pathogen combination and found it to be biologically feasible, registerable and commercially practical. The fungus is marketed as living dry spores for use on a comparatively small area weed problem and in a crop-management programme that relies heavily on chemical pesticides. Extension of this concept to control other weeds or other pests seems possible and practical.     

Antagonistic yeasts with potential to control Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. and Colletotrichum acutatum J.H. Simmonds on avocado fruits

Avocado is an economically important fruit that is severely affected by anthracnose disease due to the infection with Colletotrichum spp. In this study, avocado fruits with anthracnose symptoms were collected in Morelos, Mexico. Two phytopathogenic fungi were isolated from these fruits and identified as Colletotrichum gloeosporioides and Colletotrichum acutatum using ITS sequences. Further, eleven yeasts from avocado (three from fruits, four from leaves and four from rhizospheric soil) were isolated; of which three showed in vitro antagonistic activity against C. gloeosporioides and C. acutatum. ITS sequence analysis of the isolated yeasts revealed that the strains obtained from fruits belonged to Candida intermedia while those isolated from leaves belonged to Wickerhamomyces anomalus. C. intermedia reduced disease incidence caused only by C. gloeosporioides, whereas, W. anomalus caused a significant reduction in the incidence and severity of disease caused by both C. gloeosporioides and C. acutatum. To the best of our knowledge, this is the first report on the antagonistic activity of W. anomalus or C. intermedia against Colletotrichum acutatum. Thus, W. anomalus is a potential natural alternative for controlling anthracnose infection and associated loss in avocado crops.     

Management of the root-knot nematode Meloidogyne incognita on tomato with combinations of different biocontrol organisms

The nematicidal effect of Pseudomonas fluorescens, Paecilomyces lilacinus, Pichia guilliermondii and Calothrix parietina singly or in combination was tested against root-knot nematode, Meloidogyne incognita. Treatments with P. fluorescens and P. lilacinus caused mortality of M. incognita as 45% and 30% of juveniles after 48 h of exposures, respectively compared to water control in vitro. Under greenhouse conditions, all treatments reduced the disease severity and enhanced plant growth compared to untreated control. Application of P. fluorescens, P. lilacinus and P. guilliermondii Moh 10 was more effective compared to C. parietina. There was a negative interaction between C. parietina and either P. lilacinus or P. guilliermondii. Fresh and dry weight of shoots and roots of plants were significantly reduced as a result of infection with M. incognita, however application of biocontrol agents singly or in mix recovered this reduction. Moreover, they enhanced the growth parameters compared with the control. Our results proved that application of different biocontrol agents (P. fluorescens, P. lilacinus and P. guilliermondii) not only has a lethal effect on nematode, but also enhances the plant growth, supplying many nutritional elements and induction the systemic resistance in plants. Presence of C. parietina as a soil inhabitant cyanobacterium could antagonize biocontrol agents leading to the reduction of their practical efficiency in soil.     

The efficacy of combined application of edible coatings and thyme oil in inducing resistance components in avocado (Persea americana Mill.) against anthracnose during post-harvest storage

Avocado fruit has high economic value; however, major post-harvest losses are encountered throughout the supply chain mostly due to anthracnose disease caused by the fungus Colletotrichum gloeosporioides. Increasing consumer concern regarding food safety and demand for organically produced fruits makes it necessary to search for natural environmentally friendly alternative products and processes for the fruit industry; particularly in disease control. Antifungal effects of Gum Arabic (GA) (10%), Aloe vera (AL) (2%), chitosan (CH) (1%) alone or in combination with thyme oil (1%) were investigated in vitro. CH + thyme oil and AL + thyme oil [1:1 or 3:1 v/v] showed fungicidal effects while AL, CH, GA and GA + thyme oil [3:1 v/v] showed fungistatic effects on mycelial growth of C. gloeosporioides in vitro. CH and AL coatings alone or in combination with thyme oil [3:1 v/v], either as preventative or curative treatments in comparison with commercial treatment (prochloraz, 0.05%) and untreated control were evaluated on incidence and severity (lesion diameter) of anthracnose in vivo. Preventative CH + thyme oil treatments significantly reduced the severity of anthracnose (8.9 mm) compared to thyme oil (12.7 mm), AL + TO (14.4 mm), CH (17.8 mm), AL (20.6 mm), PZ (18.3 mm) and untreated samples (34.8 mm). As curative method, the CH + thyme oil combination also reduced the severity of anthracnose by at least 4 mm compared to the other treatments. The total phenols, polyphenol oxidase, phenylalanine ammonia-lyase, β-1,3-glucanase, chitinase, catalase and superoxide dismutase activities, firmness and flesh colour were also determined. Results showed an increase in peroxidase, phenylalanine ammonia-lyase, β-1,3-glucanase, chitinase, catalase and superoxide dismutase activities and total phenolics with reduced loss of firmness and flesh colour following CH + thyme oil treatments. This investigation recommends CH + thyme oil [3:1 v/v] combination treatment as a suitable alternative to the currently adopted prochloraz applications in controlling anthracnose disease in avocado fruit during storage.     

Induction of systemic resistance and defense-related enzymes after elicitation of resistance by rhizobacteria and silicon application against Ralstonia solanacearum in tomato (Solanum lycopersicum)

Bacterial wilt caused by Ralstonia solanacearum is a devastating plant disease affecting several economically important crops. To trigger plant-mediated systemic resistance against the pathogen, silicon and the rhizobacteria strain Bacillus pumilis were applied as elicitors. The impact of each elicitor in the induction of systemic resistance was evaluated by individual or dual application in pathogen inoculated tomato genotypes. Application of silicon and B. pumilis significantly reduced bacterial wilt incidence by 50.7 and 26.7%, respectively, in the King Kong 2 genotype (moderately resistant) and by 31.1 and 22.2%, respectively, in the L390 genotype (susceptible) but the dual application of the elicitors did not. The pathogen responsive lipoxygenase (LOX) activity was decreased in the silicon-treated tomato plants and no significant difference was detected in B. pumilis treated-pathogen inoculated plants compared to the control. In addition, a non-significant increases of peroxidase (PO) activity and phenylalanine ammonia lyase (PAL) activity were observed in either silicon/B. pumilis treated and pathogen inoculated plants. However, the PO, PAL and LOX activities declined during the dual application due to antagonistic effect of the two pathways triggered by the two elicitors. Therefore, separate application of each elicitor is recommended as the best strategy for induction of systemic resistance against bacterial wilt in tomato with silicon being a stronger inducer than B. pumilis.     

Effect of biocontrol agent Bacillus amyloliquefaciens and 1-methyl cyclopropene on the control of postharvest diseases and maintenance of fruit quality

Efficacy of biocontrol agent Bacillus amyloliquefaciens PPCB004 was evaluated on the control of anthracnose and phomopsis rot in ‘Solo’ papaya pre-treated with 1-methyl cyclopropene (100 μl) (1-MCP) during storage. This treatment was compared to the untreated control, commercial treatment (washing in chlorinated water), stand alone 1-MCP and PPCB004 treatment. Although fruit pre-treated with 1-MCP delayed the ripening (100% yellow) after cold storage by 9-10 d, it showed higher incidence and severity of anthracnose and phomopsis rot than the fruit subjected to commercial treatment. Application of PPCB004 after 1-MCP pre-treatment (1-MCP + PPCB004) reduced the anthracnose and phomopsis incidence and severity after cold storage (10 °C, 85% RH for 14 d) and ripening at 25 °C. The 1-MCP + PPCB004 treatment helped to retain the fruit firmness, overall quality and uniform yellow skin (100%) and flesh colour after ripening. The PPCB004 was effectively recovered from stand alone PPCB004 and 1-MCP + PPCB004 treated fruit after cold storage and ripening. The PPCB004 population showed an increase by 1 log units after ripening in 1-MCP + PPCB004 treated fruit. After ripening the recovery of PPCB004 population was higher (0.7 log units) in 1-MCP + PPCB004. The total recovery of fungal population on the fruit surface after ripening was lower in 1-MCP + PPCB004 and stand alone PPCB004 treated fruit. It can be concluded that application of B. amyloliquefaciens PPCB004 with 1-MCP pre-treated papaya (at 25-30% skin yellow stage) can significantly reduce disease incidence associated with 1-MCP treatment. This treatment has the potential for commercial application in the ‘organic’ papaya industry.     

Evaluation of the Ugandan sorghum accessions for grain mold and anthracnose resistance

Sorghum accessions from Uganda were evaluated for grain mold and anthracnose resistance during the 2005 and 2006 growing seasons at the Texas A&M University Research Farm, near College Station, TX. Accession PI534117 and SC719-11E exhibited the lowest grain mold severities of 2.4, whereas, accessions PI534117, PI534144, PI576337, PI297199, PI533833, and PI297210, with SC748-5 were highly resistant to anthracnose in both years. Accessions PI534117, PI297134, PI297156 exhibited low grain mold severities in 2006. Significant negative correlation was recorded between grain mold and percent germination and high temperature in both years. In 2006, daily precipitation recorded significant positive correlation with grain mold. The seed mycoflora was analyzed across sorghum lines and treatments. In 2005, Curvularia lunata and Fusarium thapsinum were the most frequently recovered fungal species with 31 and 21% incidence, respectively, followed by Alternaria spp. (19%) and F. semitectum (13%). In 2006, predominant colonizers were F. thapsinum (58%), followed by Alternaria and F. semitectum with 15 and 10, percent respectively, while C. lunata had a 6% incidence. In this study, PI534117 holds promise for multiple disease resistance, as it had the lowest disease severity of grain mold and was highly resistant to anthracnose in both years. It also has a high germination rate, a high seed weight, and its short stature is more advantageous for the new A-line conversion program.     

Antifungal activity of polyacetylenes isolated from Cirsium japonicum roots against various phytopathogenic fungi

Antifungal substances from a methanol extract of Cirsium japonicum roots were purified and characterized, and their antifungal activities against various plant pathogens were evaluated. Three polyacetylene substances were isolated from roots of C. japonicum using repeated column chromatography; these were identified as ciryneol A, ciryneol C and 1-heptadecene-11,13-diyne-8,9,10-triol by mass and nuclear magnetic resonance spectral analyses. In vitro antifungal activity of the three substances varied according to compound and target species. Magnaporthe oryzae, Colletotrichum coccodes, Colletotrichum acutatum, Pythium ultimum and Botrytis cinerea were relatively sensitive to the three polyacetylenes, with IC₅₀ values below 50 μg mL⁻¹. In vivo, they all showed similar and broad antifungal spectra against the seven plant diseases tested. At 500 μg mL⁻¹, all three compounds effectively suppressed the development of rice blast, rice sheath blight, tomato late blight, wheat leaf rust and red pepper anthracnose, with control values over 90%. They were highly active especially against wheat leaf rust; they controlled the development of this disease more than 88% even at a concentration of 125 μg mL⁻¹. In addition, ciryneol C effectively suppressed barley powdery mildew. This is the first report on the antifungal activities of the three polyacetylenes from roots of C. japonicum against plant pathogenic fungi. Polyacetylenes from roots of C. japonicum may contribute to the development of environmentally safer alternatives to protect crops from various phytopathogenic fungi.     

Antifungal activity of Vitex agnus-castus extract against Pythium ultimum in tomato

Vitex agnus-castus methanolic extract showed strong antifungal activity against Pythium ultimum in tomato under both in vitro and in vivo conditions. The 0.2% extract delayed the mycelial growth of the fungus and showed significant antifungal activity against P. ultimum on tomato seedlings with an efficacy comparable to that of the synthetic fungicide. To determine the involvement both of plant extract and pathogenic fungus in PR gene induction, tomato plants were treated with V. agnus-castus extract and/or inoculated with P. ultimum. The expression of four PR genes (PR-1, PR-2, PR-5, PR-6) was monitored at five time points within 48 h of the extract treatment and fungal inoculation. The PR-1 and PR-4 genes were activated directly by V. agnus-castus extract up to 12 h after treatments; at 24 h, the direct activation by plant extract disappeared and a synergistic inducing effect of extract and pathogen applied simultaneously on the plant was observed. The PR-6 gene was not activated directly by the V. agnus-castus extract but only when applied together with the pathogen; activation of the PR-6 gene occurred 24 h after treatments and the gene expression increased at 48 h. There was no activation of PR-5 gene by the plant extract. The ability of V. agnus-castus extract to enhance plant defence responses upon pathogen inoculation might be further investigated. The activation of various PR genes suggests that induction of defence responses by V. agnus-castus extract in tomato may be regulated by more than one signalling pathway.     

Integration of Pseudomonas fluorescens and acibenzolar-S-methyl to control bacterial spot disease of tomato

Xanthomonas axonopodis pv. vesicatoria was isolated from infected tomato seedlings grown in an open field in Egypt. All the tested isolates infected tomato plants but with different degrees of disease severity. In an attempt to manage this disease, tomato seeds and/or seedlings were treated with an antagonistic local isolate of Pseudomonas fluorescens as a suspension or its formulation or acibenzolar-S-methyl (BTH). When the above three treatments were applied to tomato seeds under laboratory conditions, they improved seed germination and seedlings vigour relative to control seeds treated with sterile distilled water and pathogen but P. fluorescens culture was the most effective. Under greenhouse and field conditions, combinations of the above treatments were used. All treatments significantly reduced disease severity of bacterial spot in tomato relative to the infected control. The biggest disease reduction compared to seedlings inoculated with the pathogen alone resulted from a foliar application of P. fluorescens. Combined application of P. fluorescens or its formulation with BTH reduced the pathogen population and increased seedling biomass and tomato yield relative to control seedlings.     

Races of Colletotrichum graminicola pathogenic to maize in Brazil

Anthracnose caused by the fungus Colletotrichum graminicola (Ces.) Wilson is the most important disease of maize in Brazil, especially in no-tillage without crop rotation. In this system, maize debris from earlier plantings increases the fungal inoculum potential over time. The use of genetic resistance is the most appropriate and advantageous strategy for anthracnose control. However, the effectiveness and durability of this practice depends on knowledge concerning the genetic variability of C. graminicola. In this study, fifteen maize genotypes were tested against 190 single spore of C. graminicola collected from infected leaves of maize plants cultivated in seven different Brazilian ecogeographic areas. Five races of C. graminicola were identified, and eleven maize genotypes were susceptible to all isolates. Results indicated that both the number of pathogen isolates and the number of genotypes to be tested are pivotal for an accurate identification of C. graminicola races. This is the first study showing the occurrence of C. graminicola races in Brazil.     

Insecticidal effects of Ungernia severtzovii bulb extracts against the grain aphid Schizaphis graminum (Rondani)

Insecticidal effect of Ungernia severtzovii bulbs extract has been investigated against the grain aphid Schizaphis graminum. LC₅₀ value of the ethanolic extract of U. severtzovii against the female aphids was 0.235% (2.35 g/l). There was 100% mortality of the subsequent nymphs of the treated females at 0.25-1.0%. The extract was equally active against the nymphs at the highest concentrations. Hexane and ethanolic extracts were the most active against the aphids of all the extracts. Fractionation of the hexane extract by TLC yielded three fractions. Fraction II was the most active (>90% mortality) against the aphids. Xanthoxylin was identified as the major constituent in fraction II of the hexane extract and may be responsible for the insecticidal effect of U. severtzovii.