Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions

The effect of relative humidity (43%, 75%, 86% and > 98%) on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25ºC. Survival of larvae inside eggs was clearly affected by the lowest humidity (43%) tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia and hyphal bodies tested. This fungus affected larvae inside eggs and has potential as a control agent of this important vector in breeding sites with high moisture.

Effect of two dosages of Metarhizium anisopliae var. acridum against Rhammatocerus schistocercoides Rehn

The fungus Metarhizium anisopliae var. acridum, strain CG 423, was tested under field conditions against the gregarious grasshopper Rhammatocerus schistocercoides (Rehn) (Orthoptera: Acrididae). Conidia formulated in a racemic mixture of soybean oil and kerosene were sprayed under field conditions using an ultralow-volume hand-held atomizer Ulva Plus adjusted to deliver 2.9 L/ha. Bands composed of 2nd instar nymphs were treated with either 5.0x10(12) or 1.0x10(13) viable conidia/ha. The number of insects in each band was estimated at day one following spraying and by the end of the field trial (15 to 16 days post-treatment). Reductions in population size reached, in average, 65.8% and 80.4% for bands treated with the higher and lower dosage, respectively. For both dosages, total mortality rates of insects collected at two days post-application, and kept in cages for 14 days under lab conditions, showed no significant differences as compared to that obtained with insects collected immediately after spraying. Healthy insects were fed to native grasses sprayed on the field with 1.0x10(13) viable conidia/ha. Mortality levels of the nymphs fed on grasses collected two and four days post-application were not affected when compared to nymphs fed on grasses collected immediately following application.

Necrotrophic fungi associated with epidermal microcracking caused by chilling injury in pickling cucumber fruit

The objective of this work was to visualize the association between microcracking and other epidermal chilling injury symptoms, and to identify rots in cucumber fruit (Cucumis sativus L.) by scanning electron microscopy (SEM). Depressed epidermal areas and surface cracking due to damages of subepidermal cells characterized the onset of pitting in cucumber fruit. The germination of conidia of Alternaria alternata, with some of them evident on the fractures in the cultivar Trópico, occurred after damaging on the epidermis. Before, the chilling injury symptoms became visible, Stemphylium herbarum conidia germinated, and mycelium penetrated through the hypodermis using the microcracks as pathway. In the cultivar Perichán 121 the fungus was identified as Botrytis cinerea.

Biological control of white mold by Trichoderma harzianum in common bean under field conditions

Abstract: The objective of this work was to evaluate Trichoderma harzianum isolates for biological control of white mold in common bean (Phaseolus vulgaris). Five isolates were evaluated for biocontrol of white mold in 'Perola' common bean under field conditions, in the 2009 and 2010 crop seasons. A commercial isolate (1306) and a control treatment were included. Foliar applications at 2x109 conidia mL-1 were performed at 42 and 52 days after sowing (DAS), in 2009, and at 52 DAS in 2010. The CEN287, CEN316, and 1306 isolates decreased the number of Sclerotinia sclerotiorum apothecia per square meter in comparison to the control, in both crop seasons. CEN287, CEN316, and 1306 decreased white mold severity during the experimental period, when compared to the control.

Occurence of thrips on Niagara table grape and its control with the insecticides thiacloprid and methiocarb associated with Metarhizium anisopliae

Thrips are reported as important pests on table grapes in United States and several countries of Europe. Damage caused by thrips, particulary Frankliniella occidentalis, was observed on niagara table grape crop in Limeira-SP, Brazil. During the blooming period, high thrips densities were observed feeding on pollen and small berries. The symptoms left were more visible after the development of the berries and were characterized by dark scars and suberized surface on berries, sometimes causing the berry to crack, and the seed to prolapse. The effect of insecticides thiacloprid or methiocarb, associated or not with the entomopathogenic fungus Metarhizium anisopliae were evaluated during the blooming period. For evaluation of thrips damage on fruits, the treatments were applied three additional times, 7, 14 and 21 days after the first application. The treatments were: a) M. anisopliae (strain 1037) 1x10(7) conidia/mL; b) thiacloprid 20mL/100L; c-d) methiocarb 100 and 150mL/100L; e) methiocarb 100mL/100L + M. anisopliae 1x10(7) conidia/mL. Only methiocarb, associated or not with the fungus, was effective in reducing thrips infestation, and no phytotoxic damage was observed. The efficiency of methiocarb 150mL/100L and the insecticide associated with the fungus for the control of the thrips population was 84.2 and 95.5%, respectively. In both cases, there was a reduction of approximately 70% in the number of berries with scars symptoms. For control of thrips on table grapes, chemical insecticides associated or not with M. anisopliae should be applied during the blooming period of the crop.

Histopathology of Colletotrichum gloeosporioides on guava fruits (Psidium guajava L.)

Anthracnose, caused by Colletotrichum gloeosporioides, produces brown lesions on guava fruits, causing severe losses on postharvest. In this study, the infection and colonization of guava fruits by C. gloeosporioides has been examined using scanning and transmission electron microscopy. Fruits at the physiologically mature stage were inoculated with a 10(5) conidia/mL spore suspension. Afterward, fruits were incubated at 25 °C in a wet chamber for periods of 6, 12, 24, 48, 96 and 120 h to allow examination of the infection and colonization process. Conidia germination and appressoria formation occurred six hours after inoculation (h.a.i). Penetration occurred directly via penetration pegs from appressoria, which penetrated the host cuticle 48 h.a.i. Notably, the appressoria did not produce an appressorial cone surrounding the penetration pore. Infection vesicles were found in epidermal cells 96 h.a.i. The same fungal structures were found in epidermal and parenchymal cells of the host 120 h.a.i. Colonization strategy of C. gloeosporioides on guava fruit was intracellular hemibiotrophic.

Synthesis and antifungal activity of halogenated aromatic bis-γ-lactones analogous to avenaciolide

Here we describe the total syntheses and characterization by elemental analyses, infrared and NMR spectroscopy of three new compounds analogous to avenaciolide, a bis-γ-lactone isolated from Aspergillus avenaceus that possesses antifungal activity, where the octyl group of the natural product was replaced by aromatic groups containing chlorine and fluorine atoms. The effects of the avenaciolide, the novel compounds and their synthetic precursors on mycelia development and conidia germination of Colletotrichum gloeosporioides and Fusarium solani were evaluated in vitro. The title compounds were almost as active as avenaciolide. The absolute structures of the chlorinated analogs were determined by X-ray diffraction analysis.

In vitro activity of essential oils of Lippia sidoides and Lippia gracilis and their major chemical components against Thielaviopsis paradoxa, causal agent of stem bleeding in coconut palms

Essential oils of Lippia sidoides, Lippia gracilis and their main chemical components were investigated for in vitro control of Thielaviopsis paradoxa. Mycelial growth and a number of pathogen conidia were inhibited by the essential oil of L. sidoides at all concentrations tested (0.2; 0.5; 1.0; 3.0 µL mL-1). L. sidoides oil contained 42.33% thymol and 4.56% carvacrol, while L. gracilis oil contained 10% thymol and 41.7% carvacrol. Mycelial growth and conidial production of T. paradoxa were completely inhibited by thymol at a 0.3 µL m-1 concentration. The results suggest that thymol could potentially be used for controlling coconut stem bleeding.

Cytogenetics of Colletotrichum lindemuthianum (Glomerella cingulata f. sp. phaseoli)

Cytogenetic and morphological studies were conducted with Colletotrichum lindemuthianum (Glomerella cingulata f. sp. phaseoli), the pathogen responsible for anthracnose of common bean (Phaseolus vulgaris). In this species, there is some evidence of genomic variation but it is unknown whether the process occurs in a manner similar to other fungal genetic models. Six isolates from bean plants were used and sexual reproduction was observed in vitro. Meiosis and ascospore formation were investigated by cytogenetical approaches and light microscopy. To study the nucleus and chromosome numbers, a mixture of carmine and orcein propionic dyes was used. Nucleus divisions as well as ascospore maturation were asynchronous. Meiosis was observed in three isolates. In the asexual form, chromosomal polymorphism in conidia was also observed microscopically and the mitosis process was described.

Detection and transmission of Drechslera avenae from oat seed

Detection, symptoms and symptomless transmission of Drechslera avenae (teleomorph Pyrenophora avenae) from seed were investigated. The present study reported that the D. avenae is frequently found in oat (Avena sativa) seed in Argentina. The prevalence of the seed lot infected was 54%. The incidence of seeds infected by D. avenae in the various seed lots from different regions ranged from 0 to 52% (overall mean of 7%). Besides conidia on conidiophore and immature pseudothecia, D. avenae produced small, spherical to pear-shape picnidia containing tiny conidia, on the seed surface. The pathogen was efficiently transmitted at a rate of 55% from seed to coleoptile tips in symptomless seedlings and at 12.5% to plumules. The importance of the infected seed and its epidemiological role are discussed.

Soybean pod blight and root rot caused by lineages of the Fusarium graminearum and the production of mycotoxins

Surveys of soybean (Glycine max) seed grown in South Brazil revealed infection with Fusarium graminearum. To determine if members of this complex were pathogenic to soybean, six strains derived from soybean were added to soil at a rate of 10³ macroconidia/ ml or individual pods were inoculated with 10(4) macroconidia/ml. Seedlings grown in infested soil developed small necrotic lesions in the crown and upper roots. Pods inoculated with conidia developed large (>1 cm), dark brown, necrotic lesions. Younger pods inoculated with the fungus blighted and dropped from the plant. Strains of the F. graminearum complex recovered from lesions on the crown, roots and pods of soybean plants were identified as lineage 1, 2 or 8 by obtaining the DNA sequence from the EF1-alpha gene and comparing it to strains of the known lineage. Two strains of F. graminearum lineage 7 from the U.S. caused similar symptoms of the disease on soybean. Mycotoxin tests on soybean and wheat (Triticum aestivum) indicate that most Brazilian strains produce nivalenol as the major trichothecene mycotoxin rather than deoxynivalenol. In addition, strains from lineages 2 and 8 produce the novel trichothecene, 3-acetylnivalenol.

New hosts of Oidiopsis haplophylli in the solanaceae family in Brazil

A powdery mildew disease was observed on leaves of Solanum gilo, S. melongena, S. tuberosum S. chacoense, Nicotiana rustica and N. tabacum in Brasília (Federal District), Brazil. Symptoms were mainly characterized by adaxial yellow areas in the leaves corresponding to white fungal colonies on the abaxial surface. Profuse sporulation was often observed. Light microscopy of the fungal colonies revealed the presence of conidiophores emerging through stomata with some having two or three branches. Ellipsoidal, subhyaline conidia were predominantly born singly and terminally on the conidiophore. All morphometrical characteristics agreed with those of Oidiopsis haplophylli (Syn. O. sicula). The teleomorph (Leveillula taurica) was not observed. Inoculation tests indicated that O. haplophylli isolates obtained from S. gilo, S. melongena, S. tuberosum, S. chacoense, Nicotiana rustica and N. tabacum were also pathogenic to sweet pepper (Capsicum annuum) and tomato (Lycopersicon esculentum). This is apparently the first report of these Solanaceae species as hosts of O. haplophylli in Brazil. This disease may become important in these crops, especially in greenhouses, and in hot and dry areas where drip irrigation is employed.

Potential of non-pathogenic Fusarium oxysporum isolates for control of Fusarium wilt of tomato

This study was done to evaluate the efficiency of non-pathogenic Fusarium oxysporum isolates (141/3, 233, 233/1, 245, 245/1, 251, 251/2, 251/5, and 257) in controlling vascular wilt caused by F. oxysporum f. sp. lycopersici, race 2 (isolates C-21A, TO11, and TO245) in tomato (Lycopersicon esculentum) cv. Viradoro seedlings. In order to determine the effect of non-pathogenic F. oxysporum isolates in tomato plants, the root system of 30-day-old seedlings was immersed in conidial suspensions (10(6) ml-1) of each isolate and the seedlings were transplanted to a cultivation substrate. Thirty-five days after transplanting it was observed that the non-pathogenic F. oxysporum isolates were not pathogenic to the cv. Viradoro nor did they affect seedling development. The efficiency of the non-pathogenic F. oxysporum isolates in controlling Fusarium wilt was determined by immersing the tomato seedling roots in the conidial suspension (10(6) ml-1) of each isolate and then transplanting them into substrates previously infested with isolates of F. oxysporum f.sp. lycopersici, race 2 (10(5) conidia ml-1 of substrate). Evaluations were performed 35 days after transplanting, for severity in scale with 1=healthy plant to 6=dead plant or plant showing vessel browning and wilted leaves up to the leader shoot and seedling height. The non-pathogenic F. oxysporum isolates were efficient in reducing the severity of the disease and maintaining normal plant development. These results provide evidence of the antagonistic activity of non-pathogenic F. oxysporum isolates in controlling vascular wilt caused by F. oxysporum f. sp. lycopersici race 2 in tomato.

Susceptibility of fruits of the 'Valência' and 'Natal' sweet orange varieties to Guignardia citricarpa and the influence of the coexistence of healthy and symptomatic fruits

The importance of the presence of symptomatic fruits with citrus black symptoms (CBS) on the disease severity level in subsequent crop production was assessed in an orchard planted with Valência and Natal sweet orange varieties. Additionally, the period of susceptibility of the fruits of these varieties was evaluated. Fruits were covered with paper bags at the stage of 75% fallen petals and were then exposed to natural infection at weekly intervals, from October 2000 to April 2001. This process was carried out in plants where the fruits from the previous harvest had been picked as well as in plants where fruits remained until natural drop. The evaluation of disease severity used a scale that varied from 0 (absence of symptoms) to 6 (severe symptoms). It was observed that, for the Valência and Natal varieties, conidia of Phyllosticta citricarpa that had formed on the lesions of fruits from the previous harvest did not significantly increase the severity of disease on the fruits of the subsequent harvest period. In this study, the protection of the fruits until 10 weeks after petal drop did not affect the number of lesions, indicating that ascospore discharges after that date were, probably, responsible for disease severity. Fruits exposed between the 20th and 24th week after 75% of the petals had fallen were symptomatic, indicating that, at this stage, the fruits were still susceptible to the pathogen.

Use of spectrophotometry as a tool to quantify the sporulation of Penicillium allii in garlic lesions

A technique to measure the concentration of Penicillium allii conidia in damp chamber experiments by spectrophotometry was developed. A negative linear correlation (R²=0.56) was observed between transmittance at 340 nm and the concentration of P. allii conidia in water agar 0.05%. The equation that relates transmittance (T) with concentration (conidia mL-1) (y) is: y = 9.3 10(6) - 86497 T. The method was assayed by inoculating 43 P. allii strains in two garlic cultivars. The method proved to be more rapid than the traditional use of a hemocytometer with an improved accuracy. The CV of the number of conidia per hemocytometer reticule was of 35.04%, while the transmittance CV was of 2.73%. The extreme values chosen for T were 40 and 80 because the sensitivity of the method decreased when concentrations of conidia were out of this range.