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.

Pathogenicity evaluation of Cytospora eucalypticola isolated from Eucalyptus spp: cankers in Uruguay

Cytospora eucalypticola has been frequently associated with twig and stem cankers and as endophyte of Eucalyptus globulus and E. grandis in Uruguay. Mycelium discs of two C. eucalypticola isolates obtained from actively growing colonies were inoculated, both superficially and on experimentally wounded stems of E. globulus and E. grandis. No inoculated and control plants have shown any discoloration, gumosis or necrosis nor did they display lesions ten months after inoculation. Callus tissue was formed, partially or wholly occluding the wounds. The ability to penetrate healthy tissues and the inability to produce lesions evidenced that the presence of C. eucalypticola in twig and stem cankers could result from saprotrophic expansion of the endophytic mycelium in dying tissues, cankers probably being produced by different environmental stress conditions.

Development of virus resistant transgenic papayas expressing the coat protein gene from a Brazilian isolate of Papaya ringspot virus

Translatable and nontranslatable versions of the coat protein (cp) gene of a Papaya ringspot virus (PRSV) isolate collected in the state of Bahia, Brazil, were engineered for expression in Sunrise and Sunset Solo varieties of papaya (Carica papaya). The biolistic system was used to transform secondary somatic embryo cultures derived from immature zygotic embryos. Fifty-four transgenic lines, 26 translatable and 28 nontranslatable gene versions, were regenerated, with a transformation efficiency of 2.7%. Inoculation of cloned R0 plants with PRSV BR, PRSV HA or PRSV TH, Brazilian, Hawaiian and Thai isolates, respectively, revealed lines with mono-, double-, and triple-resistance. After molecular analysis and a preliminary agronomic evaluation, 13 R1 and R2 populations were incorporated into the papaya-breeding program at Embrapa Cassava and Tropical Fruits, in Cruz das Almas, Bahia, Brazil.

Effect of extracts from citric biomass, rusted coffee leaves and coffee berry husks on Phoma costarricencis of coffee plants

Phoma leaf spot, caused by Phoma costarricensis poses a serious threat to coffee (Coffea arabica) production, especially in the highlands of the state of Minas Gerais, Brazil. Extracts of citric biomass, coffee berry husks and coffee leaves severely affected by rust caused by Hemileia vastatrix, were evaluated against P. costarricensis. In an in vitro assay, aqueous extracts of rusted leaves and berry husks plus the commercial extracts based on citric biomass named Ecolife® and Agromil® were tested at various dilutions on the mycelial growth inhibition of P. costarricensis. In vivo, coffee seedlings maintained in glasshouse, were sprayed with these extracts seven days before inoculation of P. costarricensis. Only extracts from citric biomass had inhibitory effects on the fungus. In vivo, Ecolife® (5 ml/l), Agromil® (5 g/l) and the aqueous extract of rusted coffee leaves (dilution 1:6) reduced Phoma leaf spot. Both, Ecolife® and the extract of rusted coffee leaves were significantly more effective in reducing the area under the lesion progress curve when applied at lower doses, indicating a possible effect on the induction of resistance.

Genetic diversity of begomoviruses infecting soybean, bean and associated weeds in Northwestern Argentina

The subtropical Northwestern region of Argentina (provinces of Tucumán, Salta, Jujuy, Santiago del Estero and Catamarca) suffers from a high incidence of the whitefly Bemisia tabaci, and the detection of begomoviruses is also common. The Northwest is the main bean-growing region of the country, and approximately 10% of Argentina's soybean crop is grown in this area. We have used a PCR-based assay to establish the identity and genetic diversity of begomoviruses associated with bean and soybean crops in Northwestern Argentina. Universal begomovirus primers were used to direct the amplification of a fragment encompassing the 5' portion of the capsid protein gene. Amplified fragments were cloned, sequenced and subjected to phylogenetic analysis to determine the sequence identity to known begomoviruses. The data indicated the presence of four distinct begomoviruses, all related to other New World begomoviruses. The prevalent virus, which was present in 94% of bean and soybean samples and also in two weed species, is closely related to Sida mottle virus (SiMoV). A virus with high sequence identity with Bean golden mosaic virus (BGMV) was found in beans. The two remaining viruses displayed less than 89% identity with other known begomoviruses, indicating that they may constitute novel species. One of these putative novel viruses was detected in bean, soybean and tomato samples.

Annona squamosa, a new host of Ceratocystis fimbriata

Mango branch blight disease, caused by Ceratocystis fimbriata, is endemic to the municipality of São Fidelis in northern Rio de Janeiro State. In addition to mango, C. fimbriata was found associated with sugar apple trees (Annona squamosa) showing symptoms of branch blight in São Fidelis. Sugar apple and mango isolates from the same region had the same morphology and showed similar ITS-rDNA sequences. These sequences were also similar to other Brazilian isolates of C. fimbriata sensu stricto. Cross inoculation of such isolates obtained from diseased sugar apple and mango resulted in diseased symptoms on both plant species. This is the first record of A. squamosa as a host for C. fimbriata.

Natural variability in Arabidopsis thaliana germplasm response to Xanthomonas campestris pv. campestris

This work aimed to study the interaction between the model plant Arabidopsis thaliana and Xanthomonas campestris pv. campestris (Xcc), the pathogen responsible for black rot of crucifers. The response of 32 accessions of A. thaliana to the Brazilian isolate of Xcc CNPH 17 was evaluated. No immunity-like response was observed. "CS1308", "CS1566" and "CS1643" grown in continuous light were among the accessions that showed strongest resistance when inoculated with 5 x 10(6) CFU/mL. In contrast, "CS1194" and "CS1492" were among the most susceptible accessions. Similar results were obtained when plants were grown under short-day conditions. To quantify the differences in disease symptoms, total chlorophyll was extracted from contrasting accessions at different time points after inoculation. Chlorophyll levels from controls and Xcc inoculated plants showed a similar reduction in resistant accessions, whereas Xcc-inoculated susceptible accessions showed a greater reduction compared to controls. To test the specificity of resistance, accessions CS1308, CS1566, CS1643 and CS1438 (which showed partial resistance to CNPH 17), were inoculated with a more aggressive isolate of Xcc (CNPH 77) and Ralstonia solanacearum. Among the accessions tested, "CS1566" was the most resistant to Xcc CNPH 77 and also displayed resistance to R. solanacearum. Accessions CS1308, CS1566 and CS1643 were also inoculated with a high titer of Xcc CNPH 17 (5 x 10(8) CFU/mL). No collapse of tissue was observed up to 48 h after inoculation, indicating that a hypersensitive response is not involved in the resistance displayed by these accessions.

Resistance to Meloidogyne mayaguensis in Psidium spp. Accessions and their grafting compatibility with P. guajava cv. Paluma

Meloidogyne mayaguensis has been reported in some states of Brazil causing severe damage on commercial guava (Psidium guajava L.). Accessions of Psidium spp. were selected from a collection maintained in Embrapa Clima Temperado (Pelotas, Rio Grande do Sul State). Plants of different accessions were grown from seed in plastic bags and, when they reached 15-20 cm in height, were inoculated with 10,000 eggs/plant of M. mayaguensis. Eight months after inoculation, the different accessions were evaluated for resistance to M. mayaguensis. Three accessions of P. guajava were highly susceptible (RF=59.2) to this nematode. Psidium friedrichsthalianium was considered to be moderately resistant (RF=1.9). Three accessions of P. cattleyanum were immune to M. mayaguensis (RF = 0). When used as rootstocks P. cattleyanum and P. friedrichsthalianium were compatible with P. guajava cv. Paluma. Considering these results, the use of resistant rootstocks provides a promising control method for M. mayaguensis in commercial guava crop.

Is it necessary to soak rice grains to prepare Rhizoctonia solani AG-4 inoculum?

A study was conducted to evaluate the effects on the development of root rot on common bean, cv. 'Dufrix' after treatment with four volumes of water (0, 30%, 60%, and 90%, v/w) added to rice grains previously immersed in water for 24 hours before autoclaving and colonization of grains by Rhizoctoniasolani AG-4. Colonized rice grains and non-infested rice grains were mixed in pots with sterilized soil and sand (2:1), where beans were sown. Based on results of area under plant emergence curve, plant height, plant dry weight, and disease severity, we conclude that inoculum is more effective in causing disease when no water is added to the rice grains before autoclaving.

Quantitative control of Lettuce mosaic virus fitness and host defence inhibition by P1-HCPro

Two Lettuce mosaic virus isolates capable of overcoming the resistance afforded by the resistance gene mo1² in lettuce, LMV-AF199 from Brazil, and LMV-E, an European isolate, were evaluated for the rapidity and severity of symptoms induced on the lettuce variety Salinas 88 (mo1²). The mosaic symptoms on Salinas 88 plants inoculated with LMV-AF199 appeared 7 days post-inoculation (dpi) and 15 dpi for LMV-E. The symptoms induced by LMV-AF199 in this cultivar were also more severe than those induced by LMV-E. In order to identify the region of the viral genome responsible for this phenotype, recombinant viruses were constructed between these isolates and the phenotype of each recombinant was analysed. The region encoding proteins P1 and HcPro from LMV-AF199 was associated with the increased virulence in Salinas 88.

Effect of Potato virus X on total phenol and alkaloid contents in Datura stramonium leaves

The present paper reports results of the effect of Potato virus X (PVX) on the contents of total phenols and alkaloids in leaves of Datura stramonium. A significant decrease in the contents of phenols and alkaloids was observed in leaves inoculated with PVX (X-I). However, there was an increase in the percentage of phenols in leaves rubbed with phosphate buffer (C1-I) and in leaves from the nodes immediately above, possibly induced by mechanical injury. Gas chromatography/mass spectroscopy revealed amounts of scopolamine in samples submitted to all treatments, except X-I, in which the amount of this alkaloid was low. High amounts of an unidentified compound (molecular ion m/z 302 and a prominent peak at m/z 129) were noted in extracts from leaves X-I, C1-I and leaves from the nodes immediately above the leaves inoculated with PVX. It is suggested that the synthesis and accumulation of the unidentified compound is a result of stress from mechanical injury and virus inoculation.

Characterization of Xanthomonas axonopodis pv. phaseoli isolates

A simple, quick and easy protocol was standardized for extraction of total DNA of the bacteria Xanthomonas axonopodis pv. phaseoli. The DNA obtained by this method had high quality and the quantity was enough for the Random Amplified Polymorphic DNA (RAPD) reactions with random primers, and Polymerase Chain Reaction (PCR) with primers of the hypersensitivity and pathogenicity gene (hrp). The DNA obtained was free of contamination by proteins or carbohydrates. The ratio 260nm/380nm of the DNA extracted ranged from 1.7 to 1.8. The hrp gene cluster is required by bacterial plant pathogen to produce symptoms on susceptible hosts and hypersensitive reaction on resistant hosts. This gene has been found in different bacteria as well as in Xanthomonas campestris pv. vesicatoria (9). The primers RST21 and RST22 (9) were used to amplify the hrp gene of nine different isolates of Xanthomonas axonopodis pv. phaseoli from Botucatu, São Paulo State, Brazil, and one isolate, "Davis". PCR amplified products were obtained in all isolates pathogenic to beans.

Semi-selective medium for Fusarium graminearum detection in seed samples

Fungi of the genus Fusarium cause a variety of difficult to control diseases in different crops, including winter cereals and maize. Among the species of this genus Fusarium graminearum deserves attention. The aim of this work was to develop a semi-selective medium to study this fungus. In several experiments, substrates for fungal growth were tested, including fungicides and antibiotics such as iprodiona, nystatin and triadimenol, and the antibacterial agents streptomycin and neomycin sulfate. Five seed samples of wheat, barley, oat, black beans and soybeans for F. graminearum detection by using the media Nash and Snyder agar (NSA), Segalin & Reis agar (SRA) and one-quarter dextrose agar (1/4PDA; potato 50g; dextrose 5g and agar 20g), either unsupplemented or supplemented with various concentrations of the antimicrobial agents cited above. The selected components and concentrations (g.L-1) of the proposed medium, Segalin & Reis agar (SRA-FG), were: iprodiona 0.05; nystatin 0,025; triadimenol 0.015; neomycin sulfate 0.05; and streptomycin sulfate, 0.3 added of ¼ potato sucrose agar. In the isolation from seeds of cited plant species, the sensitivity of this medium was similar to that of NSA but with de advantage of maintaining the colony morphological aspects similar to those observed in potato-dextrose-agar medium.

Use of botanical extracts, cassava wastewater and nematicide for the control of root-knot nematode on carrot

Aqueous extracts of several plant species have shown promising in controlling root-knot nematode, Meloidogyne incognita (Kofoid & White), one of the most limiting agents for carrot cultivation. The current study evaluated the effect of aqueous extracts from seven botanical species applied to 40, 50, 60, 70 and 80 days after sowing 'Nantes' carrots in soil infested with root-knot nematode. Three other treatments included cassava wastewater, distilled water (control), which were applied in the same periods of the extracts application, in addition to carbofuran 50G (80Kg/ha), which was applied once at 60 days after carrot sowing. Evaluations were performed at 90 days after inoculation to determine shoot and root fresh weight, as well as the diameter and the length of principal roots and the number of galls on primary and secondary roots. Plants treated with cassava wastewater, extracts of Ricinus communis L. seeds, Crotalaria juncea L. seeds, R. communis leaves + branches + fruits, Chenopodium ambrosioides L. leaves + branches + inflorescences and Azadirachta indica A. Juss. seeds showed the highest rates of total weight (root + shoot) and shoot weight. The extract of R. communis leaves + branches + fruits provides the highest total root weight and principal root diameter. Cassava wastewater and extracts of R. communis seeds provided the highest principal root weight. The extract of R. communis seeds and cassava wastewater can be considered promising for the alternative control of M. incognita.

Adjuvant concentrations and uredospore densities on Phakopsora pachyrhizi infection efficiency in soybean

Experiments were carried out in a growth chamber to evaluate the effect of spreader and uredospore concentrations on the efficiency of infection by Phakopsora pachyrhizi, the causal agent of Asian soybean rust. CD 214 RR soybean cultivar was inoculated with the following polyoxyethylene sorbitane monolaurate concentrations: 0, 30, 60, 120, 240, 480 and 960 µL.L-1 water, as well as a fixed uredospore concentration of 2 x 10(4) spores.mL-1. In a second phase, the inoculum concentrations of 0, 5 x 10³, 1 x 10(4), 2 x 10(4), 4 x 10(4), 8 x 10(4) and 16 x 10(4) uredospores.mL-1 were evaluated, and the spreader concentration of 240 µL.L-1, selected in the previous experiment, was fixed. The spreader concentration of 240 µL.L-1 can be used in artificial inoculation studies, as well as up to 4 x 10(4) uredospores.mL-1. In this work, there was a correlation between uredia and lesion density. Thus, the use of lesion density is recommended to assess disease intensity for its accuracy and less time consuming. There was also a positive correlation between uredia and lesion density.