Identification of Rhizoctonia solani associated with soybean in Brazil by rDNA-ITS sequences

The aim of this study was to identify isolates of Rhizoctonia solani causing hypocotyl rot and foliar blight in soybean (Glycine max) in Brazil by the nucleotide sequences of ITS-5.8S regions of rDNA. The 5.8S rDNA gene sequence (155 bp) was highly conserved among all isolates but differences in length and nucleotide sequence of the ITS1 and ITS2 regions were observed between soybean isolates and AG testers. The similarity of the nucleotide sequence among AG-1 IA isolates, causing foliar blight, was 95.1-100% and 98.5-100% in the ITS1 and ITS2 regions, respectively. The nucleotide sequence similarity among subgroups IA, IB and IC ranged from 84.3 to 89% in ITS1 and from 93.3 to 95.6% in ITS2. Nucleotide sequence similarity of 99.1% and 99.3-100% for ITS1 and ITS2, respectively, was observed between AG-4 soybean isolates causing hypocotyl rots and the AG-4 HGI tester. The similarity of the nucleotide sequence of the ITS-5.8S rDNA region confirmed that the R. solani Brazilian isolates causing foliar blight are AG-1 IA and isolates causing hypocotyl rot symptoms are AG-4 HGI. The ITS-5.8S rDNA sequence was not determinant for the identification of the AG-2-2 IIIB R. solani soybean isolate.

Avaliação de danos causados por duas estirpes do Soybean mosaic virus em duas cultivares de soja

Este estudo objetivou determinar os efeitos, em duas cultivares de soja (Glycine max), causados por duas estirpes do Soybean mosaic virus (SMV) e da idade das plantas na inoculação em relação a parâmetros ligados ao rendimento. Dois experimentos foram conduzidos em casa de vegetação, nos quais as cultivares BRS 183 e UFV-16 Capinópolis foram inoculadas, mecânica e isoladamente, com as estirpes G1 e G5 do SMV, em três diferentes idades após a emergência. Os experimentos foram instalados segundo um delineamento inteiramente casualizado com cinco repetições. Foram avaliados diferentes características relacionadas à produção e qualidade de grãos e constatou-se que: (i) as duas estirpes causam danos sobre diferentes características ligadas ao rendimento e manchas nas sementes; (ii) a estirpe G1 é mais severa que a estirpe G5; (iii) a idade da planta na inoculação do SMV afeta o nível de danos; (iv) a porcentagem de sementes manchadas e o grau de mancha ocorrem com diferentes intensidades de acordo com o ambiente, cultivar, estirpe e idade da planta na inoculação. As características em cada combinação de peso de matéria seca e peso de grãos, peso de raiz e volume de raiz, porcentagem de sementes manchadas e grau de manchas, apresentaram correlações significativas nas duas cultivares, permitindo que se faça opção por apenas uma característica de cada combinação, para avaliar as perdas em plantas infetadas.

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.

Differential responses of nematode communities to soybean genotypes resistant and susceptible to Heterodera glycines race 3

Heterodera glycines and Helicotylenchus dihystera were the two most abundant plant-parasitic nematodes found in two H. glycines race 3-infested fields, Chapadão do Céu, MS and Campo Alegre, MG. These fields had been planted with resistant (R) and susceptible (S) plants to cyst nematodes. In the first field, soybean (Glycine max) FT-Cristalina (S) was susceptible to H. glycines but resistant to H. dihystera, while GOBR93 122243 (R) was just the opposite. In the second field, M-Soy 8400 (R) was more resistant to the spiral nematode than M-Soy8411 (S), but the resistance to the cyst nematode was not different between the two genotypes. The total abundance of nematodes was not different between the susceptible and resistant plants in the two fields, suggesting that H. dihystera and/or bacterial feeders and other trophic groups replaced the reduced abundance of the cyst nematodes in resistant plants. Bacterial feeders acted as a compensatory factor to plant-parasitic nematodes in ecological function. The populations of fungal feeders were higher in GOBR93 122243 (R) than in susceptible FT-Cristalina (S) in Chapadão do Céu, but lower in M-Soy 8400 (R) than in M-Soy 8411 (S) in Campo Alegre. This is being attributed to the different periods of soil samplings that were made at the florescent period in the first field, and at the final growing cycle in the second field. Only four nematodes, H. glycines, H. dihystera, Acrobeles sp. and Panagrolaimus sp. dominated the nematode resistant community GOBR93 122243 (R) in Chapadão do Céu, but dominance was shared by ten genera in Campo Alegre, which explains why the five diversity indexes (S, d, Ds, H' and T) were higher in resistant plants than in susceptible plants in field two.

Detection and partial characterization of a carlavirus causing stem necrosis of soybean in Brazil

Stunting and stem necrosis were noticed in soybeans (Glycine max) grown in 2000/2001 in West Central Brazil the same condition was also observed in the following year in plantations as far as 2,000 km from the initial area. Based on transmission (mechanical, graft, insect vector), purification and serology, electron microscopy and molecular studies the causal agent was determined to be a whitefly-borne carlavirus which is possibly related to Cowpea mild mottle virus (CpMMV).

Diagrammatic scale for assessment of soybean rust severity

A diagrammatic scale to assess soybean (Glycine max) rust severity, caused by the fungus Phakopsora pachyrhizi, was developed in this study. Leaflets showing different severity levels were collected for determination of the minimum and maximum severity limits; intermediate levels were determined according to "Weber-Fechner's stimulus-response law". The proposed scale showed the levels of 0.6; 2; 7; 18; 42, and 78.5%. Scale validation was performed by eight raters (four inexperienced and four experienced), who estimated the severity of 44 soybean leaflets showing rust symptoms, with and without the use of the scale. Except for rater number eight, all showed a tendency to overestimate severity without the aid of the diagrammatic scale. With the scale, the raters obtained better accuracy and precision levels, although the tendency to overestimate was maintained. Experienced raters were more accurate and precise than inexperienced raters, and assessment improvements with the use of the scale were more significant for inexperienced raters.

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.

Powders of kudzu, velvetbean, and pine bark added to soil increase microbial population and reduce Southern blight of soybean

Southern blight (Sclerotium rolfsii) of soybean (Glycine max) is an important disease throughout the world. Some soil amendments can reduce disease levels by improving soil microbial activity. The main goals of this study were to investigate the effects of soil amendments such as dried powders of kudzu (Pueraria lobata), velvetbean (Mucuna deeringiana), and pine bark (Pinus taeda), on soil microbial population and disease caused by S. rolfsii on soybean. Pine bark, velvetbean (mucuna) and kudzu (25 g kg-1) added to soil were effective in reducing disease incidence [non-amended (NA) ~ 39%; amended (A) ~ 2 to 11%)]. Bacillus megaterium was the bacteria most frequently isolated in soils with velvetbean or kudzu (NA ~ log 5.7 CFU g-1 of dried soil; A ~ log 6.2). Soils with velvetbean and kudzu stimulated increase in population of Enterobacter aerogenes (NA ~ log 3; A ~ log 5.1-5.8). Pseudomonas putida population was higher in A than in NA (NA ~ log 4; A ~ log 5.5), and was negatively correlated (r = -0.83, P = 1%) to disease incidence. Soil amended with kudzu and pine bark stimulated increases in populations of Trichoderma koningii (NA ~ log 1.6; A ~ log 2.9) and Penicillium citreonigrum (NA ~ log 1.3; A ~ log 2.6), respectively. Penicillium herquei soil population increased with addition of kudzu (NA ~ log 1.2; A, ~ log 2.5). These microorganisms are antagonists of soil-borne pathogens. Powders of velvetbean, kudzu, and pine bark can increase antagonistic population in soil and reduce disease.

Efficiency of phylloplane bacteria in controlling aerial tomato diseases under field conditions

The capacity of two bacteria isolated from the tomato phylloplane to control late blight (Phytophthora infestans) was investigated in the field, and compared against the effectiveness of spraying with the fungicide chlorothalonil (1.5 g a.i. L-1) or water (control). A 55% reduction in late blight intensity was observed in the leaves of the middle of the plant and 62% in those of the upper leaves when using the antagonist UFV-STB 6 (Novosphingobium capsulatum) as compared to the control. Isolate UFV-IEA 6 (Bacillus cereus) was able to reduce disease intensity by 55%, but only in the upper leaves of the tomato plants. Treatment with isolate UFV-STB 6 also led to a significant reduction in the percentage of fruits with late blight symptoms. The results demonstrate the potential of these two bacteria in controlling this disease.

Management of crotalaria and pigeon pea for control of yam nematode diseases

Management of plant-parasitic nematodes with the use of nematicides has not been recommended for small farmers that grow yam in the Northeastern region of Brazil, due to its high cost and residue toxicity. The use of plants with antagonistic effect to nematodes and green manure which improves soil chemical, physical and biological characteristics can be a viable and low cost alternative to control parasitic nematodes. This work aimed to evaluate the effect of crotalaria (Crotalaria juncea) and pigeon pea (Cajanus cajan) plants on the control of yam nematodes. Three experiments were carried out. The first was conducted under in vitro conditions to evaluate the nematostatic and nematicide effect of extracts from fresh and dry matter of the above ground parts of crotalaria, pigeon pea, and the combination of both. The second experiment was carried out under greenhouse conditions to evaluate the effect of soil amendment with crotalaria, pigeon pea, and the combination of both in the infectivity of Scutellonema bradys, using tomato plants as the host plant. The third experiment was conducted under field conditions to evaluate the effect of crotalaria, pigeon pea, and the combination of both, cultivated between yam planting rows and incorporated to soil surface, on yam nematodes. The aqueous extract obtained form fresh matter of crotalaria had a nematicide effect of 100% for S. bradys. Extracts from dry matter of both crotalaria and pigeon pea did not have any nematicide effect, but had a nematostatic effect. Incorporation of crotalaria to soil inhibited infectivity of S. bradys in tomato seedlings. These results showed that planting crotalaria alone or in combination with pigeon pea, between the yam planting rows, is an efficient method for controlling S. bradys and Rotylenchulus reniformis associated with yams. Crotalaria can be used for controlling these plant-parasitic nematodes in soil.

Decomposition of corn and soybean residues under field conditions and their role as inoculum source

Necrotrophic parasites of above-ground plant parts survive saprophytically, between growing seasons in host crop residues. In an experiment conducted under field conditions, the time required in months for corn and soybean residues to be completely decomposed was quantified. Residues were laid on the soil surface to simulate no-till farming. Crop debris of the two plant species collected on the harvesting day cut into pieces of 5.0cm-long and a 200g mass was added to nylon mesh bags. At monthly intervals, bags were taken to the laboratory for weighing. Corn residues were decomposed within 37.0 months and those of soybean, within 34.5 months. Hw main necrotrophic fungi diagnosed in the corn residues were Colletotrichum gramicola, Diplodia spp. and Gibberella zeae, and those in soybeans residues were Cercospora kikuchii, Colletotrichum spp, Glomerella sp. and Phomopsis spp. Thus, those periods shoulb be observed in crop rotation aimed at to eliminating contaminated residues and, consequently, the inoculum from the cultivated area.

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.

Progress of Asian soybean rust and airborne urediniospores of Phakopsora pachyrhizi in southern Brazil

Asian soybean rust, caused by the fungus Phakopsora pachyrhizi, was reported at epidemic levels in 2003/2004 and is the main soybean disease in Brazil. The aim of this study was to investigate the spread of Asian soybean rust and to quantify airborne urediniospores in the region of Campo Mourão, Paraná State, Brazil. Three experiments were conducted under field conditions during the 2007/08 and 2008/09 crop seasons. Using the disease gradient method, provided by the application of increasing levels of the fungicide tebuconazole, four Asian soybean rust epidemics at different intensities were obtained in each experiment. To quantify the urediniospores, weathercock-type spore collectors were installed during and between the two crop seasons. Disease progress curves were plotted for each epidemic, and maximum severity was estimated. The curves were fit to the logistic model, which provided higher coefficients of determination and more randomly distributed residuals plotted over time. Analyses of the area under the disease progress curve showed that the largest epidemics occurred in the 2007/2008 crop season and that the progress rates were higher for severity, even among plants protected with the fungicide. The number of urediniospores collected in the air was related to the presence of soybean plants in the cultivated crops. The quantity of urediniospores was also positively correlated to the disease severity and incidence, as well as to cumulative rainfall and favorable days for P. Pachyrhizi infection.