Avaliação do potencial antifúngo de extratos de espécies vegetais bioativas no crescimento micelial e produção de escleródios de Sclerotium rolfsii Sacc. e Sclerotinia sclerotiorum (Lib.) de Bary.

2015

Controle do mofo branco (Sclerotinia sclerotiorum) do feijoeiro com o fungicida procimidone aplicado em pulverização e fungigação

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The pgip family in soybean and three other legume species: evidence for a birth-and-death model of evolution

Background Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat (LRR) plant cell wall glycoproteins involved in plant immunity. They are typically encoded by gene families with a small number of gene copies whose evolutionary origin has been poorly investigated. Here we report the complete characterization of the full complement of the pgip family in soybean (Glycine max [L.] Merr.) and the characterization of the genomic region surrounding the pgip family in four legume species. Results BAC clone and genome sequence analyses showed that the soybean genome contains two pgip loci. Each locus is composed of three clustered genes that are induced following infection with the fungal pathogen Sclerotinia sclerotiorum (Lib.) de Bary, and remnant sequences of pgip genes. The analyzed homeologous soybean genomic regions (about 126 Kb) that include the pgip loci are strongly conserved and this conservation extends also to the genomes of the legume species Phaseolus vulgaris L., Medicago truncatula Gaertn. and Cicer arietinum L., each containing a single pgip locus. Maximum likelihood-based gene trees suggest that the genes within the pgip clusters have independently undergone tandem duplication in each species. Conclusions The paleopolyploid soybean genome contains two pgip loci comprised in large and highly conserved duplicated regions, which are also conserved in bean, M. truncatula and C. arietinum. The genomic features of these legume pgip families suggest that the forces driving the evolution of pgip genes follow the birth-and-death model, similar to that proposed for the evolution of resistance (R) genes of NBS-LRR-type.

Qualidade fisiológica e sanitária de sementes de feijoeiro (Phaseolus vulgaris L.) provenientes do estado de Goiás

The common beans (Phaseolus vulgaris L.) is a fabacea sufficiently spread out in all domestic territory. However, the quality of its seeds represents one of the main causes of low productivity in the beans farmings in Brazil. The objective of this work was to evaluate physiological and sanitary seed qualities of eleven bean cultivars. The physiological seed quality was evaluated trough standard germination and vigor tests. The sanitary seed quality was evaluated through two tests: blotter test was employed to evaluate fungi incidence and Koch & Menten method was employed to observe Sclerotinia sclerotiorum (Lib) de Bary occurrence. Xamego, BRS Valente, Bambu and Pérola had the best results of physiological tests. Jalo Precoce, Roxo 90, Corrente and Aporé had no good results of vigor and germination, besides presenting the lowest indices of died seeds. Fusarium sp., Aspergillus spp., Penicillium sp., Phoma sp., Rhizopus sp. and Botrytis sp. were the fungi detected in the sanity tests.

Fungigação no controle do mofo branco e produtividade do feijoeiro em condições de cerrado brasileiro

The aim of this research was to evaluate the white mold severity (Sclerotinia sclerotiorum (Lib.) of Bary), bean production components and yield (Phaseolus vulgaris L.), variety Perola, according to the application of procimidone fungicide (Sialex 500), through fungigation (center pivot) and automotive sprayer (Uniport). The study was carried under field production commercial conditions, in Primavera do Leste - MT - Brazil. The experiment consisted of 5 treatments (with 4 repetitions of 4 ha each), all with two procimidone applications (1.2 kg ha-1 each application, same as, 0.6 kg a.i. per hectare) to the 42 and 52 days after seeding. The water depths of 5.5 and 11.0 mm were tested in the application through central pivot (this had your checked uniformity), providing volumes of 55.000 and 110.000 L ha-1, respectively, and the volumes of 120 and 200 L ha-1 in the automotive sprayer. The severity of disease was evaluated by the percentage of the area affected by plant damage using diagramatic grade scale of white mold severity, as described by Azevedo (1998). The values were used to calculate the area under the disease progress curve (AUDPC). They were also analyzed, the number of the fungus apothecia during the crop cycle and the residual sclerotias weight in harvest. On this occasion, it was also evaluated the crop yield parameters: number of plants per plot (final stand), pods per plant, grains per pod, medium weight of 200 grains and productivity of grains. The AUDPC values, apothecia to 42, 49 and 56 days after seeding, sclerotias in 2 soil kg and the crop productivity parameters were submitted to the variance analysis and Tukey Test at 0.05 of probability. This test was also applied in the comparison among the different fungicide application methods, independent of spray volumes in each one. The statistical processing was accomplished by STAT program. The results showed that weren't differences among application techniques studied in relation to productivity parameters, however, best white mold control, smaller apothecia number to 49 and 56 days after seeding and smaller weight of residual sclerotias in the harvest were obtained with the fungigation, independently of the spray volume used.

Desempenho de fosfitos de potássio no manejo de mofo branco em soja

Soybean plays an important role in the Brazilian agriculture being one of the products most exported by the country. Its yield may be affected by diseases such as white mold, caused by the fungus Sclerotinia sclerotiorum Lib. de Bary, which, under favorable field conditions prevents the crop of expressing all its productive potential. The fungus is cosmopolitan and infects more than 400 species of plants. This disease is difficult to control, and the use of chemicals has not been sufficient to avoid significant losses, thus, this products are expensive and may cause environmental damage. Alternative methods, such as foliar fertilizers based on potassium phosphite, can also be used in the management of this disease. In this context, this work aimed to study different sources of potassium phosphite and its effects in the control of white mold in soybeans, as well as the time of application in culture, its action in inducing plants defense responses and/or its influence over the seeds quality. The effect of phosphites, over the pathogen, was evaluated in vitro, on mycelial inhibition, the mass of dry mycelium and germination of sclerotia. In all tests, the following phosphites were utilized: Phosphite A (P2O5-40%; K2O-20% - 1 L/ha); Phosphite B (P2O5-40%; K2O-28% - 1 L/ha); Phosphite C (P2O5-40%; K2O-20% - 1 L/ha) e Phosphite D (P2O5-30%; K2O-20% - 2,4 L/ha). At the induction of resistance tests were evaluated the synthesis of phytoalexin in soybean cotyledons and the enzymes FAL and POX evaluated in seedlings in growing chamber, sprayed with phosphites and the fungicide fluazinam. Field experiment was carried out at Coronel Domingos Soares-PR, in the 2012/2013 season, in an area with natural infestation of the pathogen. Soybean cultivar BMX Active was no-till seeded with 0,5m between rows. The experimental was laid out as a factorial 5 x 4 scheme (treatment x application time). Phosphites sources were used, as described above, and water was sprayed in the control treatment. Treatments were applied at four different growth stages: V4, V4 + R1, R1 and R2 at the rates recommended by the manufacturer. Soybean yield components and seeds and health and physiological quality were evaluated after harvesting. None of the tested phosphites affected mycelial growth and sclerotia germination or influenced phytoalexin synthesis. Phosphites C and D stood out due to an increasing in the phenylalanine ammonia-lyase activity 48 hours after its inoculation. These same products also induced the synthesis and peroxidases and phosphite C kept the levels of this enzyme elevated up to 72 hours after inoculation. At the field trials, phosphites C and D stood out in the control of white mold. There was no significant interaction of potassium phosphite on physiological and sanitary quality of the seeds.

Sequential sampling of soybean and beans seeds for Sclerotinia sclerotiorum detection (Lib.) DeBary

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cell death control : the interplay of apoptosis and autophagy in the pathogenicity of sclerotinia sclerotiorum

Programmed cell death is characterized by a cascade of tightly controlled events that culminate in the orchestrated death of the cell. In multicellular organisms autophagy and apoptosis are recognized as two principal means by which these genetically determined cell deaths occur. During plant-microbe interactions cell death programs can mediate both resistant and susceptible events. Via oxalic acid (OA), the necrotrophic phytopathogen Sclerotinia sclerotiorum hijacks host pathways and induces cell death in host plant tissue resulting in hallmark apoptotic features in a time and dose dependent manner. OA-deficient mutants are non-pathogenic and trigger a restricted cell death phenotype in the host that unexpectedly exhibits markers associated with the plant hypersensitive response including callose deposition and a pronounced oxidative burst, suggesting the plant can recognize and in this case respond, defensively. The details of this plant directed restrictive cell death associated with OA deficient mutants is the focus of this work. Using a combination of electron and fluorescence microscopy, chemical effectors and reverse genetics, we show that this restricted cell death is autophagic. Inhibition of autophagy rescued the non-pathogenic mutant phenotype. These findings indicate that autophagy is a defense response in this necrotrophic fungus/plant interaction and suggest a novel function associated with OA; namely, the suppression of autophagy. These data suggest that not all cell deaths are equivalent, and though programmed cell death occurs in both situations, the outcome is predicated on who is in control of the cell death machinery. Based on our data, we suggest that it is not cell death per se that dictates the outcome of certain plant-microbe interactions, but the manner by which cell death occurs that is crucial.

Tipping the balance : sclerotinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment

Sclerotinia sclerotiorum is a necrotrophic ascomycete fungus with an extremely broad host range. This pathogen produces the non-specific phytotoxin and key pathogenicity factor, oxalic acid (OA). Our recent work indicated that this fungus and more specifically OA, can induce apoptotic-like programmed cell death (PCD) in plant hosts, this induction of PCD and disease requires generation of reactive oxygen species (ROS) in the host, a process triggered by fungal secreted OA. Conversely, during the initial stages of infection, OA also dampens the plant oxidative burst, an early host response generally associated with plant defense. This scenario presents a challenge regarding the mechanistic details of OA function; as OA both suppresses and induces host ROS during the compatible interaction. In the present study we generated transgenic plants expressing a redox-regulated GFP reporter. Results show that initially, Sclerotinia (via OA) generates a reducing environment in host cells that suppress host defense responses including the oxidative burst and callose deposition, akin to compatible biotrophic pathogens. Once infection is established however, this necrotroph induces the generation of plant ROS leading to PCD of host tissue, the result of which is of direct benefit to the pathogen. In contrast, a non-pathogenic OA-deficient mutant failed to alter host redox status. The mutant produced hypersensitive response-like features following host inoculation, including ROS induction, callose formation, restricted growth and cell death. These results indicate active recognition of the mutant and further point to suppression of defenses by the wild type necrotrophic fungus. Chemical reduction of host cells with dithiothreitol (DTT) or potassium oxalate (KOA) restored the ability of this mutant to cause disease. Thus, Sclerotinia uses a novel strategy involving regulation of host redox status to establish infection. These results address a long-standing issue involving the ability of OA to both inhibit and promote ROS to achieve pathogenic success.

Comparative growth and pathogenicity of geographical isolates of Sclerotinia sclerotiorum on lentil genotypes

Isolates of Sclerotinia sclerotiorum were collected from infected lentil plants from 2 agro-ecological zones of Syria and used to study their comparative growth on culture media and pathogenicity on different lentil genotypes. The growth studies were carried out on Potato Dextrose Agar (PDA) growth media under laboratory conditions. Mycelial radial growth and sclerotial production were the parameters used to compare the isolates. Pathogenicity studies were carried out with selected isolates on 10 lentil genotypes, infected as detached shoots and as whole potted-plants in the plastic house. The isolates showed considerable variation in cultural characteristics through mycelial growth, mycelial pigmentation and sclerotial production in the media plates. There were significant differences in the growth and sclerotial production of most of the isolates, but no apparent correlation between mycelial growth and sclerotial production among the isolates. Genotype by isolate interactions was significant for the isolates tested for pathogenicity. These interactions, however, appeared to be caused by differences in virulence of the isolates and did not suggest the occurrence of distinct pathogenic races of the pathogen isolates.

Ocorrência de doenças causadas por Sclerotinia sclerotiorum em girassol e soja.

2005

Podridão vermelha da raiz (Fusarium solani) e mofo branco (Sclerotinia sclerotiorum) na cultura da soja.

2008