SdhC-I86F Mutation in Phakopsora pachyrhizi Is Stable and Can Be Related to Fitness Penalties.

Succinate dehydrogenase inhibitors (SDHIs) fungicides are used to control Asian soybean rust (Phakopsora pachyrhizi), and the SdhC-I86F mutation is related to pathogen resistance. The objective of this study was to determine whether fitness penalties are associated with SDHI resistance (SdhC-I86F mutation) in P. pachyrhizi populations. Moreover, the study investigated whether the SdhC-I86F mutation remained stable after the fungus propagation both in the absence and presence of fungicide. The populations used in this study presented mutations for all genes analyzed (Cyp51, Cytb, and SdhC), except for a wild-type population (WTSdhC) found with no SdhC-I86F mutation. The frequencies of the SdhC-I86F mutant populations were stable after 36 generations in the absence of fungicide. However, in the case of the WTSdhC population, the SdhC-I86F mutation was further detected after one generation of the fungus in the presence of the SDHI fungicide, according to the results of a detached leaf assay. Three tests were performed to evaluate fitness components and sensitivity to fungicides (half maximal effective concentration). SdhC-I86F mutant populations were more sensitive to osmotic and oxidative stress than the WTSdhC population; however, the sensitivity to ultraviolet radiation was similar for both populations. All mutated populations were less sensitive than the WTSdhC when using SDHI (azoxystrobin + benzovindiflupyr), but more sensitive to mancozeb. The presence of fitness penalties, the mutation stability, and the sensitivity to mancozeb presented by the SdhC-I86F mutant populations can be relevant to the management of the disease in the field.

Spatio-Temporal Modeling and Simulation of Asian Soybean Rust Based on Fuzzy System.

Mato Grosso, Brazil, is the largest soy producer in the country. Asian Soy Rust is a disease that has already caused a lot of damage to Brazilian agribusiness. The plant matures prematurely, hindering the filling of the pod, drastically reducing productivity. It is caused by the Phakopsora pachyrhizi fungus. For a plant disease to establish itself, the presence of a pathogen, a susceptible plant, and favorable environmental conditions are necessary. This research developed a fuzzy system gathering these three variables as inputs, having as an output the vulnerability of the region to the disease. The presence of the pathogen was measured using a diffusion-advection equation appropriate to the problem. Some coefficients were based on the literature, others were measured by a fuzzy system and others were obtained by real data. From the mapping of producing properties, the locations where there are susceptible plants were established. And the favorable environmental conditions were also obtained from a fuzzy system, whose inputs were temperature and leaf wetness. Data provided by IBGE, INMET, and Antirust Consortium were used to fuel the model, and all treatments, tests, and simulations were carried out within the Matlab® environment. Although Asian Soybean Rust was the chosen disease here, the model was general in nature, so could be reproduced for any disease of plants with the same profile.

A Microbial Fermentation Product Induces Defense-Related Transcriptional Changes and the Accumulation of Phenolic Compounds in Glycine max.

With the progressive loss of fungicide efficacy against Phakopsora pachyrhizi, the causal agent of Asian soybean rust (ASR), alternative methods to protect soybean crops are needed. Resistance induction is a low impact alternative and/or supplement to fungicide applications that fortifies innate plant defenses against pathogens. Here, we show that a microbial fermentation product (MFP) induces plant defenses in soybean, and transcriptional induction is enhanced with the introduction of ASR. MFP-treated plants exhibited 1,011 and 1,877 differentially expressed genes (DEGs) 12 and 60 h after treatment, respectively, compared with water controls. MFP plants exposed to the pathogen 48 h after application and sampled 12 h later (for a total of 60 h) had 2,401 DEGs compared with control. The plant defense genes PR1, PR2, IPER, PAL, and CHS were induced with MFP application, and induction was enhanced with ASR. Enriched pathways associated with pathogen defense included plant-pathogen interactions, MAPK signaling pathways, phenylpropanoid biosynthesis, glutathione metabolism, flavonoid metabolism, and isoflavonoid metabolism. In field conditions, elevated antioxidant peroxidase activities and phenolic accumulation were measured with MFP treatment; however, improved ASR control or enhanced crop yield were not observed. MFP elicitation differences between field and laboratory grown plants necessitates further testing to identify best practices for effective disease management with MFP-treated soybean.

Sensitivity assessment and SDHC-I86F mutation frequency of Phakopsora pachyrhizi populations to benzovindiflupyr and fluxapyroxad fungicides from 2015 to 2019 in Brazil.

BACKGROUND: Fungicides of the succinate dehydrogenase inhibitors (SDHIs) group have been used in soybean to control Asian soybean rust (ASR) caused by Phakopsora pachyrhizi. Fungal populations with less sensitivity to SDHI fungicides have been reported since 2015. RESULTS: In this study, fungal sensitivity to benzovindiflupyr (BZV) and fluxapyroxad (FXD) was assessed using a total of 770 P. pachyrhizi populations sampled over four soybean growing seasons. Cross-resistance, intrinsic activity, and frequency of SDHC-I86F mutation were also analyzed. The average effective concentration to inhibit 50% (EC50 ) and SDHC-I86F frequency increased over the 2015/2016, 2016/2017, 2017/2018 and 2018/2019 soybean-seasons. Fourteen P. pachyrhizi populations had the EC50 value above 10 mg L-1 for both carboxamides. No difference was found in intrinsic active to BZV and FXD fungicides for sensitive P. pachyrhizi populations. For P. pachyrhizi classified as less sensitive BZV showed the highest fungitoxicity effect. High frequency of the C-I86F mutation was observed in samples collected in volunteer soybean plants. The maximum frequency of SDHC-I86F mutation in the population was 50% and resulting in ASR populations with low sensitivity to SDHIs. A low correlation between bioassay and SDHC-I86F mutation was observed possible due to the dikaryotic nature of rust fungi or other mutations in the other succinate dehydrogenase subunits. CONCLUSION: The present work provides an overview of a large sampling size of P. pachyrhizi populations and their performance over the four crop seasons. The high frequency of SDHC-I86F mutation and low sensitivity to SDHIs are widely distributed in the main soybean growing regions in Brazil and present in volunteer plants in the soybean-free period. Further detailed studies are needed to identify novel point mutations affecting the effectiveness of SDHIs. © 2021 Society of Chemical Industry.

Untargeted Metabolomics Analysis by UHPLC-MS/MS of Soybean Plant in a Compatible Response to Phakopsora pachyrhizi Infection.

Phakopsora pachyrhizi is a biotrophic fungus, causer of the disease Asian Soybean Rust, a severe crop disease of soybean and one that demands greater investment from producers. Thus, research efforts to control this disease are still needed. We investigated the expression of metabolites in soybean plants presenting a resistant genotype inoculated with P. pachyrhizi through the untargeted metabolomics approach. The analysis was performed in control and inoculated plants with P. pachyrhizi using UHPLC-MS/MS. Principal component analysis (PCA) and the partial least squares discriminant analysis (PLS-DA), was applied to the data analysis. PCA and PLS-DA resulted in a clear separation and classification of groups between control and inoculated plants. The metabolites were putative classified and identified using the Global Natural Products Social Molecular Networking platform in flavonoids, isoflavonoids, lipids, fatty acyls, terpenes, and carboxylic acids. Flavonoids and isoflavonoids were up-regulation, while terpenes were down-regulated in response to the soybean-P. pachyrhizi interaction. Our data provide insights into the potential role of some metabolites as flavonoids and isoflavonoids in the plant resistance to ASR. This information could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.

In vivo sensitivity of Phakopsora pachyrhizi to fungicides / Sensibilidade in vivo de Phakopsora pachyrhizi a fungicidas

Asian soybean rust is one of the most destructive diseases that can be found in this crop. It can be largely controlled by fungicide application. The objective was to assess the sensitivity of P. pachyrhizi isolates to fungicides. The tests were performed in a completely randomized design, with six replicates. The sensitivity of twelve isolates to site-specific and multisite fungicides at concentrations of 0.1; 1.0; 10.0, and 100.0 mg L-1, plus a control with absence of fungicide (0.0 mg L-1) was assessed. Soybean leaflets were immersed in the appropriate fungicide solutions, disposed in wet chambers in plastic boxes, and inoculated using each uredinia suspension of P. pachyhrizi (5.0 x 104 uredospores mL-1), separately. Boxes were incubated for 20 days at a temperature of 23°C and a 12-hour photoperiod. Next, the number of uredinia per cm2 on the abaxial face of each leaflet was evaluated. The active ingredients prothioconazole, trifloxystrobin, fluxapiroxade, trifloxystrobin + prothioconazole, trifloxystrobin + bixafen + prothioconazole, azoxystrobin + benzovindiflupyr, and azoxystrobin + benzovindiflupyr + diphenoconazole were highly fungitoxic for the majority of the isolates, with EC50 lower than 1.0 mg L-1. Diphenoconazole, azoxystrobin, and fenpropimorph were considered moderately fungitoxic for nine of the twelve isolates, with EC50 between 1 and 10 mg L-1. The multisites mancozeb and copper oxychloride presented EC50 responses classified as low toxic for the twelve isolates and eight for chlorothalonil (EC50 between 10 mg L-1 and 50 mg L-1). Site-specific fungicides showed high-to-moderate fungitoxicity to P. pachyrhizi isolates, even as the multisites presented moderate-to-less toxic activity.(AU)

A ferrugem-asiática da soja é uma das doenças mais destrutivas que ocorre na cultura. Seu controle é baseado, principalmente, na aplicação de fungicidas. O objetivo foi avaliar a sensibilidade de isolados de P. pachyrhizi a fungicidas. Os ensaios foram realizados em delineamento inteiramente casualizado, com seis repetições. Por meio de teste de folíolos destacados de soja, foram avaliadas as sensibilidades de doze isolados do fungo a fungicidas sítio-específicos e multissítios, nas concentrações de 0,1; 1,0; 10,0 e 100,0 mg L-1, mais uma testemunha sem fungicida (0,0 mg L-1). Os folíolos de soja foram imersos nas devidas soluções fungicida, dispostos em câmaras úmidas em caixas gerbox e inoculados com as devidas suspensões de esporos de P. pachyhrizi (5,0x104 uredosporos mL-1). As caixas foram incubadas durante 20 dias, em temperatura de 23 oC e fotoperíodo de 12 h. Em seguida, avaliou-se o número de urédias cm-2 da face abaxial de cada folíolo. Os ingredientes ativos protioconazol, trifloxistrobina, fluxapiroxade, trifloxistrobina + protioconazol, trifloxistrobina + bixafem + protioconazol, azoxistrobina + benzovindiflupir e azoxistrobina + benzovindiflupir + difenoconazol foram altamente fungitóxicos para a maioria dos isolados, com CE50 menor do que 1,0 mg L-1. Difenoconazol, azoxistrobina e fenpropimorfe foram considerados medianamente fungitóxicos para nove dos doze isolados, com CE50 entre 1 e 10 mg L-1. Os multissítios mancozebe e oxicloreto de cobre apresentaram respostas de CE50 classificadas como pouco tóxicas para os doze isolados do fungo e o clorotalonil para oito deles (CE50 entre 10 e 50 mg L-1). Os fungicidas sítio-específico apresentaram alta a moderada fungitoxicidade aos isolados de P. pachyrhizi oriundos dos distintos locais, enquanto os multissítios apresentaram atividade moderada a pouco tóxica.(AU)

In vivo sensitivity of Phakopsora pachyrhizi to fungicides / Sensibilidade in vivo de Phakopsora pachyrhizi a fungicidas

ABSTRACT: Asian soybean rust is one of the most destructive diseases that can be found in this crop. It can be largely controlled by fungicide application. The objective was to assess the sensitivity of P. pachyrhizi isolates to fungicides. The tests were performed in a completely randomized design, with six replicates. The sensitivity of twelve isolates to site-specific and multisite fungicides at concentrations of 0.1; 1.0; 10.0, and 100.0 mg L-1, plus a control with absence of fungicide (0.0 mg L-1) was assessed. Soybean leaflets were immersed in the appropriate fungicide solutions, disposed in wet chambers in plastic boxes, and inoculated using each uredinia suspension of P. pachyhrizi (5.0 x 104 uredospores mL-1), separately. Boxes were incubated for 20 days at a temperature of 23°C and a 12-hour photoperiod. Next, the number of uredinia per cm2 on the abaxial face of each leaflet was evaluated. The active ingredients prothioconazole, trifloxystrobin, fluxapiroxade, trifloxystrobin + prothioconazole, trifloxystrobin + bixafen + prothioconazole, azoxystrobin + benzovindiflupyr, and azoxystrobin + benzovindiflupyr + diphenoconazole were highly fungitoxic for the majority of the isolates, with EC50 lower than 1.0 mg L-1. Diphenoconazole, azoxystrobin, and fenpropimorph were considered moderately fungitoxic for nine of the twelve isolates, with EC50 between 1 and 10 mg L-1. The multisites mancozeb and copper oxychloride presented EC50 responses classified as low toxic for the twelve isolates and eight for chlorothalonil (EC50 between 10 mg L-1 and 50 mg L-1). Site-specific fungicides showed high-to-moderate fungitoxicity to P. pachyrhizi isolates, even as the multisites presented moderate-to-less toxic activity.

RESUMO: A ferrugem-asiática da soja é uma das doenças mais destrutivas que ocorre na cultura. Seu controle é baseado, principalmente, na aplicação de fungicidas. O objetivo foi avaliar a sensibilidade de isolados de P. pachyrhizi a fungicidas. Os ensaios foram realizados em delineamento inteiramente casualizado, com seis repetições. Por meio de teste de folíolos destacados de soja, foram avaliadas as sensibilidades de doze isolados do fungo a fungicidas sítio-específicos e multissítios, nas concentrações de 0,1; 1,0; 10,0 e 100,0 mg L-1, mais uma testemunha sem fungicida (0,0 mg L-1). Os folíolos de soja foram imersos nas devidas soluções fungicida, dispostos em câmaras úmidas em caixas gerbox e inoculados com as devidas suspensões de esporos de P. pachyhrizi (5,0x104 uredosporos mL-1). As caixas foram incubadas durante 20 dias, em temperatura de 23 oC e fotoperíodo de 12 h. Em seguida, avaliou-se o número de urédias cm-2 da face abaxial de cada folíolo. Os ingredientes ativos protioconazol, trifloxistrobina, fluxapiroxade, trifloxistrobina + protioconazol, trifloxistrobina + bixafem + protioconazol, azoxistrobina + benzovindiflupir e azoxistrobina + benzovindiflupir + difenoconazol foram altamente fungitóxicos para a maioria dos isolados, com CE50 menor do que 1,0 mg L-1. Difenoconazol, azoxistrobina e fenpropimorfe foram considerados medianamente fungitóxicos para nove dos doze isolados, com CE50 entre 1 e 10 mg L-1. Os multissítios mancozebe e oxicloreto de cobre apresentaram respostas de CE50 classificadas como pouco tóxicas para os doze isolados do fungo e o clorotalonil para oito deles (CE50 entre 10 e 50 mg L-1). Os fungicidas sítio-específico apresentaram alta a moderada fungitoxicidade aos isolados de P. pachyrhizi oriundos dos distintos locais, enquanto os multissítios apresentaram atividade moderada a pouco tóxica.

Chemical control of foliar diseases in soybean depends on cultivar and sowing date / O controle químico de doenças foliares na cultura da soja depende do cultivar e da época de plantio

The aim was to study the effect of early application of two fungicide mixtures on the control of Asian rust and powdery mildew, as well as on grain yield in five soybean cultivars at two sowing dates. The experiment was carried in the experimental area of FAMV/UPF. The cultivars A 4910 RG (super-early), BMX Apolo RR, A 6001 RR and Fundacep 55 RR (early) and Coodetec 214 RR (moderate), established at 13/11/2007 and 3/12/2007 were used. Trifolia of these cultivars were treated twice with the fungicides ciproconazol + azoxystrobin (Priori Xtra®, 0.3 L ha-1 plus Nimbus® adjuvant 0.6 L ha- 1) and epoxiconazol + pyraclostrobin (Opera®, 0.5 L ha-1), in three moments 1: pre-closure of the space between the lines and 21 days after; 2: pre-closure of space between rows and R5.1 stage (10% of beginning seed); and 3: R1 (beginning bloom) and R5.1 stages. The severity (% of affected leaf area) of powdery mildew and number of lesions cm2 of Asian rust were evaluated. After that plants were harvested, the thousand grain weight (g) and grain yield (kg ha-1) were quantified. The experimental design was a split-plot with four replicates, where the main plot were the five cultivars, and the subplots were the control treatment (untreated plants) and the six chemical control programs. Tukey and t-Student tests (p0.05) were used for separation of means of treatments and seasons, respectively. In general terms, foliar application of fungicides controlled both Asian rust and powdery mildew, as well as generated an increase on the thousand grain weight and grain yield. However, there seems to be an interaction among soybean genotypes and sowing date. Thus, our results allow us to infer that the chemical control of leaf diseases in the soybean crop depends of the cultivar and sowing date.

O objetivo foi estudar o efeito da aplicação antecipada de duas misturas de fungicidas no controle da ferrugem asiática e do oídio, bem como no rendimento de grãos em cinco cultivares de soja em duas épocas de semeadura. O experimento foi conduzido no campo experimental da FAMV/UPF. Utilizaram-se as cultivares A 4910 RG (super precoce), BMX Apolo RR, A 6001 RR e Fundacep 55 RR (precoces) e Coodetec 214 RR (moderado), estabelecidas em 13/11/2007 e 3/12/2007. Os fungicidas piraclostrobina + epoxiconazole e azoxistrobina + ciproconzole foram pulverizados duas vezes em 1: pré-fechamento do espaço entre as linhas e 21 dias após, 2: pré-fechamento do espaço entre as linhas e R5.1 (10% de granação), e 3: R1 (início da floração) e R5.1. Avaliaram-se a severidade (% da área foliar afetada) do oídio e o número de lesões cm2 da ferrugem asiática. Logo após as plantas foram colhidas, o peso de mil grãos (g) e rendimento de grãos (kg ha-1) foram quantificados. O delineamento experimental utilizado foi o de parcelas divididas com quatro repetições, onde a parcela principal foram as cinco cultivares, e as subparcelas foram o tratamento controle (plantas não tratadas) e os seis programas de controle químico. Para a separação das médias dos tratamentos e épocas, foram utilizados os testes de Tukey e t-Student (p0.05), respectivamente. De modo geral, a aplicação foliar de fungicidas controlou tanto a ferrugem asiática como o oídio, assim como também gerou um aumento do peso de mil grãos e rendimento de grãos. No entanto, parece existir uma interação entre os genótipos e data de semeadura. Assim, nossos resultados nos permitem inferir que o controle químico das doenças foliares na cultura da soja depende do cultivar e da data de semeadura.

Sensitivity of two isolates of Phakopsora pachyrhizi to dithiocarmamate, chloronitril, triazoles, strobilurins, and carboxamides fungicides / Sensibilidade de Phakopsora pachyrhizi aos fungicidas ditiocarbamatos, cloronitrilas, triazóis, estrobilurinas e carboxamidas

Asian soybean rust (ASR), caused by the phytopathogen Phakopsora pachyrhizi Sydow & Sydow, is a worldwide distributed fungal disease responsible for causing damages in soybean crops [Glycine max (L.) Merril] of up to 90% of its productive potential. So far, due to limited availability of resistant varieties, fungicide application is the most widely used strategy for ASR control, although some populations of the pathogen have shown reduced sensitivity to certain active ingredients. Several methods have been described to measure the sensitivity of a fungus to a given fungicide, or even the fungitoxicity of a chemical. The most used tests are spore germination in wateragar medium and evaluation of disease severity in detached soybean leaves. Experiments were carried out with Brazilian populations of the pathogen: one from Uberlândia - MG and the other from Chapadão do Sul - MS, following the mentioned methodologies. The results showed the reduction of benzovindiflupyr efficiency in relation to spore germination and disease severity for the MS population, as also did fluxapyroxad, cyproconazole, and tebuconazole. Multisite fungicides (chlorothalonil, copper oxychloride, and mancozeb) may be used in the management of ASR resistance in the fields of Brazil associated with strobilurins, triazoles, and carboxamides. The efficiency of the main active ingredient tested depends on the formulation and others molecules used in the commercial product.

A ferrugem asiática da soja (FAS), causada pelo fitopatógeno Phakopsora pachyrhizi Sydow & Sydow, é uma doença fúngica distribuída mundialmente e responsável por causar danos na cultura da soja [Glycine max (L.) Merril], em até 90% do potencial produtivo. Até agora, devido à limitada disponibilidade de variedades resistentes, a aplicação de fungicida é a estratégia mais utilizada para controlar a FAS, embora algumas populações do patógeno tenham demonstrado menor sensibilidade a determinados ingredientes ativos. Vários métodos foram descritos para medir a sensibilidade de um fungo a um dado fungicida, ou para monitorar sua redução ou perda de eficácia, ou mesmo a fungitoxicidade de um produto químico. Os testes mais utilizados são a germinação de esporos in vitro e a severidade da doença (%) em folhas de soja destacadas. Baseando-se nessas metodologias, foram realizados ensaios com populações do patógeno oriundas de Uberlândia - MG e do Chapadão do Sul - MS. Os resultados mostraram a redução da eficiência de benzovindiflupyr em relação à germinação e à severidade de doença para a população de Chapadão do Sul - MS. O mesmo comportamento foi obtido para os ingredientes ativos fluxapyroxad, cyproconazole e tebuconazole. Os fungicidas multissítios (clorotalonil, oxicloreto de cobre e mancozeb) podem ser utilizados no manejo da resistência à FAS nas lavouras do Brasil associados às estrobilurinas, triazóis e carboxamidas. A eficiência do principal ingrediente ativo estudado depende dos outros ingredientes ativos ou fungicidas associados na formulação comercial.

Competitive Fitness of Phakopsora pachyrhizi Isolates with Mutations in the CYP51 and CYTB Genes.

Soybean rust (Phakopsora pachyrhizi) in Brazil is mainly controlled with applications of fungicides, including demethylation inhibitors (DMI) and quinone outside inhibitors (QoI). Isolates with less sensitivity to DMI and QoI have been reported, and these have been found to have mutations in the CYP51 and CYTB genes, respectively. There have been no reports of fitness costs in isolates with mutations in CYP51 and CYTB, and the aim of this work was to compare the competitive ability of isolates with lower DMI or QoI sensitivities with that of sensitive (wild-type) isolates. Urediniospores of sensitive wild-type isolates and isolates with different CYP51 or CYTB alleles were mixed and inoculated on detached soybean leaves. After 3 weeks, urediniospores were harvested and used as inoculum for the next disease cycle. Frequencies of relevant target site mutations were monitored using the pyrosequencing method over four disease cycles. Isolates with lower DMI sensitivity and different CYP51 alleles had competitive disadvantages compared with a DMI-sensitive, wild-type CYP51 isolate. In contrast, the isolate with the F129L mutation in the CYTB gene competed equally well with a QoI-sensitive, wild-type CYTB isolate under the conditions of this experiment. The CYP51 and CYTB alleles were stable in all isolates over four disease cycles when cultivated alone.

Detection of the F129L mutation in the cytochrome b gene in Phakopsora pachyrhizi.

BACKGROUND: Asian soybean rust, caused by Phakopsora pachyrhizi, is mostly controlled by demethylation inhibitor (DMI) and quinone outside inhibitor (QoI) fungicides. Mutations in the cytochrome b (CYTB) gene can lead to pathogen resistance to QoIs. The occurrence of the mutations in codons 129, 137 and 143 in the CYTB gene was investigated, and a pyrosequencing assay was developed for rapid and quantitative detection of the F129L mutation. RESULTS: Molecular analysis of the CYTB gene showed the presence of the F129L mutation in field samples and monouredinial isolates, while other mutations (G143A and G137R) were not found. The pyrosequencing was an effective method for quantitative detection of the F129L mutation, and many of the P. pachyrhizi samples showed high frequency of F129L. CONCLUSION: This is the first report of the occurrence of the F129L mutation in P. pachyrhizi. The practical relevance of this mutation for field efficacy of QoIs needs further investigation. © 2015 Society of Chemical Industry.

Dynamics of fungicide in chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes / Dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos

The objective of this study was to evaluate the dynamics of chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes. The experiment was conducted in the experimental area of Phytus Institute, in Itaara city, central region of Rio Grande do Sul, in 2012/2013 harvest. The experimental design was completely randomized, with four replications in a factorial arrangement (2x6). The first factor was composed of two water regimes: 1) water deficit (5060% of field capacity); 2) no water deficit (90100% of field capacity). The B factor was four time intervals between fungicide application and rainfall simulation (0, 30, 60 and 120 min), a witness to application without rain and a control without fungicide application. In application the active ingredients trifloxystrobin  + prothioconazole (60.0  + 70.0  g i.a.ha1) with the addition of Aureo® at a dose of 0.375 Lpc ha1 were used. The parameters evaluated were the number of days until the first pustule preview of Asian rust, disease severity, yield and grain yield. It was found that the fungicide application in plants under water deficit provides the greatest number of days until the first pustule preview. A  rainfall simulation has great effect in removing fungicides no matter the water regime. The control of P. pachyrhizi in plants under water deficit was effective, resulting in increased productivity.

Objetivou-se com este trabalho avaliar a dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos. O experimento foi conduzido na área experimental do Instituto Phytus, no município de Itaara, região central do Rio Grande do Sul, na safra 2012/2013. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, em arranjo fatorial (2x6). O fator A foi composto por dois regimes hídricos: 1) déficit hídrico (5060% da capacidade de campo); 2) sem déficit hídrico (90100% da capacidade de campo). O fator B consistiu em quatro intervalos de tempo entre a aplicação de fungicidas e a simulação de chuva (0, 30, 60 e 120 min), uma testemunha com aplicação sem chuva e mais uma testemunha sem aplicação de fungicida. Na aplicação foram utilizados os ingredientes ativos trifloxis trobina + protioconazol (60,0 + 70,0 g i.a.ha1) com adição de Aureo® na dose de 0,375 L.p.c ha1. Os parâmetros avaliados foram o tempo decorrido entre a aplicação e o surgimento da primeira pústula de ferrugem asiática, a severidade da doença, a produtividade e a massa de grãos. Verificou-se que a aplicação de fungicida em plantas sob déficit hídrico proporciona o maior número de dias para o aparecimento da primeira pústula. A simulação de chuva tem grande efeito na remoção de fungicidas independentemente do regime hídrico. O controle de P. pachyrhizi em plantas sob déficit hídrico foi eficaz, resultando em incrementos de produtividade.

Dynamics of fungicide in chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes / Dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos

The objective of this study was to evaluate the dynamics of chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes. The experiment was conducted in the experimental area of Phytus Institute, in Itaara city, central region of Rio Grande do Sul, in 2012/2013 harvest. The experimental design was completely randomized, with four replications in a factorial arrangement (2x6). The first factor was composed of two water regimes: 1) water deficit (5060% of field capacity); 2) no water deficit (90100% of field capacity). The B factor was four time intervals between fungicide application and rainfall simulation (0, 30, 60 and 120 min), a witness to application without rain and a control without fungicide application. In application the active ingredients trifloxystrobin  + prothioconazole (60.0  + 70.0  g i.a.ha1) with the addition of Aureo® at a dose of 0.375 Lpc ha1 were used. The parameters evaluated were the number of days until the first pustule preview of Asian rust, disease severity, yield and grain yield. It was found that the fungicide application in plants under water deficit provides the greatest number of days until the first pustule preview. A  rainfall simulation has great effect in removing fungicides no matter the water regime. The control of P. pachyrhizi in plants under water deficit was effective, resulting in increased productivity.(AU)

Objetivou-se com este trabalho avaliar a dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos. O experimento foi conduzido na área experimental do Instituto Phytus, no município de Itaara, região central do Rio Grande do Sul, na safra 2012/2013. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, em arranjo fatorial (2x6). O fator A foi composto por dois regimes hídricos: 1) déficit hídrico (5060% da capacidade de campo); 2) sem déficit hídrico (90100% da capacidade de campo). O fator B consistiu em quatro intervalos de tempo entre a aplicação de fungicidas e a simulação de chuva (0, 30, 60 e 120 min), uma testemunha com aplicação sem chuva e mais uma testemunha sem aplicação de fungicida. Na aplicação foram utilizados os ingredientes ativos trifloxis trobina + protioconazol (60,0 + 70,0 g i.a.ha1) com adição de Aureo® na dose de 0,375 L.p.c ha1. Os parâmetros avaliados foram o tempo decorrido entre a aplicação e o surgimento da primeira pústula de ferrugem asiática, a severidade da doença, a produtividade e a massa de grãos. Verificou-se que a aplicação de fungicida em plantas sob déficit hídrico proporciona o maior número de dias para o aparecimento da primeira pústula. A simulação de chuva tem grande efeito na remoção de fungicidas independentemente do regime hídrico. O controle de P. pachyrhizi em plantas sob déficit hídrico foi eficaz, resultando em incrementos de produtividade.(AU)

Dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos / Dynamics of fungicide in chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes

Objetivou-se com este trabalho avaliar a dinâmica do controle químico de Phakopsora pachyrhizi em plantas de soja submetidas a diferentes regimes hídricos. O experimento foi conduzido na área experimental do Instituto Phytus, no município de Itaara, região central do Rio Grande do Sul, na safra 2012/2013. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, em arranjo fatorial (2x6). O fator A foi composto por dois regimes hídricos: 1) déficit hídrico (50-60% da capacidade de campo); 2) sem déficit hídrico (90-100% da capacidade de campo). O fator B consistiu em quatro intervalos de tempo entre a aplicação de fungicidas e a simulação de chuva (0, 30, 60 e 120 min), uma testemunha com aplicação sem chuva e mais uma testemunha sem aplicação de fungicida. Na aplicação foram utilizados os ingredientes ativos trifloxistrobina + protioconazol (60,0 + 70,0 g i.a.ha-1) com adição de Aureo(r) na dose de 0,375 L.p.c ha-1. Os parâmetros avaliados foram o tempo decorrido entre a aplicação e o surgimento da primeira pústula de ferrugem asiática, a severidade da doença, a produtividade e a massa de grãos. Verificou-se que a aplicação de fungicida em plantas sob déficit hídrico proporciona o maior número de dias para o aparecimento da primeira pústula. A simulação de chuva tem grande efeito na remoção de fungicidas independentemente do regime hídrico. O controle de P. pachyrhizi em plantas sob déficit hídrico foi eficaz, resultando em incrementos de produtividade.(AU)

The objective of this study was to evaluate the dynamics of chemical control of Phakopsora pachyrhizi in soybean plants subjected to different water regimes. The experiment was conducted in the experimental area of Phytus Institute, in Itaara city, central region of Rio Grande do Sul, in 2012/2013 harvest. The experimental design was completely randomized, with four replications in a factorial arrangement (2x6). The first factor was composed of two water regimes: 1) water deficit (50-60% of field capacity); 2) no water deficit (90-100% of field capacity). The B factor was four time intervals between fungicide application and rainfall simulation (0, 30, 60 and 120 min), a witness to application without rain and a control without fungicide application. In application the active ingredients trifloxystrobin + prothioconazole (60.0 + 70.0 g i.a.ha-1) with the addition of Aureo(r) at a dose of 0.375 Lpc ha-1 were used. The parameters evaluated were the number of days until the first pustule preview of Asian rust, disease severity, yield and grain yield. It was found that the fungicide application in plants under water deficit provides the greatest number of days until the first pustule preview. A rainfall simulation has great effect in removing fungicides no matter the water regime. The control of P. pachyrhizi in plants under water deficit was effective, resulting in increased productivity.(AU)

Use of AFLP markers to estimate molecular diversity of Phakopsora pachyrhizi

Background Asian soybean rust (SBR) caused by Phakopsora pachyrhizi Syd. & Syd., is one of the main diseases affecting soybean and has been reported as one of the most economically important fungal pathogens worldwide. Knowledge of the genetic diversity of this fungus should be considered when developing resistance breeding strategies. We aimed to analyze the genetic diversity of P. pachyrhizi combining simple sampling with a powerful and reproducible molecular technique. Results We employed Amplified Fragment Length Polymorphism (AFLP) technique for the amplification of P. pachyrhizi DNA extracted from naturally SBR-infected plants from 23 production fields. From a total of 1919 markers obtained, 77% were polymorphic. The high percentage of polymorphism and the Nei's genetic diversity coefficient (0.22) indicated high pathogen diversity. Analysis of molecular variance showed higher genetic variation within countries than among them. Temporal analysis showed a higher genetic variation within a year than between years. Cluster, phylogenetic and principal co-ordinate analysis showed that samples group by year of collection and then by country sampled. Conclusions The study proposed combining a simple collection of urediniospore with a subsequent analysis by AFLP was useful to examine the molecular polymorphism of samples of P. pachyrhizi collected and might have a significant contribution to the knowledge of its genetic diversity. Also, AFLP analysis is an important and potent molecular tool for the study of genetic diversity and could be useful to carry out wider genetic diversity studies.

Differential expression of four soybean bZIP genes during Phakopsora pachyrhizi infection.

Asian soybean rust (ASR), caused by the obligate biotrophic fungus Phakopsora pachyrhizi, is one of most important diseases in the soybean (Glycine max (L.) Merr.) agribusiness. The identification and characterization of genes related to plant defense responses to fungal infection are essential to develop ASR-resistant plants. In this work, we describe four soybean genes, GmbZIP62, GmbZIP105, GmbZIPE1, and GmbZIPE2, which encode transcription factors containing a basic leucine zipper (bZIP) domain from two divergent classes, and that are responsive to P. pachyrhizi infection. Molecular phylogenetic analyses demonstrated that these genes encode proteins similar to bZIP factors responsive to pathogens. Yeast transactivation assays showed that only GmbZIP62 has strong transactivation activity in yeast. In addition, three of the bZIP transcription factors analyzed were also differentially expressed by plant defense hormones, and all were differentially expressed by fungal attack, indicating that these proteins might participate in response to ASR infection. The results suggested that these bZIP proteins are part of the plant defense response to P. pachyrhizi infection, by regulating the gene expression related to ASR infection responses. These bZIP genes are potential targets to obtain new soybean genotypes resistant to ASR.

Phakopsora pachyrhizi and Puccinia kuehnii urediniospore germination under different adjuvants / Germinação de uredinosporos de Phakopsora pachyrhizi e Puccinia kuehnii sob diferentes adjuvantes

In Brazil Phakopsora pachyrhizi and Puccinia kuehnii are important pathogens of soybeans and sugar cane, respectively. In vitro experiments were carried out in order to evaluate P. pachyrhizi and P. kuehnii urediniospore germination under different adjuvants used in fungicide sprays to control rust diseases. Petri dishes (80 mm diameter) containing 10 mL of agarwater (1.7%) were used. In each plate 1 mL of a spore suspension with 5x105 urediniospores and 1 mL solution of adjuvants with twice the concentratation for field sprays was added. Assist®, Veget Oil®, Natur Oil®, Break Thru®, Aureo®, Silwet® and Nimbus® were evaluated. There was a control check treatment with water. P. pachyrhizi urediniospore germination was negatively affected by all adjuvants after 2 hours of exposition. Urediniospores of the Asian soybean rust exposed to Assist®, Break Thru®, Aureo® and Nimbus® had lower germination rates. Germination of P. kuehnii urediniospores was lower in all adjuvant treatments after 4 hours of exposition. At 8 hours of Veget Oil® and Break Thru® exposure urediniospores of the orange rust pathogen had 96.6 and 94.8% reduction in germination compared to control check treatment.

No Brasil, Phakopsora pachyrhizi e Puccinia kuehnii são importantes patógenos das culturas da soja e cana-de-açúcar, respectivamente. Avaliou-se a germinação in vitro de uredinosporos de P. pachyrhizi e P. kuehnii sob diferentes adjuvantes comumente utilizados em caldas fungicidas. Foram utilizadas placas de Petri com 80 mm de diâmetro, contendo 10 mL de solução ágar-água (1,7%). Adicionou-se em cada placa 1 mL de suspensão de esporos, na concentração de 5x105 esporos mL-1, e 1 mL de solução de adjuvantes, no dobro da concentração da dose recomendada pelos fabricantes. Os adjuvantes testados foram Assist®, Veget Oil®, Natur Oil®, Break Thru®,Aureo®, Silwet® e Nimbus®; em adição, o tratamento Testemunha constou apenas da exposição dos uredinosporos em água destilada esterilizada. A partir de uma hora de exposição, todos os adjuvantes reduziram a germinação de uredinosporos de P. pachyrhizi. Uredinosporos do agente causal da ferrugem asiá- tica da soja expostos aos adjuvantes Assist®, Break Thru®, Aureo® e Nimbus® tiveram menor germinação a partir de duas horas de exposição. A inibição da germinação dos uredinosporos de P. kuehnii pelos adjuvantes em relação ao tratamento Testemunha aconteceu a partir de 4 horas de exposição. Às 8 horas, uredinosporos de ferrugem alaranjada da cana-de-açúcar que foram expostos aos adjuvantes Veget Oil® e Break Thru® apresentaram redução na germinação de 96,6 e 94,8%, respectivamente, quando comparados com o tratamento Testemunha.

Germinação de uredinosporos de Phakopsora pachyrhizi e Puccinia kuehnii sob diferentes adjuvantes

ABSTRACT In Brazil Phakopsora pachyrhizi and Puccinia kuehnii are important pathogens of soybeans and sugar cane, respectively. In vitro experiments were carried out in order to evaluate P. pachyrhizi and P. kuehniiurediniospore germination under different adjuvants used in fungicide sprays to control rust diseases. Petri dishes (80 mm diameter) containing 10 mL of agar-water (1.7%) were used. In each plate 1 mL of a spore suspension with 5x105 urediniospores and 1 mL solution of adjuvants with twice the concentratation for field sprays was added. Assist(r), Veget Oil(r), Natur Oil(r), Break Thru(r)Aureo(r), Silwet(r) and Nimbus(r) were evaluated. There was a control check treatment with water. P. pachyrhiziurediniospore germination was negatively affected by all adjuvants after 2 hours of exposition. Urediniospores of the Asian soybean rust exposed to Assist(r), Break Thru(r), Aureo(r) and Nimbus(r) had lower germination rates. Germination of P. kuehnii urediniospores was lower in all adjuvant treatments after 4 hours of exposition. At 8 hours of Veget Oil(r) and Break Thru(r) exposure urediniospores of the orange rust pathogen had 96.6 and 94.8% reduction in germination compared to control check treatment.

RESUMO No Brasil, Phakopsora pachyrhizi e Puccinia kuehnii são importantes patógenos das culturas da soja e cana-de-açúcar, respectivamente. Avaliou-se a germinação in vitro de uredinosporos de P. pachyrhizi eP. kuehnii sob diferentes adjuvantes comumente utilizados em caldas fungicidas. Foram utilizadas placas de Petri com 80 mm de diâmetro, contendo 10 mL de solução ágar-água (1,7%). Adicionou-se em cada placa 1 mL de suspensão de esporos, na concentração de 5x105 esporos mL-1, e 1 mL de solução de adjuvantes, no dobro da concentração da dose recomendada pelos fabricantes. Os adjuvantes testados foram Assist(r), Veget Oil(r), Natur Oil(r), Break Thru(r), Aureo(r), Silwet(r) e Nimbus(r); em adição, o tratamento Testemunha constou apenas da exposição dos uredinosporos em água destilada esterilizada. A partir de uma hora de exposição, todos os adjuvantes reduziram a germinação de uredinosporos de P. pachyrhizi . Uredinosporos do agente causal da ferrugem asiática da soja expostos aos adjuvantes Assist(r), Break Thru(r), Aureo(r) e Nimbus(r) tiveram menor germinação a partir de duas horas de exposição. A inibição da germinação dos uredinosporos de P. kuehnii pelos adjuvantes em relação ao tratamento Testemunha aconteceu a partir de 4 horas de exposição. Às 8 horas, uredinosporos de ferrugem alaranjada da cana-de-açúcar que foram expostos aos adjuvantes Veget Oil(r) e Break Thru(r) apresentaram redução na germinação de 96,6 e 94,8%, respectivamente, quando comparados com o tratamento Testemunha.

Germinação de uredinosporos de Phakopsora pachyrhizi e Puccinia kuehnii sob diferentes adjuvantes

ABSTRACT In Brazil Phakopsora pachyrhizi and Puccinia kuehnii are important pathogens of soybeans and sugar cane, respectively. In vitro experiments were carried out in order to evaluate P. pachyrhizi and P. kuehniiurediniospore germination under different adjuvants used in fungicide sprays to control rust diseases. Petri dishes (80 mm diameter) containing 10 mL of agar-water (1.7%) were used. In each plate 1 mL of a spore suspension with 5x105 urediniospores and 1 mL solution of adjuvants with twice the concentratation for field sprays was added. Assist(r), Veget Oil(r), Natur Oil(r), Break Thru(r)Aureo(r), Silwet(r) and Nimbus(r) were evaluated. There was a control check treatment with water. P. pachyrhiziurediniospore germination was negatively affected by all adjuvants after 2 hours of exposition. Urediniospores of the Asian soybean rust exposed to Assist(r), Break Thru(r), Aureo(r) and Nimbus(r) had lower germination rates. Germination of P. kuehnii urediniospores was lower in all adjuvant treatments after 4 hours of exposition. At 8 hours of Veget Oil(r) and Break Thru(r) exposure urediniospores of the orange rust pathogen had 96.6 and 94.8% reduction in germination compared to control check treatment.

RESUMO No Brasil, Phakopsora pachyrhizi e Puccinia kuehnii são importantes patógenos das culturas da soja e cana-de-açúcar, respectivamente. Avaliou-se a germinação in vitro de uredinosporos de P. pachyrhizi eP. kuehnii sob diferentes adjuvantes comumente utilizados em caldas fungicidas. Foram utilizadas placas de Petri com 80 mm de diâmetro, contendo 10 mL de solução ágar-água (1,7%). Adicionou-se em cada placa 1 mL de suspensão de esporos, na concentração de 5x105 esporos mL-1, e 1 mL de solução de adjuvantes, no dobro da concentração da dose recomendada pelos fabricantes. Os adjuvantes testados foram Assist(r), Veget Oil(r), Natur Oil(r), Break Thru(r), Aureo(r), Silwet(r) e Nimbus(r); em adição, o tratamento Testemunha constou apenas da exposição dos uredinosporos em água destilada esterilizada. A partir de uma hora de exposição, todos os adjuvantes reduziram a germinação de uredinosporos de P. pachyrhizi . Uredinosporos do agente causal da ferrugem asiática da soja expostos aos adjuvantes Assist(r), Break Thru(r), Aureo(r) e Nimbus(r) tiveram menor germinação a partir de duas horas de exposição. A inibição da germinação dos uredinosporos de P. kuehnii pelos adjuvantes em relação ao tratamento Testemunha aconteceu a partir de 4 horas de exposição. Às 8 horas, uredinosporos de ferrugem alaranjada da cana-de-açúcar que foram expostos aos adjuvantes Veget Oil(r) e Break Thru(r) apresentaram redução na germinação de 96,6 e 94,8%, respectivamente, quando comparados com o tratamento Testemunha.

Phakopsora pachyrhizi and Puccinia kuehnii urediniospore germination under different adjuvants / Germinação de uredinosporos de Phakopsora pachyrhizi e Puccinia kuehnii sob diferentes adjuvantes

In Brazil Phakopsora pachyrhizi and Puccinia kuehnii are important pathogens of soybeans and sugar cane, respectively. In vitro experiments were carried out in order to evaluate P. pachyrhizi and P. kuehnii urediniospore germination under different adjuvants used in fungicide sprays to control rust diseases. Petri dishes (80 mm diameter) containing 10 mL of agarwater (1.7%) were used. In each plate 1 mL of a spore suspension with 5x105 urediniospores and 1 mL solution of adjuvants with twice the concentratation for field sprays was added. Assist®, Veget Oil®, Natur Oil®, Break Thru®, Aureo®, Silwet® and Nimbus® were evaluated. There was a control check treatment with water. P. pachyrhizi urediniospore germination was negatively affected by all adjuvants after 2 hours of exposition. Urediniospores of the Asian soybean rust exposed to Assist®, Break Thru®, Aureo® and Nimbus® had lower germination rates. Germination of P. kuehnii urediniospores was lower in all adjuvant treatments after 4 hours of exposition. At 8 hours of Veget Oil® and Break Thru® exposure urediniospores of the orange rust pathogen had 96.6 and 94.8% reduction in germination compared to control check treatment.(AU)

No Brasil, Phakopsora pachyrhizi e Puccinia kuehnii são importantes patógenos das culturas da soja e cana-de-açúcar, respectivamente. Avaliou-se a germinação in vitro de uredinosporos de P. pachyrhizi e P. kuehnii sob diferentes adjuvantes comumente utilizados em caldas fungicidas. Foram utilizadas placas de Petri com 80 mm de diâmetro, contendo 10 mL de solução ágar-água (1,7%). Adicionou-se em cada placa 1 mL de suspensão de esporos, na concentração de 5x105 esporos mL-1, e 1 mL de solução de adjuvantes, no dobro da concentração da dose recomendada pelos fabricantes. Os adjuvantes testados foram Assist®, Veget Oil®, Natur Oil®, Break Thru®,Aureo®, Silwet® e Nimbus®; em adição, o tratamento Testemunha constou apenas da exposição dos uredinosporos em água destilada esterilizada. A partir de uma hora de exposição, todos os adjuvantes reduziram a germinação de uredinosporos de P. pachyrhizi. Uredinosporos do agente causal da ferrugem asiá- tica da soja expostos aos adjuvantes Assist®, Break Thru®, Aureo® e Nimbus® tiveram menor germinação a partir de duas horas de exposição. A inibição da germinação dos uredinosporos de P. kuehnii pelos adjuvantes em relação ao tratamento Testemunha aconteceu a partir de 4 horas de exposição. Às 8 horas, uredinosporos de ferrugem alaranjada da cana-de-açúcar que foram expostos aos adjuvantes Veget Oil® e Break Thru® apresentaram redução na germinação de 96,6 e 94,8%, respectivamente, quando comparados com o tratamento Testemunha.(AU)