Alternativas de control biorracionales sobre Phytophthora infestans, fitopatógeno causante de la gota en papa / Biorational control alternatives on Phytophthora infestans, phytopathogen causing gout in potatoes

Resumen La gota de la papa o tizón tardío es una de las enfermedades más agresivas que atacan los cultivos de papa y en poco tiempo los destruye, ocasionando enormes pérdidas económicas, es producida por el fitopatógeno Phytophthora infestans, oomyceto que ha tomado gran importancia debido a sus efectos devastadores y las dificultades para lograr su erradicación. Los fungicidas de naturaleza química representan un problema debido al uso inadecuado, lo que hace muy difícil eliminar la enfermedad por la aparición de nuevas especies resistentes. Existen nuevas alternativas para su control, basadas en el uso de sustancias de naturaleza vegetal, y la aplicación de nuevas herramientas capaces de realizar edición de genes, reprogramar o eliminar secuencias de ADN/ARN, favoreciendo así, la obtención de cultivos libres de sustancias tóxicas. En esta revisión, se presentan los métodos existentes de biocontrol: como aceites esenciales, metabolitos microbianos, herramientas moleculares y el uso de sustancias biodegradables que favorecen el manejo y prevención de plagas, que ayudan a mitigar la problemática ambiental generada por el uso de fungicidas de naturaleza química.

Abstract The drop of the potato or late blight is one of the most aggressive diseases that attack the potato crops and in a short time destroys them, causing great economic losses, it is produced by the phytopathogen Phytophthora infestans, oomyceto that has taken great importance due to its devastating effects and difficulties in achieving its eradication. Fungicides of a chemical nature represent a problem due to the affected use, which makes it very difficult to eliminate the disease due to the appearance of new resistant species. There are new alternatives for its control, based on the use of substances of the plant nature, and the application of new specific tools to perform gene editing, reprogram or eliminate DNA / RNA sequences, thus favoring the obtaining of cultures free of toxic substances . In this review, specific biocontrol methods are presented, such as essential oils, microbial metabolites, molecular tools and the use of biodegradable substances that favor the management and prevention of pests, which help mitigate the environmental problems generated by the use of fungicides from chemical nature.

Media preferences, micro-morphometric analysis, and cardinal growth temperature determination for Phytophthora infestans sensu lato isolated from different hosts in Colombia / Preferências de mídia, análise micro-morfométricos e determinação da temperature cardeal de crescimento para Phytophthora infestans sensu lato isolada de diferentes hospedeiros na Colômbia

Abstract During the last years Phytophthora infestans sensu lato (Mont. De Bary) has caused epidemics in Colombia in Andean fruit crops such as Solanum quitoense and Solanum betaceum. Establishment of new or modified experimental procedures to study this pathogen is a mandatory subject for scientists. Twelve isolates of Phytophthora spp. obtained from six different Solanum hosts in Colombia were used to evaluate the effect of five different solid media for growth and ability to produce sporangia and liberate zoospores. Determination of the best media culture and optimal growth temperature were necessary to perform measurements and correlate the provenance of isolates with phenotypic traits. Modifications were made to use ingredients available in local markets on the following media: lime bean agar (LBA), Tree tomato or tree tomato agar (TA), carrot agar (AZ), Rye A modified agar and 32% non-clarified V8 agar. Cardinal temperature determination was performed at 10, 15, 20, and 25 °C. Morphometric traits were measured once the optimal media culture and temperature were defined. Correlation analysis showed that there is a relationship between the host and isolate's preferences for media culture and optimal growth temperature. In addition, the production of characteristic sporangia, sporangiophore and mycelia was related with the media type used and host from which the isolate was collected. In this work useful information was provided to make studies about the biology and development of isolates gathered from cultivated and wild non-traditional hosts.

Resumo Durante os últimos anos Phytophthora infestans sensu lato causou epidemias na Colômbia em lavouras de frutos andinos, como Solanum quitoense e Solanum betaceum. Estabelecimento de procedimentos experimentais novos ou modificados para estudar este patógeno é um assunto obrigatório para os cientistas. Doze isolados de Phytophthora spp. obtidos de seis diferentes hosts Solanum na Colômbia foram usados para avaliar o efeito de cinco diferentes mídias sólidas para o crescimento e a capacidade de produzir esporângios e libertar zoósporos. Determinação da melhor cultura de mídia e temperatura de crescimento ideal foram necessárias para realizar medições e correlacionar a proveniência de isolados com traços fenotípica. Foram feitas modificações para usar os ingredientes disponíveis nos mercados locais nos seguintes meios: ágar do feijão de cal (LBA), tomate da árvore ou ágar do tomate da árvore (TA), ágar da cenoura (AZ), centeio um ágar modificado e 32% de ágar-V8 não-esclarecido. A determinação da temperatura Cardeal foi realizada em 10, 15, 20 e 25 °C. Traços morfométricos foram medidos uma vez que a cultura de mídia ideal e temperatura foram definidos. Análise de correlação mostrou que há uma relação entre o hospedeiro e isolar as preferências para a cultura de mídia e temperatura de crescimento ideal. Além disso, a produção de esporângios característica, esporangióforo e mycelia foi relacionada com o tipo de mídia utilizado e hospedeiros a partir do qual o isolado foi coletado. Neste trabalho foram fornecidas informações úteis para fazer estudos sobre a biologia e o desenvolvimento de isolados recolhidos de hospedeiros não-tradicionais cultivados e selvagens.

Evaluación de la resistencia de genotipos de tomate frente a aislados de Phytophthora infestans provenientes de Guatemala / Evaluation of the resistance of tomato genotypes against isolates of Phytophthora infestans from Guatemala

El oomicete Phytophthora infestans (Mont) DeBary es el patógeno responsable de causar la enfermedad denominada comúnmente como tizón tardío. Dicho patógeno afecta cultivos de importancia económica para el país, entre ellos el tomate, cultivo en el cual puede generar pérdidas totales en la producción. Dada la agresividad del patógeno, los programas de mejoramiento desarrollan nuevos genotipos de tomate resistentes a esta enfermedad mediante la incorporación de genes de resistencia, como los genes Ph. Sin embargo, las nuevas cepas de P. infestans, producto de la recombinación genética, pueden sobrepasar la resistencia de los genotipos mejorados. En este estudio se evaluó la resistencia genética de 13 genotipos de tomate mejorados, ante cinco aislados de P. infestans obtenidos de un estudio previo. Mediante inoculaciones in vitro y con la variable de respuesta de área bajo la curva del progreso de la enfermedad (AUDPC), se determinó que existió diferencia estadísticamente significativa (p < 0.05) entre el comportamiento de la resistencia, la agresividad de los aislados del patógeno y la interacción entre ambos factores. Además, se sugiere el uso de algunas líneas para ser incorporadas en programas de mejoramiento genético y así desarrollar nuevos genotipos aptos para el país y, por último, la profundización para establecer las bases genéticas de la resistencia de los dos cultivares evaluados en este estudio.

The oomycete Phytophthora infestans (Mont) DeBary causes late blight disease. This pathogen affects economically important crops for the country, including tomato, a crop in which it can causes total losses in production. Given the aggressiveness of the pathogen, tomato breeding programs develop new genotypes with resistance to P. infestans, being a source of resistance Ph genes. However, the new strains of P. infestans, product of the genetic variability of their populations, can overcome the resistance of the genotypes. In this study, the genetic resistance of 13 tomato materials was evaluated against 5 isolates of P. infestans obtained from a previous study. Through in vitro inoculations and with the area under the disease progress curve (AUDPC), it was determined that there was a statistically significant difference (p < 0.05) between the behavior of the genetic resistance, the aggressiveness of the isolates of the pathogen and the interaction between both factors. It is also suggested the use of some lines to be incorporated into tomato breeding programs and develop new genotypes suitable for the country and, finally, to establish the genetic basis of the resistance of the two cultivars evaluated in this study.

Controle da requeima e pinta preta da batata por fungicidas e seu reflexo sobre a produtividade e a qualidade de tubérculos / Control of potato late blight and early blight by fungicides and its effects on yield and quality of tubers

A requeima (Phytophthora infestans) e a pinta preta (Alternaria solani) estão entre as doenças mais frequentes e destrutivas da cultura da batata. Com o objetivo de avaliar a ação de diferentes grupos fungicidas no controle dessas doenças, foram realizados dois experimentos em cultivo comercial de batata (cultivares Agata e Monalisa), localizados em Pilar do Sul, São Paulo, nas safras 2008 e 2009. O delineamento experimental utilizado foi o de blocos ao acaso, com 4 repetições, sendo cada parcela de 20 m2. As aplicações foram realizadas com um pulverizador costal, munido de barra de aplicação e pressão constante de 3 Bar. O volume de aplicação variou de 300 a 500 L.ha-1 em função do desenvolvimento da cultura. As variáveis avaliadas para as duas doenças foram: severidade em folhas, área abaixo da curva de progresso da doença (AACPD), produtividade total e comercial, e classificação de tubérculos. Os menores níveis de doença e aumentos significativos na produtividade comercial e qualidade de tubérculos foram observados nas parcelas tratadas com fluopicolide + propamocarbe, dimetomorfe + ametoctradina, mandipropamida + clorotalonil, mandipropamida, fenamidona + propamocarbe, bentiavalicarbe + fluazinam, seguidos de dimetomorfe + clorotalonil, mefenoxam + clorotalonil e famoxadona + cimoxanil + mancozebe para requeima, e azoxistrobina + difenoconazol, picoxistrobina, piraclostrobina + metconazol, trifloxistrobina + tebuconazol, azoxistrobina, boscalida + piraclostrobina, iprodiona + pirimetanil e ciprodinil para pinta preta.(AU)

Late blight (Phytophthora infestans) and early blight (Alternaria solani) are among the most current and destructive diseases of potato. With the aim of evaluating the efficiency of different fungicides in the control of both diseases, two experiments were carried out in commercial potato crops (cultivars Agata and Monalisa) in Pilar do Sul, São Paulo State, Brazil, during 2008 and 2009 cropping season. Randomized block designs were used, with 4 replications and each parcel of 20 m2. Applications were carried out using coastal sprayer with spray-bar under constant pressure of 3 Bar. The application volume varied from 300 to 500 L per hectare according to the crop development. The evaluated variables for both diseases were: severity on leaves, area under disease progress curve (AUDPC), total and commercial yield, and classification of tubers. For late blight, the lower severity levels and significant increases in commercial yield and quality tubers were observed on the rows treated with fluopicolide + propamocarb, dimetomorph + ametoctradin, mandipropamid + chlorothalonil, mandipropamid, fenamidone + propamocarb, benthiavalicarb + fluazinam, followed by dimetomorph + chlorothalonil, mefenoxam + chlorothalonil e famoxadone + cymoxanil + mancozeb and, for early blight, azoxystrobin + difenoconazole, picoxystrobin, pyraclostrobin +metconazole, trifloxystrobin + tebuconazole, azoxystrobin, boscalid + pyraclostrobin, iprodione + pyrimethanil and cyprodinil.(AU)

Ametoctradin: a new fungicide for potato late blight control / Ametoctradina: um novo fungicida para controle da requeima da batata

Late blight (Phytophthora infestans ) is among the most important and destructive diseases in potato cultivation. Aiming to evaluate the efficacy of the new fungicide ametoctradin, mixed with dimethomorph and metiram, an experiment was carried out in commercial potato crops (Ágata cultivar) in Pilar do Sul, São Paulo state, Brazil, during the growing season in 2013. A randomized block design was used, with 4 replications, on plots of 15 m2. Applications were carried out using a backpack sprayer with a spray-bar under 3 bar of constant pressure. The application volume varied from 300 L to 600 L.ha-1 based on crop development. The variables evaluated were leaf severity (0 to 100%), area under the disease progress curve, and yield. Ametoctradin + dimethomorph (1.25 L.ha-1) provided significant control of late blight, exceeding benthiavalicarb + fluazinam, dimethomorph + metiram, dimethomorph + pyraclostrobin, cymoxanil + mancozeb, metalaxyl-M + mancozeb, ametoctradin + metiram, and pyraclostrobin + metiram, although similar to ametoctradin + dimethomorph (1.00 L.ha-1), fluopicolide + propamocarb, and fenamidone + propamocarb. Ametoctradin + metiram resulted in moderate control, which was always similar to the standard. Ametoctradin and its associations represent a new alternative for the management of potato late blight.(AU)

A requeima (Phytophthora infestans) é uma das mais importantes e destrutivas doenças da cultura da batata. Com o objetivo de avaliar a ação do novo fungicida ametoctradina em mistura com dimetomorfe e metiram, foi realizado um experimento em cultivo comercial de batata (cultivar Ágata), localizado em Pilar do Sul, São Paulo, na safra 2013. O delineamento experimental utilizado foi o de blocos ao acaso, com 4 repetições, sendo cada parcela de 15 m2. As aplicações foram realizadas com um pulverizador costal, munido de barra de aplicação e pressão constante de 3 Bar. O volume de aplicação variou de 300 a 600 L.ha-1 em função do desenvolvimento da cultura. As variáveis avaliadas foram: severidade em folhas (0 a 100%), área abaixo da curva de progresso da doença e produtividade. Ametoctradina + dimetomorfe (1,25 L.ha-1) proporcionou controle significativo da requeima, sendo superior a bentiavalicarbe + fluazinam, dimetomorfe + metiram, dimetomorfe + piraclostrobina, cimoxanil + mancozebe, metalaxil-M + mancozebe, ametoctradina + metiram, piraclostrobina + metiram e semelhante a ametoctradina + dimetomorfe (1,00 L.ha-1), fluopicolide + propamocarbe e fenamidona + propamocarbe. Ametoctradina + metiram apresentou comportamento intermediário, sendo sempre similar aos padrões. Ametoctradina e suas associações representam uma nova alternativa para o manejo da requeima na cultura da batata.(AU)

Data mining and influential analysis of gene expression data for plant resistance gene identification in tomato (Solanum lycopersicum)

Background Molecular mechanisms of plant-pathogen interactions have been studied thoroughly but much about them is still unknown. A better understanding of these mechanisms and the detection of new resistance genes can improve crop production and food supply. Extracting this knowledge from available genomic data is a challenging task. Results Here, we evaluate the usefulness of clustering, data-mining and regression to identify potential new resistance genes. Three types of analyses were conducted separately over two conditions, tomatoes inoculated with Phytophthora infestans and not inoculated tomatoes. Predictions for 10 new resistance genes obtained by all applied methods were selected as being the most reliable and are therefore reported as potential resistance genes. Conclusion Application of different statistical analyses to detect potential resistance genes reliably has shown to conduct interesting results that improve knowledge on molecular mechanisms of plant resistance to pathogens.

Expresión diferencial durante la interacción Solanum tuberosum - Phytophthora infestans / Differential expression during Solanum tuberosum- Phytophthora infestans interaction

La papa (Solanum tuberosum L.) es el cuarto cultivo más importante a nivel mundial y es el producto agrícola con mayor demanda de fungicidas, insecticidas y fertilizantes quí­micos. Las pérdidas mundiales ocasionadas por Phytophthora infestans (Mont.) de Bary en este cultivo, ascienden a 6,7 billones de dólares al año y su control quí­mico genera un aumento en los costos, perjudica la salud humana y el ambiente. Todo esto justifica la búsqueda constante de mecanismos alternativos para el control de la enfermedad, entre ellos la obtención de variedades resistentes mediante cisgenesis usando genotipos silvestres. Como un aporte en este sentido, y dada la falta de conocimiento de lo que controla y constituye la diferencia entre una respuesta compatible e incompatible, en el presente estudio se compararon los perfiles de expresión génica obtenidos mediante Despliegue Diferencial de variedades resistentes y susceptibles durante su interacción con P. infestans. Los resultados evidenciaron diferencias en la expresión génica, tanto a distintos tiempos post-inoculación como en el tipo de cambio de expresión, incluyendo la presencia y ausencia de bandas diferenciales y el aumento o disminución en su intensidad. Al analizar las secuencias de fragmentos diferencialmente expresados, se encontró que algunos fragmentos sobre-expresados en las variedades susceptibles, tení­an homología con secuencias que codifican para una serina-acetiltranferasa y para la subunidad Β de la RNA polimerasa. Por su parte, fragmentos sobre-expresados en la variedad resistente, tení­an homología con una secuencia codificante para un dominio transmembranal.

Potato (Solanum tuberosum L.) is the fourth most important crop worldwide; also, is the agriculture product with most fungicides, insecticides and chemical fertilizers requirement. Worldwide losses caused by Phytophthora infestans (Mont.) de Bary in this crop, amount to 6,7 billion dollars per year and its chemical control increase production costs, harming human health and environment. For these reasons, is necessary constant research for alternative mechanisms to control disease, including development of resistant varieties using cis-genesis from wild genotypes. As a contribution in this way, and the lack of knowledge of what controls and is the difference between compatible and incompatible interaction, in this study we compared gene expression profiles obtained by Differential Display from resistant and susceptible varieties, during their interaction with P. infestans. The results showed differences in gene expression between resistant and susceptible varieties, at different times post-inoculation as well as exchange expression rate, including the presence and absence of differential bands and increase or decrease in their intensity. After analyzing the sequences of differential expressed fragments, we found that some overexpressed fragments from susceptible varieties had homology with an encoded sequence for a serine-acetyltransferase and for a RNA Polymerase Β subunit. On the other hand, overexpressed fragments from resistant variety, had homology with an encoded sequence for a transmembrane domain.

Resistance Induction and Enhanced Tuber Production by Pre-inoculation with Bacterial Strains in Potato Plants against Phytophthora infestans

Efficacy of resistance induction by the bacterial isolates Pseudomonas putida (TRL2-3), Micrococcus luteus (TRK2-2) and Flexibacteraceae bacterium (MRL412), which were isolated from the rhizosphere of plants growing in Jeju Mountain, were tested in a greenhouse. The disease severity caused by Phytophthora infestans was effectively reduced in the potato plants pre-inoculated with bacterial isolates compared with those of the untreated control plants growing in a greenhouse. In order to estimate the level of protection by the bacterial isolates, Mancozeb WP (Diesen M(R), Kyong nong) and DL-3-amino butyric acid (BABA) were pre-treated, whereas Dimethomorph WP (Forum(R), Kyong nong) and phosphonic acid (H3PO3) were post-treated the challenge inoculation with the pathogen. Disease severities of chemical pre-treated as well as post-treated plants were reduced compare to those of the untreated. The disease reduction in the plants pre-treated with Mancozeb WP was the highest, whereas that of post-treated with Dimethomorph WP was the lowest. The yields of plants pre-inoculated with three bacterial isolates were greatly increased than those of control plants. These results suggest that biological control by bacterial isolates might be an alternative strategy against late blight disease in potato plants growing in greenhouse.

Identification of DNA Markers Linked to Metalaxyl Insensitivity Loci in Phytophthora infestans

A total of 24 isolates of Phytophthora infestans were tested and analyzed for their resistance to metalaxyl fungicides. Sensitivity to metalaxyl was determined by growing isolates on 20% V8 medium amended with 0, 5, and 100 microg/ml metalaxyl. Four isolates among the 24 tested were resistant to metalaxyl. Eleven isolates were intermediate and nine isolates were sensitive. Amplified fragment length polymorphism (AFLP) assay was used to identify the amplification products of resistant isolates. As a result, selected fragments were cloned, sequences and primer pairs were developed which linked to metalaxyl insensitivity in P. infestans using competitive PCR.

Screening for Antifungal Endophytic Fungi Against Six Plant Pathogenic Fungi

A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 11 locations in Korea. Their antifungal activities were screened in vivo by antifungal bioassays after they were cultured in potato dextrose broth and rice solid media. Antifungal activity against plant pathogenic fungi such as Magnaporthe grisea (rice blast), Corticium sasaki (rice sheath blight), Botrytis cinerea (tomato gray mold), Phytophthora infestans (tomato late blight), Puccinia recondita (wheat leaf rust), and Blumeria graminis f. sp. hordei (barley powdery mildew) was determined in vivo by observing the inhibition of plant disease development. Twenty (11.7%) endophytic fungi fermentation broths were able to control, by more than 90%, at least one of the six plant diseases tested. Among 187 liquid broths, the F0010 strain isolated from Abies holophylla had the most potent disease control activity; it showed control values of more than 90% against five plant diseases, except for tomato late blight. On the other hand, fourteen (7.5%) solid culture extracts exhibited potent disease control values of more than 90% against one of six plant diseases. The screening results of this study strongly suggested that metabolites of plant endophytic fungi could be good potential sources for screening programs of bioactive natural products.