Avaliação da atividade antimicrobiana das defensinas recombinantes de ervilha (drr230a) e café (cd1) produzidas em Pichia pastoris

The inefficiency of chemical pesticides to control phytopathogenic fungi in agriculture and the frequent incidence of human diseases caused by bacteria which are resistant to antibiotics lead to the search for alternative antimicrobial compounds. In this context, plant defensins are a promising tool for the control of both plant and human pathogenic agents. Plant defensins are cationic peptides of about 50 amino acid residues, rich in cysteine and whose tridimensional structure is considerably conserved among different plant species. These antimicrobial molecules represent an important innate component from plant defense response against pathogens and are expressed in various plant tissues, such as leaves, tubers, flowers, pods and seeds. The present work aimed at the evaluation of the antimicrobial activity of two plant defensins against different phytopathogenic fungi and pathogenic bacteria to humans. The defensin Drr230a, whose gene was isolated from pea (Pisum sativum), and the defensin CD1,whose gene was identified within coffee (Coffea arabica) transcriptome, were subcloned in yeast expression vector and expressed in Pichia pastoris. The gene cd1 was subcloned as two different recombinant forms: CD1tC, containing a six-histidine sequence (6xHis) at the peptide C-terminal region and CD1tN, containing 6xHis coding sequence at the N-terminal region. In the case of the defensin Drr230a, the 6xHis coding sequence was inserted only at the N-terminal region. Assays of the antimicrobial activity of the purified recombinant proteins rDrr230a and rCD1 against Phakopsora pachyrhizi, causal agent of soybean Asian rust, were performed to analyze the in vitro spore germination inhibition and disease severity caused by the fungus in planta. Both recombinant defensins were able to inhibit P. pachyrhizi uredospore germination, with no difference between the antimicrobial action of either CD1tC or CD1tN. Moreover, rDrr230a and rCD1 drastically reduced severity of soybean Asian rust, as demonstrated by in planta assays. In spite of the fact that rCD1 was not able to inhibit proliferation of the human pathogenic bacteria Staplylococcus aureus and Klebsiella pneumoniae, rCD1 was able to inhibit growth of the phytopathogenic fungus Fusarium tucumaniae, that causes soybean sudden death syndrome. The obtained results show that these plant defensins are useful candidates to be used in plant genetic engineering programs to control agriculture impacting fungal diseases.

Reação de genótipos de cafeeiro à Hemileia vastatrix e à Cercospora coffeicola

Coffee plants were introduced in Brazil in the Northern State of Para around 1727. Two major diseases have affected coffee trees in the country. One is rust, caused by fungus Hemileia vastatrix and accountable for production losses up to 50%. The other one is Cercospora leaf spot, caused by fungus Cercospora coffeicola endemic to all Brazilian coffee farms and, therefore, economically critical due to production losses both in the plant nursery and in the field. Availability of resistant varieties has been a constant challenge for breeders. Research programs play an important role in the search for new resistant and/or tolerant genotypes, since over time plants can become susceptible to new, genetically variable races of pathogens. This study aimed to evaluate the incidence and severity of such diseases, the resistance of different coffee genotypes to H. vastatrix and C. coffeicola pathogens, as well as the productivity of said genotypes in dense planting system. The experimental design consisted of randomized blocks, with twelve genotypes (treatments) and two replications (blocks). SISVAR® program was used to analyze data and compare them building on Scott-Knott test and Tukey’s test with a probability of 5%. Disease incidence and severity percentage were assessed for both Cercospora leaf spot and rust. Means were used to calculate the area under the disease progress curve (AUDPC) of both diseases. As to rust, the most resistant genotypes were H586-6, IBC 12, and H556-7 H567-6. As to Cercospora leaf spot and productivity, no statistical differences were found across genotypes. The dense planting system did not impair plant development, but favored disease evolution given the microclimate it produces.

Aplicación biotecnológica de bacterias rizosféricas: elicitación de sistemas defensivos sistémicos en relación con la producción de compuestos con interés farmacológico y alimentario

Esta memoria se basa en la hipótesis del papel fundamental que el sistema microbiano asociado al sistema radical de las plantas tiene sobre el metabolismo vegetal y las consiguientes aplicaciones de los productos derivados del mismo. El sistema rizosférico microbiano, el microbioma rizosférico cumple un papel fundamental para que la planta consiga mejorar sus capacidades de adaptación a un ambiente cambiante. En el capítulo 2 se exponen de manera enlazadas hipótesis sucesivas que pretenden evidenciar el potencial de aplicación de las bacterias asociadas a la planta y los distintos enfoques de aplicación biotecnológicos. En primer lugar, se plantea el uso de la rizosfera como fuente de microorganismos especialmente adaptados a la interacción del sistema planta/microorganismos. Un sistema en el que la presión selectiva definida por la planta condiciona el tipo de microorganismos, su diversidad y en definitiva la estructura de las comunidades microbianas que se desarrollan en este ecosistema. Sobre la hipótesis de la capacidad de selección de microorganismos por la planta, primero se busca una planta que aporte una serie de factores de presiónselección. La planta se elige en base a criterios filogenéticos y metabólicos (metabolismo secundario muy activo). Nicotiana glauca, es una Solanacea, de la misma familia que especies con gran interés alimentario, como el tomate, Solanum lycopersicum, la patata, Solanum tuberosum o el pimiento, Capsicum annum. Se selecciona esta planta como sujeto de muestreo rizosférico, en busca de un microbioma cultivable y con aplicaciones por sus aportaciones beneficiosas en la interacción. A continuación, sobre las casi mil cepas aisladas de la rizosfera, a lo largo de dos años, en tres suelos de características muy diferentes, se realiza un ensayo previo de actividades con potencial para incidir favorablemente sobre la salud de la planta. Las cepas se seleccionan sobre la base de un screening a gran escala en el que se intenta absorber la máxima variabilidad genética de los microorganismos que se desarrollan en el sistema rizosférico de Nicotiana glauca y pasan a estudiarse por su capacidad para inducir resistencia sistémica y efectos sobre el crecimiento en plantas de tomate, especie elegida como modelo de trabajo...

Pollen from 50 lake surface samples in Brandenburg, Germany, in 2009

Past changes in plant and landscape diversity can be evaluated through pollen analysis, however, pollen based diversity indexes are potentially biased by differential pollen production and deposition. Studies examining the relationship between pollen and landscape diversity are therefore needed. The aim of this study is to evaluate how different pollen based indexes capture aspects of landscape diversity. Pollen counts were obtained from surface samples of 50 small to medium sized lakes in Brandenburg (Northeast Germany) and compiled into two sets, with one containing all pollen counts from terrestrial plants and the second restricted to wind-pollinated taxa. Both sets were adjusted for the pollen production/dispersal bias using the REVEALS model. A high resolution biotope map was used to extract the density of total biotopes and different biotopes per area as parameters describing landscape diversity. In addition tree species diversity was obtained from forest inventory data. The Shannon index and the number of taxa in a sample of 10 pollen grains are highly correlated and provide a useful measure of pollen type diversity which corresponds best to landscape diversity within one km of the lake and the proportion of non-forested area within seven km. Adjustments of the pollen production/dispersal bias only slightly improve the relationships between pollen diversity and landscape diversity for the restricted dataset as well as for the forest inventory data and corresponding pollen types. Using rarefaction analysis, we propose the following convention: pollen type diversity is represented by the number of types in a small sample (low count e.g. 10), pollen type richness is the number of types in a large sample (high count e.g. 500) and pollen sample evenness is characterized by the ratio of the two. Synthesis. Pollen type diversity is a robust index that captures vegetation structure and landscape diversity. It is ideally suited for between site comparisons as it does not require high pollen counts. In concert with pollen type richness and evenness, it helps evaluating the effect of climate change and human land use on vegetation structure on long timescales.

Calidad fisiológica de semillas de berenjena, morrón y tomate tratadas con tiametoxam, un producto bioactivador

Considerando la escasa información referente al efecto del bioactivador tiametoxam y los potenciales beneficios que este compuesto puede proporcionar en producción de semillas, el objetivo de este estudio fue evaluar la influencia del producto en el desempeño fisiológico de semillas de berenjena, morrón y tomate. Semillas de tres lotes de cada una de las especies fueron tratadas con seis dosis crecientes de tiametoxam (0; 0,2; 0,4; 0,6; 0,8 y 1,0 mL del producto por cada mil semillas) y sometidas a germinación, primer conteo de germinación, test de frío, test de envejecimiento acelerado, longitud total de las plántulas y emergencia de plántulas en invernadero. El bioactivador tuvo efecto positivo sobre los parámetros analizados, estimulando el desarrollo fisiológico de las semillas, con intensidades variables de acuerdo con el lote. En berenjena, el aumento medio porcentual de la germinación en el primer conteo tuvo un incremento de aproximadamente 11 %, en morrón de 13 % y para los lotes de tomate aproximadamente de 17 %. La dosis de 0,6 mL cada mil semillas resultó ser la más efectiva para aumentar el vigor de las semillas de las tres especies.

Relations between zero-inflated variables in trials with horticultural crops

Certain characteristics of some vegetable crops allow multiple harvests during the production cycle; however, to our knowledge, no study has described the behavior of fruit production with progression of the production cycle in vegetable crops with multiple harvests that present data overdispersion. We aimed to characterize the data overdispersion of zero-inflated variables and identify the behavior of these variables during the production cycle of several vegetable crops with multiple harvests. Data from 11 uniformity trials were used without applying treatments; these comprise the database from the Experimental Plants Group at the Federal University of Santa Maria, Brazil. The trials were conducted using four horticultural species grown during different cultivation seasons, cultivation environments, and experimental structures. Although at each harvest, a larger number of basic units with harvest fruit was observed than units without harvest fruit, the basic unit percentage without fruit was high, generating an overdispersion within each individual harvest. The variability within each harvest was high and increased with the evolution of the production cycle of Capsicum annuum, Solanum lycopersicum var. cerasiforme, Phaseolus vulgaris, and Cucurbita pepo species. However, the correlation coefficient between the mean weight and number of harvest fruits tended to remain constant during the crop production cycle. These behaviors show that harvest management should be done individually, at each harvest, such that data overdispersion is reduced.

Control de la salinización del suelo mediante sistemas de drenes en producciones intensivas de cultivos bajo cubierta

Los suelos en los cuales se desarrolla la actividad hortícola en el Gran La Plata presentan como limitante natural restricciones en el drenaje debido a la presencia de elevada cantidad de arcillas desde superficie (expansivas en alta proporción). Estas características edáficas, junto a la calidad del agua de riego, y al excesivo aporte de enmiendas y fertilizantes, conducen a la ocurrencia de procesos degradativos tales como, salinización, alcalinización, anegamiento, mayor incidencia de plagas, y efectos sobre el cultivo. Se tuvo como objetivo estudiar la eficiencia de drenes subsuperficiales en el control de la salinidad-alcalinidad de un suelo Hapludert típico con cultivos protegidos, que presentaba en promedio pH 8,25; CE 4,45 dS.m-1; RAS 22, aunque con amplia variabilidad. Se comparan dos tratamientos con drenes (T1 10cm y T2 15cm de diámetro), un tratamiento con yeso (T3) y un testigo. Se evaluó el suelo y el cultivo de pimiento "Capsicum annuum" durante 3 años (2009-2011). Los drenes subsuperficiales condujeron a una homogenización de todo el sitio de estudio y evitaron la ocurrencia de pulsos salinos característicos de estos sistemas productivos, sin ocasionar cambios significativos en la RAS, la cual se mantuvo en los valores dados por el agua de riego. El diámetro del dren no influyó de una manera significativa. El yeso presentó mejoras temporales en la salinidad-alcalinidad, pero no condujo a una mejora permanente del sitio. El desarrollo fenológico y rendimiento del cultivo en los tratamientos fue mejor que en el testigo en los años evaluados. Los tratamientos con drenes se diferenciaron significativamente del yeso en 2009 (P 0,01). El distanciamiento entre drenes (5m), resultó suficiente como para mejorar todo el invernáculo (460m2).

Environmental and genetic effects on production traits of ewes originally from distinct environments.

Wool production and reproductive performance components of similar genotypes, brought from distinct production areas, were evaluated during five years trial at similar environments, such as, joining season and stocking rate on winter improved pasture. The least squares means revealed that the origin (breed) effect concentrated upon the Corriedale ewes wool production, whereas in Romney females it affected the reproductive performance.

Efeitos da quantidade de lã no escroto sobre características reprodutivas de carneiros Corriedale no decorrer do ano.

O presente trabalho foi delineado para se avaliar possíveis modificações nas características reprodutivas de carneiros em função da quantidade de lã no escroto e época do ano.

Efeito da quantidade de lã no escroto sobre características reprodutivas de carneiros Corriedale no decorrer do ano.

1984

Caracterização da incidência e evolução de pragas e doenças em agroecossistemas cafeeiros sob diferentes manejos.

A presente pesquisa tem o objetivo de avaliar a incidência e evolução das principais pragas e doenças do cafeeiro em agroecossistemas sob manejo convencional, organo-mineral, orgânico e agroflorestal nos municípios sul mineiros de Machado e Poço-Fundo. Foram realizados monitoramentos mensais do bicho-mineiro (Leucoptera coffeella), broca-do-café (Hypothenemus hampei), ferrugem (Hemileia vastatrix) e da cercosporiose (Cercospora coffeicola) por um período de um ano (dez/2007 a nov/2008) em lavouras da variedade Mundo Novo sob diferentes manejos. O monitoramento do bichomineiro e das doenças do cafeeiro foi realizado através da coleta de dez folhas do terceiro ou quarto par em todos os lados do cafeeiro, sendo amostradas vinte plantas por agroecossistema, totalizando duzentas folhas coletadas para avaliação dessas moléstias em cada sistema. Para o monitoramento da brocado- café selecionou-se 32 plantas, seis pontos diferentes em cada planta (10 frutos agrupados por ponto estabelecido) e realizou-se avaliação nãodestrutiva. Em todos os agroecossistemas monitorados, ambos manejados pela agricultura familiar, a infestação da broca-do-café e do bicho-mineiro não atingiram nível de dano econômico em nenhuma avaliação. Com relação às doenças avaliadas, observou-se que tanto a ferrugem como a cercosporiose atingiram nível de dano econômico em todos os sistemas de manejo. No entanto, nos sistemas organo-mineral, orgânico (monocultivo) e agroflorestal (diversificado) a ocorrência da ferrugem foi crítica, atingindo índices elevados nos meses de abril a outubro de 2008, registrando-se valores acima de 60% de incidência em quatro meses de avaliação. A incidência da cercosporiose foi relativamente menor no sistema convencional e orgânico (monocultivo), cujos picos não ultrapassaram 41% e 55%, consecutivamente. Já no sistema organomineral e agroflorestal (orgânico) a ocorrência da cercosporiose atingiu níveis muito elevados, chegando a 76% e 67%, respectivamente, no mês de julho. Apesar dos elevados níveis de incidência da ferrugem e cercosporiose encontrados nos agroecossistemas cafeeiros organo-mineral e orgânico, a produtividade média alcançada por esses sistemas nos últimos três anos foi superior ao convencional, mesmo esse último sistema investindo pesado em agrotóxicos e fertilizantes químicos.

Progeny tests for common bean isolines resistant to diseases with the aid molecular markers.

The aim of this study was progeny tests in segregating populations for resistance genes to these three diseases.

Asian soybean rust in Brazil: past, present, and future.

ABSTRACT: Asian soybean rust, caused by the fungus Phakopsora pachyrhizi, is the most severe disease of the crop and can cause yield losses of up to 90%. The disease was first reported in Brazil in 2001. Epidemics of the disease are common in the country, where the fungus can survive year?round. Regulatory measures to reduce the inoculum between seasons and avoid late-season soybean have been adopted to manage the disease. Disease control has relied mainly on chemical control, but a lower sensibility of the fungus to fungicides has been reported in Brazil. Major?resistance genes have been mapped and incorporated into the cultivars. With the reduced efficacy of the fungicides, the adoption of integrated measures to control the disease will be important for the sustainability of the crop. This review presents the main changes in the soybean crop system caused by the introduction of the fungus in Brazil, the current management strategies adopted to avoid losses, and the new trends that, together with biotechnological strategies, can improve management in the future. RESUMO: A ferrugem?asiática da soja, causada pelo fungo Phakopsora pachyrhizi, é a doença mais severa da cultura e pode causar perdas de produtividade de até 90%. A doença foi relatada pela primeira vez no Brasil em 2001. Epidemias da doença são comuns no País, onde o fungo pode sobreviver durante todo o ano. Medidas regulatórias para reduzir o inóculo entre safras e evitar a semeadura tardia de soja têm sido adotadas para manejar a doença. O controle da doença tem se baseado principalmente no controle químico, mas uma menor sensibilidade do fungo aos fungicidas tem sido relatada no Brasil. Genes de resistência têm sido mapeados e incorporados às cultivares. Por causa da redução da eficiência dos fungicidas, a adoção de medidas integradas para o controle da doença será importante para a sustentabilidade da cultura. Este artigo de revisão apresenta as principais mudanças no sistema de produção da soja causadas pela introdução do fungo no Brasil, as medidas de controle atualmente usadas para evitar perdas, e as novas tendências que, juntas com estratégias biotecnológicas, podem melhorar o manejo da doença no futuro.

Book of abstracts...

2016

A pigeonpea gene confers resistance to Asian soybean rust in soybean.

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.