Genetic variation for tolerance to the downy mildew pathogen Peronospora variabilis in genetic resources of quinoa (Chenopodium quinoa).

BACKGROUND: Quinoa (Chenopodium quinoa Willd.) is an ancient grain crop that is tolerant to abiotic stress and has favorable nutritional properties. Downy mildew is the main disease of quinoa and is caused by infections of the biotrophic oomycete Peronospora variabilis Gaüm. Since the disease causes major yield losses, identifying sources of downy mildew tolerance in genetic resources and understanding its genetic basis are important goals in quinoa breeding. RESULTS: We infected 132 South American genotypes, three Danish cultivars and the weedy relative C. album with a single isolate of P. variabilis under greenhouse conditions and observed a large variation in disease traits like severity of infection, which ranged from 5 to 83%. Linear mixed models revealed a significant effect of genotypes on disease traits with high heritabilities (0.72 to 0.81). Factors like altitude at site of origin or seed saponin content did not correlate with mildew tolerance, but stomatal width was weakly correlated with severity of infection. Despite the strong genotypic effects on mildew tolerance, genome-wide association mapping with 88 genotypes failed to identify significant marker-trait associations indicating a polygenic architecture of mildew tolerance. CONCLUSIONS: The strong genetic effects on mildew tolerance allow to identify genetic resources, which are valuable sources of resistance in future quinoa breeding.

First report of Leptolegnia chapmanii (Peronosporomycetes: Saprolegniales) affecting mosquitoes in central Brazil.

Numerous isolates of an oomycete 'fungus', Leptolegnia chapmanii, are reported from Brazil for the first time. This aquatic pathogen was baited with Aedes aegypti sentinel larvae from stagnant, temporary bodies of water in selected locations under secondary tropical forest in and near the central Brazilian city of Goiânia and from more distant sites in the western and northern regions of the state of Goiás. Isolates were identified based on their morphological and developmental characters, comparative sequence data for the ITS and TEF loci, as well as their rapid activity against A. aegypti larvae. Taxonomic issues affecting the application of the name L. chapmanii and its typification are rectified. This study contributes to a better understanding of the presence and distribution of this oomycete in Brazil, its sequence-based identification, and of its potential as a biological agent against mosquito vectors.

Danos na soja causada por míldio / Damage to soybean caused by downy mildew

Downy mildew is widespread throughout the world. However, the damage that it causes has not been studied in Brazil yet. The objective of this work was to evaluate which components of soybean yield are affected by downy mildew and to determine the coefficient of damage. Two field experiments were conducted in Castro (2006/07) and Ponta Grossa (2007/08) in the state of Parana, Brazil. The experimental design consisted of completely randomized blocks of (i) six treatments and five replications for the 2006/07 and (ii) eight treatments and four replications for the 2007/08 field experiments. Potassium phosphite (750g a.i. ha-1), propamocarb + fenamidone (900g a.i. ha-1) and mancozeb (2400g a.i. ha-1) were applied. In all of the treatments, the maximum severity of downy mildew infection occurred at growth stage R5.3, and the percentage of severity ranged between 0 and 43%. The disease gradient was obtained only on the second season. Equations of damage were generated for the yield and grain weight. The number of grains per pod and the number of pods per plant were not affected by downy mildew but, the weight of 1000 seeds per plant was reduced linearly with increasing of mildew severity.(AU)

O míldio da soja é disseminado em todo mundo. Entretanto, o dano que ele causa não tem sido estudado no Brasil. O objetivo deste trabalho foi avaliar quais componentes de rendimento da soja são afetados por essa doença e determinar o coeficiente de dano. Dois experimentos foram conduzidos, um em Castro (2006/07) e outro em Ponta Grossa (2007/08), no Estado do Paraná, Brasil. O delineamento experimental foi de blocos ao acaso com (i) seis tratamentos e cinco repetições para os experimentos de campo, safras 2006/07, e (ii) oito tratamentos e quatro repetições para 2007/08. Foram aplicados fosfito de potássio (750g ia ha-1), propamocarb + fenamidona (900g ia ha-1) e mancozeb (2,400g ia ha-1). Em todos os tratamentos, a severidade máxima da infecção pelo míldio ocorreu no estádio R5.3 e a porcentagem de severidade variou entre 0 e 43%. O gradiente da doença foi obtido apenas na segunda safra. Equações de danos foram geradas para o peso de grãos e produtividade. O número de grãos por vagem e o número de vagens por planta não foram afetados pelo míldio mas o peso de 1000 sementes por planta foi reduzido linearmente com o aumento da severidade do míldio.(AU)

Characterization of a TIR-NBS-LRR gene associated with downy mildew resistance in grape.

Grapevine downy mildew, caused by Plasmopara viticola, is a devastating disease that results in considerable economic losses as well as environmental damage through the repeated application of fungicides. The nucleotide-binding site leucine-rich repeat gene family functions in plant immunoactivity against various pathogens and pests. In this study, the 5' and 3' ends of the resistance gene homology fragment RGA5 were obtained by rapid amplification of cDNA ends. The 4282-base pair full-length cDNA was obtained using gene-specific primers, and the corresponding 1335-amino acid protein sequence contained characteristic nucleotide-binding site leucine-rich repeat domains of plant resistance proteins, including the toll-interleukin receptor type region. Expression of RGA5 during P. viticola infection and abiotic stress was investigated using quantitative real-time polymerase chain reaction. The results showed that treatment with P. viticola and 4 abiotic stimuli (salicyclic acid, methyl-jasmonate, abscisic acid, H2O2) significantly induced RGA5 within 12 days of inoculation. Therefore, RGA5 may play a critical role in protecting grapevines against P. viticola via signaling pathways involving these molecules.

Mortality from Alzheimer's disease in Brazil, 2000-2009 / Enfermedad de Alzheimer: estudio de la mortalidad en Brasil, 2000-2009 / Doença de Alzheimer: estudo da mortalidade no Brasil, 2000-2009

Alzheimer's disease is the most prevalent type of dementia in the elderly worldwide. To evaluate the mortality trend from Alzheimer's disease in Brazil, a descriptive study was conducted with the Mortality Information System of the Brazilian Ministry of Health (2000-2009). Age and sex-standardized mortality rates were calculated in Brazil's state capitals, showing the percentage variation by exponential regression adjustment. The state capitals as a whole showed an annual growth in mortality rates in the 60 to 79 year age bracket of 8.4% in women and 7.7% in men. In the 80 and older age group, the increase was 15.5% in women and 14% in men. Meanwhile, the all-cause mortality rate declined in both elderly men and women. The increase in mortality from Alzheimer's disease occurred in the context of chronic diseases as a proxy for increasing prevalence of the disease in the population. The authors suggest healthcare strategies for individuals with chronic non-communicable diseases.

La enfermedad de Alzheimer es la demencia más frecuente entre adultos mayores en el mundo. Para evaluar la evolución de la mortalidad por la enfermedad de Alzheimer en Brasil, se ha desarrollado un estudio con datos del Sistema de Información sobre Mortalidad del Ministerio de Salud, durante el período 2000-2009. Se calcularon las tasas de mortalidad estandarizadas por edad y sexo en las capitales brasileñas y se registró la variación porcentual mediante ajuste de la regresión exponencial. El conjunto de las capitales presentó un aumento anual de las tasas de mortalidad en el grupo de edad de 60 a 79 años, de un 8,4% en mujeres y un 7,7% en hombres. En el grupo de 80 o más años, el aumento fue de un 15,5% en mujeres y un 14% en hombres. No obstante, hubo una disminución en la tasa de mortalidad por todas las causas entre los adultos mayores de ambos sexos. Se destaca un aumento de la mortalidad por enfermedad de Alzheimer en el contexto de las enfermedades crónicas como un proxy para la prevalencia de la enfermedad en la población, y se indican estrategias de asistencia en el cuidado de pacientes con enfermedades de larga duración.

A doença de Alzheimer é o tipo de demência que mais prevalece entre os idosos no mundo. Para avaliar a evolução da mortalidade por doença de Alzheimer no Brasil foi desenvolvido um estudo descritivo com os dados do Sistema de Informações sobre Mortalidade do Ministério da Saúde, no período de 2000 a 2009. Calcularam-se as taxas de mortalidade padronizadas por idade e sexo nas capitais brasileiras e se observou a variação percentual por meio de ajuste por regressão exponencial. Para o conjunto das capitais houve um crescimento anual nas taxas de mortalidade na faixa etária de 60 a 79 anos de 8,4% entre as mulheres e 7,7% entre os homens. No grupo etário de 80 anos e mais, o aumento foi de 15,5% entre as mulheres e 14% entre os homens. Contrariamente, verificou-se declínio da taxa de mortalidade por todas as causas entre os idosos em ambos os sexos. Destaca-se o aumento da mortalidade por doença de Alzheimer no contexto das doenças crônicas como um indicador aproximado da prevalência da doença na população, e são apontadas estratégias de assistência ao cuidado dos portadores de doenças de longa duração.

Development of a disease risk prediction model for downy mildew (Peronospora sparsa) in boysenberry.

Downy mildew caused by Peronospora sparsa has resulted in serious production losses in boysenberry (Rubus hybrid), blackberry (Rubus fruticosus), and rose (Rosa sp.) in New Zealand, Mexico, and the United States and the United Kingdom, respectively. Development of a model to predict downy mildew risk would facilitate development and implementation of a disease warning system for efficient fungicide spray application in the crops affected by this disease. Because detailed disease observation data were not available, a two-step approach was applied to develop an empirical risk prediction model for P. sparsa. To identify the weather patterns associated with a high incidence of downy mildew berry infections (dryberry disease) and derive parameters for the empirical model, classification and regression tree (CART) analysis was performed. Then, fuzzy sets were applied to develop a simple model to predict the disease risk based on the parameters derived from the CART analysis. High-risk seasons with a boysenberry downy mildew incidence >10% coincided with months when the number of hours per day with temperature of 15 to 20°C averaged >9.8 over the month and the number of days with rainfall in the month was >38.7%. The Fuzzy Peronospora Sparsa (FPS) model, developed using fuzzy sets, defined relationships among high-risk events, temperature, and rainfall conditions. In a validation study, the FPS model provided correct identification of both seasons with high downy mildew risk for boysenberry, blackberry, and rose and low risk in seasons when no disease was observed. As a result, the FPS model had a significant degree of agreement between predicted and observed risks of downy mildew for those crops (P = 0.002).

Molecular detection of Peronospora variabilis in quinoa seed and phylogeny of the quinoa downy mildew pathogen in South America and the United States.

Quinoa (Chenopodium quinoa) is an important export of the Andean region, and its key disease is quinoa downy mildew, caused by Peronospora variabilis. P. variabilis oospores can be seedborne and rapid methods to detect seedborne P. variabilis have not been developed. In this research, a polymerase chain reaction (PCR)-based detection method was developed to detect seedborne P. variabilis and a sequencing-based method was used to validate the PCR-based method. P. variabilis was detected in 31 of 33 quinoa seed lots using the PCR-based method and in 32 of 33 quinoa seed lots using the sequencing-based method. Thirty-one of the quinoa seed lots tested in this study were sold for human consumption, with seed originating from six different countries. Internal transcribed spacer (ITS) and cytochrome c oxidase subunit 2 (COX2) phylogenies were examined to determine whether geographical differences occurred in P. variabilis populations originating from Ecuador, Bolivia, and the United States. No geographical differences were observed in the ITS-derived phylogeny but the COX2 phylogeny indicated that geographical differences existed between U.S. and South American samples. Both ITS and COX2 phylogenies supported the existence of a Peronospora sp., distinct from P. variabilis, that causes systemic-like downy mildew symptoms on quinoa in Ecuador. The results of these studies allow for a better understanding of P. variabilis populations in South America and identified a new causal agent for quinoa downy mildew. The PCR-based seed detection method allows for the development of P. variabilis-free quinoa seed, which may prove important for management of quinoa downy mildew.

Altas diluições de sulfato de zinco sobre o manejo de tripes e rendimento de cebola em sistema orgânico / High dilutions of zinc sulfate on management of thrips and yield of onion in an organic

O cultivo da cebola convencional apresenta alto custo de produção devido ao uso de agroquímicos, e com os consequentes riscos à saúde dos agricultores e contaminação ambiental. A Epagri tem atuado com pesquisa e extensão para a implantação de sistema de produção de cebola orgânico em Santa Catarina. O objetivo deste experimento foi avaliar o efeito de altas diluições de sulfato de zinco sobre a incidência e danos de tripes, maturação fisiológica e rendimento de cebola em sistema orgânico de produção. O estudo foi conduzido durante três anos, 2011, 2012 e 2013, na Epagri, Estação Experimental de Ituporanga, SC. Os tratamentos foram altas diluições de sulfato de zinco em pulverizações foliares nas diluições 6cH, 12cH e 30cH e testemunha sem aplicação. O delineamento experimental foi o de blocos ao caso com quatro repetições. A incidência, danos de tripes, índice de clorofila, produtividade e maturação fisiológica não foram influenciados pelos tratamentos. A conservação pós-colheita de bulbos de cebola com as altas diluições de sulfato de zinco foi ajustada pelo seguinte modelo, em 2011, y= 0,014.x3 -0,626.x2 +5,894.x +56,21 (R2 = 0,56, p= 0,02). (AU)

The cost of onion cultivation in conventional systems in Santa Catarina (SC), Brazil, is high due to the use of agrochemicals, which pose risks to the farmers’ health and contaminate the nvironment. The aim of the present study was to evaluate the effect of high dilutions (HD) of zinc sulfate on the incidence of and damage by thrips, physiological maturation and yield of onion in an organic production system. The study was conducted over three years from 2011to 2013 at Ituporanga Experimental Station, Epagri, SC. Treatments consisted in zinc sulfate HD administered in foliar sprays in dilutions 6cH, 12cH and 30cH and untreated control. The experiment was performed in randomized blocks and four replications. The incidence of and damage by thrips, chlorophyll index and yield were not affected by the treatments. The postharvest storage of onion bulbs with HD of zinc sulfate was fit to the following model in 2011: y= 0.014.x3 -0.626.x2 +5.894.x +56.21 (R2 = 0.56,p= 0.02). (AU)

Inheritance of downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) resistance in grapevines.

Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two of the major diseases of most grapevine (Vitis vinifera L.) cultivars grown in Thailand. Therefore, breeding grapevines for improved downy mildew and anthracnose resistance is crucial. Factorial crosses were made between three downy mildew and/or anthracnose resistant lines ('NY88.0517.01', 'NY65.0550.04', and 'NY65.0551.05'; male parents) and two or three susceptible cultivars of V. vinifera ('Black Queen', 'Carolina Black Rose', and/or 'Italia'; female parents). F1 hybrid seedlings were evaluated for downy mildew and anthracnose resistance using a detached/excised leaf assay. For both diseases, the general combining ability (GCA) variance among male parents was significant, while the variance of GCA among females and the specific combining ability (SCA) variance were not significant, indicating the prevalence of additive over non-additive gene actions. The estimated narrow sense heritabilities of downy mildew and anthracnose resistance were 55.6 and 79.2%, respectively, suggesting that downy mildew/anthracnose resistance gene(s) were highly heritable. The 'Carolina Black Rose x NY65.0550.04' cross combination is recommended for future use.

Produtividade, incidência de tripes e perdas pós-colheita da cebola sob adubação orgânica e uso de biofertilizantes / Yield, thrips incidence and post harvest under organic fertilization and use of biofertilizers in onion crop

O rendimento, a sanidade e a qualidade de bulbos de cebola (Allium cepa L.) são infl uenciados pela nutrição das plantas, principalmente pelo nitrogênio (N), que além de ser requerido em grandes quantidades, é facilmente lixiviado do solo. O experimento teve por objetivo comparar o sistema manejo de adubação e fitossanitário convencional ao uso de métodos alternativos na produtividade, incidência de tripes e conservação de bulbos de cebola em pós-colheita. Com este propósito foi conduzido um experimento de campo, na safra 2010, num Cambissolo Háplico, em Ituporanga, SC. Foram estabelecidos quatro tratamentos: sistema de cultivo convencional (adubação mineral e agrotóxicos); adubação orgânica e fertirrigação de biofertilizante líquido (BF) a base de esterco de peru; adubação orgânica e fertirrigação com BF a base de esterco bovino (EB); e adubação orgânica e pulverização de BF a base de EB a 3%. Utilizou-se a cultivar Epagri 352 Bola Precoce. A semeadura, transplante e colheita foram realizados em 03/05, 12/07 e 06/12/2010, respectivamente. Conclui-se que independente do manejo adotado (orgânico ou convencional) não houve a redução dos danos causados por tripes; o uso de biofertilizantes não aumentou as perdas em pós-colheita; no sistema orgânico (adubação orgânica com o uso parcelado de biofertilizantes) houve a redução em 43% da produtividade em relação ao convencional.(AU)

The yield, health and quality of onion (Allium cepa L.) are infl uenced by plant nutrition mainly by nitrogen (N), which besides being required in large amounts, is easily leached from the soil. The experiment aimed to compare the system of fertilization and pest management to the conventional use of alternative methods on yield, thrips incidence, and conservation of onion bulbs post-harvest. For this purpose an experiment was conducted in 2010, in Cambisol, Ituporanga, SC. The treatments were conventional (mineral fertilizer and pesticides), organic fertilizer and fertigation of liquid biofertilizer (BF) the basis of turkey manure, organic fertilizer and irrigation with BF based manure EB), and organic fertilizer with spraying EB biofertilizer at 3%. The cultivar used was Epagri 352 Bola Precoce. Sowing, transplanting and harvesting were performed in 03/05, 12/07 and 06/12/2010, respectively. The conventional and organic management with biofertilizers did not reduce the damage caused by thrips, the use of biofertilizers not increased losses postharvest, the yield in the organic system (organic fertilization with biofertilizers) was reduced in 43 % when compared to conventional. postharvest conservation (AU)

Produtividade, incidência de tripes e perdas pós-colheita da cebola sob adubação orgânica e uso de biofertilizantes / Yield, thrips incidence and post harvest under organic fertilization and use of biofertilizers in onion crop

O rendimento, a sanidade e a qualidade de bulbos de cebola (Allium cepa L.) são infl uenciados pela nutrição das plantas, principalmente pelo nitrogênio (N), que além de ser requerido em grandes quantidades, é facilmente lixiviado do solo. O experimento teve por objetivo comparar o sistema manejo de adubação e fitossanitário convencional ao uso de métodos alternativos na produtividade, incidência de tripes e conservação de bulbos de cebola em pós-colheita. Com este propósito foi conduzido um experimento de campo, na safra 2010, num Cambissolo Háplico, em Ituporanga, SC. Foram estabelecidos quatro tratamentos: sistema de cultivo convencional (adubação mineral e agrotóxicos); adubação orgânica e fertirrigação de biofertilizante líquido (BF) a base de esterco de peru; adubação orgânica e fertirrigação com BF a base de esterco bovino (EB); e adubação orgânica e pulverização de BF a base de EB a 3%. Utilizou-se a cultivar Epagri 352 Bola Precoce. A semeadura, transplante e colheita foram realizados em 03/05, 12/07 e 06/12/2010, respectivamente. Conclui-se que independente do manejo adotado (orgânico ou convencional) não houve a redução dos danos causados por tripes; o uso de biofertilizantes não aumentou as perdas em pós-colheita; no sistema orgânico (adubação orgânica com o uso parcelado de biofertilizantes) houve a redução em 43% da produtividade em relação ao convencional.

The yield, health and quality of onion (Allium cepa L.) are infl uenced by plant nutrition mainly by nitrogen (N), which besides being required in large amounts, is easily leached from the soil. The experiment aimed to compare the system of fertilization and pest management to the conventional use of alternative methods on yield, thrips incidence, and conservation of onion bulbs post-harvest. For this purpose an experiment was conducted in 2010, in Cambisol, Ituporanga, SC. The treatments were conventional (mineral fertilizer and pesticides), organic fertilizer and fertigation of liquid biofertilizer (BF) the basis of turkey manure, organic fertilizer and irrigation with BF based manure EB), and organic fertilizer with spraying EB biofertilizer at 3%. The cultivar used was Epagri 352 Bola Precoce. Sowing, transplanting and harvesting were performed in 03/05, 12/07 and 06/12/2010, respectively. The conventional and organic management with biofertilizers did not reduce the damage caused by thrips, the use of biofertilizers not increased losses postharvest, the yield in the organic system (organic fertilization with biofertilizers) was reduced in 43 % when compared to conventional. postharvest conservation

Association of RGA-SSCP markers with resistance to downy mildew and anthracnose in grapevines.

Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two major diseases that severely affect most grapevine (Vitis vinifera) cultivars grown commercially in Thailand. Progress of conventional breeding programs of grapevine for improved resistance to these diseases can be speeded up by selection of molecular markers associated with resistance traits. We evaluated the association between 13 resistance gene analog (RGA)-single-strand conformation polymorphism (SSCP) markers with resistance to downy mildew and anthracnose in 71 segregating progenies of seven cross combinations between susceptible cultivars and resistant lines. F(1) hybrids from each cross were assessed for resistance to downy mildew and anthracnose (isolates Nk4-1 and Rc2-1) under laboratory conditions. Association of resistance traits with RGA-SSCP markers was evaluated using simple linear regression analysis. Three RGA-SSCP markers were found to be significantly correlated with anthracnose resistance, whereas significant correlation with downy mildew resistance was observed for only one RGA-SSCP marker. These results demonstrate the usefulness of RGA-SSCP markers. Four candidate markers with significant associations to resistance to these two major diseases of grapevine were identified. However, these putative associations between markers and resistance need to be verified with larger segregating populations before they can be used for marker-assisted selection.

NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions.

Plant defensins are small cysteine-rich peptides that inhibit the growth of a broad range of microbes. In this article, we describe NmDef02, a novel cDNA encoding a putative defensin isolated from Nicotiana megalosiphon upon inoculation with the tobacco blue mould pathogen Peronospora hyoscyami f.sp. tabacina. NmDef02 was heterologously expressed in the yeast Pichia pastoris, and the purified recombinant protein was found to display antimicrobial activity in vitro against important plant pathogens. Constitutive expression of NmDef02 gene in transgenic tobacco and potato plants enhanced resistance against various plant microbial pathogens, including the oomycete Phytophthora infestans, causal agent of the economically important potato late blight disease, under greenhouse and field conditions.

Universally Primed-PCR indicates geographical variation of Peronospora farinosa ex. Chenopodium quinoa.

In the Andean region of South America downy mildew, caused by Peronospora farinosa, is the most important disease of quinoa (Chenopodium quinoa). Peronospora farinosa, a highly polyphyletic species, occurs on quinoa and wild relatives on all continents. However, very little is known about the geographic diversity of the pathogen. As the interest in quinoa as a novel crop is increasing worldwide, geographical differences in the population structure of the downy mildew pathogen must be taken into consideration in order to design appropriate control strategies under a variety of circumstances. As a step towards understanding the geographic diversity of P. farinosa from quinoa, 40 downy mildew isolates from the Andean highlands and Denmark were characterized using universally primed PCR (UP-PCR). Eight UP-PCR primers were tested. A combined analysis of markers separated the Danish and Andean isolates in two distinct clusters. This study raises new questions about the origin and spread of P. farinosa on quinoa, its geographic diversity and host specificity.

Morphological and molecular characterization of the causal agent of downy mildew on Quinoa (Chenopodium quinoa).

Downy mildew is an economically important and widespread disease in quinoa (Chenopodium quinoa) growing areas. Although in many studies Peronospora farinosa is most commonly regarded as the causal agent of the disease, identification and classification of the pathogen remain still uncertain due to its taxonomic confusion. Thirty-six Peronospora isolates from quinoa with different geographic origins including Argentina, Bolivia, Denmark, Ecuador, and Peru were morphologically and molecularly compared with Peronospora species from other Chenopodium species. The morphology of three herbarium specimens was similar to that of P. variabilis, which originated from C. album, characterized by flexuous to curved ultimate branchlets and pedicellated conidia. Phylogenetic analysis based on ITS rDNA sequences also placed the quinoa pathogen within the same clade as P. variabilis. Within the ITS rDNA sequences of the quinoa pathogens, two base substitutions were found, which separated the majority of the Danish isolates from isolates from South America, but no sequence difference was found among the isolates from different cultivars of quinoa. The present results indicate that the pathogen responsible for the quinoa downy mildew is identical to Peronospora variabilis and that it should not be lumped with P. farinosa as claimed previously by most studies.

Tobacco blue mould disease caused by Peronospora hyoscyami f. sp. tabacina.

Blue mould [Peronospora hyoscyami f. sp. tabacina (Adam) Skalicky 1964] is one of the most important foliar diseases of tobacco that causes significant losses in the Americas, south-eastern Europe and the Middle East. This review summarizes the current knowledge of the mechanisms employed by this oomycete pathogen to colonize its host, with emphasis on molecular aspects of pathogenicity. In addition, key biochemical and molecular mechanisms involved in tobacco resistance to blue mould are discussed. TAXONOMY: Kingdom: Chromista (Straminipila); Phylum: Heterokontophyta; Class: Oomycete; Order: Peronosporales; Family: Peronosporaceae; Genus: Peronospora; Species: Peronospora hyoscyami f. sp. tabacina. DISEASE SYMPTOMS: The pathogen typically causes localized lesions on tobacco leaves that appear as single, or groups of, yellow spots that often coalesce to form light-brown necrotic areas. Some of the leaves exhibit grey to bluish downy mould on their lower surfaces. Diseased leaves can become twisted, such that the lower surfaces turn upwards. In such cases, the bluish colour of the diseased plants becomes quite conspicuous, especially under moist conditions when sporulation is abundant. Hence the name of the disease: tobacco blue mould. INFECTION PROCESS: The pathogen develops haustoria within plant cells that are thought to establish the transfer of nutrients from the host cell, and may also act in the delivery of effector proteins during infection. RESISTANCE: Several defence responses have been reported to occur in the Nicotiana tabacum-P. hyoscyami f. sp. tabacina interaction. These include the induction of pathogenesis-related genes, and a correlated increase in the activities of typical pathogenesis-related proteins, such as peroxidases, chitinases, beta-1,3-glucanases and lipoxygenases. Systemic acquired resistance is one of the best characterized tobacco defence responses activated on pathogen infection.

Genetic resistance to Peronospora tabacina in Nicotiana langsdorffii, a South American wild tobacco.

Several accessions of Nicotiana langsdorffii, a wild tobacco relative native to South America, express an incompatible interaction in response to infection by Peronospora tabacina, an oömycete that causes blue mold disease of tobacco (N. tabacum) and many other species of Nicotiana. In resistant accessions of N. langsdorffii, such as S-4-4, incompatibility takes the form of necrotic lesions that appear 48 h after pathogen inoculation, restricting pathogen growth, and suppressing subsequent asexual sporulation. Significantly elevated levels of salicylic acid and expression of a defense-related gene (HSR203J) were observed in S-4-4 leaves following blue mold infection. Genetic segregation analysis in F(2) and modified backcross populations showed that blue mold resistance is determined by a single dominant gene (NlRPT) present in S-4-4. Further characterization of this unique host-pathogen interaction has revealed that (i) necrotic lesion resistance is due to the hypersensitive response (HR), (ii) HR-mediated resistance is present in 7 of 10 N. langsdorffii accessions examined, but not in closely related species, (iii) in some accessions of N. langsdorffii, resistance is expressed in cotyledon tissue and seedling leaves as well as in adult plants, and (iv) several resistant accessions including S-4-4 express an unregulated ("runaway") HR in response to P. tabacina infection.

EIL2 transcription factor and glutathione synthetase are required for defense of tobacco against tobacco blue mold.

In order to identify tobacco (Nicotiana megalosiphon) genes involved in broad-spectrum resistance to tobacco blue mold (Peronospora hyoscyami f. sp. tabacina), suppression subtractive hybridization was used to generate cDNA from transcripts that are differentially expressed during an incompatible interaction. After differential screening by membrane-based hybridization, clones corresponding to 182 differentially expressed genes were selected, sequenced, and analyzed. The cDNA collection comprised a broad repertoire of genes associated with various processes. Northern blot analysis of a subset of these genes confirmed the differential expression patterns between the compatible and incompatible interaction. Subsequent virus-induced gene silencing (VIGS) of four genes that were found to be differentially induced was pursued. While VIGS of a lipid transfer protein gene or a glutamate decarboxylase gene in Nicotiana megalosiphon did not affect blue mold resistance, silencing of an EIL2 transcription factor gene and a glutathione synthetase gene was found to compromise the resistance of Nicotiana megalosiphon to P. hyoscyami f. sp. tabacina. Potentially, these genes can be used to engineer resistance in blue mold-susceptible tobacco cultivars.