Rhizoctonia solani as a potato pathogen : Variation of isolates in Finland and host response

Rhizoctonia solani is a soil inhabiting basidiomycetous fungus able to induce a wide range of symptoms in many plant species. This genetically complex species is divided to 13 anastomosis groups (AG), of which AG-3 is specialized to infect potato. However, also a few other AGs are able to infect or live in close contact with potato. On potato, R. solani infection causes two main types of diseases including stem canker observed as a dark brown lesions on developing stems and stolons, and black scurf that develops on new tubers close to the time of harvest. These disease symptoms are collectively called a ‘Rhizoctonia disease complex’. Between the growing seasons R. solani survives in soil and plant debri as sclerotia or as the sclerotia called black scurf on potato tubers which when used as seed offer the main route for dispersal of the fungus to new areas. The reasons for the dominance of AG-3 on potato seem to be attributable to its highly specialization to potato and its ability to infect and form sclerotia efficiently at low temperatures. In this study, a large nationwide survey of R. solani isolates was made in potato crops in Finland. Almost all characterized isolates belonged to AG-3. Additionally, three other AGs (AG-2-1, AG-4 and AG-5) were found associated with symptoms on potato plants but they were weaker pathogens on potato than AG-3 as less prone to form black scurf. According to phylogenetic analysis of the internal transcribed sequences (ITS) of the ribosomal RNA genes the Finnish AG-3 isolates are closely related to each other even though a wide variation of physiological features was observed between them. Detailed analysis of the ITS regions revealed single nucleotide polymorphism in 14 nucleotide positions of ITS-1 and ITS-2. Additionally, compensatory base changes on ITS-2 were detected which suggests that potato-infecting R. solani AG-3 could be considered as a separate species instead of an AG of R. solani. For the first time, molecular defence responses were studied and detected during the early phases of interaction between R. solani AG-3 and potato. Extensive systemic signalling for defence exploiting several known defence pathways was activated as soon as R. solani came into close contact with the base of a sprout. The defence response was strong enough to protect vulnerable sprout tips from new attacks by the pathogen. These results at least partly explain why potato emergence is eventually successful even under heavy infection pressure by R. solani.

Microbial L-phenylalanine ammonia-lyase. Purification, subunit structure and kinetic properties of the enzyme from Rhizoctonia solani

Phenylalanine ammonia-lyase (EC 4.3.1.5) was purified to homogeneity from the acetone-dried powders of the mycelial felts of the plant pathogenic fungus Rhizoctonia solani. 2. A useful modification in protamine sulphate treatment to get substantial purification of the enzyme in a single-step is described. 3. The purified enzyme shows bisubstrate activity towards L-phenylalanine and L-tyrosine. 4. It is sensitive to carbonyl reagents and the inhibition is not reversed by gel filtration. 5. The molecular weight of the enzyme as determined by Sephadex G-200 chromatography and sucrose-density-gradient centrifugation is around 330000. 6. The enzyme is made up of two pairs of unidentical subunits, with a molecular weight of 70000 (alpha) and 90000 (beta) respectively. 7. Studies on initial velocity versus substrate concentration have shown significant deviations from Michaelis-Menten kinetics. 8. The double-reciprocal plots are biphasic (concave downwards) and Hofstee plots show a curvilinear pattern. 9. The apparent Km value increases from 0.18 mM to as high as 5.0 mM with the increase in the concentration of the substrate and during this process the Vmax, increases by 2-2.5-fold. 10. The value of Hill coefficient is 0.5. 11. Steady-state rates of phenylalanine ammonia-lyase reaction in the presence of inhibitors like D-phenylalanine, cinnamic, p-coumaric, caffeic, dihydrocaffeic and phenylpyruvic acid have shown that only one molecule of each type of inhibitor binds to a molecule of the enzyme. These observations suggest the involvement of negative homotropic interactions in phenylalanine ammonia-lyase. 12. The enzyme could not be desensitized by treatment with HgCl2, p-chloromercuribenzoic acid or by repeated freezing and thawing.

Reacción de dos lineas de catimor y tres variedades de Coffea arabica L. al ataque de Rhizoctonia solani kuhn

Con el objetivo de evaluar la reacción de tres variedades de café (Coffea arabíca L.); Caturra Rojo, Catuaí Amarillo y Pacamara y dos lineas de Catimor (T-8667 y T-5175) al ataque de Rhízoctonía solani Kühn, fueron realizados experimentos en semillero y vivero, durante mayo de 1994 a enero de 1995, en las áreas del Centro Experimental de Café del Norte-UNICAFE. En semillero, se estudio el efecto de los substratos suelo y arena, además fueron evaluadas las variables: emergencia, índice de severidad y peso seco. En vivero se estudiaron los substratos suelo y suelo + pulpa en proporción 7:3, evaluandose incidencia, índice de severidad, altura, número de hojas y el peso seco. Los resultados mostraron que el Catimor T-5175 presentó resistencia horizontal tanto en semillero como en vivero, mientras Catuaí Amarillo fue altamente susceptible en semillero y vivero. Las variedades Caturra Rojo, Pacamara y Catimor T-8667 fueron susceptibles al patógeno. En semillero, la enfermedad se desarrolló con la misma intensidad tanto en suelo como arena. En Catuaí Amarillo, la pulpa de café disminuyó el crecimiento de la enfermedad, alcanzando la máxima incidencia a los 91 días después del transplante, mientras que en suelo a los 63 días después del trasplante.

Evaluación de tres fungicidas químicos para el control del añublo de la vaina (Rhizoctonia solani Kunh) en el cultivo del arroz (Oryza sativa L.)

El presente ensayo se estableció con el objetivo de determinar el efecto de 5 tratamientos (4 tratamientos químicos y un testigo absoluto) sobre el control del Añublo de la vaina bongo (Rhizoctonia Solani K.) en el cultivo del arroz ( Oryza sativa L. ) variedad Altamira 9, bajo las condiciones ecológicas de la finca Las Vegas, ubicada en el Valle de Sébaco. El ensayo se estableció en la época de postrera de 1995 (5 de Agosto hasta el 5 de Noviembre), utilizando un diseño de Bloques Completos al Azar con 5 tratamientos y 4 repeticiones. Las variables evaluadas fueron: Incidencia del hongo a los 60, 80 y 120 DDG; altura relativa de la lesión a los 60, 80 y 120 DDG; altura de la planta al momento de la cosecha; manchado de grano; peso mil granos expresado en gramos y rendimiento expresado en kg/ha.; de las variables evaluadas, solamente la incidencia del hongo y el rendimiento presentaron diferencias significativas. De los 4 tratamientos químicos evaluados, el que controló mejor la enfermedad fue el tratamiento 1 a base de Propiconazol aplicado en un solo momento ( 70 DDG) en dosis de 0.427ltlba.. Así mismo., el Análisis Económico determino que este tratamiento presento la mejor rentabilidad

Evaluación de quimicos, solarización y agua caliente contra Rhizoctonia solani kuhn en semilleros de cafe (Coffea arabica L.)

Se condujo un experimento durante el período de mayo a octubre de 1995, en las instalaciones del Centro Experimental del Café del Norte. (CECN- UNICAFE), Matagalpa -Nicaragua, con el objetivo de evaluar a nivel de semilleros de café (Cojfea arabica L.), dos métodos fisicos de desinfección de suelo (solarización y agua caliente), seis productos químicos desinfectantes de suelo, de los cuales cuatro son fungicidas (PCNB, clorotalonil, óxido de cobre y carboxin + captan), dos biocidas (dazornet y metarn sodio) y un testigo sin aplicación, establecidos en dos tipos de substratos (suelo y arena), para prevenir ataques de Rhizoctonia solani Külm. Los tratamientos fueron arreglados en bloques con cinco repeticiones donde se evaluaron las variables emergencia, incidencia e índice de severidad de la enfermedad y fitotoxicidad. El análisis de varianza (P = 0.05) no detectó diferencias significativas para estas variables excepto para la emergencia en substrato suelo siendo los mejores tratamientos agua caliente, dazomet y testigo según Tukey al 5 %. Se detectaron diferencias significativas en la interacción substrato * tratamiento para la variable emergencia; sin embargo Tukey al 5 % no detecta diferencias significativas. Se encontró diferencias significativas entre substratos, presentándose el mayor número de plántulas emergidas y los menores índices de incidencia y severidad de la enfermedad en los semilleros establecidos en substrato suelo (P = 0.05). El análisis económico de presupuesto parcial demostró que los tratamientos que presentaron el menor costo variable y mayor ingreso neto fueron el testigo seguido del óxido de cobre en ambos sustratos, donde el análisis de dominancia demuestra que el testigo domina a los demás tratamientos

Evaluación de pentacloronitrobenceno (PCNB) en el control de la pudrición radicular causada por Rhizoctonia solani kuhn en frijol común (Phaseolus vulgaris L.)

Este experimento fue conducido en La Compañía, Estación Experimental del Instituto Superior de Ciencias Agropecuarias (ISCA), ubicado en el departamento de Carazo, Nicaragua, Fue realizado en época de primera (junio a septiembre), 1987), con los siguientes objetivos: Determinar la dosis más adecuada de PCNB contra R solani en frijol. Evaluar el efecto de la mezcla de PCNB + Metalaxyl sobre el rendimiento del frijol. Determinar el porcentaje de pérdidas causadas por R. Solani en frijol. Seleccionar la mejor variedad de frijol en este experimento. Los resultados mostraron que la dosis de 6.81kg/ha de PCNB fue la mejor produciendo un rendimiento de 3. % más que el control y un 12% más que la mayor dosis de PCNB usada (11.36 kg/ha) la variedad Revolución 84 produjo el más alto rendimiento.

Phytophthora nicotianae e Rhizoctonia solani: dois novos patógenos da vinca no Brasil.

2007

Eficiencia in vitro de antagonistas de Rhizoctonia solani y Sclerotium rolfsii y análisis camparativo de distintos modelos de crecimiento

p.59-65

A PCR-based method to distinguish fungi of the rice sheath-blight complex, Rhizoctonia solani, R. oryzae and R. oryzae-sativae

Identification of Rhizoctonia solani, R. oryzae and R. oryzae-sativae, components of the rice sheath disease complex, is extremely difficult and often inaccurate and as a result may hinder the success of extensive breeding programmes throughout Asia. In this study, primers designed from unique regions within the rDNA internal transcribed spacers have been used to develop a rapid PCR-based diagnostic test to provide an accurate identification of the species on rice. Tests on the specificity of the primers concerned showed that they provide the means for accurate identification of the Rhizoctonia species responsible for sheath diseases in rice.

Biological control of Rhizoctonia solani Damping-off with a bacterium symbiotically associated with Steinernema abbasi

Rhizoctonia solani is a causal agent of damping-off of may cultivated plants. An isolate of the bacterium Pseudomonas oryzihabitans, symbiotically associated with the entomopathogenic nematode Steinernema abbasi, strongly inhibited the pathogen in vitro. The bacterium was firmly attached onto fungus mycelia and degraded the cell walls of the pathogen. In greenhouse experiments, bacterial suspension in sterile water applied in the soil, effectively controlled damping-off of radish.

Characterization and origin of infection of Rhizoctonia solani associated with Brassica oleracea crops in the UK

An extensive study was conducted to determine where in the production chain Rhizoctonia solani became associated with UK module-raised Brassica oleracea plants. In total, 2600 plants from 52 crops were sampled directly from propagators and repeat sampled from the field. Additional soil, compost and water samples were collected from propagation nurseries and screened using conventional agar isolation methods. No isolates of R. solani were recovered from any samples collected from propagation nurseries. Furthermore, nucleic acid preparations from samples of soil and compost from propagation nurseries gave negative results when tested for R. solani using real-time PCR. Conversely, R. solani was recovered from 116 of 1300 stem bases collected from field crops. All the data collected suggested R. solani became associated with B. oleracea in the field rather than during propagation. Parsimony and Bayesian phylogenetic studies of ribosomal DNA suggested the majority of further classified isolates belonged to anastomosis groups 2-1 (48/57) and AG-4HGII (8/57), groups known to be pathogenic on Brassica spp. in other countries. Many R. solani isolates were recovered from symptomless plant material and the possibilities for such an association are discussed.

Molecular tools to investigate Rhizoctonia solani distribution in soil

Real-time PCR protocols were developed to detect and discriminate 11 anastomosis groups (AGs) of Rhizoctonia solani using ribosomal internal transcribed spacer (ITS) regions (AG-1-IA, AG-1-IC, AG-2-1, AG-2-2, AG-4HGI+II, AG-4HGIII, AG-8) or beta-tubulin (AG-3, AG-4HGII, AG-5 and AG-9) sequences. All real-time assays were target group specific, except AG-2-2, which showed a weak cross-reaction with AG-2tabac. In addition, methods were developed for the high throughput extraction of DNA from soil and compost samples. The DNA extraction method was used with the AG-2-1 assay and shown to be quantitative with a detection threshold of 10-7 g of R. solani per g of soil. A similar DNA extraction efficiency was observed for samples from three contrasting soil types. The developed methods were then used to investigate the spatial distribution of R. solani AG-2-1 in field soils. Soil from shallow depths of a field planted with Brassica oleracea tested positive for R. solani AG-2-1 more frequently than soil collected from greater depths. Quantification of R. solani inoculum in field samples proved challenging due to low levels of inoculum in naturally occurring soils. The potential uses of real-time PCR and DNA extraction protocols to investigate the epidemiology of R. solani are discussed.

Distinctively variable sequence-based nuclear DNA markers for multilocus phylogeography of the soybean- and rice-infecting fungal pathogen Rhizoctonia solani AG-1 IA

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