Genetic diversity, aggressiveness and metalaxyl sensitivity of Pythium aphanidermatum populations infecting cucumber in Oman

Seventy three isolates of Pythium aphanidermatum obtained from cucumber from four different regions of Oman and 16 isolates of muskmelon from the Batinah region in Oman were characterized for aggressiveness, sensitivity to metalaxyl and genetic diversity using AFLP fingerprinting. Twenty isolates of P. aphanidermatum from diverse hosts from different countries were also included in the study. Most isolates from Oman were found to be aggressive on cucumber seedlings and all were highly sensitive to metalaxyl (EC50 < 0•80 µg mL−1). Isolates from cucumber and muskmelon were as aggressive as each other on both hosts (P > 0.05), which implies a lack of host specialization in P. aphanidermatum on these two hosts in Oman. AFLP analysis of all isolates using four primer-pair combinations resolved 152 bands, of which 61 (~40%) were polymorphic. Isolates of P. aphanidermatum from Oman and other countries exhibited high genetic similarity (mean = 94.1%) and produced 59 different AFLP profiles. Analysis of molecular variance indicated that most AFLP variation among populations of P. aphanidermatum in Oman was associated with geographical regions (FST = 0.118; P < 0.0001), not hosts (FST = -0.004; P = 0.4323). These data were supported by the high rate of recovery (24%) of identical phenotypes between cucumber and muskmelon fields in the same region as compared to the low recovery (10%) across regions in Oman, which suggests more frequent movement of Pythium inoculum among muskmelon and cucumber fields in the same region compared to movement across geographically separated regions. However, recovering clones among regions and different countries may imply circulation of Pythium inoculum via common sources in Oman and also intercontinental spread of isolates.

Molecular characterization and pathogenicity of Pythium species associated with damping-off in greenhouse cucumber (Cucumis sativus) in Oman

A study was undertaken in 2004 and 2005 to characterize pathogens associated with damping-off of greenhouse-grown cucumber seedlings in 13 districts in Oman. Identification of Pythium to the species level was based on sequences of the internal transcribed spacer (ITS) of the ribosomal DNA. Of the 98 Pythium isolates collected during the survey, Pythium aphanidermatum, P. spinosum, P. splendens and P. oligandrum accounted for 76%, 22%, 1% and 1%, respectively. Pythium aphanidermatum was isolated from all of the districts, while P. spinosum was isolated from seven districts. Pathogenicity tests showed inter- and intraspecific variation in aggressiveness between Pythium species. Pythium aphanidermatum, P. spinosum and P. splendens were found to be highly aggressive at 25°C. However, the aggressiveness of P. spinosum decreased when the temperature was raised to 30°C, which was found to correspond to the lower frequency of isolation of P. spinosum in the warmer seasons, compared to the cooler time of the year. Pythium aphanidermatum exhibited limited intraspecific variation in the sequences of the ITS region of the rDNA and showed 100% similarity to the corresponding P. aphanidermatum sequences from GenBank. The ITS sequence data, as well as morphological characteristics of P. spinosum isolates, showed a high level of similarity within and between P. spinosum and P. kunmingense, and suggested that the two species were synonymous. This study represents the first report of P. spinosum, P. splendens and P. oligandrum in Oman.

Genetic diversity, aggressiveness and metalaxyl sensitivity of Pythium spinosum infecting cucumber in Oman

A total of 24 isolates of Pythium spinosum from cucumber obtained from five regions in Oman were characterized for genetic diversity using amplified fragment length polymorphism (AFLP) fingerprinting and three isolates from the Netherlands, South Africa and Japan were included for comparison. Isolates from Oman were also characterized for aggressiveness on cucumber seedlings and sensitivity to metalaxyl. Identity of all isolates was confirmed using sequences of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA), which showed more than 99% nucleotide similarity among all isolates. Using six primer-pair combinations, AFLP fingerprinting resolved 295 AFLP markers of which 193 were polymorphic among isolates from other countries and only six were polymorphic among isolates of P. spinosum from Oman. Seven different AFLP phenotypes of P. spinosum were recovered in Oman; two of them were found to contain over 79% of isolates and one was recovered from all regions in Oman. Phenotypes from Oman showed very high (?99%) levels of genetic similarity to each other compared to moderate (mean =53%) levels of genetic similarity with phenotypes from other countries. In addition, all isolates from Oman were found to be highly sensitive to metalaxyl and all were aggressive on cucumber seedlings at 25°C. The high genetic similarity among phenotypes of P. spinosum in Oman as well as recovering two major clones across regions may suggest that P. spinosum has been recently introduced in Oman via a common source.

Molecular characterisation, pathogenesis and fungicide sensitivity of Pythium spp. from table beet (Beta vulgaris var. vulgaris) grown in the Lockyer Valley, Queensland

Table beet production in the Lockyer Valley of south-eastern Queensland is known to be adversely affected by soilborne root disease from infection by Pythium spp. However, little is known regarding the species or genotypes that are the causal agents of both pre- and post-emergence damping off. Based on RFLP analysis with HhaI, HinfI and MboI of the PCR amplified ITS region DNA from soil and diseased plant samples, the majority of 130 Pythium isolates could be grouped into three genotypes, designated LVP A, LVP B and LVP C. These groups comprised 43, 41 and 7% of all isolates, respectively. Deoxyribonucleic acid sequence analysis of the ITS region indicated that LVP A was a strain of Pythium aphanidermatum, with greater than 99% similarity to the corresponding P. aphanidermatum sequences from the publicly accessible databases. The DNA sequences from LVP B and LVP C were most closely related to P. ultimum and P. dissotocum, respectively. Lower frequencies of other distinct isolates with unique RFLP patterns were also obtained with high levels of similarity (>97%) to P. heterothallicum, P. periplocum and genotypes of P. ultimum other than LVP B. Inoculation trials of 1- and 4-week-old beet seedlings indicated that compared with isolates of the LVP B genotype, a higher frequency of LVP A isolates caused disease. Isolates with the LVP A, LVP B and LVP C genotypes were highly sensitive to the fungicide Ridomil MZ, which suppressed radial growth on V8 agar between approximately four and thirty fold at 5 μg/mL metalaxyl and 40 μg/mL mancozeb, a concentration far lower than the recommended field application rate.

Incidence and pathogenicity of Phytophthora palmivora and Pythium vexans associated with durian decline in far northern Queensland

In recent years, dieback of durian has become a major problem in mature orchards in the northern Queensland wet tropics region. A survey of 13 durian orchards was conducted during the dry season (July-September 2001) and following wet season (February-April 2002), with roots and soil from the root zone of affected trees being sampled. Phytophthora palmivora was recovered from the roots of affected trees on 12 of the 13 farms in the dry season, and all farms in the wet season. Pythium vexans was recovered from all 13 farms in both seasons. P. palmivora and P. vexans were recovered from diseased roots of 3-month-old durian seedlings cv. Monthong artificially inoculated with these organisms.

Efficacy of fungicides against damping-off in papaya seedlings caused by Pythium aphanidermatum.

Various chemical and non-chemical treatments were tested for their efficacy against damping-off in papaya seedlings caused by Pythium aphanidermatum. Three-week-old papaya seedlings were placed in a climate controlled experimental chamber and inoculated with macerated mycelium of P. aphanidermatum. Propamocarb as Previcur was found to be most effective at managing damping-off in papaya seedlings.

Management of Phytophthora fruit rot and Pythium-related root rot of papaya

Management of Phytophthora fruit rot and Pythium-related root rot of papaya.

Controlling Pythium and associated pests in ginger

This project is to identify treatments that ginger growers can use to control two serious soil-borne pathogens that have emerged and threaten the viability of the ginger industry. Pythium myriotylum, responsible for a severe rhizome rot, is the more serious of the two. It was first identified by ginger growers in the 2007/08 growing season, with some producers reporting total crop losses in some blocks. Symphylids are wingless soil-inhabiting arthropods that feed on the ginger plant's root tips and impair the plants´ ability to absorb nutrients, seriously restricting plant growth and development. Damage caused by symphylids to ginger roots is also expected to facilitate entry of Pythium into the plant.

Controlling Pythium in Ginger: Phase 2

The fungal disease Pythium Soft Rot is regarded by the ginger industry as their most serious disease threat. This project was developed to further investigate the factors contributing to the persistence and spread of Pythium Soft Rot on ginger farms and to identify measures for their control. The study demonstrated that the pathogen capable of causing Pythium Soft Rot in ginger was spread in contaminated ‘seed’, soil and water and that it can be managed through a combination of strategies that rely on early detection, reducing pathogen levels in soils, preventing water logging and restricting movement of contaminated ‘seed’, soil and water. However, in order to have an effective level of control, all strategies need to be integrated in an effective manner.

Pythium soft rot of ginger: Detection and identification of the causal pathogens, and their control

Ginger is considered by many people to be the outstanding member among 1400 other species in the family Zingiberaceae. Not only it is a valuable spice used by cooks throughout the world to impart unique flavour to their dishes but it also has a long track record in some Chinese and Indian cultures for treating common human ailments such as colds and headaches. Ginger has recently attracted considerable attention for its anti-inflammatory, antibacterial and antifungal properties. However, ginger as a crop is also susceptible to at least 24 different plant pathogens, including viruses, bacteria, fungi and nematodes. Of these, Pythium spp. (within the kingdom Stramenopila, phyllum Oomycota) are of most concern because various species can cause rotting and yield loss on ginger at any of the growth stages including during postharvest storage. Pythium gracile was the first species in the genus to be reported as a ginger pathogen, causing Pythium soft rot disease in India in 1907. Thereafter, numerous other Pythium spp. have been recorded from ginger growing regions throughout the world. Today, 15 Pythium species have been implicated as pathogens of the soft rot disease. Because accurate identification of a pathogen is the cornerstone of effective disease management programs, this review will focus on how to detect, identify and control Pythium spp. in general, with special emphasis on Pythium spp. associated with soft rot on ginger.

Manejo de mal seco (Pythium myriotylum) en quequisque (Xanthosoma sagittifolium L. Schott) mediante la siembra tardia, control de arvenses, enmiendas organicas y trichoderma spp. en Nueva Guinea, Nicaragua

El quequisque (Xanthosoma sagittifolium (L.) Schott.) pertenece a la familia Aráceas, es una de las raíces y tubérculos más importantes a nivel mundial. El principal problema que afecta el rendimiento es el mal seco ( Pythium myriotylum). Con el objetivo de evaluar el efecto de enmiendas orgánicas, Trichoderma y la siembra tardía en el manejo de mal seco ( Pythium myriotylum) en quequisque cv Blanco ( Xanthosoma sagittifolium (L.) Schott) en Nueva Guinea, se establecieron dos ensayos arreglados en diseño de bloques completos al azar. Ensayo I: Efecto de enmiendas orgánicas y Trichoderma spp. para el manejo de mal seco en vitroplantas establecidas en bancos con riego y control de arvenses, con tres bloques, seis tratamientos cada uno, ocho plantas por tratamiento por bloque. Ensayo II: Efecto de enmiendas orgánicas y trichoderma para el manejo de mal seco en vitroplantas establecidas en surcos sin riego y sin control de arvenses, con tres bloques, siete tratamientos, 15 plantas por tratamiento por bloque. Se evaluaron variables morfológicas y de rendimiento. Se realizó un análisis de varianza y prueba de separación de medias (Tukey, α = 0.05) a las variables morfológicas y de rendimiento. La siembra tardía de los ensayos I y II redujo el efecto de Pythium myriotylum sobre las plantas independientemente de los tratamientos evaluados. Las plantas tratadas con Trichoderma registraron mejor comportamiento morfológico y de rendimiento. La sobrevivencia de las plantas del Ensayo I fue de 70 % y en el Ensayo II fue de 83%, las plantas tratadas con Trichoderma registraron un porcentaje de sobrevivencia de 83% en el Ensayo I y 94 % en el Ensayo II. El testigo (sin aplicación) en el Ensayo I registró un 63% de sobrevivencia y el Ensayo II un 67%. Dry + humega y Trichoderma spp. registraron mejor comportamiento morfológico y de rendimiento.

Efecto del mal seco (Pythium myriotylum Drechs) en campo y sombreadero sobre la agromorfología de quince accesiones de quequisque (Xanthosoma spp) desarrollo de síntomas y detección microbiológica

El quequisque ( Xanthosoma spp.), familia Aráceae, es importante para el consumo interno y la exportación en Nicaragua, tiene como problema principal al mal seco ( Pythium myriotylum Drechs ) , que causa la pérdida total del rendimiento. Se establecieron cuatro ensayos para evaluar el efecto del mal seco sobre agromorfología de 15 accesiones de quequisque en suelos con (scams) y sin antecedentes de mal seco (ssams). Ensayo I: se evaluó el efecto del mal seco sobre la agromorfología de 15 accesiones en scams en Nueva Guinea. Se empleó el diseño BCA con tres bloques. Ensayo II: se estableció en DCA, se evaluó el comportamiento de 7 de las accesiones en ssams y se comparó con el rendimiento obtenido en scams. Ensayo III: se cultivó raíces de plantas infectadas en los medios de cultivo AN, V-8 y PARC y dilución en serie del scams para verificar la presencia de P myriotylum . Ensayo IV: se evaluó el desarrollo de síntomas de la enfermedad en las raíces, se establecieron 14 accesiones en scams en bolsas de polietileno (15 plantas por accesión) en sombreadero. Todas las accesiones desarrollaron mal seco, produjeron plantas débiles de 30-40 cm de altura, con tres hojas máximo y ningún rendimiento. Las accesiones en ssams reportaron 3-12 t ha -1 de rendimientos promedios. Las plantas en scams registrar on 91-100 % de pérdidas en el rendimiento. La infección en las raíces ocurre a partir de los 21 días después de la siembra, la planta genera nuevas raíces que también se infectan y muere por agotamiento de las reservas. La dilución en serie no resultó en el aislamiento del patógeno. Sembrar raíces recién infectadas resultó en la presencia de micelios del hongo en los medios V-8 y PARC.

Efecto de enmiendas organicas, trichoderma, microorganismos eficientes y metalaxil en el manejo de mal seco (Pythium myriotylum Drechs) en quequisque (Xanthosoma sagittifolium (L.) Schott) en suelo infectado en maceteras, mayo 2009-octubre 2010

El mal seco (Pythium myriotylum Drechs) reduce 90-100% la producción de quequisque. Con intención de aportar a la solución del problema se establecieron dos ensayos en maceteras, mayo 2009-octubre 2010, en esquema de diseño completo al azar (DCA). El objetivo fue evaluar el efecto de compost, humus de lombriz, dry, dry + humega, humega, trichoderma, microorganismos eficientes y metalaxil sobre el comportamiento agronómico de las plantas de quequisque (Xanthosoma sagittifolium (L.) Schott) infectadas con Pythium myriotylum. Ensayo I: Efecto de enmiendas orgánicas y trichoderma en vitroplantas (compost, humus de lombriz, dry, dry + humega, humega, trichoderma, testigo negativo (-) (suelo esterilizado a 105 oC por 24 horas) y testigo positivo (+) (suelo infectado sin ninguna aplicación)). Se emplearon 20 observaciones por tratamiento. Se utilizaron vitroplantas del cultivar Quequisque Blanco. Se empleó suelo con antecedentes de mal seco proveniente de Nueva Guinea. Ensayo II: Efecto de trichoderma, microorganismos eficientes y metalaxil en plantaspropagadas convencionalmente (trichoderma, EM, metalaxil, testigo (-) (suelo esterilizado a 220 ºC por 48 horas) y testigo (+) (suelo infectado sin ninguna aplicación)). Se emplearon 11 observaciones por tratamiento. Se evaluaron variables morfológicas, de rendimiento, raíz y sobrevivencia de las plantas. En el Ensayo I los tratamientos compost y testigo (-) fueron significativamente superiores en las variables morfológicas excepto el número de hijos. No hubo diferencias significativas entre los tratamientos en rendimiento, a excepción del largo de cormos donde humus de lombriz registró los cormos demenor longitud. Las plantas en compost, humus de lombriz y testigo (-) presentaron raíces al momento de la cosecha, las plantas de los demás tratamientos estaban muertas. En el Ensayo II el tratamiento metalaxil registró plantas significativamente superiores en altura, ancho y largo de la hoja y diámetro del pseudotallo a 32 y 74 dds. En las dos evaluaciones finales (127 y 193 dds) no hubo diferencias significativas entre los tratamientos en las variables morfológicas. Todas las plantas presentaron escaso crecimiento, producción y pocas raíces. A la cosecha las plantas presentaban un rango de sobrevivencia de 70-100%.

Controle biológico da podridão de raízes causada por Pythium spp. em cultivos hidropônicos.

2009

Involvement of phenazine-1-carboxylic acid, siderophores and hydrogen cyanide in suppression of Rhizoetonia solani and Pythium spp. damping-off by Pseudomonas oryzihabitans and Xenorhabdus nematophila

Entomopathogenic bacterial strains Pseudomonas (Flavimonas) oryzihabitans and Xenorhabdus nematophilus, both bacterial symbionts of the entomopathogenic nematodes Steinernema abbasi and S. carpocapsae have been recently used for suppression of soil-borne pathogens. Bacterial biocontrol agents (P. oryzihabitans and X nematophila) have been tested for production of secondary metabolites in vitro and their fungistatic effect,on mycelium and spore development of soil-borne pathogens. Isolates of Pythium spp. and Rhizoctonia solani, the causal agent of cotton damping-off, varied in sensitivity in vitro to the antibiotics phenazine-I-carboxylic acid (PCA), cyanide (HCN) and siderophores produced by bacterial strains shown previously to have potential for biological control of those pathogens. These findings affirm the role of the antibiotics PCA, HCN and siderophores in the biocontrol activity of these entomopathogenic strains and support earlier evidence that mechanisms of secondary metabolites are responsible for suppression of damping-off diseases. In the present studies colonies of R oryzihabitans showed production of PCA with presence of crystalline deposits after six days development and positive production where found as well in the siderophore's assay when X nematophila strain indicated HCN production in the in vitro assays. In vitro antifungal activity showed that bacteria densities of 101 to 10(6)cells/ml have antifungal activity in different media cultures. The results show further that isolates of Pythium spp. and R. solani insensitive to PCA, HCN and siderophores are present in the pathogen population and provide additional justification for the use of mixtures of entomopathogenic strains that employ different mechanisms of pathogen suppression to manage damping-off.