Effect of Soil Inoculum Density and Temperature on the Incidence of Cucumber Black Root Rot

The effects of the density of Phomopsis sclerotioides in soil and other environmental factors on black root rot of cucumber were investigated. Cucumber plants were grown in soil containing P. sclerotioides at 1, 10, 100, and 1000 CFU/g. Wilt incidence from 3 to 7 weeks after transplanting was strongly correlated with P. sclerotioides density in soil (P < 0.05). Root rot of squash rootstock occurred in soil with very low inoculum densities (0.1 CFU/g), and was strongly related to P. sclerotioides density (Y = −0.3x + 1.2, R2 = 0.743, P < 0.05) at 8 weeks after transplanting. Cucumber plants showed wilt symptoms in soil containing 1 CFU/g. Wilt symptoms in cucumber plants occurred 4 to 7 days earlier in soil at 22°C than in soil at 27 or 17°C. Root rot development could be predicted from the density of P. sclerotioides in soil and soil temperature. However, further studies on the effects of other environmental factors are required to test the linear model in commercial fields. This information is essential for determining the threshold pathogen density at which most control techniques, particularly those other than soil disinfection, will be effective.

Detection of Phomopsis sclerotioides in Commercial Cucurbit Field Soil by Nested Time-Release PCR

A polymerase chain reaction (PCR)-based molecular method to detect Phomopsis sclerotioides in soil was developed using a species-specific primer pair. To improve sensitivity of the detection, three PCR techniques were used; namely, nested PCR using the primer pair internal transcribed spacer (ITS)1 and ITS4, time-release PCR using two different DNA polymerases (recombinant Taq and AmpliTaq Gold), and fluorescent PCR to obtain fluorescent-labeled PCR products that can be analyzed by capillary electrophoresis. The latter two techniques were combined and termed nested time-release fluorescent (NTRF)-PCR. The minimum concentration of DNA required to obtain species-specific PCR products successfully was 50 fg/μg. Using the NTRF-PCR method, the fungus could be detected in sandy soil that was artificially infested at a density of 10 CFU/g. The pathogen was detected in most soil samples collected from commercial cucumber fields in which visual disease symptoms had appeared, and even in samples collected from fields where visual disease symptoms had not appeared. To prevent the invasion and establishment of root-inhabiting pathogens such as P. sclerotioides, it is critical to detect the fungus in soil as soon as possible after its introduction into a cucumber-growing region.

First Report of Cucumber Black Root Rot Caused by Phomopsis sclerotioides in Italy

During April 2002 to September 2003 in unheated plastic greenhouses located in Fondi and Sperlonga (Latium Region of central Italy), in which more than 100 ha of cucumber (Cucumis sativus L.) were cultivated, an unusual disease causing decay of roots and plant wilting was observed. Many of the most common cultivars showed susceptibility, and in some farms, severe economic losses occurred. Disease symptoms observed on young plants included stunting, wilting, black root rot, and marked reduction of root development where pseudosclerotial structures were produced. The degree of root symptoms was proportional to the wilting. During periods of high evapotranspiration, wilting was severe in plants at the early stages of disease development, and even lightly infected plants wilted rapidly. Symptoms resembled those caused by vascular wilt fungi and were generally more severe in greenhouses with poorly drained soils. Samples from each of four greenhouses were collected during different periods of the growing season. Each sampling unit consisted of five to eight root pieces that were surface disinfected in 0.1% HgCl2 for 30 s, rinsed in sterile water, placed on petri dishes containing potato dextrose agar (pH 5.5), and incubated for 7 days at 25°C. Phomopsis sclerotioides van Kesteren (1,2) (identification confirmed by R. A. Samson, Centraalbureau voor Schimmelcultures of Utrecht, the Netherlands) was consistently recovered from affected tissues. Subcultures of three isolates were prepared and evaluated for pathogenicity. The experiments were conducted in a greenhouse with a 12-h photoperiod at 25 to 32°C. Seven-week-old seedlings (20 representatives per isolate) of a susceptible hybrid were dipped for 2 min in an agar slurry suspension of the pathogen and then returned to pots. Within 4 to 5 weeks after inoculation, all plants inoculated with each P. sclerotioides isolate showed the same symptoms observed in the field and caused wilting and death of approximately 80% of the inoculated plants. P. sclerotioides was consistently reisolated from the symptomatic test plant, whereas the fungus was never isolated from control plants. Another experiment using naturally infested soil in comparison with sterilized soil confirmed the soilborne nature of the fungus and its pathogenicity. To our knowledge, this is the first report of P. sclerotioides on cucumber in Italy. According to the experience of farmers and agricultural consultants, the disease was first observed in the last 3 to 4 years in unheated plastic greenhouses. However, we cannot exclude the possibility that the disease may have been present in central Italy prior to our observations, since it can be misdiagnosed and the symptoms can be masked by symptoms of other diseases. For these reasons, an accurate monitoring of the pathogen is necessary to determine the magnitude of the problem and its impact on the industry. Management practices that include long-term crop rotation with nonsusceptible hosts, removal and destruction of infected crop debris, and steam soil sterilization are suggested to reduce the economic losses. References: (1) E. Punithalingam et al. No. 461 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, U.K., 1975. (2) H. A. Van Kesteren. Neth. J. Plant Pathol. 73:112, 1967.

The effect of disinfectants on fungal diseases of cucumber

Formaline, lobac P, sodium hypochlorite (NaOCl), Korsolin and Menno-Ter-forte were effective disinfectants in the control of damping-off (Pythium sp.) from peat substrate. Ipasept, Sanisept and Virkon S (1%) were shown ineffective against Pythium sp. in peat. Only formaline was effective in the control of black root rot (Phomopsis sclerotioides) from peat. In sand substrate P. sclerotioides could be eradicated also with sodium hypochlorite. Verticillium wilt (Verticillium dahliae) from peat substrate could be controlled with formaline, lobac P, sodium hypochlorite and Virkon S. Formaline and sodium hypochlorite were effective against Verticillium wilt in sand. Black stem rot (Didymella hryoniae) was susceptible to all disinfectants tested.

Scanning electron microscopy of hyphal interaction between Streptomyces griseoviridis and some plant pathogenic fungi

The interaction between Streptomyces griseoviridis and the pathogens Alternaria brassicicola, Botrytis cinerea, Fusarium oxysporum, Mycocentrospora acerina, Rhizoctonia solani and Sclerotinia sclerotiorum was studied by SEM both on autoclaved seeds and living seedlings of turnip rape and carrot and the fungi Phomopsis sclerotioides and Pythium ultimum on cucumber seedlings. The samples were prepared by the standard method for examination by scanning electron microscope. The hyperparasitism of S. griseoviridis was clearly shown. S. griseoviridis tightly wound around Alternaria conidia and Sclerotinia hyphae, eventually disintegrating them. It grew along the hyphae of B. cinerea, P. sclerotioides and M. acerina, dissolving them. The hypha of F. oxysporum seemed to be slightly affected, and its conidia not at all. The hyperparasite grew only loosely on the hypha of R. solani and on the mycelium and oogonia of Pythium which seemed not to sustain much injury.

Phomopsis sclerotioides. [Distribution map].

A new distribution map is provided for Phomopsis sclerotioides van Kesteren. Hosts: Cucumber (Cucumis sativus) and other Cucurbitaceae. Information is given on the geographical distribution in Asia, India, Karnataka, Madhya Pradesh, Malaysia, Sabah, Europe, Denmark, France, German Democratic Republic, German Federal Republic, Netherlands, Norway, Sweden, Switzerland, UK, England, North America, Canada, BC.