From 2006 to 2008, several similar Phytophthora isolates were obtained from roots of mature Quercus robur and other tree species (Acer platanoides, Fraxinus excelsior, Q. rubra, and Tilia cordata) in forests and parks in several areas in the Czech Republic. The trees were characterized by chlorotic and reduced foliage, crown dieback, and reduced root hairs. Several isolates of Phytophthora were obtained from necrotic roots of these trees and identified as Phytophthora plurivora Jung & Burgess (1). Isolated colonies grown on V8A medium were radiate to slightly chrysanthemum shaped with limited aerial mycelium in the center. Optimum growth was at 25°C, minimum at 5°C and maximum at 32°C. Radial growth of colonies averaged 6.4 mm/day at 20°C. The isolates were homothallic and produced abundant smooth-walled, spherical oogonia (23.3 to 29.1 μm in diameter), oospores were nearly plerotic or plerotic (21.8 to 26.9 μm in diameter), and the oospore wall was 1.2 to 1.4 μm thick. Antheridia were usually paragynous and measured 8.4 to 12 × 6.5 to 8 μm, but amphigynous antheridia were occasionally observed. Noncaducous, semipapillate sporangia formed on simple or sympodial sporangiophores, were obpyriform, ovoid, ellipsoid or irregular in shape, and occasionally distorted with more than one apex. Sporangia dimensions were 33 to 65 × 24 to 33 μm; L/B ratio 1.2 to 1.6 (–2.1). Comparison of DNA sequences of internal transcribed spacer (ITS) regions of isolates (representative strain GenBank Accession No. FJ952382) confirmed the 100% identity of P. plurivora (1). The soil infestation test was conducted using a P. plurivora isolate acquired from roots of Q. robur and 20 3-year-old plants of Q. robur. Sterilized millet seeds colonized by pathogen with the method as described (2) were used as inoculation medium and added into sterilized peat substrate at the rate of 0.5% (vol/vol). The plants were cultivated in 5.8-liter pots in a greenhouse (20°C, 16-h/8-h photoperiod). After 4 months, the roots of all plants were washed, dried, and weighed. The root biomass of 20 infected plants was significantly reduced by approximately 25% on average compared with the control 20 plants (P < 0.05, t-test, Statistica 7.1). The pathogen was consistently reisolated from the roots of infected plants but not from control plants. Stem inoculation tests were conducted with 20 replicates in each group of 2-year-old plants of oak, maple, ash, and lime and isolates acquired from the hosts. On each seedling, a 5-mm-diameter bark plug was removed 5 cm above the collar. The inoculum (5-mm-diameter V8A agar plug with actively growing mycelium) was applied to the exposed substrate. The wounds were sealed with Parafilm. Stem necrosis developed in all cases after 1 to 2 weeks, whereas control plants remained healthy. The pathogen was successfully reisolated from necrotic stem tissues. To our knowledge, this is the first report of P. plurivora causing root rot on oak, maple, ash, and lime in the Czech Republic. On the basis of the host range and distribution of P. plurivora in the Czech Republic, it can be assumed that, as elsewhere in Europe (1), this pathogen is widespread and is a common cause of decline of many tree species. References: (1) T. Jung and T. I. Burgess. Persoonia 22:95, 2009. (2) C. Robin et al. Plant Pathol. 50:708, 2001.
Susceptibility of Polish provenances and families of pedunculate oak (Quercus robur L.) to colonisation by Phytophthora cambivora
