Thirty-four strains of seven species of Trichoderma isolated from various fungal sources were compared for direct mycoparasitic activity (MPA), chitinase production and antibiotic activity (ANA) in order to choose the most appropriate partners for a strain-breeding programme. Within species genetic differences were also assesses in T. hamatum, T. harzianum and T. viride by means of random amplification of polymorphic DNA (RAPD). Endochitinase activities of the Trichoderma strains ranged between 20.4 and 1264.5 units/g dry weight of mycelium. The correlation between MPA and chitinase activity was not unambiguous and no correlation existed between MPA and ANA. The RAPD patterns of T. viride strains were highly variable, while isolates of T. harzianum proved to be more uniform; T. hamatum revealed remarkable intraspecific divergence. All these three comprised certain pairs of strains that are promising participants of a strain-improving programme, since their strong genetic affinities offer good changes for combining their contrasted biocontrol traits.

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Downy mildew of sunflower caused by Plasmopara halstedii (Farlow) Berlese et de Toni is a devastating disease worldwide. The treatment of seeds with fungicides and the use of resistant cultivars are widely employed control measures against this oomycete. Effective protection, however, may be hindered by the high genetic variability of the pathogen. There are 14 pathotypes of P. halstedii in Europe and as many as six of these were identified in Hungary before 2010 (1,4). In 2010, a new race, 704, was isolated in the eastern region of Hungary (3). Although the new pathotype was identified in two sunflower fields (near Vészto and Kondoros), it was expected to spread all over the country because of a lack of resistance against this race. The aim of our study, therefore, was to monitor the distribution of pathotype 704 in Hungary. Infected sunflower plants (2 to 5 samples/site) showing typical symptoms of downy mildew (leaf chlorosis, severe stunting) from four different sites (Árpádhalom, Rákóczifalva, Tiszasüly, and Újiráz) in the eastern region of the country were collected in mid-June 2012. Examination of isolates was carried out using a set of sunflower differential lines based on the internationally standardized method for race identification of P. halstedii (2). Inoculum of 17 isolates was increased on a susceptible cultivar (cv. Iregi Szürke Csíkos). Leaves containing zoosporangia were washed off in distilled water. The concentration of spore suspension for each isolate was adjusted to 20,000 to 30,000 viable zoosporangia per ml using a hemacytometer. Pre-germinated seeds of sunflower differential lines (20 seeds/line) with an optimal radicle length were selected and placed in separate petri dishes. They were filled with freshly prepared zoosporangial suspension of the isolates and incubated in the dark at 16°C for 6 h. Inoculated seeds were planted in trays. After 8 to 9 days when the first true leaves were ~0.5 to 1 cm long, the trays containing the plants were covered with transparent plastic bags overnight. Distilled water was sprayed into the bags to ensure a humid environment for stimulating sporulation. First disease assessment was performed immediately after incubation based on the appearance of characteristic white sporulation on cotyledons. A second evaluation was made of true leaves of 21-day-old plants. Twelve out of 17 isolates were pathotype 704, infecting either one of two commercial sunflower hybrids (NK Neoma and NK Brio) or volunteer sunflower plants. The remaining five isolates were races 700, 710, and 730, which are known to be widespread in Hungary (1). The presence of race 704 was proven in all sampling sites representing the eastern part of the country. This finding underscores the need to develop and grow improved sunflower hybrids with effective genes against this pathotype. To our knowledge, this is the first report of the wider distribution of pathotype 704 of P. halstedii in both Hungary and Central Europe. References: (1) T. J. Gulya. Adv. Downy Mildew Res. 3:121, 2007. (2) T. J. Gulya et al. Helia 14:11, 1991. (3) K. Rudolf et al. Növényvédelem 47:279, 2011. (4) F. Virányi and S. Maširević. Helia 14:7, 1991.

Downy mildew of sunflower, caused by Plasmopara halstedii (Farlow) Berlese et de Toni, is an economically important disease in Hungary and much of Europe. The known pathotypes (races) of the pathogen influence the resistance genes (Pl genes) incorporated into new sunflower hybrids to manage the disease. There are at least 36 pathotypes of P. halstedii worldwide (3), but the number of races is increasing rapidly. In 2010, race 704 was identified in Hungary for the first time (2). Race 704 has been reported to confer virulence on Pl6, a broad spectrum resistance gene that is widely used in sunflower hybrids. This has coincided with a significant increase in disease severity since 2010 in the country. Our objectives are to continuously monitor this pathogen and identify pathotypes of P. halstedii. Because of the unfavorable weather conditions for downy mildew in 2013, samples were collected at a single site (Kunszentmárton, South Hungary) in the beginning of July from NK Neoma sunflower hybrids. Disease incidence (early and late primary infection) was as high as 40%. Systemically mildewed plants showed severe stunting and leaf chlorosis, signs and symptoms consistent with downy mildew. P. halstedii was identified microscopically. Examination of isolates was carried out using a set of sunflower differential lines based on the internationally standardized method for race identification of P. halstedii (1). Inoculum of the isolates was increased on a susceptible cultivar (cv. Iregi szürke csíkos) and tested by inoculating 3-day-old seedlings of sunflower differential lines. Inoculated seedlings were planted in trays in glasshouse. After 8 to 9 days, seedlings were sprayed with distilled water, covered with black plastic bags, and left overnight to induce sporulation. Disease incidence was determined by examining cotyledons at 9 days after inoculation for sporulation and true leaves on 12 to 13 days after inoculation for secondary symptoms, such as leaf chlorosis and stunting (1). While several differential lines showed no typical susceptible/resistant reactions, i.e., the infection was much lower than 100%, it was concluded that the isolates were mixtures of different P. halstedii pathotypes. To obtain single isolates, we collected zoosporangia from the differential lines in question separately, and then inoculated the seedlings of the same genotype and a uniformly susceptible line. A single isolate caused as high as 100% infection on HA-335, containing resistance gene Pl6. Subsequent evaluation of this isolate with the entire differential set resulted in an aggregate virulence phenotype of 714. As resistance gene Pl6 is incorporated to the majority of sunflower hybrids grown in Hungary, pathotypes virulent on this gene, such as 704 and 714, are likely to spread. This underscores the need to prove the resistance to these races in the newly registered hybrids and for further research to identify P. halstedii pathotypes. It is also important to establish the identity of this new pathotype by already discovered 714 pathotypes in other countries like France and Italy and to discover the real conditions of local evolving of new pathogens. To our knowledge, this is the first report of pathotype 714 of P. halstedii in both Hungary and Central Europe. References: (1) T. J. Gulya et al. Helia 14:11, 1991. (2) K. Rudolf et al. Növényvédelem 47:279, 2011. (3) F. Virányi and O. Spring. Eur. J. Plant Pathol. 129:207, 2011.