Search

Rust infection results in decreases in photosynthesis and stress volatile emissions, but how these changes vary among host species has not been studied. We demonstrated that the impact of the obligate biotrophic fungus, Puccinia coronata f. sp. avenae, on foliage physiological processes is stronger in the primary host, Avena sativa (cultivated oat), than in the alternate host, Rhamnus frangula (alder buckthorn). Photosynthesis decreased with increasing percentage of damaged leaf area (DA) in both species, but reductions were greater in A. sativa. In A. sativa, photosynthetic reductions resulted from reductions in stomatal conductance and photosynthetic capacity; in R. frangula, reductions were due to reduced capacity. Infection reduced photosynthetic biomass and key nutrients in A. sativa, but not in R. frangula. In A. sativa, stress-elicited emissions (methyl jasmonate, green leaf volatiles, long-chain saturated aldehydes, mono- and sesquiterpenes, benzenoids, and carotenoid breakdown products) increased with increasing DA from 0% to 40%, but decreased with further increases in DA. In R. frangula, volatile emissions were slightly elicited but, surprisingly, constitutive isoprene emissions were enhanced. Different hosts had characteristic volatile fingerprints, indicating differential activation of biochemical pathways. Fungal-elicited reductions in photosynthesis scale uniformly with stress severity. In the sensitive host, biphasic scaling of volatiles indicates that heavy spread of chlorosis/necrosis leads to an overall cessation of physiological functioning.

There is limited understanding of the genetic variability in

Global warming affects the abiotic and biotic growth environment of plants, including the spread of fungal diseases such as Dutch elm disease (DED). Dutch elm disease-resistance of different Ulmus species varies, but how this is reflected in leaf-level physiological pathogen responses has not been investigated. We studied the impacts of mechanical injury alone and mechanical injury plus inoculation with the DED-causing pathogens Ophiostoma novo-ulmi subsp. novo-ulmi and O. novo-ulmi subsp. americana on Ulmus glabra, a more vulnerable species, and U. laevis, a more resistant species. Plant stress responses were evaluated for 12 days after stress application by monitoring leaf net CO2 assimilation rate (A), stomatal conductance (gs), ratio of ambient to intercellular CO2 concentration (Ca/Ci) and intrinsic water-use efficiency (A/gs), and by measuring biogenic volatile (VOC) release by plant leaves. In U. glabra and U. laevis, A was not affected by time, stressors or their interaction. Only in U. glabra, gs and Ca/Ci decreased in time, yet recovered by the end of the experiment. Although the emission compositions were affected in both species, the stress treatments enhanced VOC emission rates only in U. laevis. In this species, mechanical injury especially when combined with the pathogens increased the emission of lipoxygenase pathway volatiles and dimethylallyl diphosphate and geranyl diphosphate pathway volatiles. In conclusion, the more resistant species U. laevis had a more stable photosynthesis, but stronger pathogen-elicited volatile response, especially after inoculation by O. novo-ulmi subsp. novo-ulmi. Thus, stronger activation of defenses might underlay higher DED-resistance in this species.

Fungicides are widely used to reduce Fusarium infections and grain contamination by mycotoxins and increase the yield in cereals, but the efficacy of fungicide treatments in varying climates has not been systematically explored. Field experiments with Estonian spring barley (Hordeum vulgare L.) cv. ‘Maali’ were carried out in three successive years 2012–2014 with strongly varying weather conditions to study the effects of three fungicides, Folicur (active ingredient tebuconazole), Falcon Forte (prothioconazole, tebuconazole, spiroxamine) and Archer Top (fenpropidin, propiconazole), on the yield, incidence of Fusarium spp. and on the contamination of grain with mycotoxins DON, HT-2 and T-2. The fungicides were sprayed once a year at spring barley flowering time. The weather conditions during the three years of study were extremely different. The content of cycotoxin DON, HT2 and T2 was low. The spraying with fungicides had not a clear effect on the barley yield and 1 000 kernel weight, and the study year was primarily the main factor that affected barley yield (p

In this study, the effects of a winter cover crop (CC) and its combination with composted cattle manure (CC-M) on the development of potato late blight (Phytophthora infestans) and crop yield were ...

Application of organic fertilisers to soil prevents erosion, improves fertility and may suppress certain soil-borne plant pathogens, but it is still unclear how different trophic groups of fungi and oomycetes respond to long-term fertilisation treatment. The objective of the study was to examine the effect of different fertilisation regimes on fungal and oomycete pathogen- and mycorrhizal symbiont diversity and community structure in both soil and roots, using PacBio SMRT sequencing. The field experiment included three fertilisation treatments that have been applied since 1989: nitrogen fertilisation (WOM), nitrogen fertilisation with manure amendment (FYM) and alternative organic fertilisation (AOF), each applied at five different rates. Soil samples were collected three times during the growing season, while root samples were collected during the flowering stage. There was no influence of the studied variables on soil and root pathogen richness. Contrary to our hypothesis, pathogen relative abundance in both soil and roots was significantly higher in plots with the AOF treatment. Furthermore, richness and relative abundance of arbuscular mycorrhizal (AM) fungi decreased significantly in the AOF treatment. Permutational analysis of variance (PERMANOVA) demonstrated the effect of fertilisation treatment on pathogen community composition in both soil and roots. Our findings indicate that organic fertilisers may not always benefit soil microbial community composition. Therefore, further studies are needed to understand how fertilisation affects mycorrhizal mutualists and pathogens.

Potato crop losses can be substantial when conditions for late blight (Phytophthora infestans) development and spread are favourable. In this study, drivers of differences between the P. infestans population structures in Latvia and Lithuania, two neighbouring countries with similar potato-growing traditions, were investigated. Genotypes of P. infestans and population genetic diversity were analysed using a 12-plex simple sequence repeat (SSR) marker assay. High genetic diversity was demonstrated in both populations, with population diversity being higher in Latvia. It would appear that local populations established from soilborne oospores early in the season are well adapted to the conditions in the region. However, somewhat greater spread and survival of local clones was detected in Lithuania, suggesting that potato cropping there is more vulnerable to clonal invasion than in Latvia. For effective disease management, current strategies should be adjusted according to the specific pathogen populations in the region, considering the reproduction and survival of the pathogen. Potato growers should implement late blight preventive measures such as longer field rotation to prevent oospore infections, especially in Latvia, and should use more disease resistant cultivars and high-quality seed potatoes.

International audience; Potato is one of the most important agricultural crops in Algeria and worldwide. Each year, potato late blight, caused by Phytophthora infestans, is responsible for significant damage that leads to large production losses, and is thus a direct threat to food security in Algeria. In this study, 131 isolates of P. infestans and 92 DNA fingerprints captured on FTA cards were sampled from commercial and seed production fields in three major potato production regions (western, eastern, and central) during the main-season and late-season in Algeria over six cropping seasons (2010-2016). Genotypes of P. infestans and population genetic diversity were analysed using a 17-plex simple-sequence repeat (SSR) marker assay, and the mating type of all isolates was characterized. Both mating types (A1 and A2) were found, and often occurred in the same field. Differences in mating type proportion were observed between regions and between sampling periods. Analysis with SSR markers showed the prevalence of the EU_13_A2 lineage (70%) over EU_2_A1 (16%), EU_23_A1 (10%), and 4% of unknown multilocus lineage (MLL). The EU_13_A2 showed differentiation within the group. EU_23_A1 was found mainly in late-season crops. However, the cropping region did not influence the distribution of lineages due to the dispersal of the pathogen in Algeria by seeds. Genetic structure did not reveal a clear variation in distribution of the three lineages throughout the sampling regions. These data provide important new information on the composition and change over time of P. infestans populations in Algeria and open the way for a better understanding of the local epidemiology of this important pathogen.

The rhizosphere fungal community can play an important role in determining plant growth and health. In this study, using high-throughput sequencing, we investigated the fungal diversity and community composition in the roots and rhizosphere soil of 21 potato (Solanum tuberosum L.) cultivars. The samples were collected at three different sampling points. Furthermore, we assessed the differences in both diversity and composition of pathogen and saprotroph communities. In soil and roots, the fungal richness and relative abundance of pathogens and saprotrophs were mainly affected by sampling time. However, root fungal communities were also significantly affected by cultivar. The most substantial effect of cultivar was on root pathogen diversity. Moreover, the occurrence of most pathogens strongly varied among cultivars. Soil fungal community composition was primarily determined by sampling time; whereas in roots, the primary determinant was cultivar. Our results demonstrate changes in fungal communities over the potato growing season, as well as highlight the importance of potato cultivar on root fungal communities, and emphasise their importance in plant breeding.

The main aim of this study was to evaluate foliar late blight resistance and yield of selected foreign and Estonian potato cultivars to determine the most promising cultivars for the northern Baltic region. We hypothesized that local cultivars have higher late blight (Phytophthora infestans) resistance and yield due to higher blight resistance. Potato late blight observation field trials were carried out in 2012 and 2013 in Einola Farm, Tartu County, Estonia. In both growing seasons, 12 potato cultivars were compared, nine from the Dutch Breeding Company Agrico and three from the Estonian Crop Research Institute. They comprised four early cultivars: Ranomi, Esmee, Romie and Maret, four early-medium cultivars: Toluca, Mariska, Madeleine and Teele and four medium-late/late cultivars: Excellency, Bellefleur, Manitou and Anti. Of these, only two, Anti and Toluca, qualified as highly foliar late blight resistant. This research clearly indicates that, despite unfavourable conditions for the pathogen, late blight is able to destroy potato foliage in most of these cultivars before the end of the growing season in northern Baltic conditions. This reflects local genetic heterogeneity of populations of P. infestans, probably associated with the presence of strains adapted to a wide range of humidity conditions. In 2012 that had late blight favourable weather, the cultivar Toluca showed high foliar blight resistance, as it reached the maximum yield compared to most other cultivars which were susceptible or very susceptible to late blight and had drastically reduced yield. However, although the late blight infection was not recorded in the growing season 2013, the cultivar Toluca did not tolerate the dry weather well and its tuber yield remained significantly lower than that in all other cultivars. Additionally, in 2013, a potato early blight (Alternaria solani) outbreak was recorded with the two most susceptible cultivars being the local cultivar Teele and the Dutch cultivar Excellency.

Information about the root mycobiome may improve the overall quality of the plants and contribute to a valuable strategy to enhance sustainable agriculture. Therefore, we assessed differences in fungal community diversity and composition in the roots of potato, wheat and barley grown under mineral nitrogen fertilisation at five rates, with and without farmyard manure amendment. The same factorial combination of treatments has been used since 1989. Species richness and diversity, as well as community composition, of different fungal guilds were characterised using Illumina MiSeq sequencing of the ITS2 region. Crop species was the main factor determining overall fungal richness and diversity, with wheat showing the highest, and potato the lowest, richness and diversity. Pathogen diversity indices were highest in wheat plots amended with farmyard manure, whereas the lowest values were observed for potato roots. Fertilisation treatments and the interaction between crop species and fertilisation had the strongest impact on arbuscular mycorrhiza and saprotroph diversity. Crop species also determined the composition of the overall fungal community and that of fungal guilds, whereas fertilisation treatment had only a minor effect. This study highlights crop species as the main driver in shaping root fungal diversity and composition under the same environmental conditions.

In this study, the Estonian population of Phytophthora infestans was characterized with mating type, sensitivity to metalaxyl, virulence on 11 potato R-gene differentials and 12 SSR markers to show the outcome of potential sexual reproduction in the population. During the three years 2010–2012, 141 P. infestans isolates, collected from 23 potato fields, showed quite a high and stable frequency of the A2 mating type, 48% of the total population. In 87% of all sampled potato fields, both mating types were recorded, suggesting continuous sexual reproduction of P. infestans and possible oospore production. Metalaxyl-sensitive isolates prevailed in all three years (68 out of 99 isolates). Amongst the 95 isolates tested, 51 virulence races were found. The race structure was diverse, and most pathotypes were unique, appearing only once; the two most common pathotypes, 1.2.3.4.6.7.10.11 and 1.2.3.4.7.10.11, comprised 35% of the population. The P. infestans population was genetically highly diverse and most of the multilocus genotypes (MLGs) appeared only once. Furthermore, all of the MLGs appeared in only one of the three sampling years. Our results confirm that the high diversity in the Estonian P. infestans population is most likely the result of frequent sexual reproduction, which benefits the survival, adaptability and diversity of the pathogen in the climate of North-Eastern Europe.

The main aim of this study was to evaluate foliar late blight resistance and yield of selected foreign and Estonian potato cultivars to determine the most promising cultivars for the northern Baltic region. We hypothesized that local cultivars have higher late blight (Phytophthora infestans) resistance and yield due to higher blight resistance. Potato late blight observation field trials were carried out in 2012 and 2013 in Einola Farm, Tartu County, Estonia. In both growing seasons, 12 potato cultivars were compared, nine from the Dutch Breeding Company Agrico and three from the Estonian Crop Research Institute. They comprised four early cultivars: Ranomi, Esmee, Romie and Maret, four early-medium cultivars: Toluca, Mariska, Madeleine and Teele and four medium-late/late cultivars: Excellency, Bellefleur, Manitou and Anti. Of these, only two, Anti and Toluca, qualified as highly foliar late blight resistant. This research clearly indicates that, despite unfavourable conditions for the pathogen, late blight is able to destroy potato foliage in most of these cultivars before the end of the growing season in northern Baltic conditions. This reflects local genetic heterogeneity of populations of P. infestans, probably associated with the presence of strains adapted to a wide range of humidity conditions. In 2012 that had late blight favourable weather, the cultivar Toluca showed high foliar blight resistance, as it reached the maximum yield compared to most other cultivars which were susceptible or very susceptible to late blight and had drastically reduced yield. However, although the late blight infection was not recorded in the growing season 2013, the cultivar Toluca did not tolerate the dry weather well and its tuber yield remained significantly lower than that in all other cultivars. Additionally, in 2013, a potato early blight (Alternaria solani) outbreak was recorded with the two most susceptible cultivars being the local cultivar Teele and the Dutch cultivar Excellency.

Nowadays, in sustainable agriculture growing winter cover crops and using organic manure have been widely implemented to improve soil quality which leads to increase of microbial activity. Six-year study was performed to evaluate the effect of cropping system on soil microbial hydrolytic activity, content of soil organic carbon (SOC) and total nitrogen (Ntot) under potato cultivation by considering various management systems, while comparing the soil parameters after previous crop. The experiment consisted of five-field crop rotation with two diferent organic (Org 0 as control and Org II with winter cover crops plus added composted manure) and conventional (Conv 0 as control and Conv II with mineral N, 150 kg ha–1) farming systems. The results showed that hydrolytic activity of soil microbes decreased in every farming system Under potato cultivation. Interestingly, after potato cultivation, the lowest and highest decrease in the soil microbial hydrolytic activity were seen in Org II and Conv II systems, respectively. The highest soil microbial hydrolytic activity was measured in system Org II where incorporation of biomass from winter cover crops and cattle manure was used. Finally, SOC and Ntot were higher in organic farming systems and there no significant changes after potato cultivation.

Potato late blight, caused by the oomycete pathogen Phytophthora infestans, is one of the most important diseases of potato worldwide. This is the first study characterising Estonian P. infestans population using the SSR marker genotyping method. 70 P. infestans isolates collected during the growing season in 2004 from eight potato fields in three different regions of Estonia were characterised with nine polymorphic SSR markers. A1 and A2 mating type isolates were detected from every studied field indicating the high potential for sexual reproduction, which raises the genotypic diversity in P. infestans population. Results revealed highly diverse P. infestans population in Estonia resembling the Northern European populations. Most of the multilocus genotypes were detected only once among the collected isolates. Subpopulations collected from different geographical regions of Estonia showed no differentiation from each other but instead formed one highly diverse group.

A study was conducted which aimed to investigate the effect of farming systems (FS) (four conventional with increasing mineral N fertilizer amounts 0–150 kg of N ha−1) vs. two organic with catch crops (CC) and cattle manure (CC + M)) under the same five crop rotation system on the occurrence of tuber diseases such as common scab (Streptomyces spp.), silver scurf (Helminthosporium solani), dry rot (Fusarium spp.), and soft rot (Pectobacterium spp.). As the average of the first rotation years 2009–2011, the FS had a significant effect on the occurrence of silver scurf, dry rot and common scab (surface cover

Potato late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is one of the most serious potato (Solanum tuberosum L.) diseases, causing considerable yield loss in potato production worldwide, including Latvia. At present, the data on the population characteristics of P. infestans in Latvia is sparse. Therefore, the main aim of this study was to collect new data on Latvian isolates of P. infestans, to determine the main characteristics of the pathogen, particularly mating types, metalaxyl resistance and virulence with Black’s differential set of potato genotypes containing resistance (R) genes R1–R11. During 2010–2012, 181 isolates of P. infestans were collected from 23 potato fields, from 13 locations in Latvia. Out of 181 isolates tested, 52.5% were A1, 43.1% – A2 and 4.4% – self-fertile mating type. Of 116 isolates screened for resistance to metalaxyl, 25.9% were resistant, 19.8% – intermediate and 54.3% – sensitive. More than 80% of isolates were virulent to R1, R3, R4, R7, R10 and R11, while 33% or fewer isolates were virulent to R5, R8 and R9. The least frequent was virulence against R9 in 24% of isolates. Our study revealed that the Latvian population of P. infestans is diverse. The proportion of mating types and the occurrence of both A1 and A2 in the same field indicate the possibility of sexual recombination in Latvian fields. Thus, it is very important to keep the crop rotation system, to prevent soil contamination with long-living oospores. The Latvian population of P. infestans shares many similarities with other European populations, which suggests gene flow between populations.

This is the first characterisation of Lithuanian populations of Phytophthora infestans with mating type and virulence on a differential set of 11 R genes of Solanum demissum. The sensitivity to metalaxyl was also determined. In 2010–2012, 93 Phytophthora infestans isolates, collected from all over Lithuania, showed quite high and stable frequency of A2 mating type. On 45% of all studied potato fields, both mating types were recorded, suggesting sexual reproduction of P. infestans and possible oospore production in Lithuanian potato fields. Fourteen metalaxyl-resistant isolates were found among 71 isolates in the current study period, and sensitive isolates prevailed in all three years. Amongst the studied 70 isolates, 38 avirulence pathotypes were found. The Lithuanian race structure was highly diverse and complex (the average number of missing avirulence factors per isolate was 7.2). Most pathotypes were unique, appearing only once, and the four most common pathotypes comprised only 34% of the population.

In recent years, potato early blight [Alternaria solani (Ellis & G. Martin) L.R. Jones, A lternaria alternata (Fr.) Keissl.] has occurred with increasing frequency in European potato fields, including those in northern regions. In our study, early blight was evaluated during the two years 2010–2011 on potato plants in a conventional farming experiment. Both growing seasons were very favourable for early blight development and evaluation. Our study indicates that, despite chemical treatments, potato early blight is a great problem for potato growers in conventional farming systems in the North-East region in years when temperatures are higher than average. In 2011, early blight damage was particularly high on the untreated (N0P0K0) treatment plots where severity increased abruptly from an initial 2% to 91%, and killed the potato haulms within five weeks. In 2010, the most severely infected plants were also recorded on the untreated (N0P0K0) plots, where 70% of foliage was destroyed by the end of the growin...

Nowadays, organically produced products have become more popular than ever and interest in them is still growing fast. The early blight causal pathogen Alternaria solani has not been considered a great threat to potato in northern climate conditions in the past and has not been routinely sprayed against. During our study early blight was evaluated in 2010 and 2011 on the plants of a potato cultivar ‘Reet’ in an organic farming experiment. In our study, both growing seasons were very favourable for early blight development. Significant differences between the two cultivation technologies were found (F1,12 = 4.84, p = 0.048). In 2010, the area under disease progress curve (AUDPC) value was 303 on cover crop (CC) plots and 990 on CC + M (manure) plots that is three times higher, whereas in 2011, the AUDPC value was 967 on CC plots and 1195 on CC + M plots. Our results confirm that potato early blight has become a serious problem in North-East European organic potato fields and thus susceptible potato cultivars cannot be recommended for growing in an organic farming system. However, it is possible to influence the development severity of early blight by selecting the proper growing technology. Since, in the changing climate conditions and in susceptible cultivars, early blight is a potato disease that can cause early defoliation of plants and crop death, there is a need for resistant potato cultivars.

Altogether 365 isolates of Phytophthora infestans were sampled from potatoes propagated from seed potatoes of high (multiplied for two years in open field after meristem phase) and low (commercial certified seed multiplied for several consecutive years in open field) phytosanitary quality at different phases of epidemic progress during the growing seasons of 2001–2007 from field plots at two experimental institutes in Estonia, North-East Europe. High or low phytosanitary quality of seed potatoes had no effect on mating type ratio or response to metalaxyl in populations of P. infestans isolated from these two different groups of potato material. In contrast, the incidence of certain virulence factors, as well as the diversity of pathotypes, was very high in populations collected from potatoes propagated from low-quality seed in comparison to those from high-quality seed. The incidence of A2 mating type and fully metalaxyl sensitive strains was statistically significantly higher at the epidemic outbreak than later during epidemic progress. The incidence of most virulence factors and overall pathotype diversity were not affected by the temporal progress of the epidemic. Rare virulence factors 5 and 9 were more frequent at the outbreak of the epidemic and declined in the population during the course of epidemic.

The main aim of this research was to evaluate foliar resistance of the potato varieties of Dutch breeding company Agrico to find those most resistant to late blight in order to diminish the number of fungicide sprays required while obtaining a high yield in North-East European conditions. In the trial at the Einola Farm in 2010 and 2011, 10 potato varieties and 2 breeding lines were included. The disease scores of different varieties varied from 2.1 to 7.6 points. Nine of the varieties grown were susceptible and two varieties very susceptible to late blight. The most resistant variety was Toluca (foliar blight estimated mean value 5.9), which is immune to foliar blight in its land of origin. On the basis of two years data, we can conclude that most of the tested varieties are too susceptible to late blight to be grown without chemical control against this pathogen in North-East European conditions.

Microbial metabarcoding studies using high throughput sequencing technologies generate unprecedented amounts of DNA sequence data and make it possible to determine not only the composition of the communities but also the underlying factors powering the evolution of these communities. Despite the potential of community level studies in helping to better understand the ecology of pathogens and to manage the losses caused by them, very few oomycete addressing metabarcoding studies have been carried out and with highly variable results. The aim of this study was to develop new oomycete-specific ITS region PCR primers with improved specificity for metabarcoding and identification of oomycetes. The modified ITS1oo and the newly developed ITS3oo primers show improved in silico specificity for oomycetes and when paired with the universal ITS4 successfully amplified the DNA from all eleven tested oomycete species from six genera. High throughput sequencing of 20 soil samples from forest nurseries and bordering areas, using the primer pair ITS1oo/ITS4, recovered more than 400 oomycete OTUs, which is a significant increase over previous studies, and indicates the ability of the new method to detect various oomycete groups from complex substrates. The average fraction of oomycete reads per soil samples was 32–36%, with a maximum of 69%. The recovered oomycete OTUs represented the groups Lagenidiales, Peronosporales, Pythiales and Saprolegniales, with Pythiales dominating in all samples. In addition, the new primers were successfully used in identifying pathogens directly from infected plant tissues with Sanger sequencing. The pathogen was identified to the species or genus level in four samples out of six. In conclusion, the developed oomycete-specific primers provide a reliable method for the identification and metabarcoding of oomycetes.

Botrytis cinerea Pers.:Fr. is a major plant pathogen and a new approach is needed for its control in strawberry to minimise the increasing use of synthetic fungicides. The biofungicide Prestop-Mix, which contains Gliocladium catenulatum, is effective against Botrytis infections; however, the need for frequent applications increases the costs for farmers. Here, we demonstrate that bumble bees, Bombus terrestris L., effectively disseminate the preparation onto flowers in open field conditions. Over the course of three years, we found a highly significant decrease in the rate of Botrytis infection. Pathogen control was achieved with relatively low numbers of G. catenulatum spores per flower, even using flowers that are not highly attractive to bumble bees. An even distribution of spores was detected up to 100 m from the hives, either due to primary inoculation by bumble bees or secondary distribution by other flower visitors such as honey bees and solitary bees. We showed that the application of a biocontrol agent by bumble bees is reliable for the use of environmentally friendly pest control strategies in northern climatic conditions. This low cost technology is especially relevant for organic farming. This study provides valuable information for introducing this method into practice in open strawberry fields.

The potato late blight pathogen Phytophthora infestans was collected from a region of Eastern Estonia during a period of seven years (2001–07). In total, 180 isolates were assessed for virulence, 133 for mating type and 171 for metalaxyl resistance. The average frequencies of mating types A1 and A2 were 59% and 38%, respectively. Considerable differences in the proportion of A1 and A2 between years were found. The data indicate that, in the Estonian population of P. infestans, both mating types occurred simultaneously within fields, with the potential for sexual recombination. The relatively rare virulence factors 5 and 9 and the less frequent virulence factors 2, 6, 8 were found in all studied years. Racial diversity, calculated with normalised Shannon diversity index, was high (0.71) but did not show significant differences between different collection years. The data showed that the response of isolates to metalaxyl did not depend on the year of collection. Still, the interaction was considerable between response to metalaxyl and the year, which means that the proportion of sensitive isolates increased and the proportion of intermediate isolates decreased notably compared to other concentration variants. By contrast, metalaxyl resistant isolates remained at a similar level during the seven year period.

The characteristics of populations of Phytophthora infestans from organic farms, small conventional farms and large conventional farms were determined from isolates collected in northern Estonia in 2004 and 2005. For the population as a whole 41% were A2; all virulence factors to the 11 R genes from Solanum demissum were found; and more than 70% had high or intermediate resistance to metalaxyl. Isolates from organic farms tended to have more complex pathotypes than isolates from either large or small conventional farms, but there was a higher proportion of metalaxyl resistant isolates from large conventional farms than from small conventional farms or from organic farms.

A collection of 101 isolates of Phytophthora infestans, obtained from seven sampling sites representing central, east and south-east Estonia during 2002 and 2003 were assessed for several phenotypic and genotypic markers. All 101 isolates were assessed for virulence and resistance to metalaxyl. Virulence to each of the 11 classic resistance genes was found among the tested isolates. The mean number of virulences per isolate was 6.3, with a very low frequency of virulence against resistance genes R5 (5%) and R9 (14%). The most common pathotypes were 1.3.4.7.8.10.11 and 1.3.4.7.10.11, representing altogether 12% of the studied strains. In terms of metalaxyl resistance, 30 resistant, 52 intermediate and 19 sensitive isolates were found. A subgroup of 50 isolates was assessed for mating type, allozymes [glucose-6-phosphate isomerase (Gpi) and peptidase (Pep)], DNA fingerprints with probe RG57 and mtDNA haplotype. Of this subset, 30 were A1 and 20 were A2. Collections from three of the seven fields contained both mating types. Allozyme analysis did not reveal any polymorphism. However, 19 diverse RG57 fingerprints were detected, and two mitochondrial DNA haplotypes, Ia and IIa, were detected. By combining the mating type, mtDNA haplotype and RG57 fingerprint data, 26 multilocus genotypes were identified, of which 18 were detected only once. Genotypic diversity measured by the normalised Shannon diversity index was high (0.76). The large number of multilocus genotypes and the presence of both mating types in some fields indicate that sexual reproduction may take place in Estonian populations of P. infestans.