The structure of mating type,virulence, metalaxyl resistance of Phytophthora infestans in a long-term phenotypic study in distinct location in Eastern Estonia

Journal of Plant Diseases and Protection - The potato late blight pathogen Phytophthora infestans was collected from a region of Eastern Estonia during a period of seven years (2001–07). In...     

Prevalence, species composition, genetic variation and pathogenicity of clover rot (Sclerotinia trifoliorum) and Fusarium spp. in red clover in Finland

The species composition of a total of 173 red clover root fungal isolates from red clover roots from two established organic fields, a field in a transitional phase to organic and from two conventional fields was investigated based on morphology and molecular methods. Fusarium avenaceum was the most common Fusarium species overall but it occurrred less frequently in older organic fields. Gliocladium spp., Trichoderma spp. and Rhizoctonia spp. isolates were more common in the established organic clover fields, which had been under organic management for more than ten years and in one conventional field, than in a field still in the transitional phase. The taxonomical status of certain Fusarium, Alternaria and Sclerotinia isolates difficult to identify by morphological traits alone could be confirmed by species-specific primers and by comparing their ITS (internal transcribed spacer region) sequences to known sequences. The fingerprinting patterns of RAPD-PCR products could be used for the identification of fungal isolates and for studying the genetic variation among the isolates. Only one of the Fusarium isolates originating from apparently healthy red clover roots was clearly pathogenic to germinated red clover seedlings. In detached leaf experiments, the cvs Jokioinen and Ilte were more susceptible to one of the Sclerotinia trifoliorum isolates than cvs Betty and Bjursele, while all of them were equally susceptible to two other S. trifoliorum isolates. In further greenhouse experiments with intact plants it was possible to slow down the development of clover rot to some extent by means of one of the biological agents tested (Bacillus subtilis 10-VIZR, commercial name Alirin B), and almost completely by chemical control.     

Symptoms and yield reduction caused by Dickeya spp. strains isolated from potato and river water in Finland

Biochemical characterisation of Dickeya strains isolated from potato plants and river water samples in Finland showed that the majority of the strains were biovar 3. They thus resembled the strains recently isolated from potato in the Netherlands, Poland and Israel and form a new clade within the Dickeya genus. About half of the Finnish isolates resembling strains within this new clade were virulent and caused wilting, necrotic lesions and rotting of leaves and stems. Similar symptoms were caused by D. dianthicola strains isolated from one potato sample and from several river water samples. Frequently, the rotting caused by the Dickeya strains was visible in the upper parts of the stem, while the stem base was necrotic from the pith but hard and green on the outside, resulting in symptoms quite different from the blackleg caused by Pectobacterium atrosepticum. The presence of Dickeya in the symptomatic plants in the field assay was verified with a conventional PCR and with a real-time PCR test developed for the purpose. The virulent Dickeya strains reduced the yield of individual plants by up to 50% and caused rotting of the daughter tubers in the field and in storage. Management of Dickeya spp. in the potato production chain requires awareness of the symptoms and extensive knowledge about the epidemiology of the disease.     

Fusarium oxysporum,F. proliferatum and F. redolens associated with basal rot of onion in Finland

In recent years in Finland, Fusarium infections in onions have increased, both in the field and in storage, and Fusarium species have taken the place of Botrytis as the worst pathogens causing post‐harvest rot of onion. To study Fusarium occurrence, samples were taken from onion sets, harvested onions and also from other plants grown in the onion fields. Isolates of five Fusarium species found in the survey were tested for pathogenicity on onion. Fusarium oxysporum was frequently found in onions and other plants, and, of the isolates tested, 31% caused disease symptoms and 15% caused growth stunting in onion seedlings. Fusarium proliferatum, a species previously not reported in Finland, was also identified. Over 50% of the diseased onion crop samples were infected with F. proliferatum, and all the F. proliferatum isolates tested were pathogenic to onion. Thus, compared to F. oxysporum, F. proliferatum seems to be more aggressive on onion. Also some of the F. redolens isolates were highly virulent, killing onion seedlings. Comparison of the translation elongation factor 1α gene sequences revealed that the majority of the aggressive isolates of F. oxysporum f. sp. cepae group together and are distinct from the other isolates. Incidence and relative proportions of the different Fusarium species differed between the sets and the mature bulbs. More research is required to determine to what extent Fusarium infections spoiling onions originate from infected onion sets rather than the field soil.     

Phenotypic and genetic variation in Finnish potato-late blight populations, 1997–2000

This study was conducted on potato late blight samples collected for monitoring early attacks in Finland from 1997 to 2000. Most of the 1726 Phytophthora infestans isolates were collected soon after the onset of the epidemics. Most of the isolates were tested for mating type as well as metalaxyl and propamocarb hydrochloride (HCl) sensitivity, while virulence on potato R-gene differentials and mitochondrial DNA (mtDNA) haplotype were determined for a subset of the isolates. In half of the fields from which more than one isolate was tested, both mating types were detected, indicating strong potential for sexual reproduction. The mating types coexisted more often in organic fields or gardens than in conventional fields. The proportion of A1 mating type decreased from approximately 80% (1997 and 1998) to 22% by the year 2000. Simultaneously, the proportion of isolates resistant to metalaxyl decreased from approximately 40% to 16%. Resistance to metalaxyl was confined to the IIa mtDNA haplotype and clearly associated with the A1 mating type, as resistance was 10 times more common among A1 isolates than among A2 isolates. Resistance to metalaxyl therefore probably derives from common descent from an isolate with the IIa haplotype. Most of the regional variation in metalaxyl sensitivity was also linked to mating type, as both metalaxyl resistance and the A1 mating type were most common in the north and south-west of Finland. In contrast to metalaxyl, only three propamocarb-HCl-insensitive isolates were found, but propamocarb-HCl sensitivity decreased significantly during the study.     

Phenotypic variation in Nordic populations of Phytophthora infestans in 2003

A total of 743 single-lesion isolates of Phytophthora infestans were collected in summer 2003 from Denmark, Finland, Norway and Sweden. Most of the isolates were tested for mating type, and subsets were tested for sensitivity to fungicides and virulence (host specific pathogenicity). Approximately 60% of the isolates were A1 mating type in each country. Both mating types were present in 40% of the fields where more than one isolate was tested, indicating strong potential for sexual reproduction. The proportion of metalaxyl-resistant isolates dropped to under 15% from the 60% observed in the early 1990s in Norway and Finland, possibly due to lower selection pressure because of decreased use of metalaxyl. Propamocarb-HCl sensitivity remained unchanged in the Nordic countries compared to the situation in 1997–2000 in Finland. Four isolates collected from Finland and Sweden were able to sporulate in the presence of this fungicide at a concentration of 1000 mg L⁻¹. In Norway and Finland the frequencies of virulence factors and pathotypes remained nearly unchanged since the 1990s, but the mean number of virulence factors per isolate increased from 5·6 to 6·3. In Denmark and Sweden virulence factors 2 and especially 6 were more common than in Norway and Finland. In addition, in the Swedish population the frequencies of pathotypes were quite even while in other countries pathotype 1,3,4,7,10,11 was most prevalent.     

Epidemiology and Integrated Control of Potato Late Blight in Europe

Phytophthora infestans, the causal agent of late blight, is a major threat to potato production in northwestern Europe. Before 1980, the worldwide population of P. infestans outside Mexico appeared to be asexual and to consist of a single clonal lineage of A1 mating type characterized by a single genotype. It is widely believed that new strains migrated into Europe in 1976 and that this led to subsequent population changes including the introduction of the A2 mating type. The population characteristics of recently collected isolates in NW Europe show a diverse population including both mating types, sexual reproduction and oospores, although differences are observed between regions. Although it is difficult to find direct evidence that new strains are more aggressive, there are several indications from experiments and field epidemics that the aggressiveness of P. infestans has increased in the past 20?years. The relative importance of the different primary inoculum sources and specific measures for reducing their role, such as covering dumps with plastic and preventing seed tubers from becoming infected, is described for the different regions. In NW Europe, varieties with greater resistance tend not to be grown on a large scale. From the grower??s perspective, the savings in fungicide input that can be achieved with these varieties are not compensated by the higher (perceived) risk of blight. Fungicides play a crucial role in the integrated control of late blight. The spray strategies in NW Europe and a table of the specific attributes of the most important fungicides in Europe are presented. The development and use of decision support systems (DSSs) in NW Europe are described. In The Netherlands, it is estimated that almost 40% of potato growers use recommendations based on commercially available DSS. In the Nordic countries, a new DSS concept with a fixed 7-day spray interval and a variable dose rate is being tested. In the UK, commercially available DSSs are used for c. 8% of the area. The validity of Smith Periods for the new population of P. infestans in the UK is currently being evaluated.     

Impact of Phytosanitary Quality of Seed Potato and Temporal Epidemic Progress on the Phenotypic Diversity of Phytophthora infestans Populations

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.     

Carrot Pathogen ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter solanacearum’ Haplotype C Detected in Symptomless Potato Plants in Finland

‘Candidatus Liberibacter solanacearum’ (CLso) haplotype C, a bacterial pathogen transmitted by the carrot psyllid Trioza apicalis, causes yield losses in carrot production. Due to concerns that this pathogen might also threaten potato (Solanum tuberosum) production, the occurrence of CLso in cultivated and volunteer potatoes in Tavastia Proper and Satakunta regions of Finland was studied. Volunteer potato plants were found in 13 of the 27 inspected carrot fields. Of the 148 potato samples tested by PCR, eight volunteer potato plants and one cultivated potato grown at the edge of a carrot field were found to be CLso positive. The PCR products obtained from these potatoes with primers OA2/OI2c, LpFrag4-1611F/LpFrag4-480R and CL514F/CL514R all showed 100% sequence identity to CLso haplotype C. This is the first observation of CLso haplotype C in field-grown potatoes. In addition, transmission experiments were performed. Attempts to transmit CLso into potato with carrot psyllids were not successful; however, CLso haplotype C was transmitted from infected carrots to potato plants by leaf grafting and by phloem connection formed by dodder, a parasitic plant, and found to survive in the potato plants for several weeks after transmission. However, the bacterial colonisation progressed slowly in the potato phloem and the amount of bacteria detected was low. The plants produced from the daughter tubers of the CLso-positive potato plants were all CLso negative, suggesting that CLso haplotype C was not able to pass to the daughter plants. None of the CLso-positive potatoes inoculated in greenhouse or collected from fields showed symptoms characteristic of zebra chip disease, associated with CLso haplotypes A and B.     

Identifying difficulties in rapeseed root penetration in farmers’ fields in northern European conditions

Tap‐rooted rapeseed is generally considered sensitive to soil compaction. We examined 473 Finnish rapeseed fields sown in 2007 to 2009 to both spring turnip rape Brassica rapa L. (dominant crop) and oilseed rape Brassica napus L. Roots were rated late in the growing season according to penetration capacity. Roots in each field were rated from G1 (no problems with root penetration) to G5 (failure to penetrate deeper soil layers). There were significant effects of year, crop species, soil type, special soil properties and soil cultivation methods on frequency of G1 and G4+5 roots. Restricted root penetration was a common phenomenon. Typically, fields had <30% G3 and G4+5 roots, however, some fields had >70% G4+5 roots and were cases for serious concern. Yield loss was linearly related to the incidence of G4+5 plants in 2007, but there was no relationship with the other years. Oilseed rape had a greater proportion of G4+5 roots than turnip rape and twice the proportion of G4+5 roots occurred in direct‐drilled fields than in reduced or fully tilled fields. Rapeseed yield decline may at least partly be related to poor root penetration and it is a useful indicator crop to demonstrate the degree of soil compaction and should be cultivated in fields where sufficient root penetration is possible.