Asko Hannukkala

Characterization of Dickeya strains isolated from potato and river water samples in Finland

Plant pathogenic enterobacteria in the genera Pectobacterium and Dickeya (formerly classified as Erwinia) were isolated from diseased potato stems and tubers. The isolated bacteria were identified as P. atrosepticum, P. carotovorum and pathogens in the genus Dickeya with PCR tests. Furthermore, Dickeya strains were isolated from river water samples throughout the country. Phylogenetic analysis with 16S-23S rDNA intergenic spacer sequences suggested that the Dickeya strains could be divided into three groups, two of which were isolated from potato samples. Phylogenetic analysis with 16S rDNA sequences and growth at 39°C suggested that one of the groups corresponds to D. dianthicola, a quarantine pathogen in greenhouse cultivation of ornamentals, while two of the groups did not clearly resemble any of the previously characterised Dickeya species. Field trials with the strains indicated that D. dianthicola-like strains isolated from river samples caused the highest incidence of rotting and necrosis of potato stems, but some of the Dickeya strains isolated from potato samples also caused symptoms. The results showed that although P. atrosepticum is still the major cause of blackleg in Finland, virulent Dickeya strains were commonly present in potato stocks and rivers. This is the first report suggesting that Dickeya, originally known as a pathogen in tropical and warm climates, may cause diseases in potato in northern Europe.

Genetic analysis of Phytophthora infestans populations in the Nordic European countries reveals high genetic variability

Late blight, caused by the oomycete Phytophthora infestans, is the most important disease of potato (Solanum tuberosum). The pathogen is highly adaptable and to get an overview of the genetic variation in the Nordic countries, Denmark, Finland, Norway and Sweden we have analyzed 200 isolates from different fields using nine simple-sequence repeat (SSR) markers. Forty-nine alleles were detected among the nine SSR loci and isolates from all four Nordic countries shared the most common alleles across the loci. In total 169 multilocus genotypes (based on seven loci) were identified among 191 isolates. The genotypic diversities, quantified by a normalized Shannons diversity index (H(s)), were 0.95 for the four Nordic countries. The low F(ST) value of 0.04 indicates that the majority of variation is found within the four Nordic countries. The large number of genotypes and the frequency distribution of mating types (60% A1) support the hypothesis that sexual reproduction is contributing notably to the genetic variation of P. infestans in the Nordic countries.

Pests and diseases in a changing climate: a major challenge for finnish crop production

A longer growing season and higher accumulated effective temperature sum (ETS) will improve crop production potential in Finland. The production potential of new or at present underutilised crops (e.g. maize (Zea mays L.), oilseed rape (Brassica napus L.), lucerne (Medicago sativa L.)) will improve and it will be possible to grow more productive varieties of the currently grown crops (spring wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), oats (Avena sativa L.)). Also cultivation of autumn sown crops could increase if winters become milder and shorter, promoting overwintering success. Climatic conditions may on the other hand become restrictive in many ways. For example, early season droughts could intensify because of higher temperatures and consequent higher evaporation rates. Current low winter temperatures and short growing season help restrict the development and spread of pests and pathogens, but this could change in the future. Longer growing seasons, warmer autumns and milder winters may initiate new problems with higher occurrences of weeds, pests and pathogens, including new types of viruses and virus vectors. Anoxia of overwintering crops caused by ice encasement, and physical damage caused by freezing and melting of water over the fields may also increase. In this study we identify the most likely changes in crop species and varieties in Finland and the pest and pathogen species that are most likely to create production problems as a result of climate change during this century.

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.

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.

Potential and realities of enhancing rapeseed- and grain legume-based protein production in a northern climate

Crop-based protein self-sufficiency in Finland is low. Cereals dominate the field cropping systems in areas that are also favourable for legumes and rapeseed. The present paper estimated the realistic potential for expanding protein crop production taking account of climatic conditions and constraints, crop rotation requirements, field sizes, soil types and likelihood for compacted soils in different regions. The potential for current expansion was estimated by considering climate change scenarios for 2025 and 2055. By using actual regional mean yields for the 2000s, without expecting any yield increase during the expansion period (due to higher risks of pests and diseases), potential production volumes were estimated. Since rapeseed, unlike grain legumes, is a not a true minor crop, its expansion potential is currently limited. Thus, most potential is from the introduction of legumes into cropping systems. The current 100000 ha of protein crops could be doubled, and areas under cultivation could reach 350000 and 390000 ha as a result of climate warming by 2025 and 2055, respectively. Such increases result mainly from the longer growing seasons projected for the northern cropping regions of Finland. Self-sufficiency in rapeseed could soon increase from 0·25 to 0·32, and then to 0·50 and 0·60 by 2025 and 2055, respectively. If legume production expands according to its potential, it could replace 0·50–0·60 of currently imported soybean meal, and by 2025 it could replace it completely. Replacement of soybean meal is suitable for ruminants, but it presents some problems for pig production, and is particularly challenging for poultry.

High genotypic diversity found among population of Phytophthora infestans collected in Estonia

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.

Examples of alien pathogens in Finnish potato production - their introduction, establishment and consequences

Most pathogens on potato have been imported into Finland via contaminated seed more than hundred years ago. The history of migration and the consequences for potato production of potato wart, blackleg and soft rot, Potato mop-top virus (PMTV) and its vector powdery scab are reviewed as examples of economically important and biologically different potato pathogens. Potato wart spread alarmingly during 1920-1960. Plant quarantine acts and the use of resistant cultivars were successful in eradicating the disease. The pathogens causing blackleg and soft rot increased rapidly in 1960-1970. Development of seed certification schemes after the end of the 1970s decreased disease incidence and made the disease insignificant other than for seed potato production. Introduction of new strains of blackleg bacteria in 2003 caused the disease again to become a considerable threat to potato production. PMTV was imported into Finland in the 1970s where it spread rapidly, especially in starch potato production. Currently it is common in all potato production except that of seed potato. The disease cannot be eradicated but contamination of clean fields can be prevented. New diseases can spread to Finland in future but population changes of existing pathogens have recently caused more problems than species completely new to Finland.

Frequency and occurrence of the carrot pathogen ‘Candidatus Liberibacter solanacearum’ haplotype C in Finland

Occurrence of ‘Candidatus Liberibacter solanacearum’ (CLso) was studied in field-grown carrots (Daucus carota) in different regions of Finland. In addition, the frequency of CLso in carrots and in field populations of its vector, the carrot psyllid (Trioza apicalis), was studied in southwestern Finland. CLso was detected in six of the seven regions where the main carrot cultivation areas are located. The highest disease incidence was found in southwestern Finland, in the area where this carrot pathogen was originally found. In the Tavastia Proper and Southwest Finland regions, CLso was detected in 26 out of 30 randomly chosen fields inspected in 2013 and 2014, and in a third of those fields more than 10% of plants showed symptoms. Of those carrots showing both psyllid feeding-associated leaf curling and CLso infection-associated leaf discolouration symptoms, 77% were CLso positive in the PCR test. Some symptomless carrots from the affected fields also tested positive. Of the carrot psyllid individuals collected from the same area, 60% were CLso positive. Elsewhere, disease incidence was variable in South Ostrobothnia in western Finland and low but established in South Savonia in eastern Finland. CLso was not detected in the North Ostrobothnia region. Sequencing of the amplified DNA fragments confirmed that the bacteria in the carrot samples from different areas within Finland all represented CLso haplotype C. The frequent occurrence and wide distribution of this pathogen, transmitted by a psyllid that does not migrate over long distances, suggest that it is persistent in Finland.

Carrot Pathogen ‘Candidatus Liberibacter solanacearum’ Haplotype C Detected in Symptomless Potato Plants in Finland

Abstract‘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.