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Background The order Lepidoptera has an abundance of species, including both agriculturally beneficial and detrimental insects. Molecular data has been used to investigate the phylogenetic relationships of major subdivisions in Lepidoptera, which has enhanced our understanding of the evolutionary relationships at the family and superfamily levels. However, the phylogenetic placement of many superfamilies and/or families in this order is still unknown. In this study, we determine the systematic status of the family Argyresthiidae within Lepidoptera and explore its phylogenetic affinities and implications for the evolution of the order. We describe the first mitochondrial (mt) genome from a member of Argyresthiidae, the apple fruit moth Argyresthia conjugella. The insect is an important pest on apples in Fennoscandia, as it switches hosts when the main host fails to produce crops. Results The mt genome of A. conjugella contains 16,044 bp and encodes all 37 genes commonly found in insect mt genomes, including 13 protein-coding genes (PCGs), two ribosomal RNAs, 22 transfer RNAs, and a large control region (1101 bp). The nucleotide composition was extremely AT-rich (82%). All detected PCGs (13) began with an ATN codon and terminated with a TAA stop codon, except the start codon in cox1 is ATT. All 22 tRNAs had cloverleaf secondary structures, except trnS1, where one of the dihydrouridine (DHU) arms is missing, reflecting potential differences in gene expression. When compared to the mt genomes of 507 other Lepidoptera representing 18 superfamilies and 42 families, phylogenomic analyses found that A. conjugella had the closest relationship with the Plutellidae family (Yponomeutoidea-super family). We also detected a sister relationship between Yponomeutoidea and the superfamily Tineidae. Conclusions Our results underline the potential importance of mt genomes in comparative genomic analyses of Lepidoptera species and provide valuable evolutionary insight across the tree of Lepidoptera species.

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Grey mold caused by the necrotrophic fungal pathogen Botrytis cinerea can affect leaves, flowers, and berries of strawberry, causing severe pre- and postharvest damage. The defense elicitor β-aminobutyric acid (BABA) is reported to induce resistance against B. cinerea and many other pathogens in several crop plants. Surprisingly, BABA soil drench of woodland strawberry (Fragaria vesca) plants two days before B. cinerea inoculation caused increased infection in leaf tissues, suggesting that BABA induce systemic susceptibility in F. vesca. To understand the molecular mechanisms involved in B. cinerea susceptibility in leaves of F. vesca plants soil drenched with BABA, we used RNA sequencing to characterize the transcriptional reprogramming 24 h post-inoculation. The number of differentially expressed genes (DEGs) in infected vs. uninfected leaf tissue in BABA-treated plants was 5205 (2237 upregulated and 2968 downregulated). Upregulated genes were involved in pathogen recognition, defense response signaling, and biosynthesis of secondary metabolites (terpenoid and phenylpropanoid pathways), while downregulated genes were involved in photosynthesis and response to auxin. In control plants not treated with BABA, we found a total of 5300 DEGs (2461 upregulated and 2839 downregulated) after infection. Most of these corresponded to those in infected leaves of BABA-treated plants but a small subset of DEGs, including genes involved in ‘response to biologic stimulus‘, ‘photosynthesis‘ and ‘chlorophyll biosynthesis and metabolism’, differed significantly between treatments and could play a role in the induced susceptibility of BABA-treated plants.

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Denne studien ble utført på oppdrag fra Miljødirektoratet for å kartlegge antibiotikaresistens i terrestriske miljø basert på ulik eksponering av resistensdrivere. Hensikten var å få en mer helhetlig forståelse av miljøets rolle i utvikling og spredning av antibiotikaresistens. Totalt 644 prøver fra jord, rødkløver, snegler, mus/spissmus og meitemark ble samlet inn fra ulike miljøer i løpet av 2019-20 og analysert for forekomst av antibiotikaresistens, samt potensielle drivere som antibiotika, pesticider og tungmetaller i jord.

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Saprolegnia parasitica is recognized as one of the most important oomycetes pests of salmon and trout species. The amplified fragment length polymorphism (AFLP) and method sequence data of the internal transcribed spacer (ITS) were used to study the genetic diversity and relationships of Saprolegnia spp. collected from Canada, Chile, Japan, Norway and Scotland. AFLP analysis of 37 Saprolegnia spp. isolates using six primer combinations gave a total of 163 clear polymorphic bands. Bayesian cluster analysis using genetic similarity divided the isolates into three main groups, suggesting that there are genetic relationships among the isolates. The unweighted pair group method with arithmetic mean (UPGMA) and principal coordinate analysis (PCO) confirmed the pattern of the cluster analyses. ITS analyses of 48 Saprolegnia sequences resulted in five well-defined clades. Analysis of molecular variance (AMOVA) revealed greater variation within countries (91.01%) than among countries (8.99%). We were able to distinguish the Saprolegnia isolates according to their species, ability to produce oogonia with and without long spines on the cysts and their ability to or not to cause mortality in salmonids. AFLP markers and ITS sequencing data obtained in the study, were found to be an efficient tool to characterize the genetic diversity and relationships of Saprolegnia spp. The comparison of AFLP analysis and ITS sequence data using the Mantel test showed a very high and significant correlation (r2 = 0.8317).

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Shallot (Allium cepa var. aggregatum), a small bulb onion, is widely grown in the world. We previously reported a droplet-vitrification for cryopreservation of in vitro-grown shoot tips of shallot genotype ‘10603’. The present study further evaluated rooting, vegetative growth, bulb production and contents of biochemical compounds, as well as genetic stability in cryo-derived plants. The results showed no significant differences in rooting, vegetative growth, bulb production and contents of soluble sugars and flavonols between the cryo- and in vitro-derived plants. Analyses of ISSR and AFLP markers did not detect any polymorphic bands in the cryo-derived plants. These results indicate rooting and vegetative growth ability, biochemical compounds and genetic stability were maintained in cryo-derived plants. The present study provides experimental evidences that support the use of cryopreservation method for long-term preservation of genetic resources of shallots and other Allium species.

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Knowledge about population genetic structure and dispersal capabilities is important for the development of targeted management strategies for agricultural pest species. The apple fruit moth, Argyresthia conjugella (Lepidoptera, Yponomeutidae), is a pre-dispersal seed predator. Larvae feed on rowanberries (Sorbus aucuparia), and when rowanberry seed production is low (i.e., inter-masting), the moth switches from laying eggs in rowanberries to apples (Malus domestica), resulting in devastating losses in apple crops. Using genetic methods, we investigated if this small moth expresses any local genetic structure, or alternatively if gene flow may be high within the Scandinavian Peninsula (~850.000 km2, 55o - 69o N). Genetic diversity was found to be high (n = 669, mean He = 0.71). For three out of ten tetranucleotide STRs, we detected heterozygote deficiency caused by null alleles, but tests showed little impact on the overall results. Genetic differentiation between the 28 sampling locations was very low (average FST = 0.016, P < 0.000). Surprisingly, we found that all individuals could be assigned to one of two non-geographic genetic clusters, and that a third, geographic cluster was found to be associated with 30% of the sampling locations, with weak but significant signals of isolation-by-distance. Conclusively, our findings suggest wind-aided dispersal and spatial synchrony of both sexes of the apple fruit moth over large areas and across very different climatic zones. We speculate that the species may recently have had two separate genetic origins caused by a genetic bottleneck after inter-masting, followed by rapid dispersal and homogenization of the gene pool across the landscape. We suggest further investigations of spatial genetic similarities and differences of the apple fruit moth at larger geographical scales, through life-stages, across inter-masting, and during attacks by the parasitoid wasp (Microgaster politus).

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The extraction of Rhodiola rosea rhizomes using natural deep eutectic solvent (NADES) consisting of lactic acid, glucose, fructose, and water was investigated. A two-level Plackett–Burman design with five variables, followed by the steepest ascent method, was undertaken to determine the optimal extraction conditions. Among the five parameters tested, particle size, extraction modulus, and water content were found to have the highest impact on the extrability of phenyletanes and phenylpropanoids. The concentration of active compounds was analyzed by HPLC. The predicted results showed that the extraction yield of the total phenyletanes and phenylpropanoids (25.62 mg/g) could be obtained under the following conditions: extraction time of 154 min, extraction temperature of 22 °C, extraction modulus of 40, molar water content of 5:1:11 (L-lactic acid:fructose:water, mol/mol), and a particle size of rhizomes of 0.5–1 mm. These predicted values were further verified by validation experiments in predicted conditions. The experimental yields of salidroside, tyrosol, rosavin, rosin, cinnamyl alcohol and total markers (sum of phenyletanes and phenylpropanoids in mg/g) were 11.90 ± 0.02, 0.36 ± 0.02, 12.23 ± 0.21, 1.41 ± 0.01, 0.20 ± 0.01, and 26.10 ± 0.27 mg/g, respectively, which corresponded well with the predicted values from the models.

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Rhodiola rosea L. (roseroot) is an adaptogen plant belonging to the Crassulaceae family. The broad spectrum of biological activity of R. rosea is attributed to its major phenyletanes and phenylpropanoids: rosavin, salidroside, rosin, cinnamyl alcohol, and tyrosol. In this study, we compared the content of phenyletanes and phenylpropanoids in rhizomes of R. rosea from the Norwegian germplasm collection collected in 2004 and in 2017. In general, the content of these bioactive compounds in 2017 was significantly higher than that observed in 2004. The freeze-drying method increased the concentration of all phenyletanes and phenylpropanoids in rhizomes compared with conventional drying at 70 °C. As far as we know, the content of salidroside (51.0 mg g−1) observed in this study is the highest ever detected in Rhodiola spp. Long-term vegetative propagation and high genetic diversity of R. rosea together with the freeze-drying method may have led to the high content of the bioactive compounds observed in the current study.

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The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr-vnt1 effector, recognized by the potato Rpi-phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi-Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C-terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large-scale cultivation of plants containing the Rpi-Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi-phu1 gene were found, the corresponding Avr-vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.

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The apple fruit moth Argyresthia conjugella (Lepidoptera, Yponomeutidae) is a seed predator of rowan (Sorbus aucuparia) and is distributed in Europe and Asia. In Fennoscandia (Finland, Norway and Sweden), rowan fruit production is low every 2–4 years, and apple (Malus domestica) functions as an alternative host, resulting in economic loss in apple crops in inter-mast years. We have used Illumina MiSeq sequencing to identify a set of 19 novel tetra-nucleotide short tandem repeats (STRs) in Argyresthia conjugella. Such motifs are recommended for genetic monitoring, which may help to determine the eco-evolutionary processes acting on this pest insect. The 19 STRs were optimized and amplified into five multiplex PCR reactions. We tested individuals collected from Norway and Sweden (n = 64), and detected very high genetic variation (average 13.6 alleles, He = 0.75) compared to most other Lepidoptera species studied so far. Spatial genetic differentiation was low and gene flow was high in the test populations, although two non-spatial clusters could be detected. We conclude that this set of genetic markers may be a useful resource for population genetic monitoring of this economical important insect species.

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Undersøkelse av antibiotikaresistensmarkørgenet neomycin fosfotransferase II (nptII) i prøver fra 12 ville arter fra Norge I et prosjekt fra Miljødirektoratet har vi testa for tilstedeværelse av nptII genet i 219 prøver fra 12 ulike ville arter fra hele Norge. Utvalget av prøver inkluderte planter (løvetann, rødkløver og markjordbær), insekter (skogmaur, rognebærmøll og liten høstmåler), snegl (brunsnegl), fisk (ørret og rognkjeks) og pattedyr (rødrev, brunbjørn og isbjørn). Vi brukte to ulike sanntids-PCR (Real-Time-PCR) tester for å undersøke fo tilstedeværelsen av kopier av nptII-genet i de 219 prøvene. Vi fant at nesten alle prøvene var negative (99%), mens kun tre enkeltprøver (løvetann, rødkløver og skogmaur) viste et svært lavt nivå av nptII (3-4 kopier). De positive prøvene kan være naturlige varianter eller kontaminering fra forskningslaboratorier. Vi konkluderer med at der er behov for utvida undersøkelser innenen for dette fagfeltet.

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Knowledge about the reproduction strategies of invasive species is fundamental for effective control. The invasive Fallopia taxa (Japanese knotweed s.l.) reproduce mainly clonally in Europe, and preventing spread of vegetative fragments is the most important control measure. However, high levels of genetic variation within the hybrid F. × bohemica indicate that hybridization and seed dispersal could be important. In Norway in northern Europe, it is assumed that these taxa do not reproduce sexually due to low temperatures in the autumn when the plants are flowering. The main objective of this study was to examine the genetic variation of invasive Fallopia taxa in selected areas in Norway in order to evaluate whether the taxa may reproduce by seeds in their most northerly distribution range in Europe. Fallopia stands from different localities in Norway were analyzed with respect to prevalence of taxa, and genetic variation within and between taxa was studied using amplified fragment length polymorphism (AFLP). Taxonomic identification based on morphology corresponded with identification based on simple sequence repeats (SSR) and DNA ploidy levels (8× F. japonica, 6× F. × bohemica and 4× F. sachalinensis). No genetic variation within F. japonica was detected. All F. × bohemica samples belonged to a single AFLP genotype, but one sample had a different SSR genotype. Two SSR genotypes of F. sachalinensis were also detected. Extremely low genetic variation within the invasive Fallopia taxa indicates that these taxa do not reproduce sexually in the region, suggesting that control efforts can be focused on preventing clonal spread. Climate warming may increase sexual reproduction of invasive Fallopia taxa in northern regions. The hermaphrodite F. × bohemica is a potential pollen source for the male-sterile parental species. Targeted eradication of the hybrid can therefore reduce the risk of increased sexual reproduction under future warmer climate.

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Important losses in strawberry production are often caused by the oomycete Phytophthora cactorum, the causal agent of crown rot. However, very limited studies at molecular levels exist of the mechanisms related to strawberry resistance against this pathogen. To begin to rectify this situation, a PCR-based approach (NBS profiling) was used to isolate strawberry resistance gene analogs (RGAs) with altered expression in response to P. cactorum during a time course (2, 4, 6, 24, 48, 96 and 192 h post-infection). Twenty-three distinct RGA fragments of the NB-LRR type were identified from a resistance genotype (Bukammen) of the wild species Fragaria vesca. The gene transcriptional profiles after infection showed that the response of most RGAs was quicker and stronger in the resistance genotype (Bukammen) than in the susceptible one (FDP821) during the early infection stage. The transcriptional patterns of one RGA (RGA109) were further monitored and compared during the P. cactorum infection of two pairs of resistant and susceptible genotype combinations (Bukammen/FDP821 and FDR1218/1603). The 5′ end sequence was cloned, and its putative protein was characteristic of NBS-LRR R protein. Our results yielded a first insight into the strawberry RGAs responding to P. cactorum infection at molecular level.

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The apple fruit moth (Argyresthia conjugella (A. conjugella)) in Norway was first identified as a pest in apple production in 1899. We here report the first genetic analysis of A. conjugella using molecular markers. Amplified fragment length polymorphism (AFLP) analysis was applied to 95 individuals from six different locations in the two most important apple-growing regions of Norway. Five AFLP primer combinations gave 410 clear polymorphic bands that distinguished all the individuals. Further genetic analysis using the Dice coefficient, Principal Coordinate analysis (PCO) and Bayesian analyses suggested clustering of the individuals into two main groups showing substantial genetic distance. Analysis of molecular variance (AMOVA) revealed greater variation among populations (77.94%) than within populations (22.06%) and significant and high FST values were determined between the two major regions (Distance = 230 km, FST = 0.780). AFLP analysis revealed low to moderate genetic diversity in our population sample from Norway (Average: 0.31 expected heterozygosity). The positive significant correlation between the geographic and the molecular data (r2 = 0.6700) indicate that genetic differences between the two major regions may be due to geographical barriers such as high mountain plateaus (Hardangervidda) in addition to isolation by distance (IBD).

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Amplified fragment length polymorphism (AFLP) was used to study the genetic variation among 80 F. verticillioides isolates from kernels of Ethiopian maize, collected from 20 different maize growing areas in four geographic regions. A total of 213 polymorphic fragments were obtained using six EcoRI/MseI primer combinations. Analysis of the data based on all 213 polymorphic AFLP fragments revealed high level of genetic variation in the F. verticillioides entities in Ethiopia. About 58% of the fragments generated were polymorphic. The genetic similarity among F. verticillioides isolates varied from 46% to 94% with a mean Dice similarity of 73%. Unweighted Pair Group Method with Arithmetic Average (UPGMA) analysis revealed two main groups and four subgroups. The principal coordinate analysis (PCO) also displayed two main groups that agreed with the results of UPGMA analysis, and there was no clear pattern of clustering of isolates according to geographic origin. Analysis of molecular variance: (AMOVA) showed that only 1.5% of the total genetic variation was between geographic regions, while 98.5% was among isolates from the same geographic regions of Ethiopia. Eighty distinct haplotypes were recognized among the 80 isolates analyzed. Hence, breeding efforts should concentrate on quantitative resistance that is effective against all genotypes of the pathogen.

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According to the Norwegian Diversity Act, practitioners of restoration in Norway are instructed to use seed mixtures of local provenance. However, there are no guidelines for how local seed should be selected. In this study, we use genetic variation in a set of alpine species (Agrostis mertensii, Avenella flexuosa, Carex bigelowii, Festuca ovina, Poa alpina and Scorzoneroides autumnalis) to define seed transfer zones to reduce confusion about the definition of ‘local seeds’. The species selected for the study are common in all parts of Norway and suitable for commercial seed production. The sampling covered the entire alpine region (7–20 populations per species, 3–15 individuals per population). We characterised genetic diversity using amplified fragment length polymorphisms. We identified different spatial genetic diversity structures in the species, most likely related to differences in reproductive strategies, phylogeographic factors and geographic distribution. Based on results from all species, we suggest four general seed transfer zones for alpine Norway. This is likely more conservative than needed for all species, given that no species show more than two genetic groups. Even so, the approach is practical as four seed mixtures will serve the need for restoration of vegetation in alpine regions in Norway.

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Initial sources of inoculum of Phytophthora infestans were investigated in ten potato fields with early outbreaks of potato late blight. Infected plant samples and isolates from these fields were examined with respect to mating type prevalence, fungicide resistance and genotypes based on microsatellites A high proportion (91 %) of the isolates recovered were of mating type A1. However, both mating types were found in 3 of 9 fields with more than one isolate recovered, and sometimes both mating types were found on the same plant. Most of the isolates recovered from fields treated with metalaxyl-M prior to sampling had reduced sensitivity or were resistant to metalaxyl-M, and most of the isolates recovered form fields without metalaxyl treatment were sensitive. The isolates recovered from fields treated with propamocarb prior to sampling had a higher frequency of reduced sensitivity to propamocarb than isolates from fields without propamocarb treatment. We found that most plants contained more than one P. infestans SSR-genotype. Clustering analysis of the infected samples revealed that most samples clustered together according to fields. By combining information from P. infestans isolates and DNA extracts from the leaf lesions we found examples of both mating type A1 and A2 having the same multilocus genotype. This result indicates that both of these genotypes have a common ancestor, hence the inoculum originates from oospores. Although this a minor study of only 10 fields with a limited amount of isolates and plant samples, the results indicate oospores in the soil is an inoculum source. Hence the forecasting model to predict outbreaks of potato late blight should be modified to include this.       

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Initial sources of inoculum of Phytophthora infestans were investigated in ten potato fields with early outbreaks of potato late blight. Infected plant samples and isolates from these fields were examined with respect to mating type prevalence, fungicide resistance and genotypes based on microsatellites A high proportion (91 %) of the isolates recovered were of mating type A1. However, both mating types were found in 3 of 9 fields with more than one isolate recovered, and sometimes both mating types were found on the same plant. Most of the isolates recovered from fields treated with metalaxyl-M prior to sampling had reduced sensitivity or were resistant to metalaxyl-M, and most of the isolates recovered form fields without metalaxyl treatment were sensitive. The isolates recovered from fields treated with propamocarb prior to sampling had a higher frequency of reduced sensitivity to propamocarb than isolates from fields without propamocarb treatment. We found that most plants contained more than one P. infestans SSR-genotype. Clustering analysis of the infected samples revealed that most samples clustered together according to fields. By combining information from P. infestans isolates and DNA extracts from the leaf lesions we found examples of both mating type A1 and A2 having the same multilocus genotype. This result indicates that both of these genotypes have a common ancestor, hence the inoculum originates from oospores. Although this a minor study of only 10 fields with a limited amount of isolates and plant samples, the results indicate oospores in the soil is an inoculum source. Hence the forecasting model to predict outbreaks of potato late blight should be modified to include this.

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Plasmopara halstedii is a diploid oomycete plant pathogen causing downy mildew on sunflower (Helianthus annuus). Due to changes in cultural systems and the introduction of new exotic cultivars, the pathogen developed many races and have now become a serious problem affecting sunflower growing fields in Europe. The yield losses in sunflower crop caused by P. halstedii can be up to 85 %.

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Background to the research and aims. Sunflower downy mildew exhibit mixed asexual and sexual reproduction. Rare events of recombination can have a drastic effect on the reshuffling of genetic material and result in the emergence of new virulence combinations. Overview of the methods. In this study, we have used a molecular epidemiology approach to identify population sources, investigate the mating system and track pathogen movement at the field scale. Using a hierarchical sampling design, we have collected 250 P. halstedii isolates from 20 geo-referenced sites in an infected sunflower field located in Southern France. These samples were genetically characterized using 12 single nucleotide polymorphisms markers (SNP) and one microsatellite locus(Giresse et al. 2007) and, for a subset of 60 samples, the virulence profile was determined. Key results. Characterization of virulence profiles revealed the presence of 8 races within the field (100, 300, 304, 307, 703, 704, 707, 714), race 304 being predominant among the samples tested. Race were randomly distributed within the field. Among the 250 isolates that were genotyped, there were 109 different multilocus genotypes (MLG), of which four were highly represented. The significant deficit in heterozygotes observed confirmed that P. halstedii is a highly selfing species (Delmotte et al. 2008). Bayesian clustering analyses revealed that isolates belonged to two genetically differentiated groups (FST> 0.2) that were correlated with race. A low level of genetic differentiation (FST = 0.06) was observed among the 20 sites that were constituted of a mixture of samples from the two groups. Theestimation of gene flow using hierarchical FST and spatial autocorrelation analysis bring evidence a lack of spatial genetic structure that likely result from field plowing. Finally, the estimation of gene flow atvarious scales using hierarchical FST and spatial autocorrelation analysis bring evidence for race emergence through genetic recombination between differentiated genetic genotypes. Main conclusions. This study brings evidence for an absence of spatial genetic structure of P. halstedii population, that could result from field plowing. It also reveals that genetically and phenotypically (virulence) differentiated isolates of P. hasltedii do coexist at a very fine spatial scale. The coexistence of genetically and phenotypically differentiated isolates of P. hasltedii may allow the emergence of new races through recombination. Nature of the contribution to current knowledge. This study has allowed to identify and delineate pathogen populations at the field scale. By bringing new insights in to the evolutionary mechanisms responsible for the emergence of races, these results will help identifying management units for sunflower protection strategies. Keywords. Plasmopara halstedii, race, pathotype, virulence evolution, population genetics

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Rotstokkråte i jordbær ble første gang rapportert i Norge i 1992 og siden er den blitt funnet på mer enn 100 steder over hele landet. Sykdommen forårsakes av Phytophthora cactorum og karakteriseres ved at unge blader visner raskt og hele planten visner i løpet av noen dager. I løpet av en sesong kan opptil 40 % av plantene dø. P. cactorum smitter plantene gjennom rothårene ved hjelp av svermesporer (zoosporer). Sykdommen starter oftest i fuktige områder av et felt siden sporene trenger vann for å bevege seg. Når en først har fått smitten i jorda er det vanskelig å bli kvitt den siden P. cactorum danner hvilesporer som kan overleve i flere år. Ulike jordbærsorter har ulik grad av mottakelighet for sykdommen. De mest brukte kommersielle sortene er dessverre mottakelige for sykdommen. Resistensegenskaper kan styres av ett eller flere gener og man kan derfor foredle fram resistente sorter. Tradisjonell foredling er tidkrevende og overføringa av resistens til en mottakelig sort vil kreve gjentatte tilbakekrysninger slik at man ikke mister alle de positive egenskapene til denne sorten. Ved å utvikle genetiske kart med markører for resistens kan man teste planter raskere og slik komme raskere fram til en resistent sort. Kunnskap om hvor mange resistensgener som er involvert i kampen mot skadegjøreren, når disse blir slått på og hvilke proteiner disse lager er også viktig. Når en skadegjører angriper en plante lager den bl.a. proteiner som bryter ned plantecelleveggen og svekker plantens immunforsvar. Planten på sin side lager resistensproteiner som gjenkjenner proteinene laget av skadegjøreren. Denne gjenkjennelsen setter i gang en forsvarsrespons hos planten. Resistensproteinene kodes for av resistensgener (R-gener). De fleste kjente R-genene inneholder en kort bestemt nukleotidsekvens. Dette fellestrekket gjør jakten på resistensgener enklere. I jakten på resistensgener i jordbær har vi valgt å arbeide med markjordbær (Fragaria vesca) istedenfor kommersielle jordbær (Fragaria x ananassa Duch.). Markjordbær er diploid og egner seg derfor godt for molekylærbiologiske studier. For å isolere R-gener og studere hvordan de ble uttrykt ble en mottakelig kultivar og en resistent kultivar smittet med zoosporer. Vevsprøver ble høstet i en tidsserie fra tid 0 (kontroll før smitting) til maksimum 8 dager etter smitting. Resultatet så langt viser at vi har isolert fragmenter fra mange ulike resistensgener og at disse blir uttrykt gjennom hele tidsrommet fra smitting til 8 dager etterpå.

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Roseroot, Rhodiola rosea, is a perennial herbaceous plant of the family Crassulaceae. The rhizomes of 95 roseroot clones in the Norwegian germplasm collection were analysed and quantified for their content of the bioactive compounds rosavin, salidroside, rosin, cinnamyl alcohol and tyrosol using HPLC analysis. All five bioactive compounds were detected in all 95 roseroot clones but in highly variable quantities. The ranges observed for the different compounds were for rosavin 2.90-85.95mgg-1, salidroside 0.03-12.85mgg-1, rosin 0.08-4.75mgg-1, tyrosol 0.04-2.15mgg-1 and cinnamyl alcohol 0.02-1.18mgg-1. The frequency distribution of the chemical content of each clone did not reflect a certain geographic region of origin or the gender of the plant. Significant correlations were found for the contents of several of these bioactive compounds in individual roseroot clones. A low, but not significant correlation was found between AFLP markers previously used to study the genetic diversity of the roseroot clones and their production of the chemical compounds. The maximum level of rosavin, rosin and salidroside observed were higher than for any roseroot plant previously reported in literature, and the roseroot clones characterized in this study might therefore prove to be of high pharmacological value.

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Artikkelen forteller om Colletotrichum acutatum i norsk jordbærproduksjon, om vertplanter for soppen, om genetiske analyser av soppisolater fra ulike vertplanter og smitteforsøk i jordbær og kirsebær.

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Phytophthora cactorum causes crown rot in strawberry (Fragaria x ananassa Dutch.), which is characterized by wilting and eventually collapse of the plant. An efficient control measure is the use of resistant cultivars, however most commercial cultivars are susceptible to the disease. The aim of our work is to generate basic knowledge about P. cactorum resistance as well as to develop genetic markers that can be used as tools for development of resistant cultivars. The genetic complexity of the octoploid cultivated strawberry, has led to development of the diploid wild strawberry (F. vesca) as a model system for Fragaria. We have identified suitable parents after screening accessions of diploid Fragaria sp. for resistance [1], and generated a mapping population which we are currently characterizing. In order to study the plant-pathogen interaction in detail we have identified and characterized resistance genes from diploid strawberry and effector genes from P. cactorum using different transcriptional analysis techniques; nucleotide-binding site (NBS)-profiling for resistance genes, and suppression subtractive hybridization (SSH) as well as a designed effector-specific differential display (ESDD) for genes involved in pathogenicity.   This work is supported by The Research Council of Norway.   [1] Eikemo H, Brurberg MB, Davik J (2010). Resistance to Phytophthora cactorum in diploid Fragaria species. HortScience. 45:193-197.  

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Abstract Germplasm characterization is an important component contributing to the effective management of plant genetic resources. The goal of this thesis was to study the genetic diversity of two models of vegetatively propagated plant species; roseroot (Rhodiola rosea L.) and sweet potato (Ipomoea batatas (L.) Lam), based on germplasm collections. Roseroot was recently collected from natural habitats and then vegetatively propagated at the germplasm centre while sweet potato already has a long tradition as a vegetatively propagated food species. I. Roseroot (Rhodiola rosea) Roseroot, R. rosea, also commonly known as golden root or arctic root, is a perennial herbaceous plant of the Crassulaceae family. R. rosea has its origin from the cold, humid regions of the northern hemisphere and grows mostly in the mountains near the snow border. R. rosea is widely distributed in Norway. As part of an effort to identify commercially valuable genotypes characterization of a germplasm collection from Norway was initiated. Amplified Fragment Length Polymorphism (AFLP) analysis was used to estimate genetic diversity within the Norwegian R. rosea germplasm collection. AFLP analysis of 97 R. rosea clones using five primer combinations gave a total of 109 polymorphic bands. A large molecular marker variation was found among roseroot clones in Norway with an average percentage of polymorphic bands (PPB) of 82.3%. Analysis of molecular variance (AMOVA) revealed a significantly greater variation within regions (92.03%) than among regions (7.97%) demonstrating that there was no close genetic similarity among clones originating from the same county. A low level of genetic differentiation (FST = 0.043) was observed, indicating a high level of gene flow, which had a strong influence on the genetic structure in Norway. Our results indicate high gene flow among R. rosea clones that might be a result of seed dispersal rather than cross-pollination. Ninety five clones of the Norwegian roseroot germplasm collection were analysed and quantified for their content of the bioactive compounds rosavin, salidroside, rosin, cinnamyl alcohol and tyrosol using High Performance Liquid Chromatography (HPLC) analysis. All bioactive compounds were detected in all clones but in highly variable quantities. The frequency distribution of the chemical content of each clone was not correlated with geographic region of origin or gender of the plant. Significant correlations between the content of these bioactive compounds were observed within individual roseroot clones. Low and nonsignificant correlations were found between AFLP markers used to study genetic diversity of the roseroot clones and their content of chemical compounds. The maximum content of rosavin, rosin and salidroside observed were substantially higher than previously reported for roseroot plants, and the roseroot clones characterized in this study might therefore be of high pharmacological value. The large quantitative and qualitative variation of the chemical compounds observed in this study and the large genetic diversity observed in this germplasm constitute a firm basis for improving traits such as chemical composition in a breeding program for roseroot. This is the first report that combines the analysis of genetic diversity with information of the chemical composition of roseroot. Further studies of the roseroot populations from Norway as well as from other countries should be performed throughout the following years to identify clones with optimal chemical compositions and to maintain high genetic diversity of this species. II. Sweet potato (Ipomoea batatas (L.) Lam) Sweet potato has its origin in South America and is the 7th most important crop in the world. A Tanzanian sweet potato germplasm collection was characterized using molecular markers and morphological traits. The AFLP method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection ..

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Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.

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The ascigerous stage (formation of perithecia with viable ascospores) of Colletotrichum acutatum was recently reported to occur on fruits of highbush blueberry (Vaccinium corymbosum) in Norway. When 113 isolates of C. acutatum from various plant species were cultured on strawberry leaf agar, nine developed perithecia with viable ascospores. Four isolates originated from apple (Malus domestica) and one each from sweet cherry (Prunus avium), raspberry (Rubus idaeus), highbush blueberry (Vaccinium corymbosum), hollyberry cotoneaster (Cotoneaster bullatus), and northern dock (Rumex longifolius). Except from blueberry, we never detected the ascigerous stage on decaying fruits or any other parts of the above mentioned plant species. On potato dextrose agar, colour of the underside of the cultures forming perithecia varied from light grey-green to dark grey-green or dark brown-green. Colour of the upperside varied greatly, being dark grey-green, grey-brown, grey, and beige-pink, and only two of the isolates were beige-pink (the raspberry and blueberry isolates). Amplified fragment length polymorphism (AFLP) analysis of the isolates using six primer combinations resulted in 103 clear polymorphic bands. A dendrogram was constructed, and based on cluster analysis using genetic similarity, the isolates could be divided into several clusters. Eight of nine perithecia-forming isolates grouped together in the dendrogram, indicating genetical difference from other isolates. This was also supported by Principal Coordinate (PCO) analysis.

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Rhodiola rosea is widely distributed in Norway, but so far limited knowledge exists on the level of genetic diversity. To initiate a selective breeding program, Amplified Fragment Length Polymorphism (AFLP) analysis was used to estimate genetic diversity within the Norwegian R. rosea germplasm collection. AFLP analysis of 97 R. rosea clones using five primer combinations gave a total of 109 polymorphic bands. We detected high percentage of polymorphic bands (PPB) with a mean of 82.3% among the clones of R. rosea. Each of the 97 R. rosea clones could be unambiguously identified based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the Unweighted Pair Group Method with Arithmetic mean (UPGMA). Genetic similarity based on the AFLP data ranged from 0.440 to 0.950 with a mean of 0.631. This genetic analysis showed that there was no close genetic similarity among clones related to their original growing county. No gender-specific markers were found in the R. rosea clones. Analysis of molecular variance (AMOVA) revealed a significantly greater variation within regions (92.03%) than among regions (7.97%). A low level of genetic differentiation (F-ST=0.043) was observed, indicating a high level of gene flow, which had a strong influence on the genetic structure at different counties. Our results indicate high gene flow among R rosea clones that might be a result of seed dispersal rather than cross-pollination. Further world-wide studies are required to compare the level of genetic diversity and more studies in R. rosea detailing the consequences of different patterns of gene flow (pollen spread and dispersal of seeds and clonal plants) will be useful for characterization of roseroot. (C) 2008 Elsevier Ltd. All rights reserved.

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Det første og viktigste trinn i bekjempelse av planteskadegjørere er en korrekt identifisering av organismen som forårsaker skade. Testmetoder som er basert på deteksjon av arvestoff (DNA) til en organisme har vist seg å være svært nyttige i diagnostikken. Vi har etablert en DNA-basert test for identifisering av potetcystenematoder.

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Studies of genetic variation of the late blight pathogen among isolates collected in 2003  in the Nordic countries were presented

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Phytophthora infestans was isolated from potato leaves collected from 200 fields located in different parts of Finland, Denmark, Sweden and Norway in 2003. Sampling was carried out relatively late in the epidemic. The SSR analysis was carried out in Norway. Nine SSR markers were tested; Pi4B, Pi4G, PiG11, Pi02, Pi04, Pi16, Pi26, Pi33 and D13. Based on only 7 SSR markers 190 genotypes were found. Six genotypes occurred twice and one genotype occurred six times. The high genetic variation indicates sexual reproduction in the Nordic late blight population

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The pine wood nematode, Bursaphelenchus xylophilus, which is indigenous to North America, was introduced to Asia in the early 1900 and now causes severe damage to susceptible pine species in Japan, China, Korea and Taiwan. B. xylophilus was included in the A1 list of quarantine organisms by EPPO (European Plant Protection Organization) in 1985. B. xylophilus was reported for the first time in Europe in Portugal in 1999. There are more than 50 described species within the genus Bursaphelenchus worldwide, that are associated with coniferous and deciduous trees and spread by insect vectors. Within this genus is a group of morphologically very similar species; B. xylophilus, B. mucronatus, B. fraudulentus, B. kolymensis, B. conicaudatus and B. luxuriosae. This group of species is often referred to as the "B. xylophilus group". Due to the morphological similarity of the species, identification of Bursaphelenchus species in the B. xylophilus group is difficult. The common method of molecular identification for separating species within the B. xylophilus group is the use of ITS-RFLP (Hoyer et al. 1998). We have developed a multiplex polymerase chain reaction (PCR) method with specific primers, and the primers amplified product were 740, 340 and 300 bp for B. xylophilus, B. fraudulentus and B. mucronatus respectively. No cross reactions on the three studied species were observed. In contrast to the previously described PCR-RFLP method, this new method allows detection not only on pure isolates, but also on crude nematode suspensions from wood samples, and it could be very useful for quarantine purposes. References Hoyer U, Burgermeister W, Braasch H 1998 Identification of Bursaphelenchus species (Nematoda, Aphelenchoididae) on the basis of amplified ribosomal DNA (ITS-RFLP). Nachrichtenbl. Deut. Pflanzenschutzd. 50:273-27. Mota M M, Braasch H, Bravo M A, Penas A C, Burgermeister W, Metge K, and Sousa E 1999. First report of Bursaphelenchus xylophilus in Portugal and in Europe. Nematology 1:727-734. Smith, I. M. 1985. Pests and disease problems in European forests. FAO Plant Prot. Bull. 33:159-164. Yi C, Park J, and Chang K 1989. Occurrence of pinewood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle, and its vector, Monochamus alternatus Hope, in Korea. Pages 183-193 in: Proc. IUFRO Reg. Workshop For. Insect Pests and Tree Dis. in NE Asia. For. Prod. Res. Inst., Tsukuba, Japan.

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Studier av tørråtepopulasjonen i Norge fra 2003 bekrefter tidligere års undersøkelser at den er svært variabel både i genotype og fenotype. Andelen av metalaksyl-resistente individer har gått tilbake. Det er påvist en relativt stor andel isolater som tolererer å vokse på 100 ppm propamokarb, men det er ikke konstatert resistensproblemer i felt. Krysningstype A2 er ikike påvist blant 110 isolater fra Nord-Norge i 2004, noe som antyder at kjønna formering av tørråtesoppen er sjelden eller fraværende i denne landsdelen. Problemene med tørråte i Sør-Norge var mindre i 2004 enn i de foregående årene. Feltforsøk med ulike strategier i 2004 viste at en kunne bekjempe tørråte med godt resultat både med dynamiske fungiciddoser ved faste intervall og ved sprøyting etter ulike varslingsmodeller. Det var imidlertid vanskelig å påvise sikre forskjeller mellom de ulike strategiene.