Publikasjoner
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2008
Forfattere
Marianne Stenrød J Perceval P Benoit Marit Almvik Randi Bolli Ole Martin Eklo Tore E. Sveistrup Jens KværnerSammendrag
Det er ikke registrert sammendrag
Forfattere
Jihong Liu Clarke Carl Jonas Jorge Spetz Sissel Haugslien Shaochen Xing Merete Dees Roar Moe Dag-Ragnar BlystadSammendrag
Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated.
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Norwegian field production of lettuce has increased considerably since the early 1990s. Disease problems rarely required fungicide applications before 1996, when lettuce downy mildew (Bremia lactucae) caused severe losses. From 2002 to 2004, surveys were conducted to identify fungal diseases in Buskerud, Vestfold and Ostfold counties in the south-east and Rogaland County in the south-west, representing the main lettuce production regions of Norway. The distribution and incidence of B. lactucae was highly variable, but this pathogen was the most important due to the destructive nature of uncontrolled epidemics. Septoria lactucae caused severe damage, but was found in only one field. Sclerotinia sclerotiorum was the most widespread pathogen, found in 32% of the fields, but usually affecting less than 10% of the plants. Pythium tracheiphilum was reported from 33% of the fields in south-east Norway, but was not found in the south-west. Disease incidence was usually less than 5%, and a disease incidence of more than 10% was reported in one field only. Other pathogens of potential economic importance in Norwegian lettuce fields are Alternaria spp., Botrytis cinerea and Rhizoctonia solani, although they were sporadically distributed in relatively few fields in this survey.
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The Fusarium genus includes devastating plant pathogenic fungi that cause diseases in cereals around the world. They produce several mycotoxins, including the estrogenic compound zearalenone. To better understand the molecular mechanisms determining zearalenone production, we performed differential display RT-PCR under conditions where Fusarium graminearum and F. culmorum produced high amounts of zearalenone. We found 133 expressed sequence tags (ESTs) and 54 of these were considered to be up-regulated during high zearalenone production. Several of the ESTs were confirmed to be up-regulated by real-time qPCR, but none showed any significant down-regulation in the zearalenone negative mutant Delta PKS4-T9, or were similar to typical gene expression patterns of previously described zearalenone-related genes. Some of the up-regulated ESTs were similar to genes involved in secondary metabolite production, lipid metabolism, transcriptional activation, provision of precursors, signal transduction, transport or detoxification. Several of the ESTs were also located adjacent to one another in the genome and therefore might represent genes involved in the same biosynthetic pathway. Members of six such putative pathways could be found. All sequences were compared to the MIPS F. graminearum Genome Database to verify autocalled gene predictions experimentally and to introduce new exons and gene structures.
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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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Three unsprayed coffee farms (farm 1, 2 and 3) were studied for the natural occurrence of the insect pathogenic fungus Beauveria bassiana in Hypothenemus hampei populations throughout the rainy season of 2004 (July-November) and 2005 (July-December). B. bassiana infections were found during most sampling dates in both years, on all three farms. The B. bassiana infection levels were higher in 2005 than in 2004 with mean prevalence of 12.1 % and 2.7%, respectively. No consistent significant differences in infection level between farms were found in any of the years. B. bassiana infection levels fluctuated widely throughout the season, and peaked at 13.5% on farm 3 in 2004 and at 44.0% on farm 1 in 2005. The H. hampei population was significantly higher in 2004 than in 2005, with 6.9% of the berries infested in 2004 and only 0.7% in 2005. In both years, the H. hampei infestation level was significantly higher on farm 2. No consistent significant differences in H. hampei infestation levels were found between sampling dates on any of the farms. H. hampei infestation levels fluctuated throughout both seasons, and peaked at 15.3% on farm 2 in 2004 and 2.2% on farm 2 in 2005. No consistent density dependent correlation between H. hampei infestation level and B. bassiana infection level was found. Correlations between climatic conditions and R bassiana or H. hampei were not found. (C) 2007 Elsevier Inc. All rights reserved.
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A possible cost-effective real-time patch spraying implementation against seed-propagated broad-leaved weeds in cereals is a camera mounted in front of the tractor taking images at feasible distances in the direction of travel, on-board image analysis software and entire boom switched on and off. To assess this implementation, manual weed counts (0.25 m(2) quadrats) in a 1.5 m x 2 m grid, were used to simulate camera outputs. Each quadrat was classified into 'spray' and 'not spray' decisions based on a threshold model, and the resulting map defined the 'ground truth'. Subsequently, 'on/off' spraying at larger control areas where sizes were given by the boom width and image distance, and spraying decision controlled by weed status at the single quadrat simulating the camera's view, were simulated. These coarser maps were compared with 'ground truth', to estimate mapping error (area above threshold not sprayed), spraying error (area below threshold sprayed), total error (sum of mapping and spraying error) and the herbicide reduction. Three levels of the threshold model were tested. Results were used to fit models that predict errors from boom width and image distance. Size of control area did not on average affect the magnitude of the simulated herbicide reductions, but the bigger the control area the higher the risk that the simulated herbicide reduction deviate from the reduction in 'ground truth'. Mean simulated herbicide reductions were 42-59%, depending on threshold level. Only minor differences due to threshold level were seen for mean mapping and spraying errors at given spraying resolutions. Using original threshold level and image distance 2 m, predicted total errors for boom widths 2 m, 6 m, 20 m and 40 m would be 6%, 10%, 16% and 17%, respectively. Results indicate that control area should not exceed about 10 m 2 if acceptable total error is maximum 10%.
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Lack of automatic weed detection tools has hampered the adoption of site-specific weed control in cereals. An initial object-oriented algorithm for the automatic detection of broad-leaved weeds in cereals developed by SINTEF ICT (Oslo, Norway) was evaluated. The algorithm ("WeedFinder") estimates total density and cover of broad-leaved weed seedlings in cereal fields from near-ground red-green-blue images. The ability of "WeedFinder" to predict 'spray'/'no spray' decisions according to a previously suggested spray decision model for spring cereals was tested with images from two wheat fields sown with the normal row spacing of the region, 0.125 m. Applying the decision model as a simple look-up table, "WeedFinder" gave correct spray decisions in 65-85% of the test images. With discriminant analysis, corresponding mean rates were 84-90%. Future versions of "WeedFinder" must be more accurate and accommodate weed species recognition.
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The Gram-positive bacterium Clavibacter michiganensis subsp. sepedonicus is the causal agent of bacterial wilt and ring rot of potato. So far, only two proteins have been shown to be essential for virulence, namely a plasmid-encoded cellulase CelA and a hypersensitive response-inducing protein. We have examined the relative expression of CelA and eight putative virulence factors during infection of potato and in liquid culture, using quantitative real-time PCR. The examined putative virulence genes were celB, a cellulase-encoding gene and genes encoding a pectate lyase, a xylanase and five homologues of the Clavibacter michiganensis subsp. michiganensis pathogenicity factor Pat-1 thought to encode a serine protease. Six of the nine assayed genes were up-regulated during infection of potato, including celA, celB, the xylanase gene, and two of the pat genes. The pectate lyase gene showed only slightly elevated expression, whereas three of the five examined pat genes were down-regulated during infection in potato. Interestingly, the two up-regulated pat genes showed a noticeable sequence difference compared to the three down-regulated pat genes. These results reveal several new proteins that are likely to be involved in Clavibacter michiganensis subsp. sepedonicus pathogenicity.
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To evaluate overwintering strategies of the fungus Neozygites floridana, an important natural enemy of Tetranychus urticae, hibernating T. urticae females were investigated for the presence of fungal structures throughout one winter (October 12, 2006 to February 19, 2007) in field-grown strawberries in a cold climate in Norway ( min. ambient temp -15.3 degrees C). Neozygites floridana was present as hyphal bodies inside live, hibernating females in T. urticae populations throughout the sampling period. The lowest percentages of hibernating females with hyphal bodies were found at the two first dates of sampling at 5.5 and 0% on October 12 and 19, respectively. The prevalence then increased and peaked at 54.4% on January 14. Resting spores (immature) were also found in live hibernating females at some dates, but at lower prevalence than for hyphal bodies and predominantly only until November 8. Prevalence of resting spores in live hibernating females ranged from 2.5 to 13.8%. Total number of T. urticae was also recorded, and most mites of all four categories (nymphs, males, non-hibernating and hibernating females) were found at the first sampling date. At this date non-hibernating females were the most abundant. A sharp decrease in non-hibernating females, nymphs and males was, however, seen from mid-October to mid-November; also numbers of hibernating females decreased, but not as fast. The relative abundance of hibernating females compared to non-hibernating females increased from 32.2% at the first collection (October 12) to 97.7% at the last collection (February 2). This study confirms that N.floridana survives the winter as a semi-latent hyphal body infection, protected inside live hibernating females. It is therefore ready to develop and sporulate as soon as climatic conditions permit, resulting in early season infection of T. urticae.