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Publications

NIBIOs employees contribute to several hundred scientific articles and research reports every year. You can browse or search in our collection which contains references and links to these publications as well as other research and dissemination activities. The collection is continously updated with new and historical material.

2009

Abstract

To identify differentially expressed genes of the white-rot fungus Heterobasidion parviporum subtractive cDNA libraries were constructed using suppressive subtraction hybridization (SSH) technique with RNA extracted from an advanced stage of decay area and from colonization front next to the reaction zone of the stem of a mature Norway spruce trees. Besides cytochrome P450s and proteins with unknown function, the SSH libraries constructed contained genes involved in basic cellular processes, andcell wall degradation. To examine the role of selected candidate genes three trees, showing a variable degree of wood decay, were used for real-time RT-PCR profiling of candidate genes. In the decay transition areas the study revealed activity centers that showed remarkable similarity in the transcript profiles of monitored genes.

Abstract

The genome sequence of the conifer rot root pathogen Heterobasidion annosum was generated at JGI with 8.23 X coverage. The nuclear genome assembles in 39 scaffolds of total 33.7 Mbp estimated to cover 98.1% of the complete genome. We predicted 12,270 genes with an average length of 1,617 bp and exon number and length of 5.54 and 250 bp respectively. About 50% (5999) of the predicted genes could be validated by EST support with the 40,807 EST´s generated with in the project. The genome has a GC content of 52.0% and very little repetitive sequences with 2,895 SSR per mega base. The physical genome is congruent with the genetic linkage map, and most of the linkage groups have been possible to anchor to the 18 largest scaffolds.

Abstract

Due to the great economic losses caused by the root and butt-rot pathogen Heterobasidion annosum, development of efficient control measures is warranted. H. annosum a necrotroph colonize the Norway spruce from inside and is responsible for 100s of millions of Euros losses annually. Considerable clonal variation has been recorded for Norway spruce in resistance towards H. annosum, but the defence mechanisms contributing to host resistance against both necrotrophic and biotrophic fungi remain poorly understood. The recent genome sequencing of Populus has made the genus a model to facilitate tree genetics. Genome-wide transcript profiling of Populus tremula upon pathogen attack will now be used, and homologues of Norway spruce genes to defence genes up-regulated in Populus will be identified. Two aspen clones (23 and 72) from the SwAsp collection (Luquez 2007) were used in the present study. Plants were propagated from tissue culture and kept in greenhouse under un-manipulated conditions. To study the host defence mechanisms, the rust (Melampsora magnusiana Wagner) and a bluestain fungus were used as a biotrophic and necrotrophic fungus respectively. Melampsora spores solution was applied to the underside of the leaf. To control for sectoriality six leaves were infected on each plant. To ensure high humidity and avoid cross contamination, plastic bags were wrapped around infected leaves. Leaves above infected leaves (10, 9, and 8) were harvested respectively 1, 3 and 14 days after inoculation. Initial results from microarray data indicate a clear separation between two Aspen clone (23 and 72) lines. For line 23 the response to biotroph and necrotroph seems to be similar. Whereas the response for line 72 is divergent for the treatments as they go in opposite direction. The controls show that there is an initial difference in the 2 lines (controls are separated). What are the genes that make biotrohic and necrotrophic treatment of 72 look so different will be worked out from microarray data. Differential expression of defense genes in biotrohic and necrotrophic treatment will be verified further with quantitative real time PCR. Chemical analysis of Aspen leaves gave less phenolic compounds as plants were kept in greenhouse. HPLC will be carried out after reaching some conclusion from microarray data analysis.

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Abstract

Ophiostomatoid fungi were isolated from Scolytus ratzeburgi infesting Betula pendula and B. pubescens in Norway. Fungi were identified based on morphology, DNA sequence comparison for two gene regions and phylogenetic analyses. The most abundant fungus was Ophiostoma karelicum, suggesting a specific relationship between the fungus, the vector insect and the host tree. Our results suggest that O. karelicum occurs across the geographic range of S. ratzeburgi and its close relatedness to the Dutch elm disease fungi suggests that it could be important if introduced into other parts of the world. Other fungi, only occasionally isolated from S. ratzeburgi, were identified as O. quercus and a novel taxon, described here as O. denticiliatum sp. nov.

Abstract

The root-rot causing fungus Heterobasidion annosum senso lato is the most devastating pathogen of conifers in Europe. This pathogen enter Norway spruce trees trough the roots and colonizes the tree from within, growing as a saprophyte when established within the dead heartwood and acting as a necrotroph when in contact with living host tissue. The genome of this devastating pathogen has now been sequenced in collaboration with JGI and gene annotation is ongoing and genomic work is currently in progress (Stenlid et al. work in progress). We have worked with the host Norway spruce from a molecular perspective for more than ten years. Twenty percent of the trees in Norwegian spruce stands tend to be infected and this pathogen that can colonize ten meters up inside the trunk. The tree have defences against this pathogen and the attack can be fought off by the bark and living wood but not by the hearthwood. The tree has a unique defense against this internal attack by forming a reaction zone; in this case the host defense is directed inwardly by the still living sapwood toward the central colonized wood. We have in the last years studied the host responses to infection in Norway spruce clones at the transcriptional level and found that the speed of recognition and spatial defense signalling appears to be the hallmarks of trees with high degree of resistance. We strive to study both partners in this pathosystem from a molecular perspective, and are using suppressive subtractive hybridization (SSH) followed by Real-Time RT PCR verification to look at differentially expressed genes(Yakovlev et al. 2008). In addition the colonization profiles are followed on extracted gDNA using quantitative Real-Time PCR (Hietala et al. 2009).

Abstract

In natural conditions plants are continuously exposed to number of pathogens both biotrophs and necrotrophs. To understand their defense response at the transcript level two clones C72 and C23 with differential level of resistance from the SwAsp collection were inoculated with a biotroph (Melampsora magnusiana Wagnar) and necrotroph (Ceratocysis spp.) and compared to wounded and healthy controls. Samples were collected in leaves and areas some distance away from the inoculation site to examine the long distance (systemic) defense responses at day, day3 and day14 post treatments. We performed microarray experiments on the necrotrophic and biothrophic interaction compared with the healthy controls and found that the two clones respond in widely different fashions to the treatments applied. Clone C23 showed almost no response to biotroph and necrotroph inoculations after 24 hours while clone 72 gave a clear defense response to both pathogens. We are now in the process of verifying these results and looking at additional time-points using qRT-PCR.