| 1. |
- Berlin, Anna, et al.
(författare)
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Evolutionary history reveals two phylogenetically distinct species within Puccinia graminis
- 2013
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Ingår i: Acta Phytopathologica Sinica. - 0412-0914. ; 43, s. 486-
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- The cereal pathogen Puccinia graminis is considered to have high genetic diversity within the species. We hereby hypothesize that the different formae speciales of P. graminis have co-evolved with their various grass hosts. Elongation factor 1 alpha (EF-1α) and beta-tubulin (BT) genes, the internal transcribed spacer (ITS) region and mitochondrial cytochrome oxidase subunit I (COI) were sequenced from DNA isolated from Puccinia graminis collected from cereal and wild grasses to infer the phylogeny of the fungus. The related species Puccinia coronata, collected from both oats and wild grasses, were used as a reference throughout the analysis. Coalescence analysis showed that the time to the most recent common ancestor (TMRCA) for P. graminis and P. coronata were in all models further away in time than TMRCA for the two formae speciales. Within the species P. graminis, two main clades were formed; one including samples collected from Avena sativa, Avena fatua, Phleum pratense, and the other including samples collected from Triticum aestivum, Triticum monoccocum, Secale cereal and Hordeum vulgare, suggesting that P. graminis is to be divided into two different taxa. However, samples collected from the weed host Elytrigia repens, did not show any clear pattern, the samples equally grouped with either of the two groups. The phylogeny of P. graminis was thus congruent with its respective grass hosts, which confirm a co-evolution with the host.
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| 2. |
- Beste, Lisa, et al.
(författare)
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Synthesis of Hydroxylated Sterols in Transgenic Arabidopsis Plants Alters Growth and Steroid Metabolism
- 2011
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Ingår i: Plant Physiology. - : Oxford University Press (OUP). - 0032-0889 .- 1532-2548. ; 157:1, s. 426-440
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Tidskriftsartikel (refereegranskat)abstract
- To explore mechanisms in plant sterol homeostasis, we have here increased the turnover of sterols in Arabidopsis (Arabidopsis thaliana) and potato (Solanum tuberosum) plants by overexpressing four mouse cDNA encoding cholesterol hydroxylases (CHs), hydroxylating cholesterol at the C-7, C-24, C-25, or C-27 positions. Compared to the wild type, the four types of Arabidopsis transformant showed varying degrees of phenotypic alteration, the strongest one being in CH25 lines, which were dark-green dwarfs resembling brassinosteroid-related mutants. Gas chromatography-mass spectrometry analysis of extracts from wild-type Arabidopsis plants revealed trace levels of alpha and beta forms of 7-hydroxycholesterol, 7-hydroxycampesterol, and 7-hydroxysitosterol. The expected hydroxycholesterol metabolites in CH7-, CH24-, and CH25 transformants were identified and quantified using gas chromatography-mass spectrometry. Additional hydroxysterol forms were also observed, particularly in CH25 plants. In CH24 and CH25 lines, but not in CH7 ones, the presence of hydroxysterols was correlated with a considerable alteration of the sterol profile and an increased sterol methyltransferase activity in microsomes. Moreover, CH25 lines contained clearly reduced levels of brassinosteroids, and displayed an enhanced drought tolerance. Equivalent transformations of potato plants with the CH25 construct increased hydroxysterol levels, but without the concomitant alteration of growth and sterol profiles observed in Arabidopsis. The results suggest that an increased hydroxylation of cholesterol and/or other sterols in Arabidopsis triggers compensatory processes, acting to maintain sterols at adequate levels.
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| 3. |
- Chaudhary, Rajiv, et al.
(författare)
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Combining transcriptomics and genetic linkage based information to identify candidate genes associated with Heterobasidion-resistance in Norway spruce
- 2020
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- The Heterobasidion annosum s.l species complex comprises the most damaging forest pathogens to Norway spruce. We revisited previously identified Quantitative Trait Loci (QTLs) related to Heterobasidion-resistance in Norway spruce to identify candidate genes associated with these QTLs. We identified 329 candidate genes associated with the resistance QTLs using a gene-based composite map for Pinaceae. To evaluate the transcriptional responses of these candidate genes to H. parviporum, we inoculated Norway spruce plants and sequenced the transcriptome of the interaction at 3 and 7 days post inoculation. Out of 298 expressed candidate genes 124 were differentially expressed between inoculation and wounding control treatment. Interestingly, PaNAC04 and two of its paralogs in the subgroup III-3 of the NAC family transcription factors were found to be associated with one of the QTLs and was also highly induced in response to H. parviporum. These genes are possibly involved in the regulation of biosynthesis of flavonoid compounds. Furthermore, several of the differentially expressed candidate genes were associated with the phenylpropanoid pathway including a phenylalanine ammonia-lyase, a cinnamoyl-CoA reductase, a caffeoyl-CoA O-methyltransferase and a PgMYB11-like transcription factor gene. Combining transcriptome and genetic linkage analyses can help identifying candidate genes for functional studies and molecular breeding in non-model species.
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| 4. |
- Dalman, Kerstin, et al.
(författare)
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A genome-wide association study identifies genomic regions for virulence in Heterobasidion annosum s.s
- 2012
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Heterobasidion annosum (Fr.) Bref. sensu lato (s.l.) is a necrotrophic pathogen that causessevere damage to coniferous forests in the Northern Hemisphere. A genome-wide association studyanalysing the virulence of H. annosum sensu stricto (s.s.) on spruce (Picea abies) and pine (Pinussylvestris) using 23 homokaryotic haploid isolates was performed. The virulence of the isolates wasmeasured as lesion length in the phloem and fungal growth within the sapwood followinginoculations in the stem of 2-year-old pine and spruce seedlings. The fungal isolates were sequencedto between 2.6× and 12.6× coverage using the Illumina Genome Analyzer. This data set yielded33,018 single nucleotide polymorphisms (SNPs), with a minor allelic frequency of at least two out of23. These loci were present in all isolates. SNPs and mean values for each virulence trait were usedfor the association study. Twelve SNP markers distributed on seven contigs were found to besignificantly associated with fungal virulence (P< 0.0001). These regions were characterized forlinkage disequilibrium (LD) and gene contents. The LD blocks in these regions ranged between 1.2and 31.2 kb when present. Nine genes encoding calcineurin, acetylglutamate kinase/synthase,cytochrome P450 monooxygenase, serine carboxypeptidase, quinone oxidoreductase (ToxD), twoflavin-containing monooxygenases, exopolyphosphatase and a Swi5 transcription factor wereidentified as candidates for virulence.
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| 5. |
- Dalman, Kerstin, et al.
(författare)
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A Genome-Wide Association Study Identifies Genomic Regions for Virulence in the Non-Model Organism Heterobasidion annosum ss
- 2013
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 8, s. 1-10
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Tidskriftsartikel (refereegranskat)abstract
- The dense single nucleotide polymorphisms (SNP) panels needed for genome wide association (GWA) studies have hitherto been expensive to establish and use on non-model organisms. To overcome this, we used a next generation sequencing approach to both establish SNPs and to determine genotypes. We conducted a GWA study on a fungal species, analysing the virulence of Heterobasidion annosum s.s., a necrotrophic pathogen, on its hosts Picea abies and Pinus sylvestris. From a set of 33,018 single nucleotide polymorphisms (SNP) in 23 haploid isolates, twelve SNP markers distributed on seven contigs were associated with virulence (P,0.0001). Four of the contigs harbour known virulence genes from other fungal pathogens and the remaining three harbour novel candidate genes. Two contigs link closely to virulence regions recognized previously by QTL mapping in the congeneric hybrid H. irregulare6H. occidentale. Our study demonstrates the efficiency of GWA studies for dissecting important complex traits of small populations of non-model haploid organisms with small genomes.
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| 6. |
- Dalman, Kerstin, et al.
(författare)
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Evolutionary history of the conifer root rot fungus Heterobasidion annosum sensu lato
- 2012
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Ingår i: Proceedings e report. ; 93, s. 67-70
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- We investigated two hypotheses for the origin of the root rot fungus Heterobasidionannosum species complex: (i) that geology has been an important factor for the speciation (ii) that coevolutionaryprocesses with the hosts drove the divergence of the pathogen species. The H. annosumspecies complex consists of five species: three occur in Europe, H. annosum s.s., Heterobasidionparviporum and Heterobasidion abietinum, and two in North America, Heterobasidion irregulare andHeterobasidion occidentale; all with different but partially overlapping host preferences. Theevolution of the H. annosum species complex was studied using six partially sequenced genes,between 10 and 30 individuals of each species were analysed. Neighbour-joining trees wereconstructed for each gene, and a Bayesian tree was built for the combined data set. In addition,haplotype networks were constructed to illustrate the species relationships. For three of the genes, H.parviporum and H. abietinum share haplotypes supporting recent divergence and⁄or possible geneflow. We propose that the H. annosum species complex originated in Laurasia and that the H.annosum s.s.⁄H. irregulare and H. parviporum⁄H. abietinum⁄H. occidentale ancestral species emergedbetween 45 and 60 Ma in the Palaearctic, well after the radiation of the host genera. Our data implythat H. irregulare and H. occidentale were colonizing North America via different routes. Inconclusion, plate tectonics are likely to have been the main factor influencing Heterobasidionspeciation and biogeography.
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| 7. |
- Dalman, Kerstin, et al.
(författare)
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Evolutionary history of the conifer root rot fungus Heterobasidion annosum sensu lato
- 2010
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Ingår i: Molecular Ecology. - 0962-1083 .- 1365-294X. ; 19, s. 4979-4993
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Tidskriftsartikel (refereegranskat)abstract
- We investigated two hypotheses for the origin of the root rot fungus Heterobasidion annosum species complex: (i) that geology has been an important factor for the speciation (ii) that co-evolutionary processes with the hosts drove the divergence of the pathogen species. The H. annosum species complex consists of five species: three occur in Europe, H. annosum s.s., Heterobasidion parviporum and Heterobasidion abietinum, and two in North America, Heterobasidion irregulare and Heterobasidion occidentale; all with different but partially overlapping host preferences. The evolution of the H. annosum species complex was studied using six partially sequenced genes, between 10 and 30 individuals of each species were analysed. Neighbour-joining trees were constructed for each gene, and a Bayesian tree was built for the combined data set. In addition, haplotype networks were constructed to illustrate the species relationships. For three of the genes, H. parviporum and H. abietinum share haplotypes supporting recent divergence and/or possible gene flow. We propose that the H. annosum species complex originated in Laurasia and that the H. annosum s.s./H. irregulare and H. parviporum/H. abietinum/H. occidentale ancestral species emerged between 45 and 60 Ma in the Palaearctic, well after the radiation of the host genera. Our data imply that H. irregulare and H. occidentale were colonizing North America via different routes. In conclusion, plate tectonics are likely to have been the main factor influencing Heterobasidion speciation and biogeography.
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| 8. |
- Dalman, Kerstin
(författare)
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Heterobasidion root rot : genetic mapping of virulence and evolutionary history
- 2010
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Heterobasidion annosum (Fr.) Bref. sensu lato (s.l.) is a necrotrophic pathogen causing damage to conifers in the Northern Hemisphere. H. annosum s.l. consists of five species: three European [H. annosum sensu stricto (s.s.), H. parviporum and H. abietinum] and two North American (H. irregulare and H. occidentale); all with different but partially overlapping host preferences. A multilocus phylogenetic tree was built and the divergence times were estimated. Plate tectonics is likely to have been the main factor influencing Heterobasidion speciation and biogeography. Along with the geographical separation, the Heterobasidion species have specialized on different host genera. The H. annosum species complex originated in Laurasia and the H. annosum s.s./H. irregulare and H. parviporum/H. abietinum/H. occidentale ancestral species emerged between 45 million–60 million years ago in the Palaearctic. The data imply that H. irregulare and H. occidentale colonized North America via different routes: H. irregulare colonizing from the east via Trans Atlantic land bridges and H. occidentale colonizing from the west via the Bering Land Bridge. Alternatively H. occidentale originated from North America. Identification of virulence factors is important for understanding the Heterobasidion–conifer pathosystem. Two studies of genetic mapping of virulence were performed. Virulence traits were measured as lesion length in the phloem and fungal growth in the sapwood of pine and spruce. Quantitative trait loci (QTL) were identified and positioned on a genetic linkage map for virulence of 102 progeny isolates from a cross between H. irregulare and H. occidentale. Both virulence traits in Picea abies identified significant QTLs on linkage group (LG) 15. Another QTL was positioned on LG 15 for the lesion length measurement in Pinus sylvestris. Moreover, QTLs on two separate smaller LGs were identified for fungal growth in sapwood and lesion length, respectively. The QTLs probably represent loci important for specific as well as general aspects of virulence on P. sylvestris and P. abies. A genome-wide association study was performed for virulence on 23 H. annosum s.s. isolates. Twelve SNP markers distributed on seven contigs were significantly associated with virulence. From these, three regions were characterized, two with one marker each with the lowest p-values and one region containing six markers. The linkage disequilibrium blocks in these regions ranged between 1.2 and 31.2 kb. Seven genes were identified as candidate virulence determinants encoding calcineurin, acetylglutamate kinase/synthase, cytochrome P450 monooxygenase, serine carboxypeptidase, quinone oxidoreductase (ToxD) and two flavin-containing monooxygenases.
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| 9. |
- Dalman, Kerstin, et al.
(författare)
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Overexpression of PaNAC03, a stress induced NAC gene family transcription factor in Norway spruce leads to reduced flavonol biosynthesis and aberrant embryo development
- 2017
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Ingår i: BMC Plant Biology. - : BioMed Central. - 1471-2229. ; 17
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Tidskriftsartikel (refereegranskat)abstract
- Background: The NAC family of transcription factors is one of the largest gene families of transcription factors in plants and the conifer NAC gene family is at least as large, or possibly larger, as in Arabidopsis. These transcription factors control both developmental and stress induced processes in plants. Yet, conifer NACs controlling stress induced processes has received relatively little attention. This study investigates NAC family transcription factors involved in the responses to the pathogen Heterobasidion annosum (Fr.) Bref. sensu lato. Results: The phylogeny and domain structure in the NAC proteins can be used to organize functional specificities, several well characterized stress-related NAC proteins are found in III-3 in Arabidopsis (Jensen et al. Biochem J 426: 183-196, 2010). The Norway spruce genome contain seven genes with similarity to subgroup III-3 NACs. Based on the expression pattern PaNAC03 was selected for detailed analyses. Norway spruce lines overexpressing PaNAC03 exhibited aberrant embryo development in response to maturation initiation and 482 misregulated genes were identified in proliferating cultures. Three key genes in the flavonoid biosynthesis pathway: a CHS, a F3'H and PaLAR3 were consistently down regulated in the overexpression lines. In accordance, the overexpression lines showed reduced levels of specific flavonoids, suggesting that PaNAC03 act as a repressor of this pathway, possibly by directly interacting with the promoter of the repressed genes. However, transactivation studies of PaNAC03 and PaLAR3 in Nicotiana benthamiana showed that PaNAC03 activated PaLAR3A, suggesting that PaNAC03 does not act as an independent negative regulator of flavan-3-ol production through direct interaction with the target flavonoid biosynthetic genes. Conclusions: PaNAC03 and its orthologs form a sister group to well characterized stress-related angiosperm NAC genes and at least PaNAC03 is responsive to biotic stress and appear to act in the control of defence associated secondary metabolite production.
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| 10. |
- Dauphinee, Adrian N., et al.
(författare)
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Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators
- 2019
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Ingår i: Plant Physiology. - : AMER SOC PLANT BIOLOGISTS. - 0032-0889 .- 1532-2548. ; 181:3, s. 855-866
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Tidskriftsartikel (refereegranskat)abstract
- Autophagy is a major catabolic process in eukaryotes with a key role in homeostasis, programmed cell death, and aging. In plants, autophagy is also known to regulate agronomically important traits such as stress resistance, longevity, vegetative biomass, and seed yield. Despite its significance, there is still a shortage of reliable tools modulating plant autophagy. Here, we describe the first robust pipeline for identification of specific plant autophagy-modulating compounds. Our screening protocol comprises four phases: (1) high-throughput screening of chemical compounds in cell cultures of tobacco (Nicotiana tabacum); (2) confirmation of the identified hits in planta using Arabidopsis (Arabidopsis thaliana); (3) further characterization of the effect using conventional molecular biology methods; and (4) verification of chemical specificity on autophagy in planta. The methods detailed here streamline the identification of specific plant autophagy modulators and aid in unraveling the molecular mechanisms of plant autophagy.
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| 11. |
- Elander, Pernilla, et al.
(författare)
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Tudor staphylococcal nuclease is a docking platform for stress granule components and is essential for SnRK1 activation in Arabidopsis
- 2021
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Ingår i: The Embo Journal. - : EMBO. - 0261-4189 .- 1460-2075. ; 40
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Tidskriftsartikel (refereegranskat)abstract
- Tudor staphylococcal nuclease (TSN; also known as Tudor-SN, p100, or SND1) is a multifunctional, evolutionarily conserved regulator of gene expression, exhibiting cytoprotective activity in animals and plants and oncogenic activity in mammals. During stress, TSN stably associates with stress granules (SGs), in a poorly understood process. Here, we show that in the model plant Arabidopsis thaliana, TSN is an intrinsically disordered protein (IDP) acting as a scaffold for a large pool of other IDPs, enriched for conserved stress granule components as well as novel or plant-specific SG-localized proteins. While approximately 30% of TSN interactors are recruited to stress granules de novo upon stress perception, 70% form a protein-protein interaction network present before the onset of stress. Finally, we demonstrate that TSN and stress granule formation promote heat-induced activation of the evolutionarily conserved energy-sensing SNF1-related protein kinase 1 (SnRK1), the plant orthologue of mammalian AMP-activated protein kinase (AMPK). Our results establish TSN as a docking platform for stress granule proteins, with an important role in stress signalling.
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| 12. |
- Ghasemkhani, Marjan, et al.
(författare)
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Real-time PCR for detection and quantification, and histological characterization of Neonectria ditissima in apple trees
- 2016
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Ingår i: Trees - Structure and Function. - : Springer Science and Business Media LLC. - 0931-1890 .- 1432-2285. ; 30, s. 1111-1125
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Tidskriftsartikel (refereegranskat)abstract
- Neonectria ditissima, the causal pathogen of fruit tree canker, is a sordariomycete fungus that affects apple orchards, especially in north-western Europe. To prevent serious disease epidemics, an accurate, rapid, and sensitive method for detection of N. ditissima is needed for pathogen identification. A quantitative real-time PCR (qPCR) assay was developed for both detection and quantification of this pathogen in infected apple cultivars. Several primer sets were designed from regions of the β-tubulin gene. One primer set passed several validation tests, and the melting curve confirmed species-specific amplification of the correct product. In addition, the N. ditissima biomass could be detected at variable amounts in samples from the infection sites of six different cultivars, with ‘Aroma' having the lowest amount of N. ditissima biomass and ‘Elise' the highest. To complement the qPCR results, tissue from detached shoots and 1-year-old trees of ‘Cox's Orange Pippin' (susceptible) and ‘Santana' (partially resistant) was used in a histopathology study. In both detached shoots and trees, fungal hyphae were found in cells of all tissues. No qualitative differences in the anatomy of the infected samples were observed between the cultivars. In the detached shoot experiment, both cultivars were affected but differences in the rate of disease progression suggest that the partially resistant cultivar could resist the fungus longer. The qPCR assay developed in our study produced reproducible results and can be used for detection of N. ditissima in infected trees.
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| 13. |
- Minina, Alyona, et al.
(författare)
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Transcriptional stimulation of rate-limiting components of the autophagic pathway improves plant fitness
- 2018
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Ingår i: Journal of Experimental Botany. - : Oxford University Press (OUP). - 0022-0957 .- 1460-2431. ; 69, s. 1415-1432
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Tidskriftsartikel (refereegranskat)abstract
- Autophagy is a major catabolic process whereby autophagosomes deliver cytoplasmic content to the lytic compartment for recycling. Autophagosome formation requires two ubiquitin-like systems conjugating Atg12 with Atg5, and Atg8 with lipid phosphatidylethanolamine (PE), respectively. Genetic suppression of these systems causes autophagy-deficient phenotypes with reduced fitness and longevity. We show that Atg5 and the E1-like enzyme, Atg7, are rate-limiting components of Atg8-PE conjugation in Arabidopsis. Overexpression of ATG5 or ATG7 stimulates Atg8 lipidation, autophagosome formation, and autophagic flux. It also induces transcriptional changes opposite to those observed in atg5 and atg7 mutants, favoring stress resistance and growth. As a result, ATG5-or ATG7-over-expressing plants exhibit increased resistance to necrotrophic pathogens and oxidative stress, delayed aging and enhanced growth, seed set, and seed oil content. This work provides an experimental paradigm and mechanistic insight into genetic stimulation of autophagy in planta and shows its efficiency for improving plant productivity.
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| 14. |
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| 15. |
- Nahar, Nurun, et al.
(författare)
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Transcript profiling of two potato cultivars during glycoalkaloid-inducing treatments shows differential expression of genes in sterol and glycoalkaloid metabolism
- 2017
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 7
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Tidskriftsartikel (refereegranskat)abstract
- Steroidal glycoalkaloids (SGA) are sterol-derived neurotoxic defence substances present in several members of the Solanaceae. In the potato (Solanum tuberosum), high SGA levels may render tubers harmful for consumption. Tuber SGA levels depend on genetic factors, and can increase as a response to certain stresses and environmental conditions. To identify genes underlying the cultivar variation in tuber SGA levels, we investigated two potato cultivars differing in their SGA accumulation during wounding or light exposure; two known SGA-inducing treatments. Using microarray analysis coupled to sterol and SGA quantifications, we identified a small number of differentially expressed genes that were associated with increased SGA levels. Two of these genes, encoding distinct types of sterol Delta(24)-reductases, were by sense/antisense expression in transgenic potato plants shown to have differing roles in sterol and SGA metabolism. The results show that an increased SGA level in potato tubers during both wounding and light exposure is mediated by coordinated expression of a set of key genes in isoprenoid and steroid metabolism, and suggest that differences in this expression underlie cultivar variations in SGA levels. These results may find use within potato breeding and quality assessment.
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| 16. |
- Nemesio Gorriz, Miguel, et al.
(författare)
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Identification of Norway Spruce MYB-bHLH-WDR Transcription Factor Complex Members Linked to Regulation of the Flavonoid Pathway
- 2017
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Ingår i: Frontiers in Plant Science. - : Frontiers Media SA. - 1664-462X. ; 8
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Tidskriftsartikel (refereegranskat)abstract
- Transcription factors (TFs) forming MYB-bHLH-WDR complexes are known to regulate the biosynthesis of specialized metabolites in angiosperms through an intricate network. These specialized metabolites participate in a wide range of biological processes including plant growth, development, reproduction as well as in plant immunity. Studying the regulation of their biosynthesis is thus essential. While MYB (TFs) have been previously shown to control specialized metabolism (SM) in gymnosperms, the identity of their partners, in particular bHLH or WDR members, has not yet been revealed. To gain knowledge about MYB-bHLH-WDR transcription factor complexes in gymnosperms and their regulation of SW, we identified two bHLH homologs of AtTT8, six homologs of the MYB transcription factor 112 2 and one WDR ortholog of Atl I G1 in Norway spruce. We investigated the expression levels of these genes in diverse tissues and upon treatments with various stimuli including methyl-salicylate, methyl-jasmonate, wounding or fungal inoculation. In addition, we also identified protein-protein interactions among different homologs of MYB, bHLH and WDR. Finally, we generated transgenic spruce cell lines overexpressing four of the Norway spruce 112 2 homologs and observed differential regulation of genes in the flavonoid pathway and flavonoid contents.
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| 17. |
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| 18. |
- Reza, Salim Hossain, et al.
(författare)
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Transcriptome analysis of embryonic domains in Norway spruce reveals potential regulators of suspensor cell death
- 2018
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Ingår i: PLOS ONE. - San Francisco : Public Library of Science. - 1932-6203. ; 13:3
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Tidskriftsartikel (refereegranskat)abstract
- The terminal differentiation and elimination of the embryo-suspensor is the earliest manifestation of programmed cell death (PCD) during plant ontogenesis. Molecular regulation of suspensor PCD remains poorly understood. Norway spruce (Picea abies) embryos provide a powerful model for studying embryo development because of their large size, sequenced genome, and the possibility to obtain a large number of embryos at a specific developmental stage through somatic embryogenesis. Here, we have carried out global gene expression analysis of the Norway spruce embryo-suspensor versus embryonal mass (a gymnosperm analogue of embryo proper) using RNA sequencing. We have identified that suspensors have enhanced expression of the NAC domain-containing transcription factors, XND1 and ANAC075, previously shown to be involved in the initiation of developmental PCD in Arabidiopsis. The analysis has also revealed enhanced expression of Norway spruce homologues of the known executioners of both developmental and stress-induced cell deaths, such as metacaspase 9 (MC9), cysteine endopeptidase-1 (CEP1) and ribonuclease 3 (RNS3). Interestingly, a spruce homologue of bax inhibitor-1 (PaBI-1, for Picea abies BI-1), an evolutionarily conserved cell death suppressor, was likewise up-regulated in the embryosuspensor. Since Arabidopsis BI-1 so far has been implicated only in the endoplasmic reticulum (ER)-stress induced cell death, we investigated its role in embryogenesis and suspensor PCD using RNA interference (RNAi). We have found that PaBI-1-deficient lines formed a large number of abnormal embryos with suppressed suspensor elongation and disturbed polarity. Cytochemical staining of suspensor cells has revealed that PaBI-1 deficiency suppresses vacuolar cell death and induces necrotic type of cell death previously shown to compromise embryo development. This study demonstrates that a large number of cell-death components are conserved between angiosperms and gymnosperms and establishes a new role for BI-1 in the progression of vacuolar cell death.
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| 19. |
- Van Der Nest, Magriet, et al.
(författare)
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Distribution and evolution of het gene homologs in the basidiomycota
- 2014
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Ingår i: Fungal Genetics and Biology. - : Elsevier BV. - 1087-1845 .- 1096-0937. ; 64, s. 45-57
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Tidskriftsartikel (refereegranskat)abstract
- In filamentous fungi a system known as somatic incompatibility (SI) governs self/non-self recognition. SI is controlled by a regulatory signaling network involving proteins encoded at the het (heterokaryon incompatible) loci. Despite the wide occurrence of SI, the molecular identity and structure of only a small number of het genes and their products have been characterized in the model fungi Neurospora crassa and Podospora anserina. Our aim was to identify and study the distribution and evolution of putative het gene homologs in the Basidiomycota. For this purpose we used the information available for the model fungi to identify homologs of het genes in other fungi, especially the Basidiomycota. Putative het-c, het-c2 and un-24 homologs, as well as sequences containing the NACHT, HET or WD40 domains present in the het-e, het-r, het-6 and het-d genes were identified in certain members of the Ascomycota and Basidiomycota. The widespread phylogenetic distribution of certain het genes may reflect the fact that the encoded proteins are involved in fundamental cellular processes other than SI. Although homologs of het-S were previously known only from the Sordariomycetes (Ascomycota), we also identified a putative homolog of this gene in Gymnopus luxurians (Basidiomycota, class Agaricomycetes). Furthermore, with the exception of un-24, all of the putative het genes identified occurred mostly in a multi-copy fashion, some with lineage and species-specific expansions. Overall our results indicated that gene duplication followed by gene loss and/or gene family expansion, as well as multiple events of domain fusion and shuffling played an important role in the evolution of het gene homologs of Basidiomycota and other filamentous fungi. (C) 2013 Elsevier Inc. All rights reserved.
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| 20. |
- Van Der Nest, Magriet, et al.
(författare)
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Gene expression associated with intersterility in Heterobasidion
- 2014
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Ingår i: Fungal Genetics and Biology. - : Elsevier BV. - 1087-1845 .- 1096-0937. ; 73, s. 104-119
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Tidskriftsartikel (refereegranskat)abstract
- Intersterility (IS) is thought to prevent mating compatibility between homokaryons that belong to different species. Although IS in Heterobasidion is regulated by the genes located at the IS loci, it is not yet known how the IS genes influence sexual compatibility and heterokaryon formation. To increase our understanding of the molecular events underlying IS, we studied mRNA abundance changes during IS compatible and incompatible interactions over time. The clustering of the transcripts into expression profiles, followed by the application of Gene Ontology (GO) enrichment pathway analysis of each of the clusters, allowed inference of biological processes participating in IS. These analyses identified events involved in mating and sexual development (i.e., linked with IS compatibility), which included processes associated with cell-cell adhesion and recognition, cell cycle control and signal transduction. We also identified events potentially involved in overriding mating between individuals belonging to different species (i.e., linked with IS incompatibility), which included reactive oxygen species (ROS) production, responses to stress (especially to oxidative stress), signal transduction and metabolic biosynthesis. Our findings thus enabled detection and characterization of gene expression changes associated with IS in Heterobasidion, as well as identification of important processes and pathways associated with this phenomenon. Overall, the results of this study increase current knowledge regarding the molecular mechanisms underpinning IS in Heterobasidion and allowed for the establishment of a vital baseline for further studies. (C) 2014 Elsevier Inc. All rights reserved.
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| 21. |
- Vélëz, Heriberto, et al.
(författare)
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Transformation and gene-disruption in the apple-pathogen, Neonectria ditissima
- 2022
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Ingår i: HEREDITAS. - : Springer Science and Business Media LLC. - 1601-5223. ; 159
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Tidskriftsartikel (refereegranskat)abstract
- Background Apple production in Sweden and elsewhere is being threatened by the fungus, Neonectria ditissima, which causes a disease known as European canker. The disease can cause extensive damage and the removal of diseased wood and heavily infected trees can be laborious and expensive. Currently, there is no way to eradicate the fungus from infected trees and our knowledge of the infection process is limited. Thus, to target and modify genes efficiently, the genetic transformation technique developed for N. ditissima back in 2003 was modified. Results The original protocol from 2003 was upgraded to use enzymes currently available in the market for making protoplasts. The protoplasts were viable, able to uptake foreign DNA, and able to regenerate back into a mycelial colony, either as targeted gene-disruption mutants or as ectopic mutants expressing the green fluorescent protein (GFP). Conclusions A new genetic transformation protocol has been established and the inclusion of hydroxyurea in the buffer during the protoplast-generation step greatly increased the creation of knockout mutants via homologous recombination. Pathogenicity assays using the GFP-mutants showed that the mutants were able to infect the host and cause disease.
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