| 1. |
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| 2. |
- Andersson, Louise, et al.
(författare)
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Draft genome sequence of the sugar beet pathogen Rhizoctonia solani AG2-2IIIB strain BBA69670
- 2016
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Ingår i: Journal of Biotechnology. - : Elsevier BV. - 0168-1656 .- 1873-4863. ; 222, s. 11-12
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Tidskriftsartikel (refereegranskat)abstract
- Rhizoctonia solani is a widespread plant pathogenic fungus featuring a broad host range including several economically important crops. Accordingly, genome analyses of R. solani isolates are important to uncover their pathogenic potential. Draft genome sequences for four R. solani isolates representing three of the 14 R. solani anastomosis groups (AGs) are available. Here, we present the first draft genome sequence for an R. solani AG2-2IIIB isolate that is pathogenic on sugar beet. The fungal genome was assembled in 2065 scaffolds consisting of 5826 contigs amounting to a size of about 52 Mb which is larger than any other R. solani isolate known today. Genes potentially encoding cellulolytic, lignolytic and pectinolytic enzymes were identified. (c) 2016 Elsevier B.V. All rights reserved.
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| 3. |
- Andersson, Louise, et al.
(författare)
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Genome analysis of the sugar beet pathogen Rhizoctonia solani AG2-2IIIB revealed high numbers in secreted proteins and cell wall degrading enzymes
- 2016
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Ingår i: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 17
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Tidskriftsartikel (refereegranskat)abstract
- Background: Sugar beet (Beta vulgaris) is a crop cultivated for its high content in sugar, but it is vulnerable to many soil-borne pathogens. One of them is the basidiomycete Rhizoctonia solani. This fungal species has a compatibility system regulating hyphal fusions (anastomosis). Consequently, R. solani species are categorized in anastomosis groups (AGs). AG2-2IIIB isolates are most aggressive on sugar beet. In the present study, we report on the draft genome of R. solani AG2-2IIIB using the Illumina technology. Genome analysis, interpretation and comparative genomics of five sequenced R. solani isolates were carried out. Results: The draft genome of R. solani AG2-2IIIB has an estimated size of 56.02 Mb. In addition, two normalized EST libraries were sequenced. In total 20,790 of 21,980 AG2-2IIIB isotigs (transcript isoforms) were mapped on the genome with more than 95 % sequence identity. The genome of R. solani AG2-2IIIB was predicted to harbor 11,897 genes and 4908 were found to be isolate-specific. R. solani AG2-2IIIB was predicted to contain 1142 putatively secreted proteins and 473 of them were found to be unique for this isolate. The R. solani AG2-2IIIB genome encodes a high number of carbohydrate active enzymes. The highest numbers were observed for the polysaccharide lyases family 1 (PL-1), glycoside hydrolase family 43 (GH-43) and carbohydrate estarase family 12 (CE-12). Transcription analysis of selected genes representing different enzyme clades revealed a mixed pattern of up-and down-regulation six days after infection on sugar beets featuring variable levels of resistance compared to mycelia of the fungus grown in vitro. Conclusions: The established R. solani AG2-2IIIB genome and EST sequences provide important information on the gene content, gene structure and transcriptional activity for this sugar beet pathogen. The enriched genomic platform provides an important platform to enhance our understanding of R. solani biology.
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| 4. |
- Andersson, Louise, et al.
(författare)
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Major latex protein-like encoding genes contribute to Rhizoctonia solani defense responses in sugar beet
- 2021
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Ingår i: Molecular Genetics and Genomics. - : Springer Science and Business Media LLC. - 1617-4615 .- 1617-4623. ; 296, s. 155-164
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Tidskriftsartikel (refereegranskat)abstract
- Sugar beets are attacked by several pathogens that cause root damages. Rhizoctonia (Greek for "root killer") is one of them. Rhizoctonia root rot has become an increasing problem for sugar beet production and to decrease yield losses agronomical measures are adopted. Here, two partially resistant and two susceptible sugar beet genotypes were used for transcriptome analysis to discover new defense genes to this fungal disease, information to be implemented in molecular resistance breeding. Among 217 transcripts with increased expression at 2 days post-infection (dpi), three resistance-like genes were found. These genes were not significantly elevated at 5 dpi, a time point when increased expression of three Bet v I/Major latex protein (MLP) homologous genes BvMLP1, BvMLP2 and BvML3 was observed in the partially resistant genotypes. Quantitative RT-PCR analysis on diseased sugar beet seedlings validated the activity of BvMLP1 and BvMLP3 observed in the transcriptome during challenge by R. solani. The three BvMLP genes were cloned and overexpressed in Arabidopsis thaliana to further dissect their individual contribution. Transgenic plants were also compared to T-DNA mutants of orthologous MLP genes. Plants overexpressing BvMLP1 and BvMLP3 showed significantly less infection whereas additive effects were seen on Atmlp1/Atmlp3 double mutants. The data suggest that BvMLP1 and BvMLP3 may contribute to the reduction of the Rhizoctonia root rot disease in sugar beet. Impact on the defense reaction from other differential expressed genes observed in the study is discussed.
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| 5. |
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| 6. |
- Bejai, Sarosh, et al.
(författare)
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A novel role of PR2 in abscisic acid (ABA) mediated, pathogen-induced callose deposition in Arabidopsis thaliana
- 2013
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Ingår i: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 200, s. 1187-1199
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Tidskriftsartikel (refereegranskat)abstract
- Pathogenesis-related protein 2 (PR2) is known to play a major role in plant defense andgeneral stress responses. Resistance against the fungal pathogen Leptosphaeria maculans inArabidopsis requires abscisic acid (ABA), which promotes the deposition of callose, a b-1,3-glucan polymer. Here, we examined the role of PR2 in callose deposition in relation to ABAtreatment and challenge with L. maculans and Pseudomonas syringae. Characterization of PR2-overexpressing plants and the knockout line indicated that PR2negatively affects callose deposition. Recombinant PR2 purified from Pichia pastoris showedcallose-degrading activity, and a considerable reduction in the callose-degrading activity wasobserved in the leaf extract of the PR2 knockout line compared with the wild-type. ABA pretreatment before challenge with L. maculans concomitantly repressed PR2 andenhanced callose accumulation. Likewise, overexpression of an ABA biosynthesis geneNCED3 resulted in reduced PR2 expression and increased callose deposition. We propose that ABA promotes callose deposition through the transcriptional repression ofPR2 in Arabidopsis challenged by L. maculans and P. syringae. Callose by itself is likely to actantagonistically on salicylic acid (SA) defense signaling, suggesting that PR2 may function as amodulator of callose- and SA-dependent defense responses.
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| 7. |
- Courtois-Moreau, Charleen, Laetitia, 1978-
(författare)
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Programmed Cell Death in Xylem Development
- 2008
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Concerns about climate changes and scarcity of fossil fuels are rising. Hence wood is becoming an attractive source of renewable energy and raw material and these new dimensions have prompted increasing interest in wood formation in trees, in both the scientific community and wider public. In this thesis, the focus is on a key process in wood development: programmed cell death (PCD) in the development of xylem elements. Since secondary cell wall formation is dependent, inter alia, upon the life time of xylem elements, the qualitative features of wood will be affected by PCD in xylem, about which there is little information. This thesis focuses on the anatomical, morphological and transcriptional features of PCD during xylem development in both the stem of hybrid aspen, Populus tremula (L.) x tremuloides (Michx.) and the hypocotyl of the herbaceous model system Arabidopsis thaliana (L. Heynh.). In Populus, the progressive removal of organelles from the cytoplasm before the time of death (vacuolar bursts) and the slowness of the cell death process, illustrated by DNA fragmentation assays (such as TUNEL and Comet assays), have been ascertained in the xylem fibres by microscopic analyses. Furthermore, candidate genes for the regulation of fibre cell death were identified either from a Populus EST library obtained from woody tissues undergoing fibre cell death or from microarray experiments in Populus stem, and further assessed in an in silico comparative transcriptomic analysis of Arabidopsis thaliana. These candidate genes were either putative novel regulators of fibre cell death or members of previously described families of cell death-related genes, such as autophagy-related genes. The induction of the latter and the previous microscopic observations suggest the importance of autophagy in the degradation of the cytoplasmic contents specifically in the xylem fibres. Vacuolar bursts in the vessels were the only previously described triggers of PCD in the xylem, which induce the very rapid degradation of the nuclei and surrounding cytoplasmic contents, therefore unravelling a unique previously unrecorded type of PCD in the xylem fibres, principally involving autophagy. Arabidopsis is an attractive alternative model plant for exploring some aspects of wood formation, such as the characterisation of negative regulators of PCD. Therefore, the anatomy of Arabidopsis hypocotyls was also investigated and the ACAULIS5 (ACL5) gene, encoding an enzyme involved in polyamine biosynthesis, was identified as a key regulator of xylem specification, specifically in the vessel elements, though its negative effect on the cell death process. Taken together, PCD in xylem development seems to be a highly specific process, involving unique cell death morphology and molecular machinery. In addition, the technical challenges posed by the complexity of the woody tissues examined highlighted the need for specific methods for assessing PCD and related phenomena in wood.
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| 8. |
- Dixelius, Christina
(författare)
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Efficient plant regeneration of selected Kenyan sweetpotato (Ipomoea batatas (L.) Lam.) cultivars through somatic embryogenesis
- 2016
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Ingår i: Journal of Tissue Science & Engineering. - : OMICS Publishing Group. - 2157-7552. ; 7
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Tidskriftsartikel (refereegranskat)abstract
- Sweetpotato is an important food crop in the world as well as in Kenya. Various fungal and viral diseases are major constraints in its production and are currently threatening the sweetpotato production in sub-Saharan Africa. Genetic engineering offers significant potential for the crop’s genetic improvement. However, this is limited by the low efficiency and strong genotype dependency in tissue culture. This study aimed to establish an efficient somatic embryogenesis and plant regeneration system using shoot apical meristem explants of sweetpotato. Three sweetpotato cultivars that are widely grown in Kenya; KSP36, Kemb36 and Mweu mutheke along with an exotic model cultivar Jewel were evaluated. The maximum somatic embryogenic induction, at 96.72%, was obtained from explants cultured on Linsmaier and Skoog salts and vitamins medium supplemented with 0.5 mg/l dichlorophenoxyacetic acid and 0.2 mg/l zeatin riboside. The highest number of shoot induction (33) was observed after transfer of embryonic callus to embryo maturation medium supplemented with 2 mg/l abscisic acid. Significant differences were observed between cultivars for somatic embryogenesis and plant regeneration. Jewel showed the best response, while Mweu mutheke was the least responsive under the culture conditions tested in this study. Regenerated plants were successfully rooted and grown to maturity after hardening in soil in the greenhouse. Such a robust, successful and efficient system possesses the potential to become an important tool for crop improvement and functional studies of genes in sweetpotato.
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| 9. |
- Dixelius, Christina, et al.
(författare)
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European agricultural policy goes down the tubers
- 2012
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Ingår i: Nature Biotechnology. - : Springer Science and Business Media LLC. - 1087-0156 .- 1546-1696. ; 30, s. 492-493
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Annan publikation (övrigt vetenskapligt/konstnärligt)
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| 10. |
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| 11. |
- Dixelius, Christina
(författare)
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Induced Expression of Xerophyta viscosa XvSap1 Gene Enhances Drought Tolerance in Transgenic Sweet Potato
- 2019
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Ingår i: Frontiers in Plant Science. - : Frontiers Media SA. - 1664-462X. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- Drought stress often leads to reduced yields and is a perilous delimiter for expanded cultivation and increased productivity of sweet potato. Cell wall stabilization proteins have been identified to play a pivotal role in mechanical stabilization during desiccation stress mitigation in plants. They are involved in numerous cellular processes that modify cell wall properties to tolerate the mechanical stress during dehydration. This provides a plausible approach to engineer crops for enhanced stable yields under adverse climatic conditions. In this study, we genetically engineered sweet potato cv. Jewel with XvSap1 gene encoding a protein related to cell wall stabilization, isolated from the resurrection plant Xerophyta viscosa, under stress-inducible XvPSap1 promoter via Agrobacterium-mediated transformation. Detection of the transgene by PCR, Southern blot, and quantitative real-time PCR (qRT-PCR) analyses revealed the integration of XvSap1 in the three independent events. Phenotypic evaluation of shoot length, number of leaves, and yield revealed that the transgenic plants grew better than the wild-type plants under drought stress. Assessment of biochemical indices during drought stress showed higher levels of chlorophyll, free proline, and relative water content and decreased lipid peroxidation in transgenic plants than in wild types. Our findings demonstrate that XvSap1 enhances drought tolerance in transgenic sweet potato without causing deleterious phenotypic and yield changes. The transgenic drought-tolerant sweet potato lines provide a valuable resource as a drought-tolerant crop on arid lands of the world.
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| 12. |
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| 13. |
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| 14. |
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| 15. |
- Dixelius, Christina
(författare)
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Vårt dagliga bröd – i morgon
- 2011
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Ingår i: Genteknik som tar skruv. - 9789154060610 ; , s. 61-72
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Bokkapitel (populärvet., debatt m.m.)
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| 16. |
- Dixelius, Christina
(författare)
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Xerophyta viscosa Aldose Reductase, XvAld1, Enhances Drought Tolerance in Transgenic Sweetpotato
- 2018
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Ingår i: Molecular Biotechnology. - : Springer Science and Business Media LLC. - 1073-6085 .- 1559-0305. ; 60, s. 203-214
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Tidskriftsartikel (refereegranskat)abstract
- Sweetpotato is a significant crop which is widely cultivated particularly in the developing countries with high and stable yield. However, drought stress is a major limiting factor that antagonistically influences the crop's productivity. Dehydration stress caused by drought causes aggregation of reactive oxygen species (ROS) in plants, and aldose reductases are first-line safeguards against ROS caused by oxidative stress. In the present study, we generated transgenic sweetpotato plants expressing aldose reductase, XvAld1 isolated from Xerophyta viscosa under the control of a stress-inducible promoter via Agrobacterium-mediated transformation. Our results demonstrated that the transgenic sweetpotato lines displayed significant enhanced tolerance to simulated drought stress and enhanced recuperation after rehydration contrasted with wild-type plants. In addition, the transgenic plants exhibited improved photosynthetic efficiency, higher water content and more proline accumulation under dehydration stress conditions compared with wild-type plants. These results demonstrate that exploiting the XvAld1 gene is not only a compelling and attainable way to improve sweetpotato tolerance to drought stresses without causing any phenotypic imperfections but also a promising gene candidate for more extensive crop improvement.
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| 17. |
- Dölfors, Fredrik, et al.
(författare)
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A LysM effector protein from the basidiomycete Rhizoctonia solani contributes to virulence through suppression of chitin-triggered immunity
- 2019
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Ingår i: Molecular Genetics and Genomics. - : Springer Science and Business Media LLC. - 1617-4615 .- 1617-4623. ; 294, s. 1211-1218
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Tidskriftsartikel (refereegranskat)abstract
- Rhizoctonia solani is a fungal species that belongs to the fungal division Basidiomycota. It is a soil-borne pathogen that attacks a broad range of plant species and crops. Disease symptoms are commonly seen as damping off of seedlings and root rot, although it can infect plants at any developmental stage. Despite the severity of this disease, many aspects in R. solani infection biology remain unclear. Here we investigated the role of a LysM effector, previously predicted from the genome of a R. solani AG2-2IIIB strain that has sugar beet as a host. Gene expression analysis showed that RsLysM was highly induced upon sugar beet infection. When RsLysM was heterologously expressed in Cercospora beticola, necrotic lesion size and fungal colonization ability were increased, indicating a role in virulence. RsLysM displayed chitin-binding affinity and suppression of chitin-triggered immunity but could not protect hyphae from hydrolysis. Overall, this study is the first characterization of a LysM effector from Basidiomycota, suggesting that this necrotrophic fungal species relies on perturbation of chitin-triggered immunity to establish a successful infection.
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| 18. |
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| 19. |
- Dölfors, Fredrik, et al.
(författare)
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The RsRlpA Effector Is a Protease Inhibitor Promoting Rhizoctonia solani Virulence through Suppression of the Hypersensitive Response
- 2020
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Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 21
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Tidskriftsartikel (refereegranskat)abstract
- Rhizoctonia solani (Rs) is a soil-borne pathogen with a broad host range. This pathogen incites a wide range of disease symptoms. Knowledge regarding its infection process is fragmented, a typical feature for basidiomycetes. In this study, we aimed at identifying potential fungal effectors and their function. From a group of 11 predicted single gene effectors, a rare lipoprotein A (RsRlpA), from a strain attacking sugar beet was analyzed. The RsRlpA gene was highly induced upon early-stage infection of sugar beet seedlings, and heterologous expression in Cercospora beticola demonstrated involvement in virulence. It was also able to suppress the hypersensitive response (HR) induced by the Avr4/Cf4 complex in transgenic Nicotiana benthamiana plants and functioned as an active protease inhibitor able to suppress Reactive Oxygen Species (ROS) burst. This effector contains a double-psi beta-barrel (DPBB) fold domain, and a conserved serine at position 120 in the DPBB fold domain was found to be crucial for HR suppression. Overall, R. solani seems to be capable of inducing an initial biotrophic stage upon infection, suppressing basal immune responses, followed by a switch to necrotrophic growth. However, regulatory mechanisms between the different lifestyles are still unknown.
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| 20. |
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| 21. |
- Fahleson, Jan, et al.
(författare)
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Genetic analysis of Mycosphaerella fijiensis in the Ugandan Lake Victoria region
- 2009
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Ingår i: Plant Pathology. - : Wiley. - 0032-0862 .- 1365-3059. ; 58, s. 888-897
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Tidskriftsartikel (refereegranskat)abstract
- A collection of Mycosphaerella fijiensis, in total 71 isolates from 11 countries, of which 54 isolates were derived from the Ugandan Like Victoria region, were analysed with regard to Molecular markers and benomyl resistance to characterize the population structure. Forty-seven alleles from seven restriction fragment length polymorphism (PCR-RFLP) and five microsatellite markers were scored and utilized in the study. Using analysis of molecular variance (AMOVA), no significant genetic differentiation Could be found between isolates from different leaves compared to isolates from a single leaf (P = 0.07). Exact tests of the allelic frequencies revealed no differences between isolates from different districts or between single-leaf-derived isolates compared to isolates derived from different leaves (P > 0.05 in all cases except two, which, however, were shown to be of type-I error). Resistance to benomyl, analysed both by PCR and fungicide screening, revealed that all isolates were susceptible. Compiled information from the various datasets implied that the Ugandan M. fijiensis population in the Lake Victoria Basin constitutes a homogenous Population, probably as the result of a recent founder event.
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| 22. |
- Fogelqvist, Johan, et al.
(författare)
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Analysis of the hybrid genomes of two field isolates of the soil-borne fungal species Verticillium longisporum
- 2018
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Ingår i: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 19
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Tidskriftsartikel (refereegranskat)abstract
- Background: Brassica plant species are attacked by a number of pathogens; among them, the ones with a soilborne lifestyle have become increasingly important. Verticillium stem stripe caused by Verticillium longisporum is one example. This fungal species is thought to be of a hybrid origin, having a genome composed of combinations of lineages denominated A and D. In this study we report the draft genomes of 2 V. longisporum field isolates sequenced using the Illumina technology. Genomic characterization and lineage composition, followed by selected gene analysis to facilitate the comprehension of its genomic features and potential effector categories were performed.Results: The draft genomes of 2 Verticillium longisporum single spore isolates (VL1 and VL2) have an estimated ungapped size of about 70 Mb. The total number of protein encoding genes identified in VL1 was 20,793, whereas 21,072 gene models were predicted in VL2. The predicted genome size, gene contents, including the gene families coding for carbohydrate active enzymes were almost double the numbers found in V. dahliae and V. albo-atrum. Single nucleotide polymorphisms (SNPs) were frequently distributed in the two genomes but the distribution of heterozygosity and depth was not independent. Further analysis of potential parental lineages suggests that the V. longisporum genome is composed of two parts, A1 and D1, where A1 is more ancient than the parental lineage genome D1, the latter being more closer related to V. dahliae. Presence of the mating-type genes MAT1-1-1 and MAT1-2-1 in the V. longisporum genomes were confirmed. However, the MAT genes in V. dahliae, V. albo-atrum and V. longisporum have experienced extensive nucleotide changes at least partly explaining the present asexual nature of these fungal species.Conclusions: The established draft genome of V. longisporum is comparatively large compared to other studied ascomycete fungi. Consequently, high numbers of genes were predicted in the two V. longisporum genomes, among them many secreted proteins and carbohydrate active enzyme (CAZy) encoding genes. The genome is composed of two parts, where one lineage is more ancient than the part being more closely related to V. dahliae. Dissimilar mating-type sequences were identified indicating possible ancient hybridization events.
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| 23. |
- Fogelqvist, Johan, et al.
(författare)
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Genetic and morphological evidence for introgression between three species of willows
- 2015
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Ingår i: BMC Evolutionary Biology. - : Springer Science and Business Media LLC. - 1471-2148. ; 15
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Tidskriftsartikel (refereegranskat)abstract
- Background: Hybridization and introgression are said to occur relatively frequently in plants, and in particular among different species of willows. However, data on the actual frequency of natural hybridization and introgression is rare. Here, we report the first fine-scale genetic analysis of a contact zone shared between the three basket willow species, Salix dasyclados, S. schwerinii and S. viminalis in the vicinity of the Lake Baikal in Southern Siberia. Individuals were sampled in fourteen populations and classified as pure species or hybrids based on a set of morphological characters. They were then genotyped at 384 nuclear SNP and four chloroplast SSR loci. The STRUCTURE and NewHybrids softwares were used to estimate the frequency and direction of hybridization using genotypic data at the nuclear SNP loci. Results: As many as 19 % of the genotyped individuals were classified as introgressed individuals and these were mainly encountered in the centre of the contact zone. All introgressed individuals were backcrosses to S. viminalis or S. schwerinii and no F1 or F2 hybrids were found. The rest of the genotyped individuals were classified as pure species and formed two clusters, one with S. schwerinii individuals and the other with S. viminalis and S. dasyclados individuals. The two clusters were significantly genetically differentiated, with F-ST = 0.333 (0.282-0.382, p < 0.001). In contrast, for the chloroplast haplotypes, no genetic differentiation was observed as they were completely shared between the species. Based on morphological classification only 5 % of the individuals were classified as introgressed individuals, which was much less than what was detected using genotypic data. Conclusions: We have discovered a new willow hybrid zone with relatively high frequency of introgressed individuals. The low frequency of F1 hybrids indicates that ongoing hybridization is limited, which could be because of the presence of reproductive barriers or simply because the conditions are not favorable for hybridization. We further conclude that in order to get a complete picture of the species composition of a hybrid zone it is necessary to use a combination of morphological characters and genetic data from both nuclear and chloroplast markers.
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| 24. |
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| 25. |
- Hu, Xinyi, et al.
(författare)
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Phytophthora infestans Ago1-associated miRNA promotes potato late blight disease
- 2022
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Ingår i: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 233, s. 443-457
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Tidskriftsartikel (refereegranskat)abstract
- Phytophthora spp. cause serious damage to plants by exploiting a large number of effector proteins and small RNAs (sRNAs). Several reports have described modulation of host RNA biogenesis and defence gene expression. Here, we analysed Phytophthora infestans Argonaute (Ago) 1 associated small RNAs during potato leaf infection. Small RNAs were co-immunoprecipitated, deep sequenced and analysed against the P. infestans and potato genomes, followed by transcript analyses and transgenic assays on a predicted target. Extensive targeting of potato and pathogen-derived sRNAs to a range of mRNAs was observed, including 638 sequences coding for resistance (R) proteins in the host genome. The single miRNA encoded by P. infestans (miR8788) was found to target a potato alpha/beta hydrolase-type encoding gene (StABH1), a protein localized to the plasma membrane. Analyses of stable transgenic potato lines harbouring overexpressed StABH1 or artificial miRNA gene constructs demonstrated the importance of StABH1 during infection by P. infestans. miR8788 knock-down strains showed reduced growth on potato, and elevated StABH1 expression levels were observed when plants were inoculated with the two knock-down strains compared to the wild-type strain 88069. The findings of our study suggest that sRNA encoded by P. infestans can affect potato mRNA, thereby expanding our knowledge of the multifaceted strategies this species uses to facilitate infection.
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| 26. |
- Jahan, Sultana, et al.
(författare)
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Plant-mediated gene silencing restricts growth of the potato late blight pathogen Phytophthora infestans
- 2015
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Ingår i: Journal of Experimental Botany. - : Oxford University Press (OUP). - 0022-0957 .- 1460-2431. ; 66:9, s. 2785-2794
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Tidskriftsartikel (refereegranskat)abstract
- A host-induced gene-silencing strategy for controlling potato late blight is presented, a plant disease that conventionally requires regular application of fungicides at high rates.Phytophthora infestans is an oomycete that causes severe damage to potato, and is well known for its ability to evolve rapidly in order to overcome resistant potato varieties. An RNA silencing strategy was evaluated here to clarify if small interfering RNA homologous to selected genes in P. infestans could be targeted from the plant host to reduce the magnitude of the infection. As a proof-of-concept, a hairpin RNA (hp-RNA) construct using the GFP marker gene was designed and introduced in potato. At 72 hpi, a 55-fold reduction of the signal intensity of a corresponding GFP expressing P. infestans strain on leaf samples of transgenic plants, compared with wild-type potato, was detected. This suggests that an RNA interference construct in the potato host could be processed and target a transcript of the pathogen. Three genes important in the infection process of P. infestans, PiGPB1, PiCESA2, and PiPEC, together with PiGAPDH taking part in basic cell maintenance were subsequently tested using an analogous transgenic strategy. Out of these gene candidates, the hp-PiGPB1 targeting the G protein beta-subunit (PiGPB1) important for pathogenicity resulted in most restricted disease progress. Further, Illumina sequencing of inoculated transgenic potato leaves revealed sRNAs of 24/25 nt size homologous to the PiGPB1 gene in the transgenic plants indicating post-transcriptional silencing of the target gene. The work demonstrates that a host-induced gene-silencing approach is functional against P. infestans but is highly dependent on target gene for a successful outcome. This finding broadens the arsenal of control strategies to this important plant disease.
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| 27. |
- Jambagi, Shridhar, et al.
(författare)
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A robust hydroponic-based system for screening red clover (Trifolium pratense) for Fusarium avenaceum
- 2023
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Ingår i: Legume Science. - 2639-6181. ; 5
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Tidskriftsartikel (refereegranskat)abstract
- Red clover (Trifolium pratense) is an important forage legume crop that suffers likemost perennial crops from attacks by soil-borne pathogens.Fusariumroot rot is oneof the most serious diseases and at the same time problematic to identify resistancebecause of its hidden life in the soil. Current screening methods are laborious andhampered by limited reproducibility. To remedy this situation, we aimed to establisha simple and reliable hydroponics-based screening system to facilitate studies of redclover–Fusarium avenaceuminteractions. First, the fungal spore concentrations werebalanced toward the development of red clover plants grown hydroponically. Wefound that the optimum concentration was 30,000 spores in 2 L of hydroponicmedium to ensure infection during the plant growth period in this system. The proce-dure was scaled-up to screen plants from 25 populations to identify red clover indi-viduals with the improved resistance toF. avenaceum. Susceptible plants hadapproximately two-fold higher amounts of fungal DNA than resistant plants, demon-strating a correlation between the disease readings of the plants and pathogen DNA.We foresee this screening procedure meeting the needs of both applied breedingwork and in-depth molecular studies of responses between this pathogen and itshost plant. This method could be applied for the screening of other plant species forresistance toFusariumspp. or to other root microbes.
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| 28. |
- Jambagi, Shridhar, et al.
(författare)
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Red clover root-associated microbiota is shaped by geographic location and choice of farming system
- 2023
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Ingår i: Journal of Applied Microbiology. - 1364-5072 .- 1365-2672. ; 134
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Tidskriftsartikel (refereegranskat)abstract
- Aims: This study evaluated the red clover (Trifolium pratense) root-associated microbiota to clarify the presence of pathogenic and beneficial microorganisms in 89 Swedish field sites.Methods and results: 16S rRNA and ITS amplicon sequencing analysis were performed on DNA extracted from the red clover root samples collected to determine the composition of the prokaryotic and eukaryotic root-associated microbe communities. Alpha and beta diversities were calculated and relative abundance of various microbial taxa and their co-occurrence were analyzed. Rhizobium was the most prevalent bacterial genus, followed by Sphingomonas, Mucilaginibacter, Flavobacterium, and the unclassified Chloroflexi group KD4-96. The Leptodontidium, Cladosporium, Clonostachys, and Tetracladium fungal genera known for endophytic, saprotrophic, and mycoparasitic lifestyles were also frequently observed in all samples. Sixty-two potential pathogenic fungi were identified with a bias toward grass pathogens and a higher abundance in samples from conventional farms.Conclusions: We showed that the microbial community was mainly shaped by geographic location and management procedures. Co-occurrence networks revealed that the Rhizobiumleguminosarum bv. trifolii was negatively associated with all fungal pathogenic taxa recognized in this study.
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| 29. |
- Kuusk, Anna-Karin, et al.
(författare)
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Torröta
- 2010
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Ingår i: Faktablad Om Växtskydd, Jordbruk. - 1100-5025.
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Annan publikation (populärvet., debatt m.m.)
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| 30. |
- Liao, Zhen, 1983-
(författare)
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A small amoeba at the crossroads of the big RNAi world : MicroRNA biogenesis and Argonaute function in Dictyostelium discoideum
- 2018
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Small non-coding RNA (ncRNA) mediated gene silencing, known as RNAi, is a key regulatory mechanism of gene expression in eukaryotes. MicroRNAs (miRNA), one major type of small ncRNAs, are about 21nt long and bound by Argonaute proteins. This RNA-protein complex, called RISC, silences post-transcriptionally target mRNAs containing partial or full complementary sequence to the miRNA. MiRNAs are generated from step-wise endonucleolytic cleavages of long primary transcripts (pri-miRNAs) by RNase III nucleases. Biogenesis of miRNAs differs between uni- and multicellular eukaryotes, and also between plants and animals. In this thesis, I aimed to understand miRNA maturation in the social amoeba Dictyostelium discoideum, which stands at the crossroads between these phylogenetically distant groups. We showed that Dicer-like protein DrnB is essential for global maturation of D. discoideum miRNAs. The study of two pri-miRNAs revealed the conserved 5’ m7G-cap structures, but different 3’end formation from each other, and also from canonical miRNAs in plants and animals. In agreement with its evolutionary position, D. discoideum miRNA biogenesis showed unique and also shared features with both life groups.D. discoideum grows as a unicellular organism, but can switch to a multicellular development upon starvation. Most miRNAs, many other small ncRNAs, and Argonaute proteins, the core effectors of the RISC, are differentially expressed during development, indicative of a crucial role of RNAi mediated regulation throughout D. discoideum life cycle. Among the five Argonaute homologs in D. discoideum, I investigated the functions of three members, e.g. AgnB, C and E. Judging from their subcellular localization, the phenotypic consequences and transcriptional alteration resulting from single Argonaute gene deletion, our results suggested different roles of AgnB, C and E. Possibly AgnB associates with miRNAs and regulates gene expression post-transcriptionally; while AgnC seems to be involved in nuclear RNAi. Finally, the cytoplasmic AgnE inhibits D. discoideum cell growth and regulates developmental timing via an unknown mechanism.My thesis work expands our knowledge on D. discoideum RNAi with focuses on miRNA biogenesis and potential function of Argonaute proteins and, all together, sheds lights on the evolution of miRNA and RNAi.
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| 31. |
- Liao, Zhen, et al.
(författare)
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Genome-wide identification of Argonautes in Solanaceae with emphasis on potato
- 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
- Regulatory small RNAs (sRNAs) play important roles in many fundamental processes in plant biology such as development, fertilization and stress responses. The AGO protein family has here a central importance in gene regulation based on their capacity to associate with sRNAs followed by mRNA targeting in a sequence-complementary manner. The present study explored Argonautes (AGOs) in the Solanaceae family, with emphasis on potato, Solanum tuberosum (St). A genome-wide monitoring was performed to provide a deeper insight into gene families, genomic localization, gene structure and expression profile against the potato late blight pathogen Phytophthora infestans. Among 15 species in the Solanaceae family we found a variation from ten AGOs in Nicotiana obtusifolia to 17 in N. tabacum. Comprehensive analyses of AGO phylogeny revealed duplication of AGO1, AGO10 and AGO4 paralogs during early radiation of Solanaceae. Fourteen AGOs were identified in potato. Orthologs of AGO8 and AGO9 were missing in the potato genome. However, AGO15 earlier annotated in tomato was identified. StAGO15 differs from the other paralogs having residues of different physico-chemical properties at functionally important amino acid positions. Upon pathogen challenge StAGO15 was significantly activated and hence may play a prominent role in sRNA-based regulation of potato defense.
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| 32. |
- Liao, Zhen, et al.
(författare)
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Small talk and large impact: the importance of small RNA molecules in the fight of plant diseases
- 2021
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Ingår i: RNAi for plant improvement and protection. - UK : CABI. - 9781789248890
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- This short and general chapter summarizes how plants and pathogens communicate using not only proteins for recognition and signal transduction or other metabolites but also RNA molecules where small RNAs with sizes between 21 to 40 nt are most important. These small RNAs can move between plants and a range of interacting pathogenic organisms in both directions, that is, a 'cross-kingdom' communication process. The first reports on RNA-based communications between plants and plant pathogenic fungi appeared about 10 years ago. Since that time, we have learnt much about sRNA biology in plants and their function in different parasitic organisms. However, many questions on the processes involved remain unanswered. Such information is crucial in order to sustain high crop production. Besides giving a brief background, we highlight the interactions between the potato late blight pathogen and its plant host potato.
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| 33. |
- Martin, Thomas, et al.
(författare)
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A highly conserved NB-LRR encoding gene cluster effective against Setosphaeria turcica in sorghum
- 2011
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Ingår i: BMC Plant Biology. - : Springer Science and Business Media LLC. - 1471-2229. ; 11
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Tidskriftsartikel (refereegranskat)abstract
- Conclusions: Resistance genes to S. turcica, with a CC-NB-LRR protein domain architecture, have been found in maize and sorghum. VIGS analysis revealed their importance in the surveillance to S. turcica in sorghum. The St genes are highly conserved in sorghum, rice, foxtail millet, maize and Brachypodium, suggesting an essential evolutionary function.
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| 34. |
- Martin, Thomas, et al.
(författare)
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A Network of HMG-box Transcription Factors Regulates Sexual Cycle in the Fungus Podospora anserina
- 2013
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Ingår i: PLoS Genetics. - : Public Library of Science (PLoS). - 1553-7390 .- 1553-7404. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- High-mobility group (HMG) B proteins are eukaryotic DNA-binding proteins characterized by the HMG-box functional motif. These transcription factors play a pivotal role in global genomic functions and in the control of genes involved in specific developmental or metabolic pathways. The filamentous ascomycete Podospora anserina contains 12 HMG-box genes. Of these, four have been previously characterized; three are mating-type genes that control fertilization and development of the fruit-body, whereas the last one encodes a factor involved in mitochondrial DNA stability. Systematic deletion analysis of the eight remaining uncharacterized HMG-box genes indicated that none were essential for viability, but that seven were involved in the sexual cycle. Two HMG-box genes display striking features. PaHMG5, an ortholog of SpSte11 from Schizosaccharomyces pombe, is a pivotal activator of mating-type genes in P. anserina, whereas PaHMG9 is a repressor of several phenomena specific to the stationary phase, most notably hyphal anastomoses. Transcriptional analyses of HMG-box genes in HMG-box deletion strains indicated that PaHMG5 is at the hub of a network of several HMG-box factors that regulate mating-type genes and mating-type target genes. Genetic analyses revealed that this network also controls fertility genes that are not regulated by mating-type transcription factors. This study points to the critical role of HMG-box members in sexual reproduction in fungi, as 11 out of 12 members were involved in the sexual cycle in P. anserina. PaHMG5 and SpSte11 are conserved transcriptional regulators of mating-type genes, although P. anserina and S. pombe diverged 550 million years ago. Two HMG-box genes, SOX9 and its upstream regulator SRY, also play an important role in sex determination in mammals. The P. anserina and S. pombe mating-type genes and their upstream regulatory factor form a module of HMG-box genes analogous to the SRY/SOX9 module, revealing a commonality of sex regulation in animals and fungi.
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| 35. |
- Martin, Thomas, et al.
(författare)
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Disease severity, incidence and races of Setosphaeria turcica on sorghum in Uganda
- 2011
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Ingår i: European Journal of Plant Pathology. - : Springer Science and Business Media LLC. - 0929-1873 .- 1573-8469. ; 131, s. 383-392
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Tidskriftsartikel (refereegranskat)abstract
- In order to understand the underlaying causes of new severe turcicum leaf blight outbreaks in East Africa, a survey was undertaken in Uganda to examine the sorghum-Setosphaeria turcica interaction in terms of disease severity and incidence, the overall fungal population structure, and new resistant resources. Highest disease severities were recorded on caudatum accessions, whereas kafir genotypes were most resistant. The disease was more severe in the most humid farmlands compared to moderately dry agro-ecologies. In districts with wide adoption of the Epuripur variety a very high incidence (100%) of turcicum leaf blight was found. The two S. turcica mating type genes MAT1-1 and MAT1-2 assessed on fungal isolates deriving from both sorghum and maize diseased leaves were found in 20 of 23 districts sampled and in equal proportions. Upon cross inoculation on maize differential lines, four S. turcica isolates were identified as race 1, two as race 2, and one isolate corresponded to race 0 and race 3, respectively. The remaining 10 S. turcica isolates did not cause any disease symptoms on the maize lines assessed. Highly resistant accessions originating from a regional collection were found among the five sorghum races (kafir, guinea, caudatum, bicolor and durra), and are now implemented in new sorghum disease resistance programs.
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| 36. |
- Martin, Thomas, et al.
(författare)
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Tracing the Origin of the Fungal alpha 1 Domain Places Its Ancestor in the HMG-Box Superfamily: Implication for Fungal Mating-Type Evolution
- 2010
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 5
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Tidskriftsartikel (refereegranskat)abstract
- Background: Fungal mating types in self-incompatible Pezizomycotina are specified by one of two alternate sequences occupying the same locus on corresponding chromosomes. One sequence is characterized by a gene encoding an HMG protein, while the hallmark of the other is a gene encoding a protein with an alpha 1 domain showing similarity to the M protein of Saccharomyces cerevisiae. DNA-binding HMG proteins are ubiquitous and well characterized. In contrast, alpha 1 domain proteins have limited distribution and their evolutionary origin is obscure, precluding a complete understanding of mating-type evolution in Ascomycota. Although much work has focused on the role of the S. cerevisiae Mat alpha 1p protein as a transcription factor, it has not yet been placed in any of the large families of sequence-specific DNA-binding proteins.Methodology/Principal Findings: We present sequence comparisons, phylogenetic analyses, and in silico predictions of secondary and tertiary structures, which support our hypothesis that the alpha 1 domain is related to the HMG domain. We have also characterized a new conserved motif in alpha 1 proteins of Pezizomycotina. This motif is immediately adjacent to and downstream of the alpha 1 domain and consists of a core sequence Y-[LMIF]-x(3)-G-[WL] embedded in a larger conserved motif.Conclusions/Significance: Our data suggest that extant alpha 1-box genes originated from an ancestral HMG gene, which confirms the current model of mating-type evolution within the fungal kingdom. We propose to incorporate alpha 1 proteins in a new subclass of HMG proteins termed MAT alpha_ HMG.
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| 37. |
- Martin, Thomas, et al.
(författare)
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Two loci in sorghum with NB-LRR encoding genes confer resistance to Colletotrichum sublineolum
- 2012
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Ingår i: TAG Theoretical and Applied Genetics. - : Springer Science and Business Media LLC. - 0040-5752 .- 1432-2242. ; 124, s. 1005-1015
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Tidskriftsartikel (refereegranskat)abstract
- The aim of this work was to identify plant resistance genes to the sorghum anthracnose fungus Colletotrichum sublineolum. cDNA-AFLP transcript profiling on two contrasting sorghum genotypes inoculated with C. sublineolum generated about 3,000 informative fragments. In a final set of 126 sequenced genes, 15 were identified as biotic stress related. Seven of the plant-derived genes were selected for functional analysis using a Brome mosaic virus-based virus-induced gene silencing (VIGS) system followed by fungal inoculation and quantitative real-time PCR analysis. The candidate set comprised genes encoding resistance proteins (Cs1A, Cs2A), a lipid transfer protein (SbLTP1), a zinc finger-like transcription factor (SbZnTF1), a rice defensin-like homolog (SbDEFL1), a cell death related protein (SbCDL1), and an unknown gene harboring a casein kinase 2-like domain (SbCK2). Our results demonstrate that down-regulation of Cs1A, Cs2A, SbLTP1, SbZnF1 and SbCD1 via VIGS, significantly compromised the resistance response while milder effects were observed with SbDEFL1 and SbCK2. Expanded genome analysis revealed that Cs1A and Cs2A genes are located in two different loci on chromosome 9 closely linked with duplicated genes Cs1B and Cs2B, respectively. The nucleotide binding-leucine rich repeat (NB-LRR) encoding Cs gene sequence information is presently employed in regional breeding programs.
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| 38. |
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| 39. |
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| 40. |
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| 41. |
- Peele, Hanneke Marjolijn, et al.
(författare)
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Loss and retention of resistance genes in five species of the Brassicaceae family
- 2014
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Ingår i: BMC Plant Biology. - : Springer Science and Business Media LLC. - 1471-2229. ; 14, s. 1-11
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Tidskriftsartikel (refereegranskat)abstract
- Background: Plants have evolved disease resistance (R) genes encoding for nucleotide-binding site (NB) and leucine-rich repeat (LRR) proteins with N-terminals represented by either Toll/Interleukin-1 receptor (TIR) or coiled-coil (CC) domains. Here, a genome-wide study of presence and diversification of CC-NB-LRR and TIR-NB-LRR encoding genes, and shorter domain combinations in 19 Arabidopsis thaliana accessions and Arabidopsis lyrata, Capsella rubella, Brassica rapa and Eutrema salsugineum are presented.Results: Out of 528 R genes analyzed, 12 CC-NB-LRR and 17 TIR-NB-LRR genes were conserved among the 19 A. thaliana genotypes, while only two CC-NB-LRRs, including ZAR1, and three TIR-NB-LRRs were conserved when comparing the five species. The RESISTANCE TO LEPTOSPHAERIA MACULANS 1 (RLM1) locus confers resistance to the Brassica pathogen L. maculans the causal agent of blackleg disease and has undergone conservation and diversification events particularly in B. rapa. On the contrary, the RLM3 locus important in the immune response towards Botrytis cinerea and Alternaria spp. has recently evolved in the Arabidopsis genus.Conclusion: Our genome-wide analysis of the R gene repertoire revealed a large sequence variation in the 23 cruciferous genomes. The data provides further insights into evolutionary processes impacting this important gene family.
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| 42. |
- Persson Hodén, Kristian, et al.
(författare)
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smartPARE: An R Package for Efficient Identification of True mRNA Cleavage Sites
- 2021
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Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 22
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Tidskriftsartikel (refereegranskat)abstract
- Degradome sequencing is commonly used to generate high-throughput information on mRNA cleavage sites mediated by small RNAs (sRNA). In our datasets of potato (Solanum tuberosum, St) and Phytophthora infestans (Pi), initial predictions generated high numbers of cleavage site predictions, which highlighted the need of improved analytic tools. Here, we present an R package based on a deep learning convolutional neural network (CNN) in a machine learning environment to optimize discrimination of false from true cleavage sites. When applying smartPARE to our datasets on potato during the infection process by the late blight pathogen, 7.3% of all cleavage windows represented true cleavages distributed on 214 sites in P. infestans and 444 sites in potato. The sRNA landscape of the two organisms is complex with uneven sRNA production and cleavage regions widespread in the two genomes. Multiple targets and several cases of complex regulatory cascades, particularly in potato, was revealed. We conclude that our new analytic approach is useful for anyone working on complex biological systems and with the interest of identifying cleavage sites particularly inferred by sRNA classes beyond miRNAs.
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| 43. |
- Persson, Mattias, et al.
(författare)
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Layers of defense responses to Leptosphaeria maculans below the RLM1- and camalexin-dependent resistances
- 2009
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Ingår i: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 182, s. 470-482
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Tidskriftsartikel (refereegranskat)abstract
- Plants have evolved different defense components to counteract pathogen attacks. The resistance locus resistance to Leptosphaeria maculans 1 (RLM1) is a key factor for Arabidopsis thaliana resistance to L. maculans. The present work aimed to reveal downstream defense responses regulated by RLM1.Quantitative assessment of fungal colonization in the host was carried out using quantitative polymerase chain reaction (qPCR) and GUS expression analyses, to further characterize RLM1 resistance and the role of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) in disease development. Additional assessments of A. thaliana mutants were performed to expand our understanding of this pathosystem.Resistance responses such as lignification and the formation of vascular plugs were found to occur in an RLM1-dependent manner, in contrast to the RLM1-independent increase in reactive oxygen species at the stomata and hydathodes. Analyses of mutants defective in hormone signaling in the camalexin-free rlm1(Ler)pad3 background revealed a significant influence of JA and ET on symptom development and pathogen colonization.The overall results indicate that the defense responses of primary importance induced by RLM1 are all associated with physical barriers, and that responses of secondary importance involve complex cross-talk among SA, JA and ET. Our observations further suggest that ET positively affects fungal colonization.
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| 44. |
- Persson, Mattias, et al.
(författare)
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Studies on the mechanism of resistance to Bipolaris sorokiniana in the barley lesion mimic mutant bst1
- 2009
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Ingår i: Molecular Plant Pathology. - 1464-6722 .- 1364-3703. ; 10, s. 587-598
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Tidskriftsartikel (refereegranskat)abstract
- P>The Bipolaris sorokiniana tolerant 1 (bst1) barley mutant is derived from fast neutron-irradiated seeds of wild-type Bowman(Rph3). The induced mutation was genetically localized to a position on chromosome 5HL distal to the centromere using amplified fragment length polymorphism markers. In addition, the defence responses and related gene expression in the bst1 mutant after fungal challenge were compared with those occurring in wild-type plants. Hydrogen peroxide generation, determined by 3,3-diaminobenzidine staining, revealed a clearly reduced level of bst1, compared with the wild-type, during the entire experimental time: 8-120 h post-inoculation (hpi). At 48 hpi, the wild-type samples displayed twice as much fungal mass and three times greater H(2)O(2) production than bst1. At the same time, staining of B. sorokiniana showed less fungal growth in the spontaneous lesions of bst1 compared with the wild-type. Monitoring of defence-related genes at 48 hpi demonstrated strong expression of PR-1a, PR-2, PR-5 and PR-10 in bst1. A gene coding for a unique oxidoreductase enzyme, designated as HCP1, was expressed at much higher levels in inoculated leaves of the bst1 mutant than in those of the wild-type plant. Taken together, the results suggest that the defence to B. sorokiniana largely relies on salicylic acid-responsive pathogenesis-related (PR) genes, as well as selected reactive oxygen species and unknown HCP1-associated factors.
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| 45. |
- Rafieebanadaki, Vahidehalsadat, et al.
(författare)
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Advances in molecular interactions on the Rhizoctonia solani-sugar beet pathosystem
- 2023
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Ingår i: Fungal Biology Reviews. - : Elsevier BV. - 1749-4613 .- 1878-0253. ; 44
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Forskningsöversikt (refereegranskat)abstract
- Rhizoctonia solani is a soilborne pathogen with a broad host range. An anastomosis group (AG) system based on hyphal fusions has been established to distinguish between different R. solani subgroups in this species complex. Members of the AG2-2IIIB subgroup can cause serious problems in sugar beet production, resulting in Rhizoctonia root and crown rot. In this review, we summarize the current molecular advances in the R. solani sugar beet pa-thosystem. The draft genome of R. solani AG2-2IIIB has an estimated size of 56.02 Mb, larger than any of the R. solani AGs sequenced to date. The genome of AG2-2IIIB has been pre-dicted to harbor 11,897 protein-encoding genes, including a high number of carbohydrate-active enzymes (CAZymes). The highest number of CAZymes was observed for polysaccharide lyase family 1 (PL-1), glycoside hydrolase family 43 (GH-43), and carbo-hydrate esterase family 12 (CE-12). Eleven single-effector candidates were predicted based on AG2-2IIIB genome data. The RsLysM, RsRlpA, and RsCRP1 genes were highly induced upon early-stage infection of sugar beet seedlings, and heterologous expression in Cerco-spora beticola and model plant species demonstrated their involvement in virulence. How-ever, despite the progress achieved thus far on the molecular interactions in this pathosystem, many aspects remain to be elucidated, including the development of efficient transformation systems, important for functional studies, and the silencing of undesirable traits in the sugar beet crop.(c) 2022 The Author(s). Published by Elsevier Ltd on behalf of British Mycological Society. This is an open access article under the CC BY license (http://creativecommons.org/ licenses/by/4.0/).
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| 46. |
- Ramesh, Vetukuri, et al.
(författare)
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Can silencing of transposons contribute to variation in effector gene expression in Phytophthora infestans?
- 2012
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Ingår i: Landes Bioscience. - : Informa UK Limited. ; 2, s. 110-114
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Tidskriftsartikel (refereegranskat)abstract
- Transposable elements are ubiquitous residents in eukaryotic genomes. Often considered to be genomic parasites, they can lead to dramatic changes in genome organization, gene expression, and gene evolution. The oomycete plant pathogen Phytophthora infestans has evolved a genome organization where core biology genes are predominantly located in genome regions that have relatively few resident transposons. In contrast, disease effector-encoding genes are most frequently located in rapidly evolving genomic regions that are rich in transposons. P. infestans, as a eukaryote, likely uses RNA silencing to minimize the activity of transposons. We have shown that fusion of a short interspersed element (SINE) to an effector gene in P. infestans leads to the silencing of both the introduced fusion and endogenous homologous sequences. This is also likely to occur naturally in the genome of P. infestans, as transcriptional inactivation of effectors is known to occur, and over half of the translocated “RXLR class” of effectors are located within 2 kb of transposon sequences in the P. infestans genome. In this commentary, we review the diverse transposon inventory of P. infestans, its control by RNA silencing, and consequences for expression modulation of nearby effector genes in this economically important plant pathogen.
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| 47. |
- Ramesh, Vetukuri, et al.
(författare)
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Evidence for involvement of Dicer-like, Argonaute and histone deacetylase proteins in gene silencing in Phytophthora infestans
- 2011
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Ingår i: Molecular Plant Pathology. - 1464-6722 .- 1364-3703. ; 12, s. 772-785
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Tidskriftsartikel (refereegranskat)abstract
- Gene silencing may have a direct or indirect impact on many biological processes in eukaryotic cells, and is a useful tool for the determination of the roles of specific genes. In this article, we report silencing in Phytophthora infestans, an oomycete pathogen of potato and tomato. Gene silencing is known to occur in P. infestans, but its genetic basis has yet to be determined. Genes encoding the major components of the RNA interference (RNAi) pathway, Dicer-like (Pidcl1), Argonaute (Piago1-5) and RNA-directed RNA polymerase (Pirdr1), were identified in the P. infestans genome by comparative genomics, together with families of other genes potentially involved in gene silencing, such as histone deacetylases, histone methyltransferases, DEAD heli-cases, chromodomain proteins and a class 1 RNaseIII. Real-time reverse transcription-polymerase chain reaction demonstrated transcript accumulation for all candidate genes throughout the asexual lifecycle and plant infection, but at different levels of mRNA abundance. A functional assay was developed in which silencing of the sporulation-associated Picdc14 gene was released by the treatment of protoplasts with in vitro-synthesized double-stranded RNAs homologous to Pidcl1, Piago1/2 and histone deacetylase Pihda1. These results suggest that the components of gene silencing, namely Dicer-like, Argonaute and histone deacetylase, are functional in P. infestans. Our data demonstrate that this oomycete possesses canonical gene silencing pathways similar to those of other eukaryotes.
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| 48. |
- Ramesh, Vetukuri, et al.
(författare)
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Evidence for Small RNAs Homologous to Effector-Encoding Genes and Transposable Elements in the Oomycete Phytophthora infestans
- 2012
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 7:12, s. e51399-
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Tidskriftsartikel (refereegranskat)abstract
- Phytophthora infestans is the oomycete pathogen responsible for the devastating late blight disease on potato and tomato. There is presently an intense research focus on the role(s) of effectors in promoting late blight disease development. However, little is known about how they are regulated, or how diversity in their expression may be generated among different isolates. Here we present data from investigation of RNA silencing processes, characterized by non-coding small RNA molecules (sRNA) of 19-40 nt. From deep sequencing of sRNAs we have identified sRNAs matching numerous RxLR and Crinkler (CRN) effector protein genes in two isolates differing in pathogenicity. Effector gene-derived sRNAs were present in both isolates, but exhibited marked differences in abundance, especially for CRN effectors. Small RNAs in P. infestans grouped into three clear size classes of 21, 25/26 and 32 nt. Small RNAs from all size classes mapped to RxLR effector genes, but notably 21 nt sRNAs were the predominant size class mapping to CRN effector genes. Some effector genes, such as PiAvr3a, to which sRNAs were found, also exhibited differences in transcript accumulation between the two isolates. The P. infestans genome is rich in transposable elements, and the majority of sRNAs of all size classes mapped to these sequences, predominantly to long terminal repeat (LTR) retrotransposons. RNA silencing of Dicer and Argonaute genes provided evidence that generation of 21 nt sRNAs is Dicer-dependent, while accumulation of longer sRNAs was impacted by silencing of Argonaute genes. Additionally, we identified six microRNA (miRNA) candidates from our sequencing data, their precursor sequences from the genome sequence, and target mRNAs. These miRNA candidates have features characteristic of both plant and metazoan miRNAs.
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| 49. |
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| 50. |
- Ramesh, Vetukuri, et al.
(författare)
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Phytophthora infestans effector AVR3a is essential for virulence and manipulates plant immunity by stabilizing host E3 ligase CMPG1
- 2010
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Ingår i: Proceedings of the National Academy of Sciences. - : Proceedings of the National Academy of Sciences. - 1091-6490 .- 0027-8424. ; 107, s. 9909-9914
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Tidskriftsartikel (refereegranskat)abstract
- Fungal and oomycete plant pathogens translocate effector proteins into host cells to establish infection. However, virulence targets and modes of action of their effectors are unknown. Effector AVR3a from potato blight pathogen Phytophthora infestans is translocated into host cells and occurs in two forms: AVR3a(KI), which is detected by potato resistance protein R3a, strongly suppresses infestin 1 (INF1)-triggered cell death (ICD), whereas AVR3a(EM), which evades recognition by R3a, weakly suppresses host ICD. Here we show that AVR3a interacts with and stabilizes host U-box E3 ligase CMPG1, which is required for ICD. In contrast, AVR3a(KI/Y147del), a mutant with a deleted C-terminal tyrosine residue that fails to suppress ICD, cannot interact with or stabilize CMPG1. CMPG1 is stabilized by the inhibitors MG132 and epoxomicin, indicating that it is degraded by the 26S proteasome. CMPG1 is degraded during ICD. However, it is stabilized by mutations in the U-box that prevent its E3 ligase activity. In stabilizing CMPG1, AVR3a thus modifies its normal activity. Remarkably, given the potential for hundreds of effector genes in the P. infestans genome, silencing Avr3a compromises P. infestans pathogenicity, suggesting that AVR3a is essential for virulence. Interestingly, Avr3a silencing can be complemented by in planta expression of Avr3a(KI) or Avr3a(EM) but not the Avr3a(KI/Y147del) mutant. Our data provide genetic evidence that AVR3a is an essential virulence factor that targets and stabilizes the plant E3 ligase CMPG1, potentially to prevent host cell death during the biotrophic phase of infection.
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