| 1. |
- Andersson, Mats X., 1977, et al.
(författare)
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Involvement of the Electrophilic Isothiocyanate Sulforaphane in Arabidopsis Local Defense Responses
- 2015
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Ingår i: Plant Physiology. - : Oxford University Press (OUP). - 0032-0889 .- 1532-2548. ; 167:1, s. 251-
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Tidskriftsartikel (refereegranskat)abstract
- Plants defend themselves against microbial pathogens through a range of highly sophisticated and integrated molecular systems. Recognition of pathogen-secreted effector proteins often triggers the hypersensitive response (HR), a complex multicellular defense reaction where programmed cell death of cells surrounding the primary site of infection is a prominent feature. Even though the HR was described almost a century ago, cell-to-cell factors acting at the local level generating the full defense reaction have remained obscure. In this study, we sought to identify diffusible molecules produced during the HR that could induce cell death in naive tissue. We found that 4-methylsulfinylbutyl isothiocyanate (sulforaphane) is released by Arabidopsis (Arabidopsis thaliana) leaf tissue undergoing the HR and that this compound induces cell death as well as primes defense in naive tissue. Two different mutants impaired in the pathogen-induced accumulation of sulforaphane displayed attenuated programmed cell death upon bacterial and oomycete effector recognition as well as decreased resistance to several isolates of the plant pathogen Hyaloperonospora arabidopsidis. Treatment with sulforaphane provided protection against a virulent H. arabidopsidis isolate. Glucosinolate breakdown products are recognized as antifeeding compounds toward insects and recently also as intracellular signaling and bacteriostatic molecules in Arabidopsis. The data presented here indicate that these compounds also trigger local defense responses in Arabidopsis tissue.
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| 2. |
- Herdean, Andrei, 1984, et al.
(författare)
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A voltage-dependent chloride channel fine-tunes photosynthesis in plants
- 2016
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Ingår i: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 7:artikel nr 11654
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Tidskriftsartikel (refereegranskat)abstract
- n natural habitats, plants frequently experience rapid changes in the intensity of sunlight. To cope with these changes and maximize growth, plants adjust photosynthetic light utilization in electron transport and photoprotective mechanisms. This involves a proton motive force (PMF) across the thylakoid membrane, postulated to be affected by unknown anion (Cl−) channels. Here we report that a bestrophin-like protein from Arabidopsis thaliana functions as a voltage-dependent Cl− channel in electrophysiological experiments. AtVCCN1 localizes to the thylakoid membrane, and fine-tunes PMF by anion influx into the lumen during illumination, adjusting electron transport and the photoprotective mechanisms. The activity of AtVCCN1 accelerates the activation of photoprotective mechanisms on sudden shifts to high light. Our results reveal that AtVCCN1, a member of a conserved anion channel family, acts as an early component in the rapid adjustment of photosynthesis in variable light environments.
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| 3. |
- Fahlberg, Per, et al.
(författare)
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Involvement of lipid transfer proteins in resistance against a non-host powdery mildew in Arabidopsis thaliana
- 2019
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Ingår i: Molecular Plant Pathology. - : Wiley. - 1464-6722 .- 1364-3703. ; 20:1, s. 69-77
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Tidskriftsartikel (refereegranskat)abstract
- Non-specific lipid transfer proteins (LTPs) are involved in the transport of lipophilic compounds to the cuticular surface in epidermal cells and in the defence against pathogens. The role of glycophosphatidylinositol (GPI)-anchored LTPs (LTPGs) in resistance against non-host mildews in Arabidopsis thaliana was investigated using reverse genetics. Loss of either LTPG1, LTPG2, LTPG5 or LTPG6 increased the susceptibility to penetration of the epidermal cell wall by Blumeria graminis f. sp. hordei (Bgh). However, no impact on pre-penetration defence against another non-host mildew, Erysiphe pisi (Ep), was observed. LTPG1 was localized to papillae at the sites of Bgh penetration. This study shows that, in addition to the previously known functions, LTPGs contribute to pre-invasive defence against certain non-host powdery mildew pathogens.
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| 4. |
- Johansson, Oskar N., 1984, et al.
(författare)
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A quick and robust method for quantification of the hypersensitive response in plants
- 2015
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Ingår i: Peerj. - : PeerJ. - 2167-8359. ; 3
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Tidskriftsartikel (refereegranskat)abstract
- One of the most studied defense reactions of plants against microbial pathogens is the hypersensitive response (HR). The HR is a complex multicellular process that involves programmed cell death at the site of infection. A standard method to quantify plant defense and the HR is to measure the release of cellular electrolytes into water after infiltration with pathogenic bacteria. In this type of experiment, the bacteria are typically delivered into the plant tissue through syringe infiltration. Here we report the development of a vacuum infiltration protocol that allows multiple plant lines to be infiltrated simultaneously and assayed for defense responses. Vacuum infiltration did not induce more wounding response in Arabidopsis leaf tissue than syringe inoculation, whereas throughput and reproducibility were improved. The method was used to study HR-induced electrolyte loss after treatment with the bacterium Pseudomonas syringae pv. tomato DC3000 harboring the effector AvrRpm1, AvrRpt2 or AvrRps4. Specifically, the influence of bacterial titer on AvrRpm1-induced HR was investigated. Not only the amplitude, but also the timing of the maximum rate of the HR reaction was found to be dose-dependent. Finally, using vacuum infiltration, we were able quantify induction of phospholipase D activity after AvrRpm1 recognition in leaves labeled with (PO4)-P-33.
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| 5. |
- Johansson, Oskar N., 1984, et al.
(författare)
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Friends With Benefits: Exploring the Phycosphere of the Marine Diatom Skeletonema marinoi
- 2019
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Ingår i: Frontiers in Microbiology. - : Frontiers Media SA. - 1664-302X. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- Marine diatoms are the dominant phytoplankton in the temperate oceans and coastal regions, contributing to global photosynthesis, biogeochemical cycling of key nutrients and minerals and aquatic food chains. Integral to the success of marine diatoms is a diverse array of bacterial species that closely interact within the diffusive boundary layer, or phycosphere, surrounding the diatom partner. Recently, we isolated seven distinct bacterial species from cultures of Skeletonema marinoi, a chain-forming, centric diatom that dominates the coastal regions of the temperate oceans. Genomes of all seven bacteria were sequenced revealing many unusual characteristics such as the existence of numerous plasmids of widely varying sizes. Here we have investigated the characteristics of the bacterial interactions with S. marinoi, demonstrating that several strains (Arenibacter algicola strain SMS7, Marinobacter salarius strain SMRS, Sphingorhabdus flavimaris strain SMR4y, Sulfitobacter pseudonitzschiae strain SMR1, Yoonia vestfoldensis strain SMR4r and Roseovarius mucosus strain SMR3) stimulate growth of the diatom partner. Testing of many different environmental factors including low iron concentration, high and low temperatures, and chemical signals showed variable effects on this growth enhancement by each bacterial species, with the most significant being light quality in which green and blue but not red light enhanced the stimulatory effect on S. marinoi growth by all bacteria. Several of the bacteria also inhibited growth of one or more of the other bacterial strains to different extents when mixed together. This study highlights the complex interactions between diatoms and their associated bacteria within the phycosphere, and that further studies are needed to resolve the underlying mechanisms for these relationships and how they might influence the global success of marine diatoms.
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| 6. |
- Johansson, Oskar N., 1984, et al.
(författare)
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Phenomics reveals a novel putative chloroplast fatty acid transporter in the marine diatom Skeletonema marinoi involved in temperature acclimation
- 2019
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Diatoms are the dominant phytoplankton in temperate oceans and coastal regions and yet little is known about the genetic basis underpinning their global success. Here, we address this challenge by developing the first phenomic approach for a diatom, screening a collection of randomly mutagenized but identifiably tagged transformants. Based upon their tolerance to temperature extremes, several compromised mutants were identified revealing genes either stress related or encoding hypothetical proteins of unknown function. We reveal one of these hypothetical proteins is a novel putative chloroplast fatty acid transporter whose loss affects several fatty acids including the two omega-3, long-chain polyunsaturated fatty acids - eicosapentaenoic and docosahexaenoic acid, both of which have medical importance as dietary supplements and industrial significance in aquaculture and biofuels. This mutant phenotype not only provides new insights into the fatty acid biosynthetic pathways in diatoms but also highlights the future value of phenomics for revealing specific gene functions in these ecologically important phytoplankton.
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| 7. |
- Johansson, Oskar N., 1984
(författare)
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Plant pathogen defense: Signalling, resistance and cell death
- 2015
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Pathogenic microorganisms are present everywhere in nature and infect both animals and plants. Phytopathogenic microorganisms cause diseases on plants, and are responsible for crop loss amounting in the order billions of dollars annually. Plants have however co-evolved with these organisms and have consecutively been forced to develop mechanisms that prevent disease. The plant immune system unlike that of animals lack adaptive cells and rely on the innate immunity of each plant cell. There is however no doubt in the effectiveness of the plant immune system as most plants are healthy most of the time. The plant immune system consists of two main tiers of defense responses; the MAMP triggered immunity (MTI) and the Effector triggered immunity (ETI). MTI is triggered by recognition of microbe associated molecular patterns MAMPs. MTI strengthens the cell by producing antimicrobial substances, proteins and by fortifying the cell wall. This stops the majority of non-adapted microbes. A subset of microbes have adapted to these measures and evolved effector proteins that subdue the MTI responses. Again, plants have responded, by evolving resistance (R) proteins that recognize effector activity and mount the swift responses that are ETI. The plant responses during ETI are commonly termed the hypersensitive response (HR) and culminate in programmed cell death of the infected and sometimes surrounding cells. The thesis has approached the plant disease resistance response in four ways. The first focused on improving methods for quantifying the programmed cell death response during ETI (Paper I) and lipid analysis by chromatography (Paper II). These methods are then used in the following pappers. The second part focused on signalling during the HR. Signalling on gene regulation level (Paper III) and various parts of lipid metabolism (Paper IV, V and VI) during the HR was pursued. The main results from these studies include the high redundancy identified among Arabidopsis thaliana phospholipase D isoforms in producing the lipid phosphatidic acid, the identification and initial characterization of the enzyme (AGAP1) that is responsible for producing head group acylation of lipids in A. thaliana and the reported involvement of a chloroplast localized 13-lipoxygenase in initiating the HR related programmed cell death in A. thaliana. The third part of the thesis proposes a role in the HR in A.thaliana for two reactive molecules; indole acetonitrile (Paper VII) and sulforaphane (Paper VIII). Both compounds induce cell death when infiltrated into leaves and studies using mutants suggest that absence of these compounds result in a reduced cell death response. A redox related mechanism for these compounds is suggested. The fourth and final part of the thesis aimed to investigate if novel components could be identified in post penetration response against powdery mildew funguses. Much less is known on the relative dependence of MTI and ETI of this system, the results from Paper IX suggest that besides the known involvement of the protein EDS1, additional components are present. In conclusion, this thesis contributes with insight into different aspects of how lipid-, redox- and hormone signalling contributes to resistance and cell death in plants.
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| 8. |
- Johansson, Oskar N., 1984, et al.
(författare)
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Redundancy among phospholipase D isoforrns in resistance triggered by recognition of the Pseudomonas syringae effector AyrRpm1 in Arabidopsis thaliana
- 2014
-
Ingår i: Frontiers in Plant Science. - : Frontiers Media SA. - 1664-462X. ; 5
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Tidskriftsartikel (refereegranskat)abstract
- Plants possess a highly sophisticated system for defense against microorganisms. So called MAMP (microbe-associated molecular patterns) triggered immunity (MTI) prevents the majority of non-adapted pathogens from causing disease. Adapted plant pathogens use secreted effector proteins to interfere with such signaling. Recognition of microbial effectors or their activity by plant resistance (R)-proteins triggers a second line of defense resulting in effector triggered immunity (ETI). The latter usually comprises the hypersensitive response (HR) which includes programmed cell death at the site of infection. Phospholipase D (PLD) mediated production of phosphatidic acid (PA) has been linked to both MTI and ETI in plants. Inhibition of PLD activity has been shown to attenuate MTI as well as ETI. In this study, we systematically tested single and double knockouts in all 12 genes encoding PLDs in Arabidopsis thaliana for effects on ETI and MTI. No single PLD could be linked to ETI triggered by recognition of effectors secreted by the bacterium Pseudomonas syringae. However, repression of PLD dependent PA production by n-butanol strongly inhibited the HR following Pseudomonas syringae effector recognition. In addition some p/d mutants were more sensitive to n-butanol than wild type. Thus, the effect of mutations of PLDs could become detectable, and the corresponding genes can be proposed to be involved in the HR. Only knockout of PLEA caused a loss of MTI-induced cell wall based defense against the non-host powdery mildew Erysiphe pisi. This is thus in stark contrast to the involvement of a multitude of PLD isoforms in the HR triggered by AvrRpm1 recognition.
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| 9. |
- Johansson, Oskar N., 1984, et al.
(författare)
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Skeletonema marinoi as a new genetic model for marine chain-forming diatoms
- 2019
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 9
-
Tidskriftsartikel (refereegranskat)abstract
- Diatoms are ubiquitous primary producers in marine ecosystems and freshwater habitats. Due to their complex evolutionary history, much remains unknown about the specific gene functions in diatoms that underlie their broad ecological success. In this study, we have genetically transformed the centric diatom Skeletonema marinoi, a dominant phytoplankton species in temperate coastal regions. Transformation of S. marinoi is the first for a true chain-forming diatom, with the random genomic integration via non homologous recombination of a linear DNA construct expressing the resistance gene to the antibiotic zeocin. A set of molecular tools were developed for reliably identifying the genomic insertion site within each transformant, many of which disrupt recognizable genes and constitute null or knock-down mutations. We now propose S. marinoi as a new genetic model for marine diatoms, representing true chain-forming species that play a central role in global photosynthetic carbon sequestration and the biogeochemical cycling of silicates and various nutrients, as well as having potential biotechnological applications.
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| 10. |
- Nilsson, Anders K., 1982, et al.
(författare)
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Formation of oxidized phosphatidylinositol and 12-oxo-phytodienoic acid containing acylated phosphatidylglycerol during the hypersensitive response in Arabidopsis
- 2014
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Ingår i: Phytochemistry. - : Elsevier BV. - 0031-9422. ; 101, s. 65-75
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Tidskriftsartikel (refereegranskat)abstract
- Plant membranes are composed of a wide array of polar lipids. The functionality of these extends far beyond a pure structural role. Membrane lipids function as enzyme co-factors, establish organelle identity and as substrates for enzymes such as lipases and lipoxygenases. Enzymatic degradation or oxidation (enzymatic or non-enzymatic) of membrane lipids leads to the formation of a diverse group of bioactive compounds. Plant defense reactions provoked by pathogenic microorganisms are often associated with substantial modifications of the lipidome. In this study, we profiled changes in phospholipids during the hypersensitive response triggered by recognition of the bacterial effector protein AvrRpm1 in Arabidopsis thaliana. A simple and robust LC-MS based method for profiling plant lipids was designed to separate all the major species of glycerolipids extracted from Arabidopsis leaf tissue. The method efficiently separated several isobaric and near isobaric lipid species, which otherwise are difficult to quantify in direct infusion based profiling. In addition to the previously reported OPDA-containing galactolipids found to be induced during hypersensitive response in Arabidopsis, three OPDA-containing sulfoquinovosyl diacylglycerol species, one phosphatidylinositol species as well as two acylated OPDA-containing phosphatidylglycerol species were found to accumulate during the hypersensitive response in Arabidopsis. Our study confirms and extends on the notion that the hypersensitive response in Arabidopsis triggers a unique profile of Allene Oxide Synthase dependent oxidation of membrane lipids. Primary targets of this oxidation seem to be uncharged and anionic lipid species. (C) 2014 Elsevier Ltd. All rights reserved.
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