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  • 1.
    Danielsson, Marie
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Chemical defence in Norway spruce2011Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Norway spruce (Picea abies) responds to stress by biosynthesis of chemical substances, which can deter invading insects or pathogens. Some of these substances are volatile and can be emitted to the surroundings while others are accumulated within the tree. Information about the susceptibility of individual plants to infestation, their volatile emissions and chemical defence is of interest, for example, in selecting plants for tree breeding programs.

    The first part of this research focused on volatiles emitted by Norway spruce plants. Collection of headspace volatiles by SPME and subsequent separation and identification with GC-MS was used to investigate Norway spruce plants of different ages and stress conditions as well as trapping semiochemicals like nepetalactone emitted by the spruce shoot aphids. It was even possible to analyse the emission of single needles in vivo and obtain spatial localisation of the stress reaction to methyl jasmonate or spruce spinning mites. Seedlings of different ages showed differences in chemical composition of emitted volatiles, with the pine weevil repellent, (4S)-(-)-limonene, one of the main compounds. Wounded phloem of conventional plants emitted high amounts of monoterpenes while the phloem of mini plants emitted (3Z)-hexenal and (3Z)-hexen-1-ol. In addition, a method to separate and identify the four diastereomers of nepetalactone by GC-MS and characteristic m/z-fragments was accomplished.

    The second part of the research deals with the chemical response of Norway spruce roots to inoculation with Heterobasidion annosum. Terpene concentrations increased after inoculation or wounding but the composition was mainly associated with clone identity and not to susceptibility or treatment. In contrast, inoculation with H. annosum induced a treatment-specific alteration of phenol composition. The constitutive phenol composition differed between more and less susceptible clones. The phenols astringin and astringin dimers (piceasides) as well as the terpene α-longipinene may be suitable markers of low susceptibility for P. abies to Heterobasidion.

  • 2.
    Danielsson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Tracing induced stress sites in conifers by single needle analysesManuscript (preprint) (Other academic)
  • 3.
    Danielsson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Kännaste, Astrid
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Lindström, Anders
    School of Industrial Technology and Management, Dalarna University.
    Hellqvist, Claes
    Department of Ecology, Swedish University of Agricultural Sciences.
    Stattin, Eva
    School of Industrial Technology and Management, Dalarna University.
    Långström, Bo
    Department of Ecology, Swedish University of Agricultural Sciences.
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Mini-seedlings of Picea abies are less attacked by Hylobius abietis than conventional ones: Is plant chemistry the explanation?2008In: Scandinavian Journal of Forest Research, ISSN 0282-7581, E-ISSN 1651-1891, Vol. 23, no 4, p. 299-306Article in journal (Refereed)
    Abstract [en]

    The pine weevil, Hylobius abietis (L.), is a major pest in conifer reforestation areas in the Palaearctic region. Size and chemistry of the seedlings may explain the damage rates in plantations. The performance of 10-week containerized seedlings (mini-seedlings) was compared with 1-year-old conventional seedlings of Norway spruce, Picea abies (L.), in a field experiment in central Sweden. After 2 years the weevil damage was lower for the mini-seedlings than for the conventional seedlings (3.5 vs 55%). After 3 years, the overall survival was 82 and 75%, respectively. Weevil damage was the main cause of mortality for conventional seedlings, whereas mini-seedlings mainly died from drought. Volatiles of the two seedling types were compared by solid-phase microextraction-gas chromatography -mass spectrometry (SPME-GC-MS). Unwounded mini-seedlings and conventional seedlings differed in their compositions of monoterpenes and sesquiterpenes. Miniseedlings mainly emitted limonene, known to be repellent to the pine weevil. When wounded, green leaf volatiles were released by mini-seedlings while the pine weevil attractant alpha-pinene was released by conventional seedlings. Volatiles may partly explain the mini-seedlings' resistance against weevil attack. Further studies are needed to clarify how long this miniseedling effect remains.

  • 4.
    Danielsson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Lunden, Karl
    Elfstrand, Malin
    Hu, Jiang
    Zhao, Tao
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Arnerup, Jenny
    Ihrmark, Katarina
    Swedjemark, Gunilla
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Stenlid, Jan
    Chemical and transcriptional responses of Norway spruce genotypes with different susceptibility to Heterobasidion spp. infection2011In: BMC Plant Biology, ISSN 1471-2229, E-ISSN 1471-2229, Vol. 11, p. 154-Article in journal (Refereed)
    Abstract [en]

    Background: Norway spruce [Picea abies (L.) Karst.] is one of the most important conifer species in Europe. The wood is economically important and infections by wood-rotting fungi cause substantial losses to the industry. The first line of defence in a Norway spruce tree is the bark. It is a very efficient barrier against infection based on its mechanical and chemical properties. Once an injury or an infection is recognized by the tree, induced defences are activated. In this study we examined transcriptional response, using 454-sequencing, and chemical profiles in bark of Norway spruce trees with different susceptibility to Heterobasidion annosum s.l. infection. The aim was to find associations between the transcriptome and chemical profiles to the level of susceptibility to Heterobasidion spp. in Norway spruce genotypes. Results: Both terpene and phenol compositions were analysed and at 28 days post inoculation (dpi) high levels of 3-carene was produced in response to H. annosum. However, significant patterns relating to inoculation or to genotypes with higher or lower susceptibility could only be found in the phenol fraction. The levels of the flavonoid catechin, which is polymerized into proanthocyanidins (PA), showed a temporal variation; it accumulated between 5 and 15 dpi in response to H. annosum infection in the less susceptible genotypes. The transcriptome data suggested that the accumulation of free catechin was preceded by an induction of genes in the flavonoid and PA biosynthesis pathway such as leucoanthocyanidin reductase. Quantitative PCR analyses verified the induction of genes in the phenylpropanoid and flavonoid pathway. The qPCR data also highlighted genotype-dependent differences in the transcriptional regulation of these pathways. Conclusions: The varying dynamics in transcriptional and chemical patterns displayed by the less susceptible genotypes suggest that there is a genotypic variation in successful spruce defence strategies against Heterobasidion. However, both high levels of piceasides and flavonoids in the less susceptible genotypes suggested the importance of the phenolic compounds in the defence. Clearly an extended comparison of the transcriptional responses in the interaction with Heterobasidion between several independent genotypes exhibiting reduced susceptibility is needed to catalogue mechanisms of successful host defence strategies.

  • 5.
    Danielsson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Lundén, Karl
    Arnerup, Jenny
    Hu, Jiang
    Zhao, Tao
    Swedjemark, Gunilla
    Elfstrand, Malin
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Stenlid, Jan
    Chemical and transcriptional responses of Norway spruce clones with different susceptibility to Heterobasidion spp. infectionManuscript (preprint) (Other academic)
  • 6.
    Danielsson, Marie
    et al.
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry.
    Zhao, Tao
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Organic chemistry. Department of Science and Technology, Örebro University, Örebro, Sweden.
    Borg-Karlson, Anna-Karin
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Organic chemistry.
    Arthropod infestation sites and induced defence can be traced by emission from single spruce needles2019In: Arthropod-Plant Interactions, ISSN 1872-8855, E-ISSN 1872-8847, Vol. 13, no 2, p. 253-259Article in journal (Refereed)
    Abstract [en]

    Emissions of defence chemicals from Norway spruce seedlings can be induced by feeding arthropods or by exogenous hormonal application. Some defence chemicals may attract or repel associated arthropods. The aim of this study was to show that it is possible to detect and collect stress-induced volatiles from micro sites, such as at the scale of a single needle, in vivo by using SPME. Methyl jasmonate application on the stem of Norway spruce seedlings induced emission of (E)-beta-farnesene only from the needles closest to the application site. Emissions of (E)-beta-farnesene, (E,E)-alpha-farnesene and (E)-alpha-bisabolene were only detected from needles infested by the spider mite Oligonychus ununguis. The total volatile amount detected by SPME-GC-MS reached a considerable mass of 14 ng/needle/24 h, suggesting that emission from damaged and stressed conifers might have a larger impact on the macro climate than previously estimated.

  • 7. Lunden, Karl
    et al.
    Danielsson, Marie
    KTH, School of Chemical Science and Engineering (CHE), Chemistry.
    Durling, Mikael Brandstrom
    Ihrmark, Katarina
    Gorriz, Miguel Nemesio
    Stenlid, Jan
    Asiegbu, Frederick O.
    Elfstrand, Malin
    Transcriptional Responses Associated with Virulence and Defence in the Interaction between Heterobasidion annosum s. s. and Norway Spruce2015In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 10, no 7, article id e0131182Article in journal (Refereed)
    Abstract [en]

    Heterobasidion annosum sensu lato is a serious pathogen causing root and stem rot to conifers in the northern hemisphere and rendering the timber defective for sawing and pulping. In this study we applied next-generation sequencing to i) identify transcriptional responses unique to Heterobasidion-inoculated Norway spruce and ii) investigate the H. annosum transcripts to identify putative virulence factors. To address these objectives we wounded or inoculated 30-year-old Norway spruce clones with H. annosumand 454-sequenced the transcriptome of the interaction at 0, 5 and 15 days post inoculation. The 491860 high-quality reads were de novo assembled and the relative expression was analysed. Overall, very few H. annosum transcripts were represented in our dataset. Three delta-12 fatty acid desaturase transcripts and one Clavaminate synthase-like transcript, both associated with virulence in other pathosystems, were found among the significantly induced transcripts. The analysis of the Norway spruce transcriptional responses produced a handful of differentially expressed transcripts. Most of these transcripts originated from genes known to respond to H. annosum. However, three genes that had not previously been reported to respond to H. annosum showed specific induction to inoculation: an oxophytodienoic acid-reductase (OPR), a beta-glucosidaseand a germin-like protein (GLP2) gene. Even in a small data set like ours, five novel highly expressed Norway spruce transcripts without significant alignment to any previously annotated protein in Genbank but present in the P. abies (v1.0) gene catalogue were identified. Their expression pattern suggests a role in defence. Therefore a more complete survey of the transcriptional responses in the interactions between Norway spruce and its major pathogen H. annosumwould probably provide a better understanding of gymnosperm defence than accumulated until now.

  • 8.
    Pettersson, Marie
    KTH, School of Chemical Science and Engineering (CHE), Chemistry.
    Stress Related Emissions of Norway Spruce Plants2007Licentiate thesis, comprehensive summary (Other scientific)
    Abstract [en]

    The interactions between plants and insects are mediated by volatile molecules. Plants respond to stress by biosynthesis of chemical substances which can deter invading insects or pathogens. Some of these substances are volatile and are emitted to the surroundings and may attract or repel insects. Information about the susceptibility of individual plants to infestation, their volatile emissions and chemical defence is of interest, for example in selecting plants for tree breeding programs.

    This research was focused on finding volatile chemical markers of resistance in Norway spruce plants that do influence insects associated to conifers. Collection of headspace volatiles by SPME followed by separation and identification with GC-MS is effective in investigating biological systems with a minimum of disturbance. This method has here been used to investigate Norway spruce plants of different ages and stress conditions as well as trapping semiochemicals like nepetalactone emitted by the spruce shoot aphids. It was even possible to analyse the emission of single needles in vivo and obtain a chemical pattern of the site of the stress reaction. Seedlings of different ages showed differences in chemical composition of emitted volatiles, with the pine weevil repellent (S)-(-)-limonene as one of the main compounds. Wounded phloem of conventional plants emitted high amounts of monoterpenes while the phloem of mini plants emitted (3Z)-hexenal and (3Z)-hexen-1-ol.

    Norway spruce plants did respond to different stress elicitors with similar response, regardless of their genetic origin. The emissions from stressed Norway spruce plants mainly consist of (E)-β-farnesene, (E,E)-α-farnesene, (E)-α-bisabolene, (R)-(-)-linalool and methyl salicylate. Emissions from live spruce shoot aphids were detected during autumn periods, and a method to separate and identify the four diastereomers of nepetalactone by GC-MS and characteristic m/z-fragments was accomplished.

  • 9.
    Pettersson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Single needle analyses by SPME-GC-MS, a miniaturixed method to localize induced stress sites in conifersManuscript (Other academic)
  • 10.
    Pettersson, Marie
    et al.
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Unelius, Rikard
    Borg-Karlson, Anna-Karin
    KTH, School of Chemical Science and Engineering (CHE), Chemistry, Organic Chemistry.
    Semiochemicals related to the aphid Cinara pillicornis (Hartig) and its host, Picea abies. A method to assign diastereomers of nepetalactone2008In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1180, no 1-2, p. 165-170Article in journal (Refereed)
    Abstract [en]

    Volatiles released by seedlings of Norway spruce infested with the aphid Cinara pilicornis were analyzed using SPME–GC–MS. Among the stress-induced compounds released by the host plant, citronellol, cis–trans-nepetalactone and cis–trans-nepetalactol was found. These compounds originated from the aphids and they were assumed to be pheromone components for this aphid species. To determine the relative stereochemistry of the nepetalactone, a diagnostic method was developed. The method was based on multivariate analysis of tabulated relative intensities of mass fragments of the four nepetalactone diastereomers. In the practical method described, a few pairs of fragments in the mass spectra were compared and, in combination with the Kovat's index, were used to unambiguously identify the relative stereochemistry of the nepetalactone.

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