Ultraviolet absorbance of Sphagnum magellanicum, S. fallax and S. fuscum extracts with seasonal and species-specific variation

Bryophytes, including Sphagnum, are common species in alpine and boreal regions especially on mires, where full sunlight exposes the plants to the damaging effects of UV radiation. Sphagnum species containing UV-protecting compounds might offer a biomass source for nature-based sunscreens to replace the synthetic ones. In this study, potential compounds and those linked in cell wall structures were obtained by using methanol and alkali extractions and the UV absorption of these extracts from three common Sphagnum moss species Sphagnum magellanicum, Sphagnum fuscum and Sphagnum fallax collected in spring and autumn from western Finland are described. Absorption spectrum screening (200–900 nm) and luminescent biosensor (Escherichia coli DPD2794) methodology were used to examine and compare the protection against UV radiation. Additionally, the antioxidant potential was evaluated using hydrogen peroxide scavenging (SCAV), oxygen radical absorbance capacity (ORAC) and ferric reducing absorbance capacity (FRAP). Total phenolic content was also determined using the Folin-Ciocalteu method. The results showed that methanol extractable compounds gave higher UV absorption with the used methods. Sphagnum fallax appeared to give the highest absorption in UV-B and UV-A wavelengths. In all assays except the SCAV test, the methanol extracts of Sphagnum samples collected in autumn indicated the highest antioxidant capacity and polyphenol content. Sphagnum fuscum implied the highest antioxidant capacity and phenolic content. There was low antioxidant and UV absorption provided by the alkali extracts of these three species

Metabolic Profiling of Water-Soluble Compounds from the Extracts of Dark Septate Endophytic Fungi (DSE) Isolated from Scots Pine (Pinus sylvestris L.) Seedlings Using UPLC-Orbitrap-MS

Endophytes are microorganisms living inside plant hosts and are known to be beneficial for the host plant vitality. In this study, we isolated three endophytic fungus species from the roots of Scots pine seedlings growing on Finnish drained peatland setting. The isolated fungi belonged to dark septate endophytes (DSE). The metabolic profiles of the hot water extracts of the fungi were investigated using Ultrahigh Performance Liquid Chromatography with Diode Array Detection and Electron Spray Ionization source Mass Spectrometry with Orbitrap analyzer (UPLC-DAD-ESI-MS-Orbitrap). Out of 318 metabolites, we were able to identify 220, of which a majority was amino acids and peptides. Additionally, opine amino acids, amino acid quinones, Amadori compounds, cholines, nucleobases, nucleosides, nucleotides, siderophores, sugars, sugar alcohols and disaccharides were found, as well as other previously reported metabolites from plants or endophytes. Some differences of the metabolic profiles, regarding the amount and identity of the found metabolites, were observed even though the fungi were isolated from the same host. Many of the discovered metabolites have been described possessing biological activities and properties, which may make a favorable contribution to the host plant nutrient availability or abiotic and biotic stress tolerance

Polyamine metabolism of Scots pine under abiotic stress

Abstract Changes in climate will impose abiotic stress on plant species, and eventually affect their distributional range. This is a particular challenge especially to tree species with long generation times and slow rates of evolution. In the boreal zone, most of these species also hold high economic value. The aim of the study was to enhance understanding of the abiotic stress coping strategies of Scots pine (Pinus sylvestris L.) in its vulnerable early growth phase. In particular, the study evaluates the role of polyamine (PA) metabolism of Scots pine under abiotic stress. Polyamines (PAs) are small, ubiquitous nitrogenous compounds involved in fundamental biological processes of plants such as growth, organogenesis, embryogenesis and abiotic and biotic stress defence. The effects of different water availabilities and spring frost temperatures on shoots and roots of young seedlings were investigated in two growth chamber experiments. Proembryogenic liquid cell culture system was established and used as a controllable platform. Effects of polyethylene glycol (PEG) induced osmotic stress was investigated at the cellular level in liquid cultures. Responses to treatments were evaluated by quantifying changes in the expression of PA metabolism, stress, cell death and cell division -related genes, metabolite concentrations, as well as stress-induced physiology and morphology. The results revealed that PA metabolism is tissue specific and strictly regulated. Drought stress induced the accumulation of putrescine (Put) in photosynthesizing tissues, whereas the prime response to spring frost was a reorganization of root growth. Generally, drought and osmotic stress decreased the expression of PA catabolizing genes, leading to consistent PA levels in tissues even under severe stress. To conclude, PAs have a protective role in maintaining the growth and development of pine tissues subjected to abiotic stress.Tiivistelmä Ilmastonmuutos vaikuttaa eri kasvilajien levinneisyyteen. Nopeasti muuttuva ilmasto on haaste erityisesti metsäpuulajeille, jotka pitkän sukupolvenvälin takia kehittyvät evolutiivisesti hitaasti. Erilaisten ympäristötekijöiden aiheuttamien puolustusaineenvaihdunnan muutosten tunteminen metsäpuilla on tulevaisuuden kannalta myös taloudellisesti tärkeää. Työn tavoitteena oli selvittää millainen merkitys polyamiineilla on metsämännyn (Pinus sylvestris L.) abioottisessa stressivasteessa. Polyamiinit (PAt) ovat pienimolekyylisiä typpiyhdisteitä, joiden tiedetään olevan mukana solujen perusaineenvaihdunnassa, kasvien kasvussa ja kehityksessä ja stressivasteissa. Tutkimus koostuu kolmesta kokeesta, joissa selvitettiin PA aineenvaihduntaa osana männyn taimien ja erilaistumattomien somaattisista alkioista peräisin olevien soluviljelmien stressivasteita. Kahdessa kokeellisessa tutkimuksessa männyn sirkkataimia altistettiin erilaisille kasvualustan vesipitoisuuksille ja keväthallan lämpötilaolosuhteille. Kolmannessa kokeessa PA aineenvaihdunnan reagointia osmoottiseen stressiin tutkittiin erilaistumattomissa somaattisissa epäkypsissä männyn alkiosta peräisin olevissa solukoissa kontrolloidussa nesteviljely-ympäristössä. Tutkimuksessa tarkastelin PA aineenvaihduntaa säätelevien geenien ilmenemistä ja PA pitoisuuksien muutoksia suhteessa kasvuun, kehitykseen ja stressissä indusoituvien geenien ilmenemiseen. Tulokset osoittavat, että PA aineenvaihdunta on tiukasti säädeltyä erilaisissa solukoissa. Taimien yhteyttävät solukot kerryttivät putreskiinia (Put) kuivuudessa. Ensimmäiset vasteet keväthallaolosuhteisiin havaittiin muutoksena juurten kasvun säätelyssä. Sekä kuivuus että osmoottinen stressi laskevat PA hajottavien geenien ilmenemistä, mikä on metsämännyn tapa ylläpitää tasaiset PA pitoisuudet solukoissa stressitekijöistä huolimatta. Johtopäätöksenä voidaan todeta, että PA aineenvaihdunta on tärkeä metsämännyn kasvun ja kehityksen kannalta

Polyamine metabolism in Scots pine embryogenic cells under potassium deficiency

Abstract Polyamines (PA) have a protective role in maintaining growth and development in Scots pine during abiotic stresses. In the present study, a controlled liquid Scots pine embryogenic cell culture was used for studying the responses of PA metabolism related to potassium deficiency. The transcription level regulation of PA metabolism led to the accumulation of putrescine (Put). Arginine decarboxylase (ADC) had an increased expression trend under potassium deficiency, whereas spermidine synthase (SPDS) expression decreased. Generally, free spermidine (Spd) and spermine (Spm)/ thermospermine (t-Spm) contents were kept relatively stable, mostly by the downregulation of polyamine oxidase (PAO) expression. The low potassium contents in the culture medium decreased the potassium content of the cells, which inhibited cell mass growth, but did not affect cell viability. The reduced growth was probably caused by repressed metabolic activity and cell division, whereas there were no signs of H₂O₂-induced oxidative stress or increased cell death. The low intracellular content of K⁺ decreased the content of Na⁺. The decrease in the pH of the culture medium indicated that H⁺ ions were pumped out of the cells. Altogether, our findings emphasize the specific role(s) of Put under potassium deficiency and strict developmental regulation of PA metabolism in Scots pine

Coping with spring frost-effects on polyamine metabolism of Scots pine seedlings

Polyamines (PA) are ubiquitous polycations known to be involved in several phases of plant development as well as in tolerance to abiotic stresses. Phenols are complex secondary metabolites produced via the phenylpropanoid pathway that contain, e.g., cell wall compounds and antioxidants. Phenols are known to enhance chilling tolerance of plants. PA and phenolic pathways are connected via conjugation. In boreal coniferous forests spring frost has been considered to have severe effects on the survival of tree seedlings. Such effects are likely to increase in the future. The present study focuses on the role of PA and phenylpropanoid syntheses in the coping strategies of Scots pine exposed to cold temperatures during the vulnerable early seedling phase in late spring and early summer. We found that spring frost affects the expression of genes regulating PA metabolism and phenylpropanoid synthesis differently in above and below ground parts of the seedlings, whereas PA or phenol contents in tissues were not affected. The results suggest that Scots pine seedlings may not have time to develop metabolite level responses during a short period of freezing stress and, therefore, the originally different PA levels, especially in roots, may influence the tolerance of Scots pine seedlings to spring frost