Browsing by Subject "pigmentit"

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  • Seppälä, Jukka (Finnish Environment Institute, 2009)
    Monographs of the Boreal Environment Research 34
    To obtain data on phytoplankton dynamics (abundance, taxonomy, productivity, and physiology) with improved spatial and temporal resolution, and at reduced cost, traditional phytoplankton monitoring methods have been supplemented with optical approaches. Fluorescence detection of living phytoplankton is very sensitive and not disturbed much by the other optically active components. Fluorescence results are easy to generate, but interpretation of measurements is not straightforward as phytoplankton fluorescence is determined by light absorption, light reabsorption, and quantum yield of fluorescence - all of which are affected by the physiological state of the cells. In this thesis, I have explored various fluorescence-based techniques for detection of phytoplankton abundance, taxonomy and physiology in the Baltic Sea.In algal cultures used in this thesis, the availability of nitrogen and light conditions caused changes in pigmentation, and consequently in light absorption and fluorescence properties of cells. The variation of absorption and fluorescence properties of natural phytoplankton populations in the Baltic Sea was more complex. Physical environmental factors (e.g. mixing depth, irradiance and temperature) and related seasonal succession in the phytoplankton community explained a large part of the seasonal variability in the magnitude and shape of Chlorophyll a (Chla)-specific absorption. Subsequent variations in the variables affecting fluorescence were large; 2.4-fold for light reabsorption at the red Chla peak and 7-fold for the spectrally averaged Chla-specific absorption coefficient for Photosystem II. In the studies included in this thesis, Chla-specific fluorescence varied 2-10 fold. This variability in Chla-specific fluorescence was related to the abundance of cyanobacteria, the size structure of the phytoplankton community, and absorption characteristics of phytoplankton.Cyanobacteria show very low Chla-specific fluorescence. In the presence of eukaryotic species, Chla fluorescence describes poorly cyanobacteria. During cyanobacterial bloom in the Baltic Sea, phycocyanin fluorescence explained large part of the variability in Chla concentrations. Thus, both Chla and phycocyanin fluorescence were required to predict Chla concentration.Phycobilins are major light harvesting pigments for cyanobacteria. In the open Baltic Sea, small picoplanktonic cyanobacteria were the main source of phycoerythrin fluorescence and absorption signal. Large filamentous cyanobacteria, forming harmful blooms, were the main source of the phycocyanin fluorescence signal and typically their biomass and phycocyanin fluorescence were linearly related. It was shown that for reliable phycocyanin detection, instrument wavebands must match the actual phycocyanin fluorescence peak well. In order to initiate an operational ship-of-opportunity monitoring of cyanobacterial blooms in the Baltic Sea, the distribution of filamentous cyanobacteria was followed in 2005 using phycocyanin fluorescence.Various taxonomic phytoplankton pigment groups can be separated by spectral fluorescence. I compared multivariate calibration methods for the retrieval of phytoplankton biomass in different taxonomic groups. During a mesocosm experiment, a partial least squares regression method gave the closest predictions for all taxonomic groups, and the accuracy was adequate for phytoplankton bloom detection. This method was noted applicable especially in the cases when not all of the optically active compounds are known.Variable fluorescence has been proposed as a tool to study the physiological state of phytoplankton. My results from the Baltic Sea emphasize that variable fluorescence alone cannot be used to detect nutrient limitation of phytoplankton. However, when combined with experiments with active nutrient manipulation, and other nutrient limitation indices, variable fluorescence provided valuable information on the physiological responses of the phytoplankton community. This thesis found a severe limitation of a commercial fast repetition rate fluorometer, which couldn’t detect the variable fluorescence of phycoerythrin-lacking cyanobacteria. For these species, the Photosystem II absorption of blue light is very low, and fluorometer excitation light did not saturate Photosystem II during a measurement.This thesis encourages the use of various in vivo fluorescence methods for the detection of bulk phytoplankton biomass, biomass of cyanobacteria, chemotaxonomy of phytoplankton community, and phytoplankton physiology. Fluorescence methods can support traditional phytoplankton monitoring by providing continuous measurements of phytoplankton, and thereby strengthen the understanding of the links between biological, chemical and physical processes in aquatic ecosystems.
  • Lausala, Ronja (Helsingin yliopisto, 2021)
    Tutkielman tarkoitus oli selvittää kolmen eri valointensiteetin vaikutuksia mikrolevälajien, E.gracilis ja Selenastrum sp., kasvuun ja karotenoidituotantoon. Käytetyt valointensiteetit olivat 0 (pimeä), 200 ja 400 µmol m-2s-1. Levät kasvatettiin kiertovesikalankasvattamon jätevedessä, minkä vuoksi työssä tarkasteltiin myös ravinnepoistumaa. Kasvatukset havaittiin typpirajoitteisiksi, mikä osaltaan heikensi levien kasvua ja siten karotenoidituotantoa. Jätevesi kasvualustana aiheutti E.gracilis -levän kasvatuksiin kontaminaation vieraslevällä. Karotenoidit eristettiin kasvatuskokeen päätyttyä uuttamalla paineistetulla nesteuutolla (Accelerated Solvent Extraction, ASE) etanolin ja 2-metyylitetrahydrofuraanin liuotinyhdistelmällä. Karotenoidien jatkoanalyysiin käytettiin erittäin korkean erotuskyvyn nestekromatografiaa (UHPLC, Ultra-High Performance Liquid Chromatography) yhdistettynä sähkösumutusionisaatioon ja massaspektrometriin. Tunnistettujen karotenoidien määriä analysoitiin niiden signaalivahvuuksiin ja käytettävissä olleisiin malliaineisiin perustuen. Valointensiteeteillä havaittiin olevan vaikutus levien kasvuun, kokonaiskarotenoidi- ja klorofyllipitoisuuksiin sekä astaksantiiniin, luteiiniin ja neoksantiiniin. Kyseiset erikseen tunnistetut pigmentit olivat kaikki ksantofyllejä, joilla on liialta valointensiteetiltä suojaava tehtävä leväsoluissa. Valointensiteetin sietokyky poikkesi kuitenkin levälajeilla eivätkä sen vaikutukset olleet yksiselitteisiä. Pimeässä levien kasvu ja ravinnepoistuma olivat heikkoja. Pigmenttien säilymisen puolesta pimeä osoittautui jopa voimakkainta valointensiteettiä paremmaksi vaihtoehdoksi. Molempien levien ravinteiden kulutus oli tehokasta. Valointensiteetin lisäksi kasvuun ja karotenoidituotantoon vaikutti muutkin tekijät, kuten kokeen aikana saavutettu kasvuvaihe, jatkuva valotus, kontaminoituminen, ravinnerajoitteisuus sekä käytettyjen malliaineiden laatu. Jatkotutkimuksissa tulisi pyrkiä minimoimaan tai poissulkemaan näiden tekijöiden vaikutukset.