Light absorption and scattering by cell suspensions of some cyanobacteria and microalgae

Absorption spectra of cyanobacteria (Anacystis nidulans, Anabaena variabilis, and Chlorogloeopsis fritschii), red (Cyanidium caldarum and Porphyridium cruentum), green (Dunaliella maritima and Dunaliella salina) and diatom (Thalassiosira weisflogii) alga cell suspensions are presented; the spectra were obtained by using an approach developed earlier to compensate for scattering [1, 2]. In all species, the shapes of the absorption spectra were independent of the cell concentration. For Th. weisflogii and D. maritima, the analysis of selective and nonselective scattering was carried out. The effect of mechanical cell disruption on optical properties (absorption, scattering, and “package” effect) on D. maritima was studied. The character and dynamics of optical changes in D. salina under the influence of sodium chloride were followed.     

Estimation of biotechnological potential and clarification of taxonomic status of <Emphasis Type="Italic">Parietochloris</Emphasis> genus microalgae (Trebouxiophyceae) from the CALU collection

Morphological, biochemical, and molecular genetic studies of green microalgae from the collection of cyanobacteria, algae, and algal parasites of St. Petersburg State University (CALU) (presumably belonging to the Parietochloris genus) were conducted in order to estimate biotechnological potential and clarify the phylogenetic position. It was determined that the studied strains have a close relationship to two genera from different classes (Lobosphaera (Trebouxiaceae) and Deasonia (Actinochloridaceae)) and can be of biotechnological interest as producers of valuable polyunsaturated fatty acids (especially arachidonic, linoleic, and α-linolenic).     

Desmodesmus sp. 3Dp86E-1—a Novel Symbiotic Chlorophyte Capable of Growth on Pure CO2

A novel chlorophyte Desmodesmus sp. 3Dp86E-1 isolated from a White Sea hydroid Dynamena pumila was cultivated at CO₂ levels from atmospheric (the ‘low-CO₂’ conditions) to pure carbon dioxide (the 5, 20, and 100 % CO₂ conditions) under high (480 μE/(m² s) PAR) light. After 7 days of cultivation, the ‘100 % CO₂’ (but not 5 or 20 % CO₂) cells possessed ca. four times higher chlorophyll content per dry weight (DW) unit than the low-CO₂ culture. The rate of CO₂ fixation under 100 % CO₂ comprised ca. 1.5 L/day per L culture volume. After a lag period which depended on the CO₂ level, biomass accumulation and volumetric fatty acid (FA) content of the Desmodesmus sp. 3Dp86E-1 bubbled with CO₂-enriched gas mixtures increased and was comparable to that of the culture continuously bubbled with air. Under the low-to-moderate CO₂ conditions, the FA percentage of the algal cells increased (to 40 % DW) whereas under high-CO₂ conditions, FA percentage did not exceed 15 % DW. A strong increase in oleate (18:1) proportion of total FA at the expense of linolenate (18:3) was recorded in the ‘100 % CO₂’ cells. Electron microscopy and pulse–amplitude-modulated chlorophyll fluorescence investigation revealed no damage to or significant downregulation of the photosynthetic apparatus in ‘100 % CO₂’ cells grown at the high-PAR irradiance. Possible mechanisms of high-CO₂ tolerance of Desmodesmus sp. 3Dp86E-1 are discussed in view of its symbiotic origin and possible application for CO₂ biomitigation.     

Non-destructive optical detection of pigment changes during leaf senescence and fruit ripening

Reflectance spectra in the visible and near infra-red range of the spectrum, acquired for maple ( Acer platanoides L.), chestnut ( Aesculus hippocastanum L.), potato ( Solanum tuberosum L.), coleus ( Coleus blumei Benth.), leaves and lemon (Citrus limon L.) and apple ( Malus domestica Borkh.) fruits were studied. An increase of reflectance between 550 and 740 nm accompanied senescence-induced degradation of chlorophyll (Chl), whereas in the range 400–500 nm it remained low, due to retention of carotenoids (Car). It was found that both leaf senescence and fruit ripening affect the difference between reflectance ( R ) near 670 and 500 nm ( R ₆₇₈− R ₅₀₀), depending on pigment composition. The plant senescing reflectance index in the form ( R ₆₇₈− R ₅₀₀)/ R ₇₅₀ was found to be sensitive to the Car/Chl ratio, and was used as a quantitative measure of leaf senescence and fruit ripening. The changes in the index were followed during leaf senescence, and natural and ethylene-induced fruit ripening. This novel index can be used for estimating the onset, the stage, relative rates and kinetics of senescence/ripening processes.     

Light absorption by isolated chloroplasts and leaves: effects of scattering and ‘packing’

Light absorption was quantified in the following systems: isolated chloroplasts and leaves of spinach (Spinacea oleracea L.), a mutant of geranium (Pelargonium zonale L.) widely differing in pigment content, and coleus (Coleus blumei Benth.) at different stages of leaf ontogenesis. For these species and pea (Pisum sativum L.), scattering-compensated absorption spectra of chloroplast suspensions are presented. Comparison of leaf and chloroplast spectra showed considerable changes in the extent of the ‘package’ effect and the lengthening of the effective optical path in a leaf. The difference between leaf and isolated chloroplast absorption could be quantitatively described by adapting Duysens’s treatment of flattening. It was found that the accumulation of chlorophyll in leaves is accompanied by a monotonous enhancement of the package effect. The results are discussed with special reference to the role of light scattering in leaf optics, light utilization in photosynthesis and wavelength-dependent light gradients in a leaf.     

Application of Reflectance Spectroscopy for Analysis of Higher Plant Pigments

Nondestructive techniques developed by the authors for assessment of chlorophylls, carotenoids, and anthocyanins in higher plant leaves and fruits are presented. The spectral features of leaf reflectance in the visible and near infrared regions are briefly considered. For pigment analysis only reflectance values at several specific wavelengths are required. The chlorophyll (Chl) content over a wide range of its changes can be assessed during leaf ontogeny using reflectance near 700 nm and, in the absence of anthocyanins, at 550 nm. The approaches used for elimination of Chl interference in the analysis of carotenoids (reflectance at 520 nm) and anthocyanins (at 550 nm) are described. The suitability of reflectance spectroscopy for estimates of carotenoid/chlorophyll ratios during leaf senescence and fruit ripening is demonstrated. The algorithms developed for pigment analysis are presented, and the conditions of their applicability are considered. Further perspectives for the application of reflectance spectroscopy including remote sensing for estimation of plant pigment content and physiological states are discussed.     

Effects of illumination and nitrogen starvation on accumulation of arachidonic acid by the microalga Parietochloris incisa

Parietochloris incisa is a unicellular freshwater green alga capable of accumulating high amounts of the valuable long-chain polyunsaturated arachidonic acid (AA) in triacylglycerols (TAG) of cytoplasmic oil bodies. To find the cultivation conditions providing maximum AA yield, the effects of illumination and N-availability on the dry weight (DW), chlorophyll, carotenoid, and AA content were studied. Under nitrogen starvation, TAG accounted for over 30% of dry weight (DW) and the AA content became as high as about 55% of total fatty acids. For biomass accumulation, light intensity of ca 400 μE m^?2 s^?1 was found to be optimal for growing P. incisa on a complete medium. Lower light intensities (or a higher cell density of inoculum) resulted in a higher AA yield when the alga was cultivated on nitrogen-free media. In the absence of nitrogen, algal cells were unable to cope with high illumination and suffered from photooxidative damage, whereas the nutrientsufficient culture survived under such illumination conditions, probably due to accumulation of carotenoids. Nitrogen-deprived P. incisa cells displayed elevated sensitivity to light.     

Nondestructive estimation of anthocyanins and chlorophylls in anthocyanic leaves

The anthocyanin and chlorophyll contents in leaves provide valuable information about the physiological status of plants. Thus, there is a need for accurate, efficient, and practical methodologies to estimate these biochemical parameters of vegetation. In this study, we tested the performance and accuracy of several nondestructive, reflectance-based techniques for estimating anthocyanin and chlorophyll contents in leaves of four unrelated species, European hazel (Corylus avellana), Siberian dogwood (Cornus alba =Swida alba), Norway maple (Acer platanoides), and Virginia creeper (Parthenocissus quinquefolia), with widely variable pigment content and composition. An anthocyanin reflectance index, which uses reflectances in the green and red edge spectral bands, and a modified anthocyanin reflectance index, employing, in addition, the near-infrared (NIR) band, were able to accurately estimate leaf anthocyanin for all species taken together with no reparameterization of algorithms. Total chlorophyll content was accurately estimated by a red edge chlorophyll index that uses spectral bands in the red edge and the NIR. These approaches can be used to estimate anthocyanin and chlorophyll nondestructively and allow the development of simple handheld field instrumentation.